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["Details on projects of the"]
["SPP 2084:  µBONE: Colonization and interaction of tumor cells within the bone microenvironment "]
["SPP description","Cancer is one of the leading causes of death worldwide. Despite medical progress in cancer therapies, most patients suffer and die from metastases at distant locations. Forming bone metastases are charac-teristic features of prostate and breast cancer. Bone metastases frequently lead to fractures, pain, and a poor life quality and are also associated with a high socioeconomic burden. The key molecular steps resulting in bone metastasis, including the initial colonisation of bone by tumor cells and the early in-teraction with bone and bone marrow cells are poorly understood. Once established, bone metastases represent a point of no return, are not curable, and may be a new source for systemic relapse. There-fore, this Priority Programme aims to generate new insights into (1) the mechanisms of tumor cell col-onisation to the bone marrow and (2) the interactions of tumor cells with the bone microenvironment to identify novel therapeutic concepts for the treatment and/or prevention of bone metastases. The focus is on bone metastasis secondary to breast and prostate cancer as well as myeloma bone disease. "]
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["Proejct Title","Development and validation of a gene expression assay as prognostic and predictive biomarker in prostate cancer bone metastases ","Regulation of Tumor Cell Dormancy in Human Prostate Cancer Bone Metastasis ","The role of adipocytes in the bone tumor micro-environment ","Dissecting the pleiotropic effects of Dickkopf-1 in homing and colonization of breast cancer cells to and within bone.","Wnt5a signaling in the bone microenvironment in prostate cancer ","Characterization of the impact of bone-seeking tumors on the osteocyte network and the osteocyte-mediated regulation of bone turnover ","Central administration, scientific exchange, and clinical translation ","Dissecting the role/s of breast cancer cell membrane protrusions in invasion, colonization and transformation of the bone marrow microenvironment: From cell biology to clinical applications ","Influence of the skeletal remodeling status on tumor cell dissemination and metastatic outgrowth ","The impact of myostatin on breast cancer and multiple myeloma bone metastases ","Hypoxia pathway proteins during homing and propagation of bone metastasis in mice ","Dissecting the role of TAM receptors in myeloma-induced osteoclast activation and their immune-modulatory function ","Control of Tumor Cell – Bone Metastasis By Regulation of F-Actin Dynamics","The role of TRIM proteins in colonization of the bone and bone microenvironment modulation of metastatic prostate cancer. ","RAI2 as novel suppressor of cancer cell homing and survival in the bone marrow ","Probing the role of bone marrow niche-derived SPARC (Secreted Protein Acidic Rich in Cysteine) in the metastatic behavior of human breast cancer ","Bone marrow stromal cells affect cancer cell homing and growth ","Junctional adhesion molecule interactions at the multiple myeloma interface ","The role of tumor-associated macrophages in bone metastasis formation in zebrafish ","Role of Tgif1 in breast cancer-induced pathological bone remodeling ","Molecular Characterization of Bone Metastasis Founder Cells in Prostate Cancer ","Functional characterization of bone metastasis-initiating and radioresistant prostate tumor cells ","Molecular and functional characterization of the normal and malignant plasma cell – bone interface – from single cell mutual interaction analysis to clinical implications in myeloma bone disease and survival "]
["Subject Area","Pathology, Hematology, Oncology ","Anatomy, Hematology, Oncology","Hematology, Oncology, Radiology, Nuclear Medicine, Radiotherapy, Radiobiology, Cell Biology ","Endocrinology, Diabetology, Metabolism, Hematology, Oncology ","Endocrinology, Diabetology, Metabolism, Hematology, Oncology ","Orthopaedic, Traumatology, Reconstructive Surgery \nHematology, Oncology ","Endocrinology, Diabetology, Metabolism \nHematology, Oncology ","Cell Biology \nGynaecology and Obstetrics \nHematology, Oncology ","Orthopaedic, Traumatology, Reconstructive Surgery \nHematology, Oncology ","Hematology, Oncology ","Hematology, Oncology ","Hematology, Oncology ","Hematology, Oncology \nCell Biology ","Pathology \nHematology, Oncology ","Hematology, Oncology ","Hematology, Oncology ","Hematology, Oncology ","Hematology, Oncology ","Hematology, Oncology \nDevelopmental Biology \nCell Biology ","Orthopaedic, Traumatology, Reconstructive Surgery \nHematology, Oncology ","Pathology, Hematology, Oncology ","Hematology, Oncology \nRadiology, Nuclear Medicine, Radiotherapy, Radiobiology \nReproductive Medicine, Urology ","Hematology, Oncology "]
["PI and Institution","Dr. Verena-Wilbeth Sailer\nUniversitätsklinikum Schleswig-Holstein\nCampus Lübeck\nInstitut für Pathologie\nRatzeburger Allee 160\n23562 Lübeck","Professor Dr. Tobias Lange, Ph.D.\nUniversitätsklinikum Hamburg-Eppendorf \nZentrum für experimentelle Medizin\nInstitut für Anatomie und Experimentelle Morphologie\nMartinistraße 52\n20251 Hamburg\n+49 40 741052591\nto.lange@uke.de","Professorin Dr. Aline Bozec\nUniversitätsklinikum Erlangen AöR\nMedizinische Klinik 3 - Rheumatologie und Immunologie\nInternistisches Zentrum (INZ)\nUlmenweg 18\n91054 Erlangen\n+49 9131 8543141\naline.bozec@uk-erlangen.de\nwww.medizin3.uk-erlangen.de/​forschung/​arbeitsgruppen/​juniorprofessur-osteoimmunologie/​ ","Dr. Andy Göbel\nUniversitätsklinikum Carl Gustav Carus Dresden\nFetscherstraße 74\n01307 Dresden\n+49 351 4587595\nAndy.Goebel@uniklinikum-dresden.de","Professor Dr. Lorenz Hofbauer\nUniversitätsklinikum Carl Gustav Carus Dresden \nMedizinische Klinik und Poliklinik III \nBereich Endokrinologie, Diabetes und Knochenstoffwechselerkrankungen\nFetscherstraße 74\n01307 Dresden\n+49 351 4583173\nlorenz.hofbauer@uniklinikum-dresden.de\nwww.bone-lab.de ","Professor Dr. Björn Busse\nUniversitätsklinikum Hamburg-Eppendorf\nInstitut für Osteologie und Biomechanik (IOBM)\nMartinistraße 52\n20251 Hamburg\nb.busse@uke.uni-hamburg.de","Professor Dr. Lorenz Hofbauer (coordinator)\nUniversitätsklinikum Carl Gustav Carus Dresden \nMedizinische Klinik und Poliklinik III \nBereich Endokrinologie, Diabetes und Knochenstoffwechselerkrankungen\nFetscherstraße 74\n01307 Dresden\n+49 351 4583173\nlorenz.hofbauer@uniklinikum-dresden.de\nwww.bone-lab.de ","Dr. Denis Corbeil\nTechnische Universität Dresden \nCenter for Molecular and Cellular Bioengineering (CMCB)\nBiotechnologisches Zentrum (Biotec)\nTatzberg 47/49\n01307 Dresden\n+49 351 46340118\ndenis.corbeil@tu-dresden.de\nwww.biotec.tu-dresden.de/​corbeil ","Professor Dr. Michael Amling\nUniversitätsklinikum Hamburg-Eppendorf\nInstitut für Osteologie und Biomechanik (IOBM)\nMartinistraße 52\n20251 Hamburg\n+49 40 741056373\namling@uke.uni-hamburg.de\nwww.iobm.de ","Dr. Berno Dankbar\nUniversitätsklinikum Münster \nZentrum für Muskuloskelettale Medizin \nInstitut für Experimentelle Muskuloskelettale Medizin\nAlbert-Schweitzer-Campus 1\n48149 Münster\n+49 251 8356756\ndankbarb@uni-muenster.de","Professorin Dr. Martina Rauner\nUniversitätsklinikum Carl Gustav Carus Dresden \nMedizinische Klinik und Poliklinik III \nBereich Endokrinologie, Diabetes und Knochenstoffwechselerkrankungen\nFetscherstraße 74\n01307 Dresden\n+49 351 4584206\nmartina.rauner@uniklinikum-dresden.de","Dr. Isabel Ben Batalla, Ph.D.\nUniversitätsklinikum Hamburg-Eppendorf\nMartinistraße 52\n20251 Hamburg\n+49 40 51950\ni.ben-batall@uke.de","Professor Dr. Rolf Jessberger\nTechnische Universität Dresden \nMedizinische Fakultät Carl Gustav Carus \nInstitut für Physiologische Chemie\nFiedlerstraße 42\n01307 Dresden\n+49 351 4586446\nrolf.jessberger@tu-dresden.de\ntu-dresden.de/​med/​mf/​pch/​das-institut/​arbeitsgruppen/​rolf-jessberger/​research ","Professor Dr. Sven Perner\nUniversitätsklinikum Schleswig-Holstein\nCampus Lübeck\nInstitut für Pathologie\nRatzeburger Allee 160\n23562 Lübeck\n+49 451 50015800\nsven.perner@uksh.de","Professor Dr. Klaus Pantel\nUniversitätsklinikum Hamburg-Eppendorf \nZentrum für experimentelle Medizin \nInstitut für Tumorbiologie\nMartinistraße 52\n20251 Hamburg\n+49 40 741053503\npantel@uke.de\nwww.uke.de/​institute/​tumorbiologie/​ ","Dr. Hind Medyouf, Ph.D.\nGeorg-Speyer-Haus\nInstitut für Tumorbiologie und experimentelle Therapie\nPaul-Ehrlich-Straße 42-44\n60596 Frankfurt am Main\n+49 69 540\nmedyouf@gsh.uni-frankfurt.de\nwww.georg-speyer-haus.de/​forschung/​forschungsgruppen/​medyouf/​forschung.html ","Professorin Dr. Inaam Nakchbandi\nUniversitätsklinikum Heidelberg \nInstitut für Immunologie\nIm Neuenheimer Feld 305\n69120 Heidelberg\n+49 6221 568744\ninaam.nakchbandi@immu.uni-heidelberg.de","Dr. Andreas Brandl\nUniversitätsklinikum Würzburg\nMedizinische Klinik und Poliklinik II\nIZKF Forschergruppe für Experimentelle Stammzelltransplantation\nZinklesweg 10\n97078 Würzburg\n+49 931 20144055\nBrandl_A@ukw.de","Professorin Dr. Franziska Knopf\nTechnische Universität Dresden\nZentrum für Regenerative Therapien Dresden - CRTD\nFetscherstraße 105\n01307 Dresden\n+49 351 46382303\nFranziska.Knopf@tu-dresden.de\nwww.biotec.tu-dresden.de/​ ","Professor Dr. Eric Hesse, Ph.D.\nKlinikum der Universität München\nInstitut für Molekulare Muskuloskelettale Forschung\nFraunhoferstraße 20\n82152 Planegg\nEric.Hesse@med.uni-muenchen.de\nwww.med.uni-muenchen.de/​aktuell/​direktor_immf/​index.html ","Dr. Miodrag Guzvic, Ph.D.\nUniversität Regensburg \nFakultät für Medizin \nLehrstuhl für Experimentelle Medizin und Therapieverfahren\nFranz-Josef-Strauß-Allee 11\n93053 Regensburg\nmiodrag.guzvicukr.de","Professor Dr. Michael Baumann\nDeutsches Krebsforschungszentrum (DKFZ)\nIm Neuenheimer Feld 280\n69120 Heidelberg\n+49 6221 422854\nmichael.baumann@dkfz.de\nwww.tu-dresden.de/​medstrahlneu/​ ","Professor Dr. Franz Jakob\nJulius-Maximilians-Universität Würzburg \nOrthopädisches Zentrum für Muskuloskelettale Forschung \nLehrstuhl für Orthopädie am König-Ludwig-Haus\nBrettreichstraße 11\n97074 Würzburg\n+49 931 8031580\nf-jakob.klh@uni-wuerzburg.de\nwww.koenig-ludwig-haus.de/​startseite/​koenig-ludwig-haus/​patienten-und-besucher/​ansprechpartner/​aerzte/​index.html "]
[null,null,null,"Professor Dr. Tobias Bäuerle\nUniversitätsklinikum Erlangen AöR \nRadiologisches Institut\nMaximiliansplatz 1\n91054 Erlangen\n+49 9131 85-45521\ntobias.baeuerle@uk-erlangen.de","Dr. Tilmann Rachner\nUniversitätsklinikum Carl Gustav Carus Dresden \nMedizinische Klinik und Poliklinik III \nBereich Endokrinologie, Diabetes und Knochenstoffwechselerkrankungen\nFetscherstraße 74\n01307 Dresden\n+49 351 45813731\ntilman.rachner@uniklinikum-dresden.de","Privatdozentin Dr. Christine Hofbauer\nUniversitätsklinikum Carl Gustav Carus Dresden\nUniversitätsCentrum für Orthopädie und Unfallchirurgie\nFetscherstraße 74\n01307 Dresden\n+49 351 458-13323\nchristine.hofbauer@uniklinikum-dresden.de","Professor Dr. Kay Raum\nCharité - Universitätsmedizin Berlin \nCampus Virchow-Klinikum \nBerlin Institute of Health Center for Regenerative Therapies (BCRT)\nAugustenburger Platz 1\n13353 Berlin\n+49 30 450539503\nkay.raum@charite.de\nwww.b-crt.de/​ausbildung/​bsrt-professuren/​kay-raum/​ ",null,"Professorin Dr. Pauline Wimberger\nUniversitätsklinikum Carl Gustav Carus\nan der Technischen Universität Dresden\nKlinik und Poliklinik für Frauenheilkunde und Geburtshilfe\nFetscherstraße 74\n01307 Dresden\n+49 351 4583420\npauline.wimberger@uniklinikum-dresden.de\nwww.uniklinikum-dresden.de/​de/​das-klinikum/​kliniken-polikliniken-institute/​gyn ","Professor Dr. Thorsten Schinke, Ph.D.\nUniversitätsklinikum Hamburg-Eppendorf\nInstitut für Osteologie und Biomechanik (IOBM)\nMartinistraße 52\n20251 Hamburg\n+49 40 7410-58057\nschinke@uke.uni-hamburg.de",null,"Professor Dr. Ben Wielockx, Ph.D.\nUniversitätsklinikum Carl Gustav Carus\nan der Technischen Universität Dresden\nInstitut für Klinische Chemie und Laboratoriumsmedizin\nFetscherstraße 74\n01307 Dresden\n+49 351 45816260\nben.wielockx@tu-dresden.de\ntu-dresden.de/​die_tu_dresden/​fakultaeten/​medizinische_fakultaet/​inst/​kpb/​groups/​research_focus_wielockx ","Professorin Dr. Sonja Loges\nDeutsches Krebsforschungszentrum (DKFZ)\nDKFZ-Hector Krebsinstitut an der Universitätsmedizin Mannheim\nIm Neuenheimer Feld 280\n69120 Heidelberg\ns.loges@dkfz-heidelberg.de\nlogeslab.bplaced.net/​AG_Loges/​Home.html ",null,null,"Professorin Dr. Harriet Wikman-Kocher, Ph.D.\nUniversitätsklinikum Hamburg-Eppendorf \nZentrum für experimentelle Medizin \nInstitut für Tumorbiologie\nMartinistraße 52\n20251 Hamburg\n+49 40 741051319\nh.wikman@uke.uni-hamburg.de\nwww.uke.de/​tumorbiology/​wikman ","Dr. Lisa Sevenich\nGeorg-Speyer-Haus\nInstitut für Tumorbiologie und experimentelle Therapie\nPaul-Ehrlich-Straße 42-44\n60596 Frankfurt am Main\n+49 69 560\nsevenich@gsh.uni-frankfurt.de\nwww.georg-speyer-haus.de/​de/​forschung/​forschungsgruppen/​sevenich/​forschung.html ",null,null,null,"Dr. Hanna Taipaleenmäki, Ph.D.\nUniversitätsklinikum Hamburg-Eppendorf \nZentrum für Operative Medizin\nKlinik und Poliklinik für Unfall- Hand- und Wiederherstellungschirurgie\nMartinistraße 52\n20251 Hamburg\n+49 40 741051163\nh.taipaleenmaeki@uke.de","Professor Dr. Christoph Klein\nUniversität Regensburg \nFakultät für Medizin \nLehrstuhl für Experimentelle Medizin und Therapieverfahren\nFranz-Josef-Strauß-Allee 11\n93053 Regensburg\n+49 941 9446720\nchristoph.klein@klinik.uni-regensburg.de\nexperimentelle-medizin.de/​ ","Professorin Dr. Anna Dubrovska\nTechnische Universität Dresden\nMedizinische Fakultät Carl Gustav Carus\nOncoRay - Zentrum für Medizinische Strahlenforschung in der Onkologie\nFetscherstraße 74\n01307 Dresden\nanna.dubrovska@oncoray.de\nwww.oncoray.de "]
[{"$nulls":6},"Professorin Dr. Martina Rauner\nUniversitätsklinikum Carl Gustav Carus Dresden \nMedizinische Klinik und Poliklinik III \nBereich Endokrinologie, Diabetes und Knochenstoffwechselerkrankungen\nFetscherstraße 74\n01307 Dresden\n+49 351 4584206\nmartina.rauner@uniklinikum-dresden.de",{"$nulls":15},"Professorin Dr. Mechthild Krause\nUniversitätsklinikum Carl Gustav Carus\nan der Technischen Universität Dresden\nKlinik und Poliklinik für Strahlentherapie und Radioonkologie\nFetscherstraße 74\n01307 Dresden\n+49 351 458-5292\nmechthild.krause@uniklinikum-dresden.de\nwww.uniklinikum-dresden.de"]
