Zhou Pengcheng, Ding Xian'guang, Du Xuanlong, Wang Lianhui, Zhang Yewei
School of Medicine, Southeast University, Nanjing 210000, China.
Department of General Surgery, Affiliated Hospital of Nantong University, Nantong 226001, China.
Biomater Res. 2024 Aug 2;28:0050. doi: 10.34133/bmr.0050. eCollection 2024.
As one of the most aggressive and lethal cancers, pancreatic cancer is highly associated with cancer-associated fibroblasts (CAFs) that influence the development and progression of cancer. Targeted reprogramming of CAFs may be a promising strategy for pancreatic cancer. This study aims to construct engineered extracellular vesicles (EVs) with surface modification of integrin α5 (ITGA5)-targeting peptide and high internal expression of miR-148a-3p by endogenous modification for targeted reprogramming of pancreatic CAFs. Bone marrow mesenchymal stem cells (BMSCs) and pancreatic CAFs were cocultured to examine the effect of BMSC-derived EVs on the expression levels of CAF markers. miR-148a-3p was identified as a functional molecule. The mechanism of miR-148a-3p was elucidated using the dual-luciferase reporter assay. BMSCs were infected with TERT-encoding and miR-148a-3p-encoding lentiviruses. Subsequently, BMSCs were modified with ITGA5-specific targeting peptide. The supernatant was ultracentrifuged to obtain the engineered EVs (ITGA5-EVs), which were used to reprogram CAFs. BMSCs modulated CAF marker expressions through EVs. miR-148a-3p was up-regulated in BMSCs. The expression of miR-148a-3p in pancreatic CAFs was down-regulated when compared with that in normal fibroblasts (NFs). Mechanistically, ITGA5-EVs effectively suppressed the proliferation and migration of pancreatic CAFs by targeting ITGA5 through the TGF-β/SMAD pathway. ITGA5-EVs was associated with enhanced cellular uptake and exhibited enhanced in vitro and in vivo targeting ability. Moreover, ITGA5-EVs exerted strong reconfiguration effects in inactivating CAFs and reversing tumor-promoting effects in 3D heterospheroid and xenograft pancreatic cancer models. This targeted CAF reprogramming strategy with genetically engineered ITGA5-EVs holds great promise as a precision therapeutics in clinical settings.
作为最具侵袭性和致命性的癌症之一，胰腺癌与影响癌症发生和发展的癌症相关成纤维细胞（CAF）高度相关。对CAF进行靶向重编程可能是一种有前景的胰腺癌治疗策略。本研究旨在通过内源性修饰构建表面修饰整合素α5（ITGA5）靶向肽且高表达miR-148a-3p的工程化细胞外囊泡（EV），用于胰腺癌CAF的靶向重编程。将骨髓间充质干细胞（BMSC）与胰腺CAF共培养，以检测BMSC来源的EV对CAF标志物表达水平的影响。miR-148a-3p被鉴定为一种功能分子。使用双荧光素酶报告基因测定法阐明miR-148a-3p的作用机制。用编码TERT和miR-148a-3p的慢病毒感染BMSC。随后，用ITGA5特异性靶向肽修饰BMSC。将上清液超速离心以获得工程化EV（ITGA5-EV），用于重编程CAF。BMSC通过EV调节CAF标志物的表达。BMSC中miR-148a-3p上调。与正常成纤维细胞（NF）相比，胰腺CAF中miR-148a-3p的表达下调。机制上，ITGA5-EV通过TGF-β/SMAD途径靶向ITGA5，有效抑制胰腺CAF的增殖和迁移。ITGA5-EV与增强的细胞摄取相关，并表现出增强的体外和体内靶向能力。此外，ITGA5-EV在三维异质球体和异种移植胰腺癌模型中对CAF失活和逆转肿瘤促进作用发挥了强大的重构作用。这种用基因工程ITGA5-EV进行的靶向CAF重编程策略作为一种精准治疗方法在临床应用中具有很大前景。
