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浙江大学学报（医学版）
浙江大学学报（医学版）  2022, Vol. 51 Issue (2): 185-191    DOI: 10.3724/zdxbyxb-2022-0047
专题报道     
脂质纳米粒-mRNA递送系统及其在嵌合抗原受体T细胞治疗中的应用
叶柏新1,2,3,4,胡永仙1,2,3,4,张明明1,2,3,4,黄河1,2,3,4,*()
1.浙江大学医学院附属第一医院骨髓移植中心，浙江 杭州 310003
2.浙江大学医学中心良渚实验室，浙江 杭州 311121
3.浙江大学血液学研究所，浙江 杭州 310058
4.浙江省干细胞与细胞免疫治疗工程实验室，浙江 杭州 310058
Research advance in lipid nanoparticle-mRNA delivery system and its application in CAR-T cell therapy
YE Baixin1,2,3,4,HU Yongxian1,2,3,4,ZHANG Mingming1,2,3,4,HUANG He1,2,3,4,*()
1. Bone Marrow Transplantation Center, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China;
2. Liangzhu Laboratory, Zhejiang University Medical Center, Hangzhou 311121, China;
3. Institute of Hematology, Zhejiang University, Hangzhou 310058, China;
4. Zhejiang Provincial Laboratory for Stem Cell and Immunity Therapy, Hangzhou 310058, China
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摘要：

尽管嵌合抗原受体（CAR）T细胞治疗在血液系统恶性肿瘤患者中取得了显著的临床疗效，但需要进一步优化。脂质纳米粒（LNP）-信使核糖核酸（mRNA）递送系统作为一种非病毒性基因载体运用于CAR-T细胞治疗研究中，一方面通过LNP将密封蛋白-6 mRNA靶向递送至抗原提呈细胞，从而实现抗原提呈细胞辅助性增强密封蛋白-6靶向的CAR-T细胞的功能，以进一步诱导对实体瘤的清除；另一方面，通过LNP将成纤维细胞激活蛋白（FAP）CARmRNA靶向递送至T细胞，实现体内FAP靶向的CAR-T细胞的制备，以通过阻断心脏纤维化过程达到治疗急性心肌损伤的目的。在CAR-T细胞研究和治疗中，LNP-mRNA递送系统具有不与细胞基因组整合、价格便宜、毒副作用小及可修饰等优点，亦存在蛋白瞬时表达导致调控细胞功能的持久性不足及制备等方面的技术局限性。本文综述了LNP-mRNA递送系统及其在CAR-T细胞治疗中的应用研究。
关键词： 脂质纳米粒;  信使核糖核酸;  体内递送技术;  基因递送载体;  嵌合抗原受体T细胞;  综述    
Abstract:

Chimeric antigen receptor (CAR) T cell therapy has shown significant efficacy for hematological malignancies, however, it needs to be further optimized. Recently, the lipid nanoparticle (LNP)-mRNA delivery system as a nonviral gene transfer vector has gained rapid progress in CAR-T cell therapy. The claudin-6 (CLDN6) mRNA is delivered to antigen presenting cells (APCs) through LNP system, thereby enhancing the function of CLDN6 CAR-T cells for the clearance of solid tumor cells. For treatment of acute cardiac injury, the fibroblast activation protein (FAP) CAR mRNA can be delivered to T cells through LNP system for the in vivo production of FAP CAR-T cells, thereby blocking the process of myocardial fibrosis. The LNP-mRNA delivery system has advantages including having no integration in host genome, inexpensiveness, low toxicity and modifiability; on the other hand, it has certain disadvantages such as limited cell persistence caused by transient protein expression and limitations in preparation techniques. This article reviews the research advance in LNP-mRNA in vivo delivery system and its application in CAR-T cell therapy.
Key words: Lipid nanoparticle    Messenger RNA    In vivo delivery system    Gene transfer vector    Chimeric antigen receptor T cell    Review
收稿日期: 2022-02-17 出版日期: 2022-08-02
CLC:  R96  
基金资助: 国家自然科学基金（81730008）
通讯作者: 黄河     E-mail: huanghe@zju.edu.cn
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引用本文:

叶柏新,胡永仙,张明明,黄河. 脂质纳米粒-mRNA递送系统及其在嵌合抗原受体T细胞治疗中的应用[J]. 浙江大学学报（医学版）, 2022, 51(2): 185-191.

YE Baixin,HU Yongxian,ZHANG Mingming,HUANG He. Research advance in lipid nanoparticle-mRNA delivery system and its application in CAR-T cell therapy. J Zhejiang Univ (Med Sci), 2022, 51(2): 185-191.

链接本文:

https://www.zjujournals.com/med/CN/10.3724/zdxbyxb-2022-0047    或    https://www.zjujournals.com/med/CN/Y2022/V51/I2/185

图 1  LNP-mRNA递送系统组成及结构LNP-mRNA递送系统由外层脂质分子形成的膜性外壳及包裹了mRNA分子的含水内核组成，外层脂质分子可进行修饰，如连接相应的基团或特异性抗体等，从而实现其细胞靶向性. LNP：脂质纳米粒；mRNA：信使核糖核酸.
图 2  LNP-mRNA递送系统的作用过程LNP-mRNA与细胞膜相关分子或受体结合（A），以内吞的方式进入细胞质内（B）形成内体（C），LNP-mRNA与内体膜融合并释放mRNA分子，mRNA在细胞质内进行翻译合成功能性蛋白（D）. LNP：脂质纳米粒；mRNA：信使核糖核酸.
图 3  基于LNP-mRNA递送系统的树突状细胞激活对CAR-T细胞治疗效果的影响LNP-mRNA靶向导入树突状细胞后，在树突状细胞细胞膜表面表达靶标分子CLDN6，该靶标分子CLDN6与CLDN6靶向的CAR-T细胞结合，能辅助性功能增强CLDN6靶向的CAR-T细胞杀伤肿瘤细胞的功能. CLDN：密封蛋白；CAR：嵌合抗原受体；LNP：脂质纳米粒；mRNA：信使核糖核酸.
图 4  基于LNP-mRNA递送系统的体内嵌合抗原受体T细胞的制备LNP-mRNA递送系统通过其表面抗CD5抗体，与体内T细胞膜表面CD5结合，促使T细胞膜表面表达FAP CAR分子，从而完成体内生成FAP靶向的CAR-T细胞.FAP靶向的CAR-T细胞与心肌纤维细胞膜表面FAP分子结合，可介导心肌纤维细胞的凋亡过程. FAP：成纤维细胞激活蛋白；CAR：嵌合抗原受体；LNP：脂质纳米粒；mRNA：信使核糖核酸.
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