Transcriptomic profile of untreated DG75 lymphoma cell line clones expressing iCASP9 and untreated versus AP1903-resistant clones

In this study, we evaluate the efficacy of inserting an inducible caspase9 (iCasp9) suicide gene into B-cells, in combination with a single-chain immunoglobulin cassette that redirects B-cells against a tumor-specific antigen. We demonstrate that a single immunoglobulin (IgH) locus modification enables expression of both the iCasp9 gene and a cassette that hijacks the antigen specificity of the BCR, while preserving the functionality of the modified cells. Induction of iCasp9 using the dimerizing molecule AP1903 effectively induced the death of edited cells, even when starting from the malignant lymphoma cell line DG75. However, in cultures prolonged for 14 days in vitro and maintained under pressure with AP1903, resistant clone finally grow. This RNAseq experiment compares the transcriptomic profile of untreated clones versus AP1903-resistant clones.

本研究评估了将诱导型半胱氨酸蛋白酶9（inducible caspase9, iCasp9）自杀基因插入B细胞，并联合使用可重定向B细胞靶向肿瘤特异性抗原的单链免疫球蛋白盒的应用效果。研究证实，仅对免疫球蛋白重链（IgH）位点进行单次修饰，即可同时表达iCasp9基因与劫持B细胞受体（B cell receptor, BCR）抗原特异性的盒式结构，且可维持修饰后细胞的功能活性。使用二聚化分子AP1903诱导iCasp9，可有效触发编辑后细胞的凋亡，即便起始细胞为恶性淋巴瘤细胞系DG75。不过，在体外持续培养14天并于AP1903筛选压力下维持培养后，最终会出现耐药克隆增殖。本RNA测序（RNA sequencing, RNA-seq）实验旨在比较未处理克隆与AP1903耐药克隆的转录组谱差异。