ABSTRACT
Chimeric antigen receptor (CAR) T cell therapy has exhibited dramatic anti-tumor efficacy in clinical trials. In this study, we reported the transcriptome profiles of bone marrow cells in four B cell acute lymphoblastic leukemia (B-ALL) patients before and after CD19-specific CAR-T therapy. CD19-CAR-T therapy remarkably reduced the number of leukemia cells, and three patients achieved bone marrow remission (minimal residual disease negative). The efficacy of CD19-CAR-T therapy on B-ALL was positively correlated with the abundance of CAR and immune cell subpopulations, e.g., CD8 T cells and natural killer (NK) cells, in the bone marrow. Additionally, CD19-CAR-T therapy mainly influenced the expression of genes linked to cell cycle and immune response pathways, including the NK cell mediated cytotoxicity and NOD-like receptor signaling pathways. The regulatory network analyses revealed that microRNAs (e.g., miR-148a-3p and miR-375), acting as oncogenes or tumor suppressors, could regulate the crosstalk between the genes encoding transcription factors (TFs; e.g., JUN and FOS) and histones (e.g., HIST1H4A and HIST2H4A) involved in CD19-CAR-T therapy. Furthermore, many long non-coding RNAs showed a high degree of co-expression with TFs or histones (e.g., FOS and HIST1H4B) and were associated with immune processes. These transcriptome analyses provided important clues for further understanding the gene expression and related mechanisms underlying the efficacy of CAR-T immunotherapy.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Antigens, CD19 , Metabolism , Bone Marrow , Metabolism , CD8-Positive T-Lymphocytes , Allergy and Immunology , Gene Expression Regulation, Leukemic , Gene Regulatory Networks , Immunotherapy, Adoptive , MicroRNAs , Genetics , Metabolism , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Genetics , Allergy and Immunology , Therapeutics , RNA, Long Noncoding , Genetics , Metabolism , Receptors, Antigen, T-Cell , Transcription Factors , Metabolism , Transcriptome , GeneticsABSTRACT
Objective To evaluate the effect of cytokine-induced killer cells (CIKs) as an adoptive immunotherapy option for treatment of leukemia relapse after allo-hematopoietic stem cell transplantation (allo-HSCT).Methods Two cases of infusion of donor CIKs in patients with leukemia relapse after allo-HSCT were retrospectively analyzed.Patient one relapsed 986 days (+986d) after HLA-matched unrelated donor allo-HSCT.Applications of chemotherapy only resulted in short term remission,but allo-CIKs were successfully expanded from the patient's peripheral blood mononuclear cells of donor origin.Totally five cycles of CIKs infusion were infused as an alternative of adoptive immunotherapy.Patient two had recurrent in the + 158d after HLA-matched sibling alloHSCT.At + 204d and + 294d,two cycles of CIKs which were expanded from donor peripheral blood mononuclear cells were infused.Results One cycle of CIKs was given to patient one after the application of chemotherapy to reduce the tumor burden,and the patient successively achieved complete remission.Again after additional four cycles of CIKs infusion,consistent remission was maintained during the following seven months.Patient two who had relapsed disease posttransplantation,achieved cytological complete remission after withdrawal of immunosuppressants and undergoing chemotherapy combined with G-CSF mobilized stem cell infusion.However,at + 187d,the patient suffered from side-effect of acute graft versus host disease and extramedullary infiltration.The symptoms were alleviated markedly after one cycle of CIKs infusion at + 204d.Moreover,the pain disappeared after an additional infusion at + 294d.And up to the present,the bone marrow aspiration showed complete remission while the extramedullary disease vanished.Conclusion The use of CIKs in the treatment of leukemia relapse after allogeneic bone marrow transplantation can be feasible and well tolerated.
ABSTRACT
Objective To investingate the effect of survivin shRNA on chemotherapy resistance in GBC-SD cells. Methods They were divided into three groups of GBC-SD, GBC-SD/EGFP and GBC-SD/survivin. MTT assay was used to detect cell viability in the 3groups. mRNA and protein of survivin were tested by RT-PCR and Western blot. Then the cells were treated with proper construction DDP. The cell survival rate and IC_(50) were determined with MTT, cell apoptosis detected by FACS and the alteration of nucleus observed by TUNEL. Meanwhile, caspase-3 activity was determined using the colorimetric method. Results Cell viability was reduced remarkably in GBC-SD/survivin and survivin expression was decreased obviously. After being treated with DDP, cell survival rate and IC_(50) were decreased obviously in GBC-SD/survivin, apoptotic rate elevated remarkably compared with other groups. There were brownly nucleuses in three groups. Caspase-3 activity increased first and then decreased, but it exceeded in GBC-SD/survivin than that in other two groups. Conclusion The survivin shRNA can down-regulate the expression of survivin in GBC-SD cells remarkably and improve the sensibility to chemotherapy.
ABSTRACT
AIM: To determine the cleavage activity of anti-transforming growth factor ?1 hammerhead ribozymes which was inserted into U1 small nuclear RNA in cell-free system.METHODS: The hammerhead ribozyme targeting against transforming growth factor ?1 was designed through the analysis of computer software.The ribozyme fragments were synthesized and cloned into the U1 snRNA ribozyme vector pZeoU1EcoSpe,which contained U1 snRNA promoter/enhancer and terminator.TGF ?1 cDNA partial fragment was generated by RT-PCR,and then cloned into the T-vector at the downstream of T7 promoter.The transcripts of ribozyme and target RNA incorporated into isotope were transcribed in vitro and purified by denaturing polyacrylamide gel electrophoresis.-labeled U1 snRNA chimeric ribozyme transcripts were incubated with target-RNAs at different conditions and autoradiographed after running denaturing PAGE.RESULTS: U1snRNA chimeric ribozyme(U1Rz803) cleaved TGF?1 mRNA efficiently and specifically at 37 ℃,while the disable ribozyme(U1Rz803m) showed no cleavage activity,so these indicated the design of U1Rz803 was correct.CONCLUSION: U1Rz803 prepared in this study possesses the perfect specific catalytic cleavage activity in cell-free system.These results indicate that U1 snRNA chimeric ribozyme U1Rz803 may suppress the expression of TGF?1 in vivo,therefore it may provide a new means for exploring the role of TGF?1 in hematopoietic regulation in the future.