c-Myb通过调节能量代谢维持CAR-T细胞干性特征的作用和机制研究

项目申请人的前期研究结果显示，对于复发难治的急性B淋巴细胞白血病（B-ALL），靶向CD19的CAR-T细胞在患者体内的长期存续可显著减少治疗后的复发，探索维持CAR-T细胞干性特征的技术手段和机制对于提高CAR-T的疗效具有重要意义。转录因子c-Myb对于细胞的分化和增殖具有重要作用，我们在预实验中观察到可在患者体内长期存续的CAR-T细胞高表达c-Myb，并伴随能量代谢和线粒体相关基因的上调，提示c-Myb可能通过影响细胞能量代谢维持CAR-T细胞的干性特征。本项目计划通过干扰或过表达CAR-T细胞的c-Myb，检测细胞表型及开展体外杀伤实验和长期扩增培养实验，明确c-Myb对于CAR-T细胞的影响，并进一步在动物实验中验证其作用，最后从细胞能量代谢角度探索c-Myb维持CAR-T细胞干性的具体机制。本项目将为提高CAR-T的体内存续提供一种新方法，并为c-Myb的研究提供新的视角。

Results from applicant’s previous studies show that, for relapsed/refractory acute B lymphoblastic leukemia patients, long-term persistence of CD19-specific CAR-T cells significantly reduces the post-treatment relapse. Exploring the mechanism and strategies to maintain CAR-T cell stemness is significantly important to enhance the clinical outcomes of CAR-T therapy. Transcription factor c-Myb plays critical roles during cellular differentiation and proliferation. In our preliminary experiments, we observed elevated expression of c-Myb in CAR-T samples with longer in vivo persistence, concomitant with up-regulation of genes involved in energy metabolism and mitochondria regulation. This finding suggests that c-Myb may help maintain CAR-T stemness through the regulation of cellular energy metabolism. . In this proposal, we first plan to confirm our previous findings: how modulation of c-Myb expression levels, such as by RNA interference or overexpression, affects CAR-T cell phenotypes and in vitro target killing. Long-term in vitro expansions, the anti-tumor performance of CAR-T cells with different c-myb expression levels in tumor-bearing animal models, and the potential mechanism will be further studied. We hope to clarify the specific mechanism of how c-Myb maintains CAR-T stemness through energy metabolism. This research may provide a new approach to enhance the in vivo persistence of CAR-T cells. It also reveals a new strategy of c-Myb study.