Transcriptome and Regulatory Network Analyses of CD19-CAR-T Immunotherapy for B-ALL / 基因组蛋白质组与生物信息学报·英文版

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.

Development of Mass Spectrum Fingerprint Extraction Technology for Antibiotics in Surface Water / 分析化学

An extract method for the fingerprint feature of 49 kinds of antibiotics belonging to multiple classes in surface water was developed.Water sample was purified and concentrated by tandem dual column (MAX and HLB),and qualitatively and quantitatively analyzed by ultra-high performance liquid chromatography-tandem mass spectrometric (SPE-UPLC-MS/MS) under multiple reaction monitoring (MRM) mode.The pretreatment was optimized in types of SPE column,loading pH,eluent and redissolution for multiclass antibiotics.The results showed that the linearity of target antibiotics was good in the range of 0.001-0.5 μg/mL (0.01-5 μg/mL for streptomycin).The recoveries were from 51.7% to 94.8%,and the relative standard deviations (RSDs) ranged from 2.19% to 9.67%.The limits of detection(LOD,S/N=3) were 0.01-3.23 μg/L and 0.05-3.43 μg/L and the limits of quantification (LOQ,S/N=10) were 0.04-10.8 μg/L and 0.17-11.4 μg/L in different redissolve solutions.This method was applied to the determination of antibiotics in water samples from 9 sites of Qinhuai River and Xuanwu Lake.

Adeno-associated vector mediated intracellular biological activity of human Kallistatin / 药学学报

Human tissue kallikrein-binding protein (Kallistatin, KAL), a secretory protein that participates in the regulation of multiple signaling pathways by binding to the extracellular receptor, however, at present has not been reported about the intracellular activity, and whether it has the similar biological activity with extracellular activity. Here we constructed no signal peptide KAL (NSK) into the adeno-associated virus vector to explore the intracellular activity of KAL. Both the endothelial cell and lung cancer cells could express KAL, but not secreted after rAAV2-NSK transfection. The proliferation and migration of human umbilical vein endothelial cells (HUVECs) were inhibited, but the apoptosis rate was not affected. The proliferation rates, mobility and tubule formation of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited to different extents. This cellular study not only confirmed the intracellular activity, but also suggested it may serve as a kind of "balance factor" in multi-targeted controlling, which may provide a new train of thoughts to explain the regulatory contradiction in PI3K-Akt signaling pathways by KAL.