hsa_circ_0003183/miR-224/CD59信号通路在利妥昔单抗耐药 弥漫大B细胞淋巴瘤中的机制研究

Revealing the molecular mechanism of diffuse large B-cell lymphoma (DLBCL) developing rituximab resistance is urgent needed. Our previous study showed that high expression of CD59 was negatively correlated with the prognosis of DLBCL patients treated rituximab. Moreover, our study demonstrated that miR-224 could down-regulate of expression of CD59 in DLBCL. Our research group found that hsa_circ_0003183 could down-regulate miR-224 in DLBCL and IFN-α enhanced complement-dependent cytotoxicity (CDC) in DLBCL. Given these evidences, we could reasonably hypothesize that in the process of developing rituximab resistance, hsa_circ_0003183 was up-regulated, hence miR-224 was down-regulated and as a result CD59 was up-regulated. The latter could inhibit CDC effects induced by rituximab, and eventually drug resistance developed. IFN-α enhanced rituximab sensitivity in DLBCL probably via targeting hsa_circ_0003183/miR-224/CD59 signal pathway. To validate this, we plan to continue this research in three levels, namely molecular level, cell level and animal level. This study will unveil the underlying mechanism of rituximab resistance progression in DLBCL, providing theoretical basis for the clinical interventions and new insight for drug development of DLBCL rituximab resistance.

揭示弥漫大B细胞淋巴瘤（DLBCL）利妥昔单抗耐药分子机制对DLBCL治疗至关重要。课题组前期发现在DLBCL中CD59高表达与DLBCL利妥昔单抗治疗预后负相关；miR-224可下调CD59的表达；进一步研究发现hsa_circ_0003183负调控miR-224。预实验表明IFN-α能增加利妥昔单抗的CDC效应。课题组拟提出以下假说：利妥昔单抗耐药形成过程中，hsa_circ_0003183上调，导致miR-224表达下调，进而CD59表达上调，后者抑制利妥昔单抗的CDC效应，表现为利妥昔单抗药敏性下降和耐药；IFN-α可能通过调控hsa_circ_0003183/miR-224/CD59通路提高利妥昔单抗的敏感性。课题组拟从分子水平、细胞水平和动物水平进行研究。本课题揭示DLBCL细胞利妥昔单抗耐药的分子机制，为DLBCL耐药的临床干预提供理论基础。

本课题揭示DLBCL细胞利妥昔单抗耐药的分子机制，为DLBCL耐药的临床干预提供理论基础。主要研究内容包括a.耐药形成前后DLBCL细胞的CD59蛋白及miR-224表达差异：该部分主要探索 DLBCL细胞耐药形成前后CDC作用及相关蛋白表达异常；b.耐药形成前后环状芯片表达差异：明确耐药形成后环状RNA表达异常，并通过数据分析找出能与miR-224起到海绵体作用的环状RNA，并验证其结果；c.通过过表达载体和小干扰载体研究明确了hsa_circ_0008514/miR-224/CD59通路 对DLBCL耐药机制的影响。本项目按照标书设计，安排课题进度并完成相关实验；此外，在完成原有内容的基础上，项目执行过程中还进一步应用环状RNA芯片、荧光素酶报告基因等新技术和手段，进一步了解DLBCL耐药的机制。本项目共资助发表论文4 篇，2篇SCI论文，1篇OAT英文文章，1篇省级刊物文章。人才培养方面，本项目资助培养了6名相关专业研究生。总之，本项目的顺利进行为今后进一步探索利妥昔单抗耐药的原因提供了实验基础。