An All-in-One Nanomedicine Consisting of CRISPR-Cas9 and an Autoantigen Peptide for Restoring Specific Immune Tolerance
An All-in-One Nanomedicine Consisting of CRISPR-Cas9 and an Autoantigen Peptide for Restoring Specific Immune Tolerance
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一种由 CRISPR-Cas9 和自身抗原肽组成的一体化纳米药物，用于恢复特异性免疫耐受
DOI：
10.1021/acsami.0c10885
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发表时间：
2020
影响因子：
9.5
通讯作者：
Jun Wang
中科院分区：
文献类型：
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作者：
Ying-Li Luo;Li-Fang Liang;Yun-Jiu Gan;Jing Liu;Yue Zhang;Ya-Nan Fan;Gui Zhao;Anna Czarna;Zi-Dong Lu;Xiao-Jiao Du;Song Shen;Cong-Fei Xu;Zhe-Xiong Lian;Jun Wang
纳米技术在治疗各种疾病方面显示出巨大的前景。然而，开发可以治愈自身免疫性疾病而不引起全身免疫抑制的纳米药物仍然具有相当大的挑战性。在本文中，我们提出了一种包含自身抗原肽和CRISPR-Cas9的一体化纳米药物，以通过将树突状细胞（DC）工程化为致耐受性表型来恢复特异性免疫耐受，其可以扩增自身抗原特异性调节性T（Treg）细胞。简而言之，我们利用阳离子脂质辅助的聚（乙二醇）-b-聚（丙交酯-共-乙交酯）（PEG-PLGA）纳米颗粒来同时包封自身免疫性糖尿病相关肽（2.5mi）、CRISPR-Cas9质粒（pCas 9）和靶向共刺激分子（CD 80、CD 86和CD 40）的三种指导RNA（gRNA）。我们证明了一体化纳米药物能够有效地将这些组分共同递送到DC中，然后同时破坏三种共刺激分子并在基因组编辑的DC上呈递2.5mi肽。所得到的致耐受性DC通过在不存在共刺激信号的情况下将2.5mi肽呈递给CD 4 +T细胞来触发自身抗原特异性Treg细胞的产生和扩增。使用自身免疫性1型糖尿病（T1 D）作为典型的疾病模型，我们证明了我们的纳米药物可以预防对胰岛成分的自身免疫，并抑制T1 D的发展。我们的一体化纳米药物实现了CRISPR-Cas9和肽到DC的共递送，并且可以通过取代不同的自身抗原肽而容易地应用于其他自身免疫性疾病。
Nanotechnology has shown great promise in treating diverse diseases. However, developing nanomedicines that can cure autoimmune diseases without causing systemic immunosuppression is still quite challenging. Herein, we propose an all-in-one nanomedicine comprising an autoantigen peptide and CRISPR-Cas9 to restore specific immune tolerance by engineering dendritic cells (DCs) into a tolerogenic phenotype, which can expand autoantigen-specific regulatory T (Treg) cells. In brief, we utilized cationic lipid-assisted poly(ethylene glycol)-b-poly(lactide-co-glycolide) (PEG-PLGA) nanoparticles to simultaneously encapsulate an autoimmune diabetes-relevant peptide (2.5mi), a CRISPR-Cas9 plasmid (pCas9), and three guide RNAs (gRNAs) targeting costimulatory molecules (CD80, CD86, and CD40). We demonstrated that the all-in-one nanomedicine was able to effectively codeliver these components into DCs, followed by simultaneous disruption of the three costimulatory molecules and presentation of the 2.5mi peptide on the genome-edited DCs. The resulting tolerogenic DCs triggered the generation and expansion of autoantigen-specific Treg cells by presenting the 2.5mi peptide to CD4+T cells in the absence of costimulatory signals. Using autoimmune type 1 diabetes (T1D) as a typical disease model, we demonstrated that our nanomedicine prevented autoimmunity to islet components and inhibited T1D development. Our all-in-one nanomedicine achieved codelivery of CRISPR-Cas9 and the peptide to DCs and could be easily applied to other autoimmune diseases by substitution of different autoantigen peptides.