基于CRISPR/Cas9技术靶向敲除TRIM37泛素化连接酶基因治疗乳腺癌的实验研究

The rapid development of gene editing technology offers novel strategies for cancer gene therapy. CRISPR/Cas9 gene knockout system is a RNA guided DNA splicing technology, which could efficiently knockout targeted gene. It offers a novel method for gene edition as well as cancer gene therapy. In this study, we will targeted knockout an overexpressed oncogene TRIM37 in breast cancer by CRISPR/Cas9 technology. The constructed CRISPR/Cas9 system will be delivered by a novel biodegradable non-viral gene vector, Heparin-PEI, which was developed in our lab. The anti-cancer effects of TRIM37 knockout by CRISPR/Cas9 system will be evaluated both in vitro and in vivo, and their molecular mechanisms will be further studied. Several progresses including TRIM37 knockout and Heparin-PEI preparation have been made, and to our best known, there is no report about taking advantage of CRISPR/Cas9 technology in cancer therapy. The applicant has published 5 SCI papers as first or co-author on research journals such as ACS Nano (IF=12.033) and more than 20 papers in related area, demonstrating good experience. This study is well designed and practicable; it will contribute to the discovery of new therapeutic target and strategy for breast cancer, and is potent in further clinical study.

基因编辑技术的迅速发展，为肿瘤基因治疗提供了新策略。CRISRP/Cas9基因敲除技术是一种由RNA介导的DNA剪切技术，可以靶向地实现对目的基因的快速敲除，为肿瘤基因治疗提供了新手段。本研究基于CRISPR/Cas9技术，利用前期已构建的肝素-PEI新型可降解非病毒基因载体，对新发现的乳腺癌中高表达的TRIM37致癌基因进行特异性敲除，研究其对乳腺癌肿瘤模型的治疗效果及安全性并深入探讨相关分子机制。本课题已在TRIM37基因体外敲除和肝素-PEI的制备研究中取得一定进展，且尚未见将CRISPR/Cas9技术用于肿瘤治疗研究的相关报道。申请人在相关领域发表SCI论文20余篇，以第一和共同第一作者身份在ACS Nano（IF=12）等杂志发表论文5篇，积累了丰富经验。该题目具有良好可行性，将为建立一种新的乳腺癌基因治疗靶点和策略提供理论和实践基础，为临床应用基因治疗手段控制乳腺癌起到积极意义

基因编辑技术的迅速发展，为肿瘤基因治疗提供了新策略。CRISRP/Cas9基因敲除技术是一种由RNA介导的DNA剪切技术，可以靶向地实现对目的基因的快速敲除，为肿瘤基因治疗提供了新手段。本研究基于CRISPR/Cas9技术，利用前期已构建的肝素-PEI新型可降解非病毒基因载体，对新发现的乳腺癌中高表达的TRIM37致癌基因进行了特异性敲除，成功实现了抑制乳腺癌细胞中TRIM37致癌基因功能的作用。我们的研究结果显示：Cas9-Trim37/肝素-PEI纳米基因复合物可以通过敲除TRIM37基因而诱导乳腺癌肿瘤细胞凋亡、抑制肿瘤细胞迁移及新生血管生成，进而在体内和体外有效抑制乳腺癌肿瘤细胞及组织的生长，展示出明显的抗肿瘤活性。同时，Cas9-Trim37/肝素-PEI纳米基因复合物还具有较好的安全性。上述进展已达到了本项目的预期研究目标，相关研究成果为乳腺癌的基因治疗提供了新的策略及研究基础。