Entwicklung eines humanisierten Tumor-Mausmodells
Zusammenfassung der Projektergebnisse
Tumor xenografts in immunodeficient mice, while routinely used in cancer research, preclude studying interactions of immune and cancer cells or, if humanized by allogeneic immune cells, are of limited use for tumor-immunological questions. Here, we explored a novel way to generate cancer models with an autologous humanized immune system. We demonstrate that hematopoietic stem and progenitor cells (HSPCs) from bone marrow aspirates of nonmetastasized carcinoma patients, which are taken at specialized centers for diagnostic purposes, can be used to generate a human immune system in NOD-scid IL2rγ(null) (NSG) and HLA-I expressing NSG mice (NSG-HLA-A2/HHD) comprising both, lymphoid and myeloid cell lineages. Using NSG-HLA-A2/HHD mice, we show that responsive and self-tolerant human T cells develop and human antigen presenting cells can activate human T cells. As critical factors we identified the low potential of bone marrow HSPCs to engraft, generally low HSPC numbers in patient-derived bone marrow samples, cryopreservation and routes of cell administration. We could establish optimized protocols that use a minimum number of HSPCs, preseleceted highquality bone marrow samples defined by the number of initially isolated leukocytes and intrafemoral or intra-venous injection. In conclusion, the use of diagnostic bone marrow aspirates from non-metastasized carcinoma patients for the immunological humanization of immunodeficient mice is feasible and opens the chance for individualized analyses of antitumoral T cell responses. In addition, we were able to establish DCC-derived xenografts from non-metastasized melanoma patients by transplanting few DCCs or DCC-derived spheres into NSG-mice. All xenografts were serially transplantable and therefore can be used for further functional studies. Not all patients were able to give rise to xenografts, only those with evidence for sentinellymphnode colonization. We are planning to investigate in the future the underlying molecular causes and hope to identify thereby markers on DCCs with metastasiogenic potential that allow better, individualize prognosis and stratification for therapeutic success. Taken together, we have provided an important framework for the use of diagnostic BM-aspirates from carcinoma patients as adult HSPC-source and the establishment of DCC-derived xenografts.
Projektbezogene Publikationen (Auswahl)
- (2014). Immune humanization of immunodeficient mice using diagnostic bone marrow aspirates from carcinoma patients. PLoS One 15;9(5):e97860
Werner-Klein M, Proske J, Werno C, Schneider K, Hofmann HS, Rack B, Buchholz S, Ganzer R, Blana A, Seelbach-Göbel B, Nitsche U, Männel DN, Klein CA
(Siehe online unter https://doi.org/10.1371/journal.pone.0097860) - Genetic alterations driving metastatic colony formation are acquired outside of the primary tumour in melanoma. Nature Communications, volume 9, Article number: 595 (2018)
Sebastian Scheitler, Isabelle Hodak, Melanie Werner-Klein, Klaus Dietz, Petra Lehnert, Veronika Naimer, Bernhard Polzer, Steffi Treitschke, Christian Werno, Ulrich Hohenleutner, Christian Hafner, Mark Berneburg, Petra Rümmele, Martin Hoffmann, Anja Ulmer
(Siehe online unter https://doi.org/10.1038/s41467-017-02674-y)