["Project summary","Skeletal metastases from prostate cancer are a major health burden worldwide and severely affect a patient’s quality of life. They harbor molecular alterations distinctly different from the primary tumor. Moreover, systemic treatment itself can change the molecular landscape of metastatic disease. Thus, a tissue-based biomarker evaluating molecular changes in bone metastases rather than in the primary tumor to predict prognosis or response to –targeted– therapy could be of benefit for these patients. In this proposed research project, we will apply a gene expression assay to robustly detect differentially expressed candidate genes and patterns of pathway regulation in archival formalin-fixed, paraffin embedded (FFPE) bone metastases from prostate cancer patients, because FFPE material is usually the only available material. Ninety-six candidate genes have already been identified through our preliminary work. Gene expression will be studied using a highly sensitive novel array system, that uses color-coded barcodes to detect transcripts (NanoString nCounter®) and can be custom-built. Correlation of gene expression with clinical data will subsequently identify a panel of genes with prognostic and/or predictive potential. This gene panel will undergo retrospective as well as prospective validation in tumor tissue from bone metastases. In addition, RNA from circulating tumor cells will be tested to evaluate its potential as non-invasive biomarker (liquid biopsy) for patients, for whom metastatic tissue is not available. Functional validation using two prostate cancer cell lines (VCaP/PC-3), both derived from a prostate cancer bone metastasis, will be performed. For 5-7 overexpressed genes in the gene panel, we will perform transient knockdown using siRNA to evaluate the biological consequence of overexpression. Data generated in this proposed research project will enhance our knowledge of the transcriptome of prostate cancer bone metastases and its response to systemic treatment. It will provide a translational benefit for patients with advanced prostate cancer. We expect, that our gene expression assay can act as a biomarker either on its own or in conjunction with traditional clinical or tissue-based parameters. In addition, we will show, that robust gene expression analysis is feasible from FFPE tissue from bone metastases thus facilitating further research in other tumor types that are prone to bone metastases. ","The general aim of this project is to improve our understanding of the mechanisms that regulate outgrowth or otherwise dormancy of PCa bone metastases. This is a major clinical problem as it accounts for late metastatic relapse after unpredictable periods of minimal residual disease. Our preliminary data, which are mainly based on our newly established spontaneous bone metastasis xenograft models of human PCa, and the literature collectively suggest that the outgrowth of PCa bone metastases might be suppressed by a paracrine and/or autocrine interplay of the soluble factors Sparc and Dkk1 that apparently inhibits tumor colonization by promoting the EMT-phenotype of metastatic cells (EMT=epithelial-to-mesenchymal transition). The first aim of this project will therefore be to obtain for the first time direct functional evidence whether the bone marrow (BM) colonization changes upon genetic modification (e.g., inducible knockdown) of Sparc or Dkk1 in our spontaneous metastasis PCa xenograft models (WP1). Our preliminary work demonstrates that our models closely recapitulate the process of EMT, that they have clinical and functional relevance, and that we are able to monitor the switch from dormancy to colonization in vivo. In WP2 we will determine whether Dkk1 exerts its effects on tumor cell dormancy not only by directly targeting the tumor cells, but also the BM stromal cells. For this purpose, we will use immunodeficient mice with mutated Dkk1-binding site of the main receptor Lrp5 (Lrp5-G170V) or with knockout of the co-receptors Krm1 and/or Krm2 for cell line-based PCa xenograft experiments. In addition, we will engraft these mice with our newly established patient-derived xenograft (PDX) model 'C5' that spontaneously metastasizes to the bone and expresses Dkk1 and Sparc. WP1 and WP2 are embedded into a long-standing, close collaboration between the applicant and the Institute of Osteology and Biomechanics at UKE (Prof. Thorsten Schinke). Prof. Schinke will contribute his expertise in static, dynamic and cellular histomorphometry and provide the Lrp5-G170V and Krm1/2 k.o. mice for WP2. WP1 and WP2 might demonstrate that elevated levels of Sparc and Dkk1 and Dkk1 receptor activation suppress the switch from dormancy to colonization. The expected results would provide a rationale for future experiments on how to maintain Sparc and Dkk1 expression in the phase of minimal residual cancer to prevent metastatic relapse (presumably in a second funding period). Moreover, in order to identify novel markers of metastatic outgrowth in PCa by an unbiased approach, we will use a recently established method of bioluminescence imaging-guided in situ laser ablation of xenograft primary tumors and corresponding bone metastases and subsequent proteome analysis (WP3, cooperation with Prof. H. Schlüter, UKE). The resulting candidates will be functionally validated in a second funding period. ","Adipocytes are metabolically active and their number increases with age and obesity, being associated with a variety of metabolic pathologies. For cancer, adipocyte-supplied factors are related to development and progression, in particular for tumors that grow in or metastasize to adipocyte enriched environments like bone. Therefore, our main research question is to elucidate on the role of adipocytes in the bone marrow microenvironment of osteolytic metastasis using small animal models.Our preliminary results indicate that melanoma growth within the bone is enhanced in an adipocyte-rich bone marrow environment, which is consistent with the finding that obesity derived factors induced osteoclastogenesis through cancer cell activation. Adipocytes would therefore adopt an unprecedented role in metastatic bone disease and would represent a novel cellular target to influence the progression of bone metastasis. In this context, investigations on the function of adipocytes in metastatic bone disease during breast cancer and malignant melanoma are needed. Therefore, the aims of the project are the assessment of adipocyte-derived factors as well as the determination of their influence on bone marrow microenvironment and tumor cell growth in the metastatic niche. For this purpose, in vivo and in vitro molecular assays as well as multimodal imaging will be used to investigate processes like tumor cell proliferation, bone resorption and angiogenesis in osteolytic bone metastasis in different models of expansion and loss of bone marrow adipocytes. ","Dickkopf-1 is best known for its role as a Wnt-Inhibitor that promotes osteolytic bone lesions by inhibiting osteoblast function. Our data indicate that DKK-1 has pleiotropic, context dependent effects during the steps of establishing bone metastases secondary to breast cancer. While direct anti-tumor effects were observed following DKK-1 knockout, key steps of the metastatic process like migration or the potential to form colonies were unaltered or even increased. In the proposed project, we aim to decipher these effects in detail, explain the underlying molecular mechanisms, and help to define the therapeutic and prognostic potential of DKK-1 in the complex process of bone metastases, covering the steps from homing to the manifestation of overt bone lesions. We will apply a range of in vitro methods to specifically address the effects of DKK-1 knockout or inhibition using a neutralizing antibody on vitality, migration, adhesion, invasion and colony formation. Our preliminary data show that many of these processes are distinctly influenced by DKK-1 modulation. Furthermore, it remains unclear how high DKK-1 levels, as found in the bone microenvironment, may influence our findings. Therefor the importance of osteoblast-derived DKK-1 will be specifically addressed using co-culture and supernatant experiments.These experiments will be complemented by a range of murine models. Specifically, influence of DKK-1 on homing will be assessed using breast cancer cell lines depleted of DKK-1 using CRISPR/Cas9 and compared to the potential of these cells to establish overt bone metastases. These experiments will be expanded by the application of an inducible DKK-1 knockout that will be applied only once metastases have been established in the bone. To dissect effects of bone-derived DKK-1, our osteoblast-specific DKK-1 knockout mice will be tested.In a last step, serum samples of approx. 500 breast cancer patients will be measured for DKK-1. For these patients full data with regard to the presence of disseminated tumor cells (DTCs) in the bone marrow is available as well as full clinical follow-up. The prognostic value of DKK-1 for occurrence of bone metastases, survival and other parameters of clinical outcome will be assessed in the context of DTC presence. In addition, single cell deep sequencing data of disseminated breast cancer cells will be provided by our collaborator and assessed for their DKK-1 levels.In summary, using the above described approaches we aim to decipher the complex and pleiotropic effects of DKK-1 in the context of breast cancer bone metastases. ","Prostate cancer is one of the most common malignancies in men and has a high propensity for bone metastasis, predominantly osteosclerotic lesions. This osteotropism reflects a high degree of adaptation and osteoblast-like properties of prostate cancer cells within the bone microenvironment. However, the underlying molecular mechanisms are poorly defined. Our previous work has focused on the role of Wnt ligand Wnt5a in bone and bone marrow physiology, metabolic bone diseases, and cross-talks between bone and prostate cancer. In particular, we found that Wnt5a exerts anti-prostate cancer effects in vitro, reduces prostate cancer growth in the skeleton of mice, and is associated with longer survival in men with prostate cancer. We demonstrated that Wnt5a employs both canonical and non-canonical signaling pathways in prostate cancer cells, including Frizzled (FZD) receptor 5 and the atypical receptor tyrosine kinase RYK, but not receptor tyrosine kinase-like orphan receptors (ROR) 1 and 2.Importantly, these studies addressed the role of Wnt5a in the tumor itself. However, the bone microenvironment and especially osteoblasts produce large amounts of Wnt5a, which may govern tumor cell homing into the bone microenvironment and/or tumor-bone interactions. Thus, to date it has remained unclear which cells in the bone microenvironment operate in Wnt5a signaling in bone metastases, and how targeted deletion of Wnt5a in mice affects the colonization and interactions of prostate cancer cells with bone. The central hypothesis is that non-tumor-derived Wnt5a from osteoblastic lineage cells controls critical aspects of prostate cancer cells in their interaction with the bone microenvironment. To test this hypothesis, we will (I) characterize bone-cancer interactions secondary to prostate cancer metastasis in conditional global and osteoblast-specific Wnt5a knockout mice using combined cellular, histological, biomechanical, and imaging analyses, (II) analyze the role of exosomes and involved cargo in bone-tumor cell cross-talks, and (III) investigate the impact of tumor-adjacent stroma tissue on prostate cancer colonization of the bone microenvironment with the support of a clinician-scientist working in the field of Tumor Orthopedics. This individual will make full use of human material obtained in our bone bank and the tissue bank from the Comprehensive Cancer Center Dresden.The results of this project will yield novel mechanistic insights into the role of Wnt5a and associated pathways in prostate cancer interactions with the bone microenvironment and the contribution of tumor vs. non-tumor tissues. This may result in a refined and perhaps revised view of Wnt signaling in bone metastases and could provide a molecular basis for therapeutic intervention. ","Osteocytes are the most frequent cell type in bone. Residing in lacunae, their protrusions span an area of about 1200 m². With this intricate network, osteocytes orchestrate skeletal adaptation to mechanical stimuli and regulate bone mineral homeostasis through intense communication with osteoblasts and osteoclasts. Recent studies indicate that the bone composition is altered by prostate cancer and that material stiffness is an important factor for the metastatic process. However, little is known about the characteristics of the osteocyte-lacunar network in bone of metastatic entities and how this affects tumor cell behavior. The central hypothesis is that the osteocyte network and functionality are affected by skeletal metastasis and these alterations contribute to pathological bone cell actions and the resulting bone fragility. To test this hypothesis, we will (I) characterize the osteocyte network, local mechanical properties, and bone ultrastructure using 2D/3D imaging/microanalysis during bone metastases formation, (II) determine the impact of osteocyte functionality on the bone metastatic process, (III) assess the bidirectional communication between osteocytes and tumor cells and the role of mechanosensation therein in vitro, and (IV) analyze the adhesion molecule profile of naïve osteocytes and tumor-primed osteocytes to identify targets to modulate osteocyte-tumor cell interactions. Key findings will be validated in bone metastases biopsies obtained from breast- and prostate cancer patients using the Hamburg bone registry. This project will provide novel and detailed insights into the interaction between osteocytes and tumor cells and may provide new therapeutic strategies to inhibit the formation of bone metastases via modulation of osteocyte/tumor cell cross-talks. ","Interdisciplinary collaboration, shared expertise, scientific exchange, integration of early career scientists, and clinical translation are key ingredients for successful and dynamic work within the SPP2084. Active communication will be essential to ensure effective networking between the individual sites within the consortium. This requires regular face-to-face meetings at annual retreats, symposia, and workshops as well as everyday communication measures like webinars and skype and phone conferences. A webpage(www.microbone.de) and a proactive administrative coordinator will foster communication and the networking process. Major ambitions of the SPP2084 are strong interactions between the projects and the support of young researchers. We wish to realize this by establishing professional boot camp-type workshops and training courses. Shortterm lab rotations will also be important to share key technologies in tumor and animal models as well as imaging techniques. In addition, we will specifically promote PhD students by offering expert classes on scientific topics and soft skills and by ensuring progress and direction using thesis advisory committees (TACs). Moreover, participation at the DKTK-School of Oncology will provide specific knowledge and scientific skills in translational oncology for physicians and scientists. To guarantee the best synergistic outcome of both basic and translational research, we will further foster bidirectional networking between basic scientists and clinicians. We therefore apply for clinician-scientist positions, which will be allocated to individual projects for up to 12 months based on documented excellence with a proven track record in clinical translation. These positions will strengthen clinical translation by human validation of animal research. To make this consortium attractive for female scientists and young parents, we intend to offer an intensive mentoring program as well as a “Flying nanny” childcare service for work-related emergencies. To further support women during or immediately after pregnancy, we will specifically incorporate flexible bridge funding by applying for a technician, which will be allocated to individual approved projects within the μBONE consortium. These central measures serve the entire consortium and should help to ensure efficient and productive working conditions for all involved scientists. ","The bone metastasizing capacity of breast cancer (BC) is subtype dependent. Moreover, only little is known about the biology and molecular principles of BC metastasis and it remains elusive, how breast cancer cells modify and exploit the hematopoietic stem and progenitor cell niche in the bone. Since the underlying functional mechanisms of this phenomenon are largely unknown, we plan to dissect the implication of plasma membrane protrusions emerging from BC cells in migration, adhesion and invasion of bone marrow microenvironment. The morphological impact of factors and/or signaling pathways that modulate the metastatic capacity of cancer cells will be evaluated. As cellular representatives of bone marrow niches, mesenchymal stromal cells (and osteoblasts derived therefrom) will be used. Mechanisms of intercellular communication between BC cells and bone-derived cells will be investigated as well and a particular attention will be devoted to thin membrane processes that could act as tunneling nanotubes. For clinical translation, we will study the role of those plasma membrane protrusions and communication pathways with regard to drug response, metastatic potential and the intrinsic molecular subtypes of BC. ","The bone matrix is continuously remodeled through the balanced activities of two substantially different cell types, bone-forming osteoblasts and bone-resorbing osteoclasts. Our Institute has a long expertise in bone-specific cellular and molecular characterization of genetically modified mouse models, and we have identified several key mechanisms controlling skeletal remodeling in the last years. For the present proposal we will continue our ongoing collaboration with the group of Prof. Klaus Pantel to address the question, if and how an impaired bone remodeling status affects tumor cell dissemination and metastatic outgrowth in vivo.For that purpose we will inject human and murine breast cancer cells into mice to study skeletal metastasis formation as well as changes in the bone microenvironment by Luciferase reporter gene assays, µCT, undecalcified histology and bone-specific histomorphometry. Most importantly, and in contrast to the majority of previously published studies, we do not aim at modifying the seed (i.e. the tumor cells), but the soil (i.e. the recipient mouse). To achieve our goals we have backcrossed different mouse models into genetic backgrounds (BALB/c and NSG) allowing xenotransplantation or syngeneic transplantation of cancer cells. While three of the models (Calca-/-, Calcr-/- and Notch2+/HCS) display specific disturbances of the coupling mechanisms required to synchronize bone formation and bone resoprtion, the other four models (Lrp5+/HBM, Col1a1-Sost, Col1a1-Krm2 and Col1a1-tTA;pTet-Wnt1) display selective changes in osteoblast activity. Since all these genetically modified mouse models have been previously analyzed to define not only their bone remodeling status, but also the molecular causes of the observed phenotypes, we expect that our results will clearly identify, which bone remodeling cell type is primarily involved in the suggested detrimental crosstalk between bone and tumor cells.In addition to this in vivo approach we will perform ex vivo co-culture experiments with breast cancer and bone remodeling cells to define specific molecular interactions. Although our project principally follows an unbiased approach, there are some hypotheses that will be addressed. For instance, since we have previously found that calcitonin (CT) controls bone formation by inhibiting the release of sphingosine 1-phosphate (S1P) from osteoclasts, we will analyze if CT, S1P and/or antagonists of specific S1P receptors will influence proliferation and/or migration of cancer cells, cultured alone or together with bone remodeling cell types. ","The growth of cancers in the skeleton is associated with severe effects on bone and results in considerable morbidity and mortality. Both, breast cancer (BC) as well as multiple multiple (MM)-derived bone metastases are osteolytic and generation of metastases is mediated by the formation and activation of mature osteoclasts.There is a complex crosstalk between tumor cells and the bone microenvironment providing a variety of growth factors for cancer cells and osteoclastic factors. In this regard inhibition of TGFß or activin A, both members of the TGFß familiy, suppress osteoclastic bone resorption in mouse models of MM and BC, indicating an important role of these factors in the formation of bone metastases.In this context, we have recently discovered that myostatin, another member of the TGFß family, strongly enhances osteoclast development and that deficiency or inhibition of myostatin highly ameliorates disease severity and in particular bone erosion in arthritic mice. Interestingly, preliminary experiments revealed a high similarity between mystatin and action A. We could show that myostatin and activin A enhance RANKL-induced osteoclast formation to the same extent, indicating that both factors are equivalent promoters of osteoclast development. Moreover, both factors act via similar Smad-pathways (Smad2/3), pointing to a common mechanism in osteoclast differentiation. Beside this similarity, additional lines of evidence suggest a role for myostatin in bone metastases: 1. Myostatin is expressed in breast cancer cells, 2. ActRII, the receptor type shared by activin and myostatin, is expressed by breast cancer and myeloma cells, 3. Higher levels of phosphorylated SMAD2 are present in human BC bone metastatases.We hypothesize, that myostatin participates in the interactions of tumor cells with the bone microenvironment, promoting osteoclast development and thereby bone lesions and that inhibition of myostatin reduce bone metastases.For this purpose, we will treat two syngeneic and two xenograft models for both BC and MM with anti-myostatin antibody and assess the formation of bone metastases. Moreover, the role of myostatin in cell-cell interaction will be investigated and direct/ indirect effects of tumor cells on osteoclast differentiation and activity will be tested. Furthermore, assuming that both myostatin and activin A are involved in bone metastases, inhibiting myostatin and activin A simultaneously may provide a powerful tool for the treatment of bone metastases in BC and MM. A protein which is able to bind and antagonize myostatin as well as activin is a natural occuring antagonist called „Follistatin-related gene“ (FLRG). In preliminary experiments, we could demonstrate that FLRG completely abrogates RANKL-induced osteoclast formation either stimulated with myostatin and activin. We therefore proposed to treat BC and MM mice also with recombinant FLRG in the course of the disease and analysed as described above. ","Sufficient oxygen pressure is required for our organs to function properly. Conversely, insufficient oxygen supply (hypoxia) is a prominent feature in various pathological processes, including tumor development and metastasis. The central mediators during deprived oxygen are hypoxia inducible factors (HIF) whereas their downstream effects are tightly regulated by oxygen-dependent HIF prolyl hydroxylases (PHDs). Previously, we and others revealed that PHD2 plays a central role during different stages of tumor development, whilst this oxygen sensor is also essential during bone mineralization, and normalization of the endothelial barrier in the bone (marrow) after stress. In this project we aim to test the hypothesis that modulating hypoxia in the tumor and/or bone microenvironment via PHD2 will affect the bone metastatic process. To test this hypothesis, we will (i) define the role of PHD2 in prostate cancer cells using knock-down and overexpression of PHD2 on the formation of bone metastases and determine the impact on the bone vasculature and local bone turnover. Downstream pathways will be determined with an additional knock-down of HIF proteins. Second (ii), we want to assess the role of endothelial cell-derived PHD2 and (iii) the role of osteoblast-derived PHD2 in the bone metastatic process and the triad interactions between tumor cells, angiogenesis, and osteoblasts/bone turnover. This project will provide novel insights into the role of hypoxia pathway proteins in the interaction between tumor cells, bone vasculature and bone cells and may provide new therapeutic strategies to interrupt the malignant crosstalk and ameliorate the formation of bone metastases. ","A hallmark of multiple myeloma (MM) is the reciprocal “vicious” interaction between PC and osteoclasts (OC). This cross-talk leads to disease progression, immunosuppression and osteolytic bone disease. Despite significant advances in the treatment of MM the disease remains incurable in the majority of patients. Therefore, the discovery of novel mechanisms promoting myeloma and its associated bone disease are still urgently needed. In this context, the development of an increased understanding of the interaction between myeloma cells, immune cells and bone cells is of special interest because immune cells hold great potential to destroy malignant cells. However, the regulation and therapeutic activation of anti-myeloma immune responses in the bone marrow is understudied.Our preliminary data show that Protein S (ProS) promotes the proliferation and survival of MM cells. At the same time, it fosters the activation of OC via the receptor Mer, which belongs to the TAM receptor family. Genetic blockade of ProS reduces myeloma burden and prolongs the survival of mice bearing a systemic MM model. Importantly, MM-induced bone disease is almost completely abrogated upon neutralization of ProS. Thus, the blockade of the ProS-Mer axis holds potential to reduce myeloma burden and decrease MM-induced OC activation. In addition, we found that ProS induces an immunosuppressive phenotype of OC. Based on these data we propose a detailed functional investigation of the underlying molecular mechanisms and of the therapeutic potential of the clinically applicable small molecule Mer inhibitor R992 for treatment of MM with a special focus on bone disease. In addition, we will dissect the role of the two alternative TAM receptors Axl and Tyro 3 in osteoclastogenesis. For this purpose, we will use co-cultures and mouse models of MM with OC-specific blockade of TAMR by genetic approaches. Furthermore, we will evaluate the therapeutic effect of TAMR blockade on MM progression as well as on its associated bone disease and immunosuppression. Ultimately, we will translate our findings from the bench to the bedside by analyzing the association of ProS and TAMR with prognosis and MM-induced bone disease.The project will be in particular linked to the groups of L. Hofbauer (Dresden) and A. Garbe (Dresden), with whom we will form an osteoimmunology network. By assessing overlapping cell types and molecular mediators we expect a significant knowledge gain concerning whether the interplay between bone and immune cells is via different or overlapping mechanisms in solid cancer versus MM. Furthermore, we will build a strong bidirectional link to D. Hose (Heidelberg), who will contribute a unique MM patient cohort allowing us to assess the association of the ProS/TAMR axis with MM bone disease. Conversely, we will investigate novel mediators of MM-induced bone disease derived from this patient cohort in our functional mouse models. ","While primary tumors can be treated with increasing success, >80 % of all cancer-related deaths are caused by metastatic tumors. A diverse arsenal of new approaches to address this most significant problem for human health is required. We focus on one specific approach and one area of cancer: bone metastasis, and we propose that the control of F-actin cytoskeletal dynamics is of central importance for tumor metastasis and bone interactions. Having long-standing expertise in research on F-actin control, we are focusing on an F-actin modulatory protein, SWAP-70, that we recently found to be clearly involved in tumor cell metastasis. Deletion of SWAP-70 significantly reduced metastasis in two mouse cancer models.Thus our overall hypothesis is that SWAP-70 controlled F-actin dynamics contributes significantly to bone metastasis. Thus, we aim at understanding SWAP-70 dependent bone metastatic processes. Our long-term aim is to develop new therapeutic approaches based on preventing SWAP-70’s function in bone metastasis.Our immediate specific aims for the 1st funding period therefore are to:(1) Determine the role of SWAP-70 in tumor cell bone metastasis in vivo(2) Define SWAP-70 dependent, F-actin related features of bone metastatic tumor cells in vitroWe have all the necessary tools and methods in place, e.g. standard and conditional knockout mouse models, an orthotopic cancer model, a large variety of SWAP-70 mutants, Crispr/Cas-generated SWAP-70 deficient tumor cells, and a plethora of analysis technologies. We expect to determine important features of a pathway that has been largely overlooked in the context of tumor bone metastasis. ","The most common metastatic site of prostate cancer (PCa) is the bone associated with high morbidity and lethality of patients. With the aim to identify novel therapeutic targets which prevent or treat metastasized PCa, major efforts are being made to better understand the molecular mechanisms enabling bone metastatic transition of PCa cells. Besides well-known genetic alterations of oncogenes and tumor suppressor genes, aberrant activity of regulatory elements for gene transcription, post-translational modification and protein degradation mainly contribute to the metastatic potential of PCa cells. Amongst others, the family of tripartite motif containing proteins (TRIMs) have been identified to be crucial regulators of cellular processes driving metastasis, and to possess altered activity in diverse cancer types. In our preliminary work, we found TRIM24 to be highly overexpressed in PCa bone metastases and to predict outcome. In addition, TRIM8, TRIM11, TRIM47 and TRIM68 have been described be overexpressed in PCa or to influence mechanisms driving metastasis. Based on these data, our overall hypothesis is that these specific TRIM proteins have a crucial role in invasiveness and metastatic niche formation of PCa. To test our hypothesis, we will pursue three specific aims. First, we will characterize the expression and genetic status of TRIMs in PCa tissues including primary tumors as well as metastases. Using large and clinically well-characterized cohorts, we will correlate our molecular data with diverse clinic-pathological parameters and patients’ survival. Second, in-vitro experiments using bone metastatic PCa cells will be conducted to investigate the functional role of TRIMs. We will explore whether cellular features which are required for crucial steps during metastasis are influenced by specific TRIMs. Detailed analysis of signaling molecules involved in these processes will allow conclusions about the underlying molecular mechanisms of TRIM mediated metastatic potential of PCa. Third, co-cultivation experiments will be applied to investigate whether TRIMs influence the communication between bone metastatic PCa cells and cells forming the bone microenvironment. Results of this project will contribute to a better understanding of molecular processes promoting PCa metastasis to the bone. Due to the incurable nature of metastatic PCa to date, targeting specific molecules like specific TRIM proteins might be a therapeutic option for patients suffering from ","We and other research groups have shown that bone marrow (BM) is a common homing organ for disseminated tumor cells (DTCs, the seed of metastases) derived from various types of malignant epithelial tumors. This includes tumors, which do not typically form metastases in bone like colon cancer. In several studies, we have shown that the presence of single DTCs in the BM is an independent predictor of metastatic relapse in many different solid tumors including breast cancer. We have furthermore developed internationally accepted assays to detect and characterize DTCs as well as circulating tumor cells (CTCs) at the single cell level. Moreover, we have identified genes, expression signatures and mechanisms associated with and involved in breast cancer BM micrometastasis, including RAI2 for breast cancer. The retinoic acid–induced 2 (RAI2) was shown not only significantly associated with DTC status but also poor prognosis. BM represents a retinoic acid–rich microenvironment where it regulates differentiation and self-renewal of hematopoietic stem cells. DTCs might be exposed to the same regulatory mechanisms and therefore the RAI2 protein might also be involved in the dormancy control and of overt bone metastasis formation. Our functional studies have furthermore shown that RAI2 sustains differentiation of ER-positive tumor cells, whereas loss of RAI2 induces hormone independent growth and activation of AKT signaling as an important mediator of dormancy control. RAI2 may also regulate the expression of bone morphogenic factors (BMPs) –factors important for breast cancer- bone interaction crosstalk. Therefore, our results indicate that loss of RAI2 expression might represent a so far undiscovered key event at the onset of metastatic progression in the bone. The primary goal of this project is to better understand the precise role of RAI2 for the development of bone metastasis in breast cancer. The investigation of the RAI2-mediated crosstalk between DTCs and cells of the bone microenvironment could lead to better mechanistic insights into cancer dormancy. This could eventually lead to new therapeutic strategies to prevent outgrowth of dormant DTCs into overt bone metastases. Since BM is a reservoir for DTCs that may also recirculate to other metastatic sites, this project may also have implications beyond blocking bone metastasis formation. More specifically we plan to a) investigate the role of RAI2 in the crosstalk between tumor and bone cells, b) perform functional analyses of the role of RAI2 mediated AKT activation in survival and dormancy of DTCs in BM, c) investigate the role of TGFβ and BMP pathways in RAI2-mediated bone metastasis formation and d) establish RAI2 mouse models and assess the functional in vivo role of RAI2 in bone metastasis formation. Thus, the results obtained in the ongoing and future grant will contribute to a better understanding of cancer dormancy and metastatic development in the BM microenvironment. ","Despite major advances in the treatment of localized cancer, disseminated disease remains one of the most challenging clinical issues, with limited treatment options and poor prognosis for patients. Dormancy at metastatic sites represents a major therapeutic hurdle as standard therapy fails at targeting quiescent cells. Recent insights emphasize the critical role of stromal cells in regulating primary tumor progression and organ-specific metastasis. Besides cellular mediators, components of the extracellular matrix (ECM) have recently attracted attention. Tumor-derived SPARC expression was proposed to maintain prostate cancer cell dormancy in the bone marrow. More recently, stromal expression of SPARC was shown to associate with shorter PFS and OS in metastatic breast cancer. We therefore propose to address the role of niche-derived SPARC in metastatic dormancy using state-of-the-art PDX models, including a newly generated NSG SPARC knock-out line and a fully humanized bone marrow 3D model, both generated by our lab. We will evaluate whether SPARC deficiency triggers the exit of metastatic cells from dormancy thereby sensitizing them to therapeutic targeting using standard of care (e.g. chemotherapy, irradiation). Based on our previous findings, loss of niche-derived SPARC would concomitantly protect hematopoietic stem cells from the detrimental effects of chemotherapy. Lastly, functional CRISPR screens will be performed to unravel downstream mediators of SPARC on metastasis, thereby identifying potential novel therapeutic targets. ","Breast and prostate cancer cells home to the bone marrow, where they presumably hijack the hematopoietic stem cell niche, and develop metastatic lesions. We aim to characterize the elusive premetastatic niche by examining the role of mesenchymal stromal cells (MSCs) in cancer cell homing and growth of bone metastases. Decreasing the number of MSCs pharmacologically enhanced cancer cell homing to the bone marrow in mice. Initial characterization confirmed a relationship with a mesenchymal subpopulation. We would like to characterize this subpopulation further. This will be followed by both genetic and pharmacologic manipulation in mice in order to increase the identified subpopulation and diminish cancer cell homing. Preliminary work also suggests that this subpopulation suppresses cancer growth in an in vivo model of breast cancer metastasis in mice. We therefore aim to characterize how this subpopulation modulates the immune response towards cancer in vivo. In addition, we will also evaluate whether modifying matrix components and matrix receptors affects the immune cell response to cancer and/or growth. The goal is to take advantage of the lessons learned in this work to maximize the immune inhibitory effect on cancer. Using bone marrow samples from patients with breast and prostate carcinoma we will finally confirm the relationship between stromal population changes and cancer cell homing prospectively. In addition, we will try to confirm data obtained on the contribution of stromal cells to metastasis growth in mice on human cancer samples.","Multiple myeloma (MM) is a B-cell malignancy of clonally expanding antibody producing plasma cells that ultimately leads to osteolytic bone lesions, bone destruction, hypercalcemia, kidney failure, anemia and infections. Despite advances in conventional therapy, clinical application of novel agents and high dose chemotherapy supported by autologous stem cell transplantation MM remains an incurable and highly fatal hematologic disease. MM is characterized by dissemination of multiple tumor cells throughout the bone marrow (BM) and signaling through adhesion molecules regulates MM – BM-microenvironment interactions and might therefore be involved in the dissemination process. We postulate that family members of the junctional adhesion molecules (JAMs) play a pivotal role for MM survival, progression and dissemination by facilitating the crosstalk with BM endothelial cells and we will address the following specific aims: (I) To elucidate and visualize MM cell interactions with the BM endothelium regarding the involvement of JAM-C. (II) To analyse the expression/signaling profile in phenotypically heterogeneous MM subclones. (III) To decipher signaling pathways and their impact on drug sensitivity, homing and dissemination. To achieve these aims we have established models of orthotopic syngeneic and humanized MM, non-invasive bioluminescence imaging and advanced microscopy techniques (e.g. high-resolution 3D light-sheet fluorescence microscopy). Imaging and microscopy will direct further cellular (e.g. multi-parameter flow cytometry, functional in vitro assays) and molecular in vitro analyses. Our studies will help to refine therapeutic interventions, in particularly immunotherapy of MM bone disease. Ultimately, we envision that a better understanding of the pathophysiology of JAMs in MM will result in optimal combinatorial treatment strategies to overcome drug resistance in patients suffering from MM and help to establish semi-personalized anti-MM strategies. ","The aim of this project is to better understand cancer cell colonisation into bone and to unravel unknown interactions between tumour, immune and bone cells in the bone niche. To elucidate such interactions, in vivo imaging of transplanted breast cancer cells in larval zebrafish will be used. In general, in vivo imaging is a powerful tool in order to demonstrate interplay of cells, also in cancer research. In mice, intravital imaging of tumors has revealed that there is a physical connection of tumor-associated macrophages (TAM) with cancer cells, both during initial invasion of tissues and intravasation of cancer cells, i.e. their entry into the blood stream. During these stages mutual dependence by way of positive feedback loops between cancer cells and TAM has been shown. However, the role of TAM for the cancer cells' exit from the blood stream (extravasation), as well as during colonisation of bone is less well understood. By monitoring the interaction of tumor cells (breast cancer cell lines, CTCs), macrophages and bone cells in living zebrafish bone tissue, the following questions will be addressed: i. How do TAM help cancer cells to extravasate and to initially grow in the bone niche? ii. How do TAM contribute to altered osteoblast and osteoclast function during metastasis? To answer these questions, spinning disc confocal microscopy of transgenic zebrafish, in which the respective cell types are fluorescently labelled, will be used. In these fish, interaction of macrophages with bone and cancer cells can be monitored. This approach will be combined with in vivo bone staining techniques and subsequent bone composition analyses in order to characterize bone lesions at the site of metastasis. Furthermore, changes in macrophage number and function will unravel the molecular links of TAM and cancer cells. Using these tools, the role of TAM in bone metastasis formation will be further elucidated which will help to modulate","Breast cancer is one the most prevalent malignancies worldwide, frequently leading to distant metastases even decades after treatment of the primary tumor and a subsequent disease-free interval. Bone is a common site for breast cancer metastases, causing an incurable stage of the disease and ultimately osteolytic skeletal destruction with pathological fractures and pain. In the bone microenvironment, breast cancer cells severely alter the balanced osteoblast-dependent bone formation and osteoclast-mediated bone resorption by the production of secreted molecules such as Parathyroid hormone-related protein (PTHrP), which stimulates osteoblasts to secrete receptor activator of NFκB ligand (RANKL) and other cytokines. RANKL increases osteoclast activity and subsequent bone resorption during which matrix-derived growth factors are released into the metastatic microenvironment. These factors further stimulate cancer cell proliferation, which perpetuates metastatic bone destruction. At the stage of bone metastases, anti-resorptive drugs like bisphosphonates and an anti-RANKL antibody are used in the clinics to restrict osteolytic bone disease. However, more insights into the cellular and molecular mechanisms of the interaction between breast cancer cells and bone cells within the bone microenvironment and the process of cancer cell colonization into bone is needed, to better understand the process of metastases progression. Using a comprehensive set of in vitro and in vivo approaches we determined that PTHrP and RANKL induce the expression of the homeodomain protein TG-interacting factor 1 (Tgif1) in osteoblasts and osteoclasts, respectively, and firmly established Tgif1 as a novel regulator of osteoclast function and bone resorption. In addition, formation of breast cancer metastases in bones of Tgif1-deficient mice was greatly impaired. These findings strongly suggest a functional role of Tgif1 in the bone microenvironment in the context of breast cancer-induced bone disease. Furthermore, our results show that Tgif1 protein abundance can be reduced by delivery of synthetic microRNA-23a/b (miR-23a/b) mimics, an approach that could be of translational value for the treatment of metastatic bone disease. We therefore hypothesize that Tgif1 affects the cellular cross talk in breast cancer-induced pathological bone remodeling. In addition, we propose that Tgif1 may serve as a therapeutic target to reduce osteolytic bone disease, at least in preclinical models. By combining in vitro experiments, innovative in vivo studies and patient samples we aim to i) investigate the role of Tgif1 in cancer-induced bone destruction in vitro and in vivo, ii) target Tgif1 by miR-23a/b replacement therapy to alleviate metastatic bone disease, and ii) analyze miR-23a/b-Tgif1 signaling in breast cancer patients. This research proposal is original, highly innovative and very likely to expand our knowledge on the mechanisms of breast cancer metastases to bone. ","Disseminated prostate cancer cells (DCCs) in bone marrow (BM) comprise the founder cells of later arising lethal bone metastases and understanding the biology of BM-DCCs is paramount for an improved clinical management of prostate cancer (PC) patients. We found that DCCs in prostate cancer displayed significant genetic heterogeneity. Notably, preliminary analysis of the transcriptome indicates that DCCs in PC express a mixed epithelial/haematopoietic phenotype, suggesting a complex patter of adaptation to the ectopic BM microenvironment. We have collected genomes and transcriptomes from almost 400 PC BM-DCCs. In this project we will use novel single cell sequencing technologies to characterize genomes and transcriptomes of BM-DCCs. Furthermore, we will correlate the molecular data with the clinical outcome, and in particular look for (i) genetic aberrations and mutations enabling metastatic progression, and (ii) pathways responsible for quiescence or proliferation of DCCs. For this part of the project, we will collaborate with the team of Ana Dubrovska, and exchange the molecular data obtained from DCCs and circulating tumor cells specific for different stages of the disease. In collaboration with the team of Gelinsky/Alblas we will use their in vitro culture system for BM to model the biology of DCC. We will use prostate cancer and patient-derived metastatic cell lines to establish conditions for expansion of early metastasis founder cells. The resulting in vitro model of minimal residual prostate cancer will enable us to perform further functional assays and drug screens aiming to eradicate the lethal seed of metastasis before clinical manifestation. ","Once prostate tumor spreads to the bone, it typically cannot be cured. The supportive sites in bone maintain survival and proliferation of cancer cells by special molecular components such as C-X-C motif chemokine ligand 12 (CXCL12) which also mediates tumor cell homing to bone. In turn, metastatic PCa cells in bone express different proteins that regulate both, bone remodeling and tumorigenesis including Endo 180 collagen receptor. Prostate tumors have a hierarchical organization where cancer stem cells (CSC) maintain and propagate the tumor mass. A number of studies showed a high radioresistance of CSCs compared to the tumor bulk, and many recent clinical studies use CSC related biomarkers for prediction of radiotherapy outcome. Tumor metastases are driven by the evolved populations of CSC at their worst. In support of this, cells representing different levels of metastatic processes including circulating tumor cells (CTC) in the blood of cancer patients and disseminated tumor cells (DTC) in their bone marrow express CSC markers, and metastasis initiating cells are sharing key CSC features including ability to form tumors and to activate CSC specific signaling mechanisms. Our studies revealed a few proteins that are involved in regulation of metastatic and radioresistant prostate tumor cells. That includes aldehyde dehydrogenase protein 1A1 (ALDH1A1) which is a marker of radioresistant CSC and has a high expression in PCa metastases. Another example is CXCL12 receptor, CXCR4, which is a marker of CSC, upregulates in CTC after radiotherapy and correlates with radiotherapy outcome in PCa. Finally, expression of collagen receptor Endo180/MRC2 which is one of the key regulators of PCa metastases is upregulated in highly tumorigenic radioresistant PCa cells. We assume that activation of certain signaling pathways detected on the genomic and transcriptomic levels in bone-marrow DTC, CTC or CSC in primary tumors can be prognostic for bone metastasis development and, on other hand, can be involved in the bone remodeling and response of metastases to radiotherapy. The first objective of this study is investigation of the role of signaling pathways mediated by CXCR4, ALDH1A1 and Endo180 for regulation of bone metastasis-initiating and radioresistant prostate tumor cells and bone remodeling, and possible interaction of these