Establishment And Application Of Mouse Models In Hematological Malignant Diseases

Objective:Based on the traditional chemotherapy and HSCT(hematopoietic stem cell transplantation), the application of targeted drugs improve the prognosis of hematological malignant diseases. But drug resistance and relapse is the major cause for treatment failure. For this reason, a model with a phenotype that more closely resembles human hematological malignant diseases will provide a good platform for the study of pathology and the exploration of antileukemic therapy. Here we will elucidate how to generate and apply the mouse models of cell lines and primary samples.Methods:Part I:We use11cell lines of hematological malignant diseases including AML, ALL, BAL, CML and Lymphoma to generate11kinds of mouse models via tail vein injection. Engraftment was monitored by weekly blood collections and FACS analysis for human CD45+cells. At the first indication of morbidity (>20%weight loss, lethargy and ruffled fur), mice were sacrificed by cervical dislocation) and ascertain the leukemic infiltration levels in tissues at this endpoint. We also apply these mouse models to verify the role of Raptor in MLL (Mixed Lingeage Leukemia) associated leukemia.Part II:Mononuclear cells of bone marrow were collected from patients with newly diagnosed hematological malignant diseases, and were inoculated into NOD/SCID mouse via traditional tail vein injection. Also, in this study, we present the first data indicating that intrasplenic inoculation of human primary samples are able to reconstitute a complete leukemic phenotype in NOD/SCID mice, sustain the self-renewal capacity of LSCs and can be used for in vivo drug treatments and predicting the immuophenotype of ALL initiation cells.Results:We successfully used11cell lines to generate different types of mouse models via tail vein injection, and elucidated the feature of each mouse model. When we knocked down the expression of Raptor in RS4;11cell line with MLL-AF4mutation and K562cell line, the survival of RS4;11mouse model but not K562mouse model were improved.We successfully established the ALL humanized NOD/SCID mouse models via tail vein injection(5/9) and intrasplenic injection(10/20). the proportion of CD150+cells declined obviously in engrafted cases compared with non-engrafted cases (P<0.01). We successfully established3AML humanized NOD/SCID mouse models with FLT3-ITD mutation, and a ANKL(Aggressive Natural Killer cell Leukemia) humanized NOD/SCID mouse model are still under monitoring.Conclusion:Intrasplenic inoculation are able to reconstitute a complete leukemic phenotype in NOD/SCID mice, and cell lines are easier than primary samples to engrafted into NOD/SCID mouse,while ALL samples are easier than AML samples. Leukemia-initiating potential of leukemic blasts is correlated negatively with the proportion of CD150+cells, the percentage of which can serve as a useful predictor for engraftment success of B-ALL to immune deficient mice. Raptor is correlated with MLL associated leukemia, and need furthermore study to verify.