Zweifel
Fakultäten » Medizinische Fakultät » Onkologie, Klinik für » Prof. Dr. Alexander Knuth » Zweifel
Completed research project
| Title / Titel | Chemotherapy-mediated recruitment of monocytes and other VEGFR-1 positive cells and the preparation of the metastatic niche | ||||
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| Abstract (PDF, 14 KB) | |||||
| Summary / Zusammenfassung | We hypothesize that VEGFR-1 positive cells such as immature monocytes, mobilized from bone marrow during recovery from hematotoxic chemotherapy, may differ from normal monocytes in terms of expression of the differentially spliced isoforms of VEGFR-1 and thus support the formation of micrometastatic niches. Peripheral blood mononuclear cells (PBMC) are isolated from the blood of patients with breast cancer, and colon cancer, presenting with low/no tumor load (neoadjuvant or adjuvant chemotherapy), and high tumor load (palliative chemotherapy). Peripheral blood monocytes (PBMo), and VEGFR-1 positive cells, respectively, are separated from PBMC by adherence assay, and FACS. The expression of the distinct mRNA splice variants for VEGFR-1 of cell lysates is analyzed by RT-PCR using isoform-specific primers and probes. Semiquantitative analysis of transmembrane and soluble VEGFR-1 protein by western blot of cell lysates, and immunoprecipitation of culture medium supernatants, respectively, is performed. Migration properties of PBMo and VEGFR-1+ cells under normoxic and hypoxic conditions toward breast carcinoma cell line MCF-7 cells, and colon carcinoma cell line HT-29 cells, respectively, are assessed using a transwell assay. Nude mice are injected with monocytes from patients blood, and later with green fluorescent protein-labeled MCF-7 breast cancer cells (MCF-7-GFP). Different organs are screened by fluorescence microscopy for the presence of tumor cells, and co-located monocytes are identified. Using siRNA in order to selectively inhibit splice variants of VEGFR-1, the potential of such treated VEGFR-1 positive cells to create (pre-)metastatic niches is functionally assessed. | ||||
| Publications / Publikationen | Differential expression of soluble and transmembrane VEGF Receptor-1 under hypoxia and acidic pH in endothelial cells. Zweifel M, Driscoll R, Ruegg C. Proceedings of the AACR 2006:1866:440 |
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| Keywords / Suchbegriffe | Angiogenesis, metastasis, breast cancer, colorectal cancer, VEGFR-1, alternative splicing, monocytes | ||||
| Project leadership and contacts / Projektleitung und Kontakte |
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| Funding source(s) / Unterstützt durch |
Foundation |
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| In collaboration with / In Zusammenarbeit mit |
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| Duration of Project / Projektdauer | Jan 2007 to Dec 2008 |