Patient-derived Xenograft Model of Chronic Lymphocytic Leukaemia Based on Mimicking Its Microenvironment

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F65269705%3A_____%2F21%3A00075084" target="_blank" >RIV/65269705:_____/21:00075084 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.ceitec.eu/ceitec-phd-conference-2021/a3988" target="_blank" >https://www.ceitec.eu/ceitec-phd-conference-2021/a3988</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Patient-derived Xenograft Model of Chronic Lymphocytic Leukaemia Based on Mimicking Its Microenvironment

Popis výsledku v původním jazyce

Chronic lymphocytic leukemia (CLL) is an incurable B-cell malignancy with characteristic dependence on human immune microenvironment. Interactions in the microenvironment are provided by various types of cells and factors, but of them only CD4-positive T-cells can promote proliferation of leukemic cells. This makes T-cells also indispensable for current CLL-derived xenograft (PDX) mouse models. However, limited availability of autologous T-cells and their rapid spontaneous growth in immunodeficient mice, led us to an idea of replacing them in in vitro and in vivo models with a novel, genetically engineered cell line. We prepared an adherent supportive cell line, expressing T-cell factors, which induces long term survival and major proliferation of primary CLL cells in vitro. Using this cell line, we developed a coculture system for testing drugs with antiproliferative effect and identified a novel inhibitor with such activity in CLL. We introduced the cell line into immunodeficient mice (NSG) to mimic human lymphoid microenvironment. We seeded collagen scaffolds, implanted them subcutaneously and injected mice with 30 350x106 purified CLL cells (containing undetectable number of T-cells). By this, we achieved engraftment of B-cells, but these were positive for Epstein-Barr virus (EBV). As CLL malignancy in vivo is not based on EBV activation and CLL cells are virtually free of EBV, subsequently we attempted to implant mice with small number of CLL cells containing statistically no healthy B-cells (healthy B cells from CLL patients can carry EBV). The first strategy to achieve that is to pre-expand small number of CLL cells (103-104) cocultured on supportive cells in vitro. The second strategy is based on direct implantation of mice with supportive cells and 105 CLL cells into spleen. Currently we are waiting for the first results of both strategies, as none of them was ever applied in CLL research so far. The established model will be used for testing of novel therapeutic combinations and studies of CLL biology.

Název v anglickém jazyce

Patient-derived Xenograft Model of Chronic Lymphocytic Leukaemia Based on Mimicking Its Microenvironment

Popis výsledku anglicky

Chronic lymphocytic leukemia (CLL) is an incurable B-cell malignancy with characteristic dependence on human immune microenvironment. Interactions in the microenvironment are provided by various types of cells and factors, but of them only CD4-positive T-cells can promote proliferation of leukemic cells. This makes T-cells also indispensable for current CLL-derived xenograft (PDX) mouse models. However, limited availability of autologous T-cells and their rapid spontaneous growth in immunodeficient mice, led us to an idea of replacing them in in vitro and in vivo models with a novel, genetically engineered cell line. We prepared an adherent supportive cell line, expressing T-cell factors, which induces long term survival and major proliferation of primary CLL cells in vitro. Using this cell line, we developed a coculture system for testing drugs with antiproliferative effect and identified a novel inhibitor with such activity in CLL. We introduced the cell line into immunodeficient mice (NSG) to mimic human lymphoid microenvironment. We seeded collagen scaffolds, implanted them subcutaneously and injected mice with 30 350x106 purified CLL cells (containing undetectable number of T-cells). By this, we achieved engraftment of B-cells, but these were positive for Epstein-Barr virus (EBV). As CLL malignancy in vivo is not based on EBV activation and CLL cells are virtually free of EBV, subsequently we attempted to implant mice with small number of CLL cells containing statistically no healthy B-cells (healthy B cells from CLL patients can carry EBV). The first strategy to achieve that is to pre-expand small number of CLL cells (103-104) cocultured on supportive cells in vitro. The second strategy is based on direct implantation of mice with supportive cells and 105 CLL cells into spleen. Currently we are waiting for the first results of both strategies, as none of them was ever applied in CLL research so far. The established model will be used for testing of novel therapeutic combinations and studies of CLL biology.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

30204 - Oncology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů