In situ repurposing of dendritic cells with CRISPR/Cas9-based nanomedicine to induce transplant tolerance
In situ repurposing of dendritic cells with CRISPR/Cas9-based nanomedicine to induce transplant tolerance
复制标题
利用基于 CRISPR/Cas9 的纳米药物原位重新利用树突状细胞以诱导移植耐受
DOI：
10.1016/j.biomaterials.2019.119302
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发表时间：
2019
期刊：
影响因子：
14
通讯作者：
Jun Wang
中科院分区：
文献类型：
--
作者：
Yue Zhang;Song Shen;Gui Zhao;Cong-Fei Xu;Hou-Bing Zhang;Ying-Li Luo;Zhi-Ting Cao;Jia Shi;Zhi-Bin Zhao;Zhe-Xiong Lian;Jun Wang
器官移植是治疗终末期器官衰竭的唯一有效方法。然而，它一直受到免疫排斥的困扰，这主要是由T细胞介导的反应引起的。树突状细胞（Dendritic cells，DC）是专职的抗原提呈细胞，阻断DC中的共刺激信号分子CD 40可抑制T细胞活化，诱导移植耐受。在这项研究中，为了缓解移植排斥，制备了Cas9 mRNA（mCas 9）和靶向共刺激分子CD 40（gCD 40）的指导RNA，并将其封装到基于聚（乙二醇）-嵌段-聚（丙交酯-共-乙交酯）（PEG-b-PLGA）的阳离子脂质辅助纳米颗粒（CLAN）中，表示为CLANmCas 9/gCD 40。CLAN在体外和体内均有效地将mCas 9/gCD 40递送到DC中，并在基因组水平上破坏DC中的CD 40。在静脉注射到急性小鼠皮肤移植模型中后，CLANmCas 9/gCD 40介导的CD 40破坏显著抑制了T细胞活化，这减少了移植物损伤并延长了移植物存活。这项工作为用携带CRISPR/Cas9系统的纳米颗粒重编程DC以减轻移植排斥提供了一种有前途的策略。
Organ transplantation is the only effective method to treat end-stage organ failure. However, it is continuously plagued by immune rejection, which is mostly caused by T cell-mediated reactions. Dendritic cells (DCs) are professional antigen-presenting cells, and blocking the costimulatory signaling molecule CD40 in DCs inhibits T cell activation and induces transplant tolerance. In this study, to relieve graft rejection, Cas9 mRNA (mCas9) and a guide RNA targeting the costimulatory molecule CD40 (gCD40) were prepared and encapsulated into poly(ethylene glycol)-block-poly(lactide-co-glycolide) (PEG-b-PLGA)-based cationic lipid-assisted nanoparticles (CLAN), denoted CLANmCas9/gCD40. CLAN effectively delivered mCas9/gCD40 into DCs and disrupted CD40 in DCs at the genomic level bothin vitroandin vivo. After intravenous injection into an acute mouse skin transplant model, CLANmCas9/gCD40-mediated CD40 disruption significantly inhibited T cell activation, which reduced graft damage and prolonged graft survival. This work provides a promising strategy for reprogramming DCs with nanoparticles carrying the CRISPR/Cas9 system to abate transplant rejection.