mechanisms during metastatic development. The second objective of this study is an unbiased and systematic analysis of genomic and transcriptomic profiling of prostate tumor cells representing different levels of metastatic processes (primary tumors, CSC, CTC, DTC, radioresistant PCa cells) to validate activation of CSC related signaling pathways. These mechanisms and phenotypes will be validated by using CRISPR/Cas9 mediated knockout of candidate genes, metastatic mice models, monitoring of bone remodeling in vitro and in vivo, and correlation of candidate genes to clinical data and patients’ response to therapy. ","Multiple Myeloma is a disease of malignant plasma cells spreading and accumulating in the bone marrow with subsequent induction of bone disease and angiogenesis. These features are in common with other malignant diseases. Multiple myeloma is different in two regards: Myeloma cells (MMC) spread to and live in the same “natural environment” as their physiological counterpart, bone marrow plasma cells (BMPC). Further, as shown by us, MMC rely at least partly on normal plasma cell functions for malignant behaviour when inducing angiogenesis and interacting with bone turnover. Malignant like normal plasma cells depend on receiving survival-support by interaction with other cell types in the bone marrow, collectively called “niche”. In vitro, we have shown in myeloma/ mesenchymal stromal cell (MSC)//osteoblast co-cultures that this interaction is prominently exerted by physical interaction and leads to changes in expression and behaviour in both cells types. We hypothesize that this physical interaction creates niches and elicits individual signatures that strengthen adhesion, support tumor cells, and disrupt osteogenesis, clinically leading to bone loss and pathological fractures. Niches may confer dormancy or resistance (minimal residual disease) features to myeloma. We have shown this molecular crosstalk being drugable exemplified by blocking the osteocyte-specific wnt antagonist sclerostin, observing increased osteoblast numbers and bone formation in myeloma bearing mice. We also showed that the contact-induced KISS1R system is a putative target as a theranostic PET tracer.In this application, we will focus on surface molecules featuring the interaction between normal and malignant plasma cells and MSC/osteoblasts. We will use expression analysis including single cell RNA-seq to address population heterogeneity, and functional assessment of interaction by atomic-force microscopy and CRISPR/CAS9 screen with subsequent collaborative in vivo validation in myeloma mouse models. In a second step, our in vitro findings will be related to a large cohort of molecularly characterized malignant plasma cell samples alongside whole-body imaging assessing bone disease, plasma cell infiltration and angiogenesis. We have shown that relation of data at nano-/micro- to large scale clinical data sets can act as a filter for relevance and will allow identification of pre-clinically applicable findings. The first is evidenced by sclerostin being expressed in osteocytes only, in contrast to malignant plasma cells from 630 MM patients; the second by our co-development of the T-cell bispecific antibody EM801/901 against the BMPC/MMC cell-surface molecule BCMA entering clinical trials in Q1/18, building the basis for a separate application for development of a theranostic BCMA-PET-Tracer. We will contribute to a collaborative network for myeloma dissemination, bone disease and angiogenesis related research questions within the µBone-consortium. "]

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["SPP 2084 : µBONE: Colonization and interaction of tumor cells within the bone microenvironment "]
["SPP Description :  Cancer is one of the leading causes of death worldwide. Despite medical progress in cancer therapies, most patients suffer and die from metastases at distant locations. Forming bone metastases are charac-teristic features of prostate and breast cancer. Bone metastases frequently lead to fractures, pain, and a poor life quality and are also associated with a high socioeconomic burden. The key molecular steps resulting in bone metastasis, including the initial colonisation of bone by tumor cells and the early in-teraction with bone and bone marrow cells are poorly understood. Once established, bone metastases represent a point of no return, are not curable, and may be a new source for systemic relapse. There-fore, this Priority Programme aims to generate new insights into (1) the mechanisms of tumor cell col-onisation to the bone marrow and (2) the interactions of tumor cells with the bone microenvironment to identify novel therapeutic concepts for the treatment and/or prevention of bone metastases. The focus is on bone metastasis secondary to breast and prostate cancer as well as myeloma bone disease. "]
["#","Project Title","Subject Area","PI Information",null,null,null,"Project Summary"]
[null,null,null,"Name","Institution","Contact","Email"]
[1,"Development and validation of a gene expression assay as prognostic and predictive biomarker in prostate cancer bone metastases ","Pathology, Hematology, Oncology ","Dr. Verena-Wilbeth Sailer","Universitätsklinikum Schleswig-Holstein","-","-","Skeletal metastases from prostate cancer are a major health burden worldwide and severely affect a patient’s quality of life. They harbor molecular alterations distinctly different from the primary tumor. Moreover, systemic treatment itself can change the molecular landscape of metastatic disease. Thus, a tissue-based biomarker evaluating molecular changes in bone metastases rather than in the primary tumor to predict prognosis or response to –targeted– therapy could be of benefit for these patients. In this proposed research project, we will apply a gene expression assay to robustly detect differentially expressed candidate genes and patterns of pathway regulation in archival formalin-fixed, paraffin embedded (FFPE) bone metastases from prostate cancer patients, because FFPE material is usually the only available material. Ninety-six candidate genes have already been identified through our preliminary work. Gene expression will be studied using a highly sensitive novel array system, that uses color-coded barcodes to detect transcripts (NanoString nCounter®) and can be custom-built. Correlation of gene expression with clinical data will subsequently identify a panel of genes with prognostic and/or predictive potential. This gene panel will undergo retrospective as well as prospective validation in tumor tissue from bone metastases. In addition, RNA from circulating tumor cells will be tested to evaluate its potential as non-invasive biomarker (liquid biopsy) for patients, for whom metastatic tissue is not available. Functional validation using two prostate cancer cell lines (VCaP/PC-3), both derived from a prostate cancer bone metastasis, will be performed. For 5-7 overexpressed genes in the gene panel, we will perform transient knockdown using siRNA to evaluate the biological consequence of overexpression. Data generated in this proposed research project will enhance our knowledge of the transcriptome of prostate cancer bone metastases and its response to systemic treatment. It will provide a translational benefit for patients with advanced prostate cancer. We expect, that our gene expression assay can act as a biomarker either on its own or in conjunction with traditional clinical or tissue-based parameters. In addition, we will show, that robust gene expression analysis is feasible from FFPE tissue from bone metastases thus facilitating further research in other tumor types that are prone to bone metastases. "]
[2,"Regulation of Tumor Cell Dormancy in Human Prostate Cancer Bone Metastasis ","Anatomy, Hematology, Oncology","Professor Dr. Tobias Lange, Ph.D.","Universitätsklinikum Hamburg-Eppendorf ","+49 40 741052591","to.lange@uke.de","The general aim of this project is to improve our understanding of the mechanisms that regulate outgrowth or otherwise dormancy of PCa bone metastases. This is a major clinical problem as it accounts for late metastatic relapse after unpredictable periods of minimal residual disease. Our preliminary data, which are mainly based on our newly established spontaneous bone metastasis xenograft models of human PCa, and the literature collectively suggest that the outgrowth of PCa bone metastases might be suppressed by a paracrine and/or autocrine interplay of the soluble factors Sparc and Dkk1 that apparently inhibits tumor colonization by promoting the EMT-phenotype of metastatic cells (EMT=epithelial-to-mesenchymal transition). The first aim of this project will therefore be to obtain for the first time direct functional evidence whether the bone marrow (BM) colonization changes upon genetic modification (e.g., inducible knockdown) of Sparc or Dkk1 in our spontaneous metastasis PCa xenograft models (WP1). Our preliminary work demonstrates that our models closely recapitulate the process of EMT, that they have clinical and functional relevance, and that we are able to monitor the switch from dormancy to colonization in vivo. In WP2 we will determine whether Dkk1 exerts its effects on tumor cell dormancy not only by directly targeting the tumor cells, but also the BM stromal cells. For this purpose, we will use immunodeficient mice with mutated Dkk1-binding site of the main receptor Lrp5 (Lrp5-G170V) or with knockout of the co-receptors Krm1 and/or Krm2 for cell line-based PCa xenograft experiments. In addition, we will engraft these mice with our newly established patient-derived xenograft (PDX) model 'C5' that spontaneously metastasizes to the bone and expresses Dkk1 and Sparc. WP1 and WP2 are embedded into a long-standing, close collaboration between the applicant and the Institute of Osteology and Biomechanics at UKE (Prof. Thorsten Schinke). Prof. Schinke will contribute his expertise in static, dynamic and cellular histomorphometry and provide the Lrp5-G170V and Krm1/2 k.o. mice for WP2. WP1 and WP2 might demonstrate that elevated levels of Sparc and Dkk1 and Dkk1 receptor activation suppress the switch from dormancy to colonization. The expected results would provide a rationale for future experiments on how to maintain Sparc and Dkk1 expression in the phase of minimal residual cancer to prevent metastatic relapse (presumably in a second funding period). Moreover, in order to identify novel markers of metastatic outgrowth in PCa by an unbiased approach, we will use a recently established method of bioluminescence imaging-guided in situ laser ablation of xenograft primary tumors and corresponding bone metastases and subsequent proteome analysis (WP3, cooperation with Prof. H. Schlüter, UKE). The resulting candidates will be functionally validated in a second funding period. "]
[3,"The role of adipocytes in the bone tumor micro-environment ","Hematology, Oncology, Radiology, Nuclear Medicine, Radiotherapy, Radiobiology, Cell Biology ","Professorin Dr. Aline Bozec","Universitätsklinikum Erlangen AöR","+49 9131 8543141","aline.bozec@uk-erlangen.de","Adipocytes are metabolically active and their number increases with age and obesity, being associated with a variety of metabolic pathologies. For cancer, adipocyte-supplied factors are related to development and progression, in particular for tumors that grow in or metastasize to adipocyte enriched environments like bone. Therefore, our main research question is to elucidate on the role of adipocytes in the bone marrow microenvironment of osteolytic metastasis using small animal models.Our preliminary results indicate that melanoma growth within the bone is enhanced in an adipocyte-rich bone marrow environment, which is consistent with the finding that obesity derived factors induced osteoclastogenesis through cancer cell activation. Adipocytes would therefore adopt an unprecedented role in metastatic bone disease and would represent a novel cellular target to influence the progression of bone metastasis. In this context, investigations on the function of adipocytes in metastatic bone disease during breast cancer and malignant melanoma are needed. Therefore, the aims of the project are the assessment of adipocyte-derived factors as well as the determination of their influence on bone marrow microenvironment and tumor cell growth in the metastatic niche. For this purpose, in vivo and in vitro molecular assays as well as multimodal imaging will be used to investigate processes like tumor cell proliferation, bone resorption and angiogenesis in osteolytic bone metastasis in different models of expansion and loss of bone marrow adipocytes. "]
[4,"Dissecting the pleiotropic effects of Dickkopf-1 in homing and colonization \nof breast cancer cells to and within bone.","Endocrinology, Diabetology, Metabolism, Hematology, Oncology ","Dr. Andy Göbel","Universitätsklinikum Carl Gustav Carus Dresden","+49 351 4587595","Andy.Goebel@uniklinikum-dresden.de","Dickkopf-1 is best known for its role as a Wnt-Inhibitor that promotes osteolytic bone lesions by inhibiting osteoblast function. Our data indicate that DKK-1 has pleiotropic, context dependent effects during the steps of establishing bone metastases secondary to breast cancer. While direct anti-tumor effects were observed following DKK-1 knockout, key steps of the metastatic process like migration or the potential to form colonies were unaltered or even increased. In the proposed project, we aim to decipher these effects in detail, explain the underlying molecular mechanisms, and help to define the therapeutic and prognostic potential of DKK-1 in the complex process of bone metastases, covering the steps from homing to the manifestation of overt bone lesions. We will apply a range of in vitro methods to specifically address the effects of DKK-1 knockout or inhibition using a neutralizing antibody on vitality, migration, adhesion, invasion and colony formation. Our preliminary data show that many of these processes are distinctly influenced by DKK-1 modulation. Furthermore, it remains unclear how high DKK-1 levels, as found in the bone microenvironment, may influence our findings. Therefor the importance of osteoblast-derived DKK-1 will be specifically addressed using co-culture and supernatant experiments.These experiments will be complemented by a range of murine models. Specifically, influence of DKK-1 on homing will be assessed using breast cancer cell lines depleted of DKK-1 using CRISPR/Cas9 and compared to the potential of these cells to establish overt bone metastases. These experiments will be expanded by the application of an inducible DKK-1 knockout that will be applied only once metastases have been established in the bone. To dissect effects of bone-derived DKK-1, our osteoblast-specific DKK-1 knockout mice will be tested.In a last step, serum samples of approx. 500 breast cancer patients will be measured for DKK-1. For these patients full data with regard to the presence of disseminated tumor cells (DTCs) in the bone marrow is available as well as full clinical follow-up. The prognostic value of DKK-1 for occurrence of bone metastases, survival and other parameters of clinical outcome will be assessed in the context of DTC presence. In addition, single cell deep sequencing data of disseminated breast cancer cells will be provided by our collaborator and assessed for their DKK-1 levels.In summary, using the above described approaches we aim to decipher the complex and pleiotropic effects of DKK-1 in the context of breast cancer bone metastases. "]
