Mechanism Exploration on the Immunoregulation of Allogeneic Heart Transplantation Rejection in Rats With Exosome miRNA and Proteins From Overexpressed IDO1 BMSCs

IF 3.2 4区医学 Q3 CELL & TISSUE ENGINEERING

Cell Transplantation Pub Date : 2024-04-17 DOI:10.1177/09636897241245796

Rui Zheng, Xinxin Wu, Si Li, Xinhao Chen, Dan Yan, Jigang He

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引用次数: 0

Abstract

Immunoregulation and indoleamine 2,3-dioxygenase 1 (IDO1) play pivotal roles in the rejection of allogeneic organ transplantation. This study aims to elucidate the immune-related functional mechanisms of exosomes (Exos) derived from bone marrow–derived mesenchymal stem cells (BMSCs) overexpressing IDO1 in the context of allogeneic heart transplantation (HTx) rejection. A rat model of allogeneic HTx was established. Exos were extracted after transfection with oe-IDO1 and oe-NC from rat BMSCs. Exos were administered