A functionalized collagen-I scaffold delivers microRNA 21-loaded exosomes for spinal cord injury repair
A functionalized collagen-I scaffold delivers microRNA 21-loaded exosomes for spinal cord injury repair
复制标题
功能化 I 型胶原蛋白支架可提供负载 microRNA 21 的外泌体，用于脊髓损伤修复
DOI：
10.1016/j.actbio.2022.10.027
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发表时间：
2022-12-07
影响因子：
9.7
通讯作者：
Suo, Guangli
中科院分区：
文献类型：
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作者：
Liu, Xingzhi;Zhang, Lulu;Suo, Guangli
基于microRNA（miRNA）的疗法在脊髓损伤（SCI）的修复中显示出巨大的潜力。microRNA 21（miR 21）已被证明对SCI具有重要的保护作用。然而，由于其易降解和无效的细胞渗透性，miRNA的应用存在一些挑战。exosomes作为天然囊泡，具有免疫原性低、固有稳定性好、组织/细胞穿透性强等优点，是理想的miRNA载体。然而，靶向性差和特异性miRNA的低能力阻碍了它们的实际应用。本研究的目的是开发一种基因工程的miR 21外泌体，它可以包埋在胶原-I（Col-I）支架中修复SCI。胶原结合结构域（CBD）融合的溶酶体相关膜糖蛋白2b（Lamp 2b）蛋白（CBD-LP）和miR 21在宿主HEK 293 T（293 T）细胞中过表达，所述宿主HEK 293 T（293 T）细胞用于产生工程化的装载miR 21的外泌体。将CBD肽与外泌体表面的Lamp 2b融合，可以稳定地将外泌体与Col-I支架连接，促进载miR 21的外泌体在病变部位的滞留，促进miR 21向细胞的持续释放。最后，我们开发了一种富含miR 21外泌体的功能化Col-I支架生物材料，它可能有助于脊髓损伤的修复。意义声明基于miRNA的治疗方法在脊髓损伤（SCI）修复中具有很好的潜力。然而，miRNAs的易降解性和无效的细胞渗透性阻碍了其应用。外泌体是递送miRNA的天然载体，但面临体内扩散的挑战。在此，在HEK 293 T细胞中过表达胶原结合结构域（CBD）融合的Lamp 2b和miR 21，以产生装载miR 21和CBD修饰的外泌体（CBD-LP-miR 21-EXO）。外泌体表面修饰的CBD可以稳定地将外泌体拴系到胶原蛋白-I支架上，以形成功能化的CBD-LP-miR 21-EXO-Col支架，其可以促进负载miR 21的外泌体的保留，促进miR 21向细胞的持续释放，并最终有利于SCI修复。此外，这种类型的功能化胶原蛋白-I材料可以通过富集负载有适当miRNA的CBD-LP-EXO而广泛应用于其他组织损伤修复。(c)2022 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
MicroRNA (miRNA)-based therapies have shown great potential in the repair of spinal cord injury (SCI). MicroRNA 21 (miR21) has been proven to have an essential protective effect on SCI. However, there are some challenges for miRNAs application due to their easy degradation and ineffective cell penetration. As natural vesicles, exosomes were considered ideal carriers for miRNAs delivery for their advantages of low immunogenicity, inherent stability and tissue/cell penetration. However, poor targeting and the low capacity of specific miRNAs impede their practical applications. This study aims to develop a type of ge-netically engineered miR21-loaded exosomes that can be entrapped in collagen-I (Col-I) scaffold to repair SCI. The collagen-binding domain (CBD)-fused lysosome-associated membrane glycoprotein 2b (Lamp2b) protein (CBD-LP) and miR21 were overexpressed in host HEK293T (293T) cells that were used to produce engineered miR21-loaded exosomes. The CBD peptide fused in Lamp2b on the exosome surface can stably tether exosomes to Col-I scaffold, facilit ate the retention of miR21-loaded exosomes in lesion sites, pro-mote the sustained release of miR21 to cells. Finally, a functionalized Col-I scaffold biomaterial enriched with miR21-loaded exosomes was developed and it could benefit the repair of SCI.Statement of significance MiRNA-based therapeutics have promising potential in spinal cord injury (SCI) repair. However, easy degradation and ineffective cell penetration impede miRNAs application. Exosomes are natural vehicles for miRNAs delivery but face the challenge of diffusion in vivo . Here, the collagen-binding domain (CBD)-fused Lamp2b and miR21 were overexpressed in HEK293T cells to produce miR21-loaded and CBD-modified exosomes (CBD-LP-miR21-EXOs). The CBD modified on the exosome surface can stably tether exosomes to collagen-I scaffold to form functionalized CBD-LP-miR21-EXO-Col scaffold that can facilitate the retention of miR21-loaded exosomes, promote the sustained release of miR21 to cells and finally benefit SCI repair. Furthermore, this type of functionalized collagen-I materials can be widely applied for other tissue injury repairs by enriching the CBD-LP-EXOs loaded with appropriate miRNAs.(c) 2022 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.