[5,"Wnt5a signaling in the bone microenvironment in prostate cancer ","Endocrinology, Diabetology, Metabolism, Hematology, Oncology ","Professor Dr. Lorenz Hofbauer","Universitätsklinikum Carl Gustav Carus Dresden","+49 351 4583173","lorenz.hofbauer@uniklinikum-dresden.de","Prostate cancer is one of the most common malignancies in men and has a high propensity for bone metastasis, predominantly osteosclerotic lesions. This osteotropism reflects a high degree of adaptation and osteoblast-like properties of prostate cancer cells within the bone microenvironment. However, the underlying molecular mechanisms are poorly defined. Our previous work has focused on the role of Wnt ligand Wnt5a in bone and bone marrow physiology, metabolic bone diseases, and cross-talks between bone and prostate cancer. In particular, we found that Wnt5a exerts anti-prostate cancer effects in vitro, reduces prostate cancer growth in the skeleton of mice, and is associated with longer survival in men with prostate cancer. We demonstrated that Wnt5a employs both canonical and non-canonical signaling pathways in prostate cancer cells, including Frizzled (FZD) receptor 5 and the atypical receptor tyrosine kinase RYK, but not receptor tyrosine kinase-like orphan receptors (ROR) 1 and 2.Importantly, these studies addressed the role of Wnt5a in the tumor itself. However, the bone microenvironment and especially osteoblasts produce large amounts of Wnt5a, which may govern tumor cell homing into the bone microenvironment and/or tumor-bone interactions. Thus, to date it has remained unclear which cells in the bone microenvironment operate in Wnt5a signaling in bone metastases, and how targeted deletion of Wnt5a in mice affects the colonization and interactions of prostate cancer cells with bone. The central hypothesis is that non-tumor-derived Wnt5a from osteoblastic lineage cells controls critical aspects of prostate cancer cells in their interaction with the bone microenvironment. To test this hypothesis, we will (I) characterize bone-cancer interactions secondary to prostate cancer metastasis in conditional global and osteoblast-specific Wnt5a knockout mice using combined cellular, histological, biomechanical, and imaging analyses, (II) analyze the role of exosomes and involved cargo in bone-tumor cell cross-talks, and (III) investigate the impact of tumor-adjacent stroma tissue on prostate cancer colonization of the bone microenvironment with the support of a clinician-scientist working in the field of Tumor Orthopedics. This individual will make full use of human material obtained in our bone bank and the tissue bank from the Comprehensive Cancer Center Dresden.The results of this project will yield novel mechanistic insights into the role of Wnt5a and associated pathways in prostate cancer interactions with the bone microenvironment and the contribution of tumor vs. non-tumor tissues. This may result in a refined and perhaps revised view of Wnt signaling in bone metastases and could provide a molecular basis for therapeutic intervention. "]
[6,"Characterization of the impact of bone-seeking tumors on the osteocyte network and the osteocyte-mediated regulation of bone turnover ","Orthopaedic, Traumatology, Reconstructive Surgery \nHematology, Oncology ","Professor Dr. Björn Busse","Universitätsklinikum Hamburg-Eppendorf","-","b.busse@uke.uni-hamburg.de","Osteocytes are the most frequent cell type in bone. Residing in lacunae, their protrusions span an area of about 1200 m². With this intricate network, osteocytes orchestrate skeletal adaptation to mechanical stimuli and regulate bone mineral homeostasis through intense communication with osteoblasts and osteoclasts. Recent studies indicate that the bone composition is altered by prostate cancer and that material stiffness is an important factor for the metastatic process. However, little is known about the characteristics of the osteocyte-lacunar network in bone of metastatic entities and how this affects tumor cell behavior. The central hypothesis is that the osteocyte network and functionality are affected by skeletal metastasis and these alterations contribute to pathological bone cell actions and the resulting bone fragility. To test this hypothesis, we will (I) characterize the osteocyte network, local mechanical properties, and bone ultrastructure using 2D/3D imaging/microanalysis during bone metastases formation, (II) determine the impact of osteocyte functionality on the bone metastatic process, (III) assess the bidirectional communication between osteocytes and tumor cells and the role of mechanosensation therein in vitro, and (IV) analyze the adhesion molecule profile of naïve osteocytes and tumor-primed osteocytes to identify targets to modulate osteocyte-tumor cell interactions. Key findings will be validated in bone metastases biopsies obtained from breast- and prostate cancer patients using the Hamburg bone registry. This project will provide novel and detailed insights into the interaction between osteocytes and tumor cells and may provide new therapeutic strategies to inhibit the formation of bone metastases via modulation of osteocyte/tumor cell cross-talks. "]
[7,"Central administration, scientific exchange, and clinical translation ","Endocrinology, Diabetology, Metabolism \nHematology, Oncology ","Professor Dr. Lorenz Hofbauer","Universitätsklinikum Carl Gustav Carus Dresden ","+49 351 4583173","lorenz.hofbauer@uniklinikum-dresden.de","Interdisciplinary collaboration, shared expertise, scientific exchange, integration of early career scientists, and clinical translation are key ingredients for successful and dynamic work within the SPP2084. Active communication will be essential to ensure effective networking between the individual sites within the consortium. This requires regular face-to-face meetings at annual retreats, symposia, and workshops as well as everyday communication measures like webinars and skype and phone conferences. A webpage(www.microbone.de) and a proactive administrative coordinator will foster communication and the networking process. Major ambitions of the SPP2084 are strong interactions between the projects and the support of young researchers. We wish to realize this by establishing professional boot camp-type workshops and training courses. Shortterm lab rotations will also be important to share key technologies in tumor and animal models as well as imaging techniques. In addition, we will specifically promote PhD students by offering expert classes on scientific topics and soft skills and by ensuring progress and direction using thesis advisory committees (TACs). Moreover, participation at the DKTK-School of Oncology will provide specific knowledge and scientific skills in translational oncology for physicians and scientists. To guarantee the best synergistic outcome of both basic and translational research, we will further foster bidirectional networking between basic scientists and clinicians. We therefore apply for clinician-scientist positions, which will be allocated to individual projects for up to 12 months based on documented excellence with a proven track record in clinical translation. These positions will strengthen clinical translation by human validation of animal research. To make this consortium attractive for female scientists and young parents, we intend to offer an intensive mentoring program as well as a “Flying nanny” childcare service for work-related emergencies. To further support women during or immediately after pregnancy, we will specifically incorporate flexible bridge funding by applying for a technician, which will be allocated to individual approved projects within the μBONE consortium. These central measures serve the entire consortium and should help to ensure efficient and productive working conditions for all involved scientists. "]
[8,"Dissecting the role/s of breast cancer cell membrane protrusions in invasion, colonizationand transformation of the bone marrow microenvironment: From cell biology to clinical applications ","Cell Biology \nGynaecology and Obstetrics \nHematology, Oncology ","Dr. Denis Corbeil","Technische Universität Dresden ","+49 351 46340118","denis.corbeil@tu-dresden.de","The bone metastasizing capacity of breast cancer (BC) is subtype dependent. Moreover, only little is known about the biology and molecular principles of BC metastasis and it remains elusive, how breast cancer cells modify and exploit the hematopoietic stem and progenitor cell niche in the bone. Since the underlying functional mechanisms of this phenomenon are largely unknown, we plan to dissect the implication of plasma membrane protrusions emerging from BC cells in migration, adhesion and invasion of bone marrow microenvironment. The morphological impact of factors and/or signaling pathways that modulate the metastatic capacity of cancer cells will be evaluated. As cellular representatives of bone marrow niches, mesenchymal stromal cells (and osteoblasts derived therefrom) will be used. Mechanisms of intercellular communication between BC cells and bone-derived cells will be investigated as well and a particular attention will be devoted to thin membrane processes that could act as tunneling nanotubes. For clinical translation, we will study the role of those plasma membrane protrusions and communication pathways with regard to drug response, metastatic potential and the intrinsic molecular subtypes of BC. "]
[9,"Influence of the skeletal remodeling status on tumor cell dissemination and metastatic outgrowth ","Orthopaedic, Traumatology, Reconstructive Surgery \nHematology, Oncology ","Professor Dr. Michael Amling","Universitätsklinikum Hamburg-Eppendorf","+49 40 741056373","amling@uke.uni-hamburg.de","The bone matrix is continuously remodeled through the balanced activities of two substantially different cell types, bone-forming osteoblasts and bone-resorbing osteoclasts. Our Institute has a long expertise in bone-specific cellular and molecular characterization of genetically modified mouse models, and we have identified several key mechanisms controlling skeletal remodeling in the last years. For the present proposal we will continue our ongoing collaboration with the group of Prof. Klaus Pantel to address the question, if and how an impaired bone remodeling status affects tumor cell dissemination and metastatic outgrowth in vivo.For that purpose we will inject human and murine breast cancer cells into mice to study skeletal metastasis formation as well as changes in the bone microenvironment by Luciferase reporter gene assays, µCT, undecalcified histology and bone-specific histomorphometry. Most importantly, and in contrast to the majority of previously published studies, we do not aim at modifying the seed (i.e. the tumor cells), but the soil (i.e. the recipient mouse). To achieve our goals we have backcrossed different mouse models into genetic backgrounds (BALB/c and NSG) allowing xenotransplantation or syngeneic transplantation of cancer cells. While three of the models (Calca-/-, Calcr-/- and Notch2+/HCS) display specific disturbances of the coupling mechanisms required to synchronize bone formation and bone resoprtion, the other four models (Lrp5+/HBM, Col1a1-Sost, Col1a1-Krm2 and Col1a1-tTA;pTet-Wnt1) display selective changes in osteoblast activity. Since all these genetically modified mouse models have been previously analyzed to define not only their bone remodeling status, but also the molecular causes of the observed phenotypes, we expect that our results will clearly identify, which bone remodeling cell type is primarily involved in the suggested detrimental crosstalk between bone and tumor cells.In addition to this in vivo approach we will perform ex vivo co-culture experiments with breast cancer and bone remodeling cells to define specific molecular interactions. Although our project principally follows an unbiased approach, there are some hypotheses that will be addressed. For instance, since we have previously found that calcitonin (CT) controls bone formation by inhibiting the release of sphingosine 1-phosphate (S1P) from osteoclasts, we will analyze if CT, S1P and/or antagonists of specific S1P receptors will influence proliferation and/or migration of cancer cells, cultured alone or together with bone remodeling cell types. "]
[10,"The impact of myostatin on breast cancer and multiple myeloma bone metastases ","Hematology, Oncology ","Dr. Berno Dankbar","Universitätsklinikum Münster ","+49 251 8356756","dankbarb@uni-muenster.de","The growth of cancers in the skeleton is associated with severe effects on bone and results in considerable morbidity and mortality. Both, breast cancer (BC) as well as multiple multiple (MM)-derived bone metastases are osteolytic and generation of metastases is mediated by the formation and activation of mature osteoclasts.There is a complex crosstalk between tumor cells and the bone microenvironment providing a variety of growth factors for cancer cells and osteoclastic factors. In this regard inhibition of TGFß or activin A, both members of the TGFß familiy, suppress osteoclastic bone resorption in mouse models of MM and BC, indicating an important role of these factors in the formation of bone metastases.In this context, we have recently discovered that myostatin, another member of the TGFß family, strongly enhances osteoclast development and that deficiency or inhibition of myostatin highly ameliorates disease severity and in particular bone erosion in arthritic mice. Interestingly, preliminary experiments revealed a high similarity between mystatin and action A. We could show that myostatin and activin A enhance RANKL-induced osteoclast formation to the same extent, indicating that both factors are equivalent promoters of osteoclast development. Moreover, both factors act via similar Smad-pathways (Smad2/3), pointing to a common mechanism in osteoclast differentiation. Beside this similarity, additional lines of evidence suggest a role for myostatin in bone metastases: 1. Myostatin is expressed in breast cancer cells, 2. ActRII, the receptor type shared by activin and myostatin, is expressed by breast cancer and myeloma cells, 3. Higher levels of phosphorylated SMAD2 are present in human BC bone metastatases.We hypothesize, that myostatin participates in the interactions of tumor cells with the bone microenvironment, promoting osteoclast development and thereby bone lesions and that inhibition of myostatin reduce bone metastases.For this purpose, we will treat two syngeneic and two xenograft models for both BC and MM with anti-myostatin antibody and assess the formation of bone metastases. Moreover, the role of myostatin in cell-cell interaction will be investigated and direct/ indirect effects of tumor cells on osteoclast differentiation and activity will be tested. Furthermore, assuming that both myostatin and activin A are involved in bone metastases, inhibiting myostatin and activin A simultaneously may provide a powerful tool for the treatment of bone metastases in BC and MM. A protein which is able to bind and antagonize myostatin as well as activin is a natural occuring antagonist called „Follistatin-related gene“ (FLRG). In preliminary experiments, we could demonstrate that FLRG completely abrogates RANKL-induced osteoclast formation either stimulated with myostatin and activin. We therefore proposed to treat BC and MM mice also with recombinant FLRG in the course of the disease and analysed as described above. "]
[11,"Hypoxia pathway proteins during homing and propagation of bone metastasis in mice ","Hematology, Oncology ","Professorin Dr. Martina Rauner","Universitätsklinikum Carl Gustav Carus Dresden ","+49 351 4584206","martina.rauner@uniklinikum-dresden.de","Sufficient oxygen pressure is required for our organs to function properly. Conversely, insufficient oxygen supply (hypoxia) is a prominent feature in various pathological processes, including tumor development and metastasis. The central mediators during deprived oxygen are hypoxia inducible factors (HIF) whereas their downstream effects are tightly regulated by oxygen-dependent HIF prolyl hydroxylases (PHDs). Previously, we and others revealed that PHD2 plays a central role during different stages of tumor development, whilst this oxygen sensor is also essential during bone mineralization, and normalization of the endothelial barrier in the bone (marrow) after stress. In this project we aim to test the hypothesis that modulating hypoxia in the tumor and/or bone microenvironment via PHD2 will affect the bone metastatic process. To test this hypothesis, we will (i) define the role of PHD2 in prostate cancer cells using knock-down and overexpression of PHD2 on the formation of bone metastases and determine the impact on the bone vasculature and local bone turnover. Downstream pathways will be determined with an additional knock-down of HIF proteins. Second (ii), we want to assess the role of endothelial cell-derived PHD2 and (iii) the role of osteoblast-derived PHD2 in the bone metastatic process and the triad interactions between tumor cells, angiogenesis, and osteoblasts/bone turnover. This project will provide novel insights into the role of hypoxia pathway proteins in the interaction between tumor cells, bone vasculature and bone cells and may provide new therapeutic strategies to interrupt the malignant crosstalk and ameliorate the formation of bone metastases. "]
[12,"Dissecting the role of TAM receptors in myeloma-induced osteoclast activation and their immune-modulatory function ","Hematology, Oncology ","Dr. Isabel Ben Batalla, Ph.D.","Universitätsklinikum Hamburg-Eppendorf","+49 40 51950","i.ben-batall@uke.de","A hallmark of multiple myeloma (MM) is the reciprocal “vicious” interaction between PC and osteoclasts (OC). This cross-talk leads to disease progression, immunosuppression and osteolytic bone disease. Despite significant advances in the treatment of MM the disease remains incurable in the majority of patients. Therefore, the discovery of novel mechanisms promoting myeloma and its associated bone disease are still urgently needed. In this context, the development of an increased understanding of the interaction between myeloma cells, immune cells and bone cells is of special interest because immune cells hold great potential to destroy malignant cells. However, the regulation and therapeutic activation of anti-myeloma immune responses in the bone marrow is understudied.Our preliminary data show that Protein S (ProS) promotes the proliferation and survival of MM cells. At the same time, it fosters the activation of OC via the receptor Mer, which belongs to the TAM receptor family. Genetic blockade of ProS reduces myeloma burden and prolongs the survival of mice bearing a systemic MM model. Importantly, MM-induced bone disease is almost completely abrogated upon neutralization of ProS. Thus, the blockade of the ProS-Mer axis holds potential to reduce myeloma burden and decrease MM-induced OC activation. In addition, we found that ProS induces an immunosuppressive phenotype of OC. Based on these data we propose a detailed functional investigation of the underlying molecular mechanisms and of the therapeutic potential of the clinically applicable small molecule Mer inhibitor R992 for treatment of MM with a special focus on bone disease. In addition, we will dissect the role of the two alternative TAM receptors Axl and Tyro 3 in osteoclastogenesis. For this purpose, we will use co-cultures and mouse models of MM with OC-specific blockade of TAMR by genetic approaches. Furthermore, we will evaluate the therapeutic effect of TAMR blockade on MM progression as well as on its associated bone disease and immunosuppression. Ultimately, we will translate our findings from the bench to the bedside by analyzing the association of ProS and TAMR with prognosis and MM-induced bone disease.The project will be in particular linked to the groups of L. Hofbauer (Dresden) and A. Garbe (Dresden), with whom we will form an osteoimmunology network. By assessing overlapping cell types and molecular mediators we expect a significant knowledge gain concerning whether the interplay between bone and immune cells is via different or overlapping mechanisms in solid cancer versus MM. Furthermore, we will build a strong bidirectional link to D. Hose (Heidelberg), who will contribute a unique MM patient cohort allowing us to assess the association of the ProS/TAMR axis with MM bone disease. Conversely, we will investigate novel mediators of MM-induced bone disease derived from this patient cohort in our functional mouse models. "]
[13,"Control of Tumor Cell – Bone Metastasis By Regulation of F-Actin Dynamics","Hematology, Oncology \nCell Biology ","Professor Dr. Rolf Jessberger","Technische Universität Dresden ","+49 351 4586446","rolf.jessberger@tu-dresden.de","While primary tumors can be treated with increasing success, >80 % of all cancer-related deaths are caused by metastatic tumors. A diverse arsenal of new approaches to address this most significant problem for human health is required. We focus on one specific approach and one area of cancer: bone metastasis, and we propose that the control of F-actin cytoskeletal dynamics is of central importance for tumor metastasis and bone interactions. Having long-standing expertise in research on F-actin control, we are focusing on an F-actin modulatory protein, SWAP-70, that we recently found to be clearly involved in tumor cell metastasis. Deletion of SWAP-70 significantly reduced metastasis in two mouse cancer models.Thus our overall hypothesis is that SWAP-70 controlled F-actin dynamics contributes significantly to bone metastasis. Thus, we aim at understanding SWAP-70 dependent bone metastatic processes. Our long-term aim is to develop new therapeutic approaches based on preventing SWAP-70’s function in bone metastasis.Our immediate specific aims for the 1st funding period therefore are to:(1) Determine the role of SWAP-70 in tumor cell bone metastasis in vivo(2) Define SWAP-70 dependent, F-actin related features of bone metastatic tumor cells in vitroWe have all the necessary tools and methods in place, e.g. standard and conditional knockout mouse models, an orthotopic cancer model, a large variety of SWAP-70 mutants, Crispr/Cas-generated SWAP-70 deficient tumor cells, and a plethora of analysis technologies. We expect to determine important features of a pathway that has been largely overlooked in the context of tumor bone metastasis. "]
[14,"The role of TRIM proteins in colonization of the bone and bone microenvironment modulation of metastatic prostate cancer. ","Pathology \nHematology, Oncology ","Professor Dr. Sven Perner","Universitätsklinikum Schleswig-Holstein","+49 451 50015800","sven.perner@uksh.de","The most common metastatic site of prostate cancer (PCa) is the bone associated with high morbidity and lethality of patients. With the aim to identify novel therapeutic targets which prevent or treat metastasized PCa, major efforts are being made to better understand the molecular mechanisms enabling bone metastatic transition of PCa cells. Besides well-known genetic alterations of oncogenes and tumor suppressor genes, aberrant activity of regulatory elements for gene transcription, post-translational modification and protein degradation mainly contribute to the metastatic potential of PCa cells. Amongst others, the family of tripartite motif containing proteins (TRIMs) have been identified to be crucial regulators of cellular processes driving metastasis, and to possess altered activity in diverse cancer types. In our preliminary work, we found TRIM24 to be highly overexpressed in PCa bone metastases and to predict outcome. In addition, TRIM8, TRIM11, TRIM47 and TRIM68 have been described be overexpressed in PCa or to influence mechanisms driving metastasis. Based on these data, our overall hypothesis is that these specific TRIM proteins have a crucial role in invasiveness and metastatic niche formation of PCa. To test our hypothesis, we will pursue three specific aims. First, we will characterize the expression and genetic status of TRIMs in PCa tissues including primary tumors as well as metastases. Using large and clinically well-characterized cohorts, we will correlate our molecular data with diverse clinic-pathological parameters and patients’ survival. Second, in-vitro experiments using bone metastatic PCa cells will be conducted to investigate the functional role of TRIMs. We will explore whether cellular features which are required for crucial steps during metastasis are influenced by specific TRIMs. Detailed analysis of signaling molecules involved in these processes will allow conclusions about the underlying molecular mechanisms of TRIM mediated metastatic potential of PCa. Third, co-cultivation experiments will be applied to investigate whether TRIMs influence the communication between bone metastatic PCa cells and cells forming the bone microenvironment. Results of this project will contribute to a better understanding of molecular processes promoting PCa metastasis to the bone. Due to the incurable nature of metastatic PCa to date, targeting specific molecules like specific TRIM proteins might be a therapeutic option for patients suffering from "]
[15,"RAI2 as novel suppressor of cancer cell homing and survival in the bone marrow ","Hematology, Oncology ","Professor Dr. Klaus Pantel","Universitätsklinikum Hamburg-Eppendorf ","+49 40 741053503","pantel@uke.de","We and other research groups have shown that bone marrow (BM) is a common homing organ for disseminated tumor cells (DTCs, the seed of metastases) derived from various types of malignant epithelial tumors. This includes tumors, which do not typically form metastases in bone like colon cancer. In several studies, we have shown that the presence of single DTCs in the BM is an independent predictor of metastatic relapse in many different solid tumors including breast cancer. We have furthermore developed internationally accepted assays to detect and characterize DTCs as well as circulating tumor cells (CTCs) at the single cell level. Moreover, we have identified genes, expression signatures and mechanisms associated with and involved in breast cancer BM micrometastasis, including RAI2 for breast cancer. The retinoic acid–induced 2 (RAI2) was shown not only significantly associated with DTC status but also poor prognosis. BM represents a retinoic acid–rich microenvironment where it regulates differentiation and self-renewal of hematopoietic stem cells. DTCs might be exposed to the same regulatory mechanisms and therefore the RAI2 protein might also be involved in the dormancy control and of overt bone metastasis formation. Our functional studies have furthermore shown that RAI2 sustains differentiation of ER-positive tumor cells, whereas loss of RAI2 induces hormone independent growth and activation of AKT signaling as an important mediator of dormancy control. RAI2 may also regulate the expression of bone morphogenic factors (BMPs) –factors important for breast cancer- bone interaction crosstalk. Therefore, our results indicate that loss of RAI2 expression might represent a so far undiscovered key event at the onset of metastatic progression in the bone. The primary goal of this project is to better understand the precise role of RAI2 for the development of bone metastasis in breast cancer. The investigation of the RAI2-mediated crosstalk between DTCs and cells of the bone microenvironment could lead to better mechanistic insights into cancer dormancy. This could eventually lead to new therapeutic strategies to prevent outgrowth of dormant DTCs into overt bone metastases. Since BM is a reservoir for DTCs that may also recirculate to other metastatic sites, this project may also have implications beyond blocking bone metastasis formation. More specifically we plan to a) investigate the role of RAI2 in the crosstalk between tumor and bone cells, b) perform functional analyses of the role of RAI2 mediated AKT activation in survival and dormancy of DTCs in BM, c) investigate the role of TGFβ and BMP pathways in RAI2-mediated bone metastasis formation and d) establish RAI2 mouse models and assess the functional in vivo role of RAI2 in bone metastasis formation. Thus, the results obtained in the ongoing and future grant will contribute to a better understanding of cancer dormancy and metastatic development in the BM microenvironment. "]
[16,"Probing the role of bone marrow niche-derived SPARC (Secreted Protein Acidic Rich in Cysteine) in the metastatic behavior of human breast cancer ","Hematology, Oncology ","Dr. Hind Medyouf, Ph.D.","Georg-Speyer-Haus","+49 69 540","medyouf@gsh.uni-frankfurt.de","Despite major advances in the treatment of localized cancer, disseminated disease remains one of the most challenging clinical issues, with limited treatment options and poor prognosis for patients. Dormancy at metastatic sites represents a major therapeutic hurdle as standard therapy fails at targeting quiescent cells. Recent insights emphasize the critical role of stromal cells in regulating primary tumor progression and organ-specific metastasis. Besides cellular mediators, components of the extracellular matrix (ECM) have recently attracted attention. Tumor-derived SPARC expression was proposed to maintain prostate cancer cell dormancy in the bone marrow. More recently, stromal expression of SPARC was shown to associate with shorter PFS and OS in metastatic breast cancer. We therefore propose to address the role of niche-derived SPARC in metastatic dormancy using state-of-the-art PDX models, including a newly generated NSG SPARC knock-out line and a fully humanized bone marrow 3D model, both generated by our lab. We will evaluate whether SPARC deficiency triggers the exit of metastatic cells from dormancy thereby sensitizing them to therapeutic targeting using standard of care (e.g. chemotherapy, irradiation). Based on our previous findings, loss of niche-derived SPARC would concomitantly protect hematopoietic stem cells from the detrimental effects of chemotherapy. Lastly, functional CRISPR screens will be performed to unravel downstream mediators of SPARC on metastasis, thereby identifying potential novel therapeutic targets. "]
[17,"Bone marrow stromal cells affect cancer cell homing and growth ","Hematology, Oncology ","Professorin Dr. Inaam Nakchbandi","Universitätsklinikum Heidelberg ","+49 6221 568744","inaam.nakchbandi@immu.uni-heidelberg.de","Breast and prostate cancer cells home to the bone marrow, where they presumably hijack the hematopoietic stem cell niche, and develop metastatic lesions. We aim to characterize the elusive premetastatic niche by examining the role of mesenchymal stromal cells (MSCs) in cancer cell homing and growth of bone metastases. Decreasing the number of MSCs pharmacologically enhanced cancer cell homing to the bone marrow in mice. Initial characterization confirmed a relationship with a mesenchymal subpopulation. We would like to characterize this subpopulation further. This will be followed by both genetic and pharmacologic manipulation in mice in order to increase the identified subpopulation and diminish cancer cell homing. Preliminary work also suggests that this subpopulation suppresses cancer growth in an in vivo model of breast cancer metastasis in mice. We therefore aim to characterize how this subpopulation modulates the immune response towards cancer in vivo. In addition, we will also evaluate whether modifying matrix components and matrix receptors affects the immune cell response to cancer and/or growth. The goal is to take advantage of the lessons learned in this work to maximize the immune inhibitory effect on cancer. Using bone marrow samples from patients with breast and prostate carcinoma we will finally confirm the relationship between stromal population changes and cancer cell homing prospectively. In addition, we will try to confirm data obtained on the contribution of stromal cells to metastasis growth in mice on human cancer samples."]
[18,"Junctional adhesion molecule interactions at the multiple myeloma interface ","Hematology, Oncology ","Dr. Andreas Brandl","Universitätsklinikum Würzburg","+49 931 20144055","Brandl_A@ukw.de","Multiple myeloma (MM) is a B-cell malignancy of clonally expanding antibody producing plasma cells that ultimately leads to osteolytic bone lesions, bone destruction, hypercalcemia, kidney failure, anemia and infections. Despite advances in conventional therapy, clinical application of novel agents and high dose chemotherapy supported by autologous stem cell transplantation MM remains an incurable and highly fatal hematologic disease. MM is characterized by dissemination of multiple tumor cells throughout the bone marrow (BM) and signaling through adhesion molecules regulates MM – BM-microenvironment interactions and might therefore be involved in the dissemination process. We postulate that family members of the junctional adhesion molecules (JAMs) play a pivotal role for MM survival, progression and dissemination by facilitating the crosstalk with BM endothelial cells and we will address the following specific aims: (I) To elucidate and visualize MM cell interactions with the BM endothelium regarding the involvement of JAM-C. (II) To analyse the expression/signaling profile in phenotypically heterogeneous MM subclones. (III) To decipher signaling pathways and their impact on drug sensitivity, homing and dissemination. To achieve these aims we have established models of orthotopic syngeneic and humanized MM, non-invasive bioluminescence imaging and advanced microscopy techniques (e.g. high-resolution 3D light-sheet fluorescence microscopy). Imaging and microscopy will direct further cellular (e.g. multi-parameter flow cytometry, functional in vitro assays) and molecular in vitro analyses. Our studies will help to refine therapeutic interventions, in particularly immunotherapy of MM bone disease. Ultimately, we envision that a better understanding of the pathophysiology of JAMs in MM will result in optimal combinatorial treatment strategies to overcome drug resistance in patients suffering from MM and help to establish semi-personalized anti-MM strategies. "]
[19,"The role of tumor-associated macrophages in bone metastasis formation in zebrafish ","Hematology, Oncology \nDevelopmental Biology \nCell Biology ","Professorin Dr. Franziska Knopf","Technische Universität Dresden","+49 351 46382303","Franziska.Knopf@tu-dresden.de","The aim of this project is to better understand cancer cell colonisation into bone and to unravel unknown interactions between tumour, immune and bone cells in the bone niche. To elucidate such interactions, in vivo imaging of transplanted breast cancer cells in larval zebrafish will be used. In general, in vivo imaging is a powerful tool in order to demonstrate interplay of cells, also in cancer research. In mice, intravital imaging of tumors has revealed that there is a physical connection of tumor-associated macrophages (TAM) with cancer cells, both during initial invasion of tissues and intravasation of cancer cells, i.e. their entry into the blood stream. During these stages mutual dependence by way of positive feedback loops between cancer cells and TAM has been shown. However, the role of TAM for the cancer cells' exit from the blood stream (extravasation), as well as during colonisation of bone is less well understood. By monitoring the interaction of tumor cells (breast cancer cell lines, CTCs), macrophages and bone cells in living zebrafish bone tissue, the following questions will be addressed: i. How do TAM help cancer cells to extravasate and to initially grow in the bone niche? ii. How do TAM contribute to altered osteoblast and osteoclast function during metastasis? To answer these questions, spinning disc confocal microscopy of transgenic zebrafish, in which the respective cell types are fluorescently labelled, will be used. In these fish, interaction of macrophages with bone and cancer cells can be monitored. This approach will be combined with in vivo bone staining techniques and subsequent bone composition analyses in order to characterize bone lesions at the site of metastasis. Furthermore, changes in macrophage number and function will unravel the molecular links of TAM and cancer cells. Using these tools, the role of TAM in bone metastasis formation will be further elucidated which will help to modulate"]
[20,"Role of Tgif1 in breast cancer-induced pathological bone remodeling ","Orthopaedic, Traumatology, Reconstructive Surgery \nHematology, Oncology ","Professor Dr. Eric Hesse, Ph.D.","Klinikum der Universität München","-","Eric.Hesse@med.uni-muenchen.de","Breast cancer is one the most prevalent malignancies worldwide, frequently leading to distant metastases even decades after treatment of the primary tumor and a subsequent disease-free interval. Bone is a common site for breast cancer metastases, causing an incurable stage of the disease and ultimately osteolytic skeletal destruction with pathological fractures and pain. In the bone microenvironment, breast cancer cells severely alter the balanced osteoblast-dependent bone formation and osteoclast-mediated bone resorption by the production of secreted molecules such as Parathyroid hormone-related protein (PTHrP), which stimulates osteoblasts to secrete receptor activator of NFκB ligand (RANKL) and other cytokines. RANKL increases osteoclast activity and subsequent bone resorption during which matrix-derived growth factors are released into the metastatic microenvironment. These factors further stimulate cancer cell proliferation, which perpetuates metastatic bone destruction. At the stage of bone metastases, anti-resorptive drugs like bisphosphonates and an anti-RANKL antibody are used in the clinics to restrict osteolytic bone disease. However, more insights into the cellular and molecular mechanisms of the interaction between breast cancer cells and bone cells within the bone microenvironment and the process of cancer cell colonization into bone is needed, to better understand the process of metastases progression. Using a comprehensive set of in vitro and in vivo approaches we determined that PTHrP and RANKL induce the expression of the homeodomain protein TG-interacting factor 1 (Tgif1) in osteoblasts and osteoclasts, respectively, and firmly established Tgif1 as a novel regulator of osteoclast function and bone resorption. In addition, formation of breast cancer metastases in bones of Tgif1-deficient mice was greatly impaired. These findings strongly suggest a functional role of Tgif1 in the bone microenvironment in the context of breast cancer-induced bone disease. Furthermore, our results show that Tgif1 protein abundance can be reduced by delivery of synthetic microRNA-23a/b (miR-23a/b) mimics, an approach that could be of translational value for the treatment of metastatic bone disease. We therefore hypothesize that Tgif1 affects the cellular cross talk in breast cancer-induced pathological bone remodeling. In addition, we propose that Tgif1 may serve as a therapeutic target to reduce osteolytic bone disease, at least in preclinical models. By combining in vitro experiments, innovative in vivo studies and patient samples we aim to i) investigate the role of Tgif1 in cancer-induced bone destruction in vitro and in vivo, ii) target Tgif1 by miR-23a/b replacement therapy to alleviate metastatic bone disease, and ii) analyze miR-23a/b-Tgif1 signaling in breast cancer patients. This research proposal is original, highly innovative and very likely to expand our knowledge on the mechanisms of breast cancer metastases to bone. "]
[21,"Molecular Characterization of Bone Metastasis Founder Cells in Prostate Cancer ","Pathology, Hematology, Oncology ","Professor Dr. Christoph Klein","Universität Regensburg ","+49 941 9446720","christoph.klein@klinik.uni-regensburg.de","Disseminated prostate cancer cells (DCCs) in bone marrow (BM) comprise the founder cells of later arising lethal bone metastases and understanding the biology of BM-DCCs is paramount for an improved clinical management of prostate cancer (PC) patients. We found that DCCs in prostate cancer displayed significant genetic heterogeneity. Notably, preliminary analysis of the transcriptome indicates that DCCs in PC express a mixed epithelial/haematopoietic phenotype, suggesting a complex patter of adaptation to the ectopic BM microenvironment. We have collected genomes and transcriptomes from almost 400 PC BM-DCCs. In this project we will use novel single cell sequencing technologies to characterize genomes and transcriptomes of BM-DCCs. Furthermore, we will correlate the molecular data with the clinical outcome, and in particular look for (i) genetic aberrations and mutations enabling metastatic progression, and (ii) pathways responsible for quiescence or proliferation of DCCs. For this part of the project, we will collaborate with the team of Ana Dubrovska, and exchange the molecular data obtained from DCCs and circulating tumor cells specific for different stages of the disease. In collaboration with the team of Gelinsky/Alblas we will use their in vitro culture system for BM to model the biology of DCC. We will use prostate cancer and patient-derived metastatic cell lines to establish conditions for expansion of early metastasis founder cells. The resulting in vitro model of minimal residual prostate cancer will enable us to perform further functional assays and drug screens aiming to eradicate the lethal seed of metastasis before clinical manifestation. "]
[22,"Functional characterization of bone metastasis- initiating and radioresistant prostate tumor cells ","Hematology, Oncology \nRadiology, Nuclear Medicine, Radiotherapy, Radiobiology \nReproductive Medicine, Urology ","Professor Dr. Michael Baumann","Deutsches Krebsforschungszentrum (DKFZ)","+49 6221 422854","michael.baumann@dkfz.de","Once prostate tumor spreads to the bone, it typically cannot be cured. The supportive sites in bone maintain survival and proliferation of cancer cells by special molecular components such as C-X-C motif chemokine ligand 12 (CXCL12) which also mediates tumor cell homing to bone. In turn, metastatic PCa cells in bone express different proteins that regulate both, bone remodeling and tumorigenesis including Endo 180 collagen receptor. Prostate tumors have a hierarchical organization where cancer stem cells (CSC) maintain and propagate the tumor mass. A number of studies showed a high radioresistance of CSCs compared to the tumor bulk, and many recent clinical studies use CSC related biomarkers for prediction of radiotherapy outcome. Tumor metastases are driven by the evolved populations of CSC at their worst. In support of this, cells representing different levels of metastatic processes including circulating tumor cells (CTC) in the blood of cancer patients and disseminated tumor cells (DTC) in their bone marrow express CSC markers, and metastasis initiating cells are sharing key CSC features including ability to form tumors and to activate CSC specific signaling mechanisms. Our studies revealed a few proteins that are involved in regulation of metastatic and radioresistant prostate tumor cells. That includes aldehyde dehydrogenase protein 1A1 (ALDH1A1) which is a marker of radioresistant CSC and has a high expression in PCa metastases. Another example is CXCL12 receptor, CXCR4, which is a marker of CSC, upregulates in CTC after radiotherapy and correlates with radiotherapy outcome in PCa. Finally, expression of collagen receptor Endo180/MRC2 which is one of the key regulators of PCa metastases is upregulated in highly tumorigenic radioresistant PCa cells. We assume that activation of certain signaling pathways detected on the genomic and transcriptomic levels in bone-marrow DTC, CTC or CSC in primary tumors can be prognostic for bone metastasis development and, on other hand, can be involved in the bone remodeling and response of metastases to radiotherapy. The first objective of this study is investigation of the role of signaling pathways mediated by CXCR4, ALDH1A1 and Endo180 for regulation of bone metastasis-initiating and radioresistant prostate tumor cells and bone remodeling, and possible interaction of these mechanisms during metastatic development. The second objective of this study is an unbiased and systematic analysis of genomic and transcriptomic profiling of prostate tumor cells representing different levels of metastatic processes (primary tumors, CSC, CTC, DTC, radioresistant PCa cells) to validate activation of CSC related signaling pathways. These mechanisms and phenotypes will be validated by using CRISPR/Cas9 mediated knockout of candidate genes, metastatic mice models, monitoring of bone remodeling in vitro and in vivo, and correlation of candidate genes to clinical data and patients’ response to therapy. "]
[23,"Molecular and functional characterization of the normal and malignant plasma cell – bone interface – from single cell mutual interaction analysis to clinical implications in myeloma bone disease and survival ","Hematology, Oncology ","Professor Dr. Franz Jakob","Julius-Maximilians-Universität Würzburg ","+49 931 8031580","f-jakob.klh@uni-wuerzburg.de","Multiple Myeloma is a disease of malignant plasma cells spreading and accumulating in the bone marrow with subsequent induction of bone disease and angiogenesis. These features are in common with other malignant diseases. Multiple myeloma is different in two regards: Myeloma cells (MMC) spread to and live in the same “natural environment” as their physiological counterpart, bone marrow plasma cells (BMPC). Further, as shown by us, MMC rely at least partly on normal plasma cell functions for malignant behaviour when inducing angiogenesis and interacting with bone turnover. Malignant like normal plasma cells depend on receiving survival-support by interaction with other cell types in the bone marrow, collectively called “niche”. In vitro, we have shown in myeloma/ mesenchymal stromal cell (MSC)//osteoblast co-cultures that this interaction is prominently exerted by physical interaction and leads to changes in expression and behaviour in both cells types. We hypothesize that this physical interaction creates niches and elicits individual signatures that strengthen adhesion, support tumor cells, and disrupt osteogenesis, clinically leading to bone loss and pathological fractures. Niches may confer dormancy or resistance (minimal residual disease) features to myeloma. We have shown this molecular crosstalk being drugable exemplified by blocking the osteocyte-specific wnt antagonist sclerostin, observing increased osteoblast numbers and bone formation in myeloma bearing mice. We also showed that the contact-induced KISS1R system is a putative target as a theranostic PET tracer.In this application, we will focus on surface molecules featuring the interaction between normal and malignant plasma cells and MSC/osteoblasts. We will use expression analysis including single cell RNA-seq to address population heterogeneity, and functional assessment of interaction by atomic-force microscopy and CRISPR/CAS9 screen with subsequent collaborative in vivo validation in myeloma mouse models. In a second step, our in vitro findings will be related to a large cohort of molecularly characterized malignant plasma cell samples alongside whole-body imaging assessing bone disease, plasma cell infiltration and angiogenesis. We have shown that relation of data at nano-/micro- to large scale clinical data sets can act as a filter for relevance and will allow identification of pre-clinically applicable findings. The first is evidenced by sclerostin being expressed in osteocytes only, in contrast to malignant plasma cells from 630 MM patients; the second by our co-development of the T-cell bispecific antibody EM801/901 against the BMPC/MMC cell-surface molecule BCMA entering clinical trials in Q1/18, building the basis for a separate application for development of a theranostic BCMA-PET-Tracer. We will contribute to a collaborative network for myeloma dissemination, bone disease and angiogenesis related research questions within the µBone-consortium. "]

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["Hematology, Oncology ",null,"Hematology, Oncology "]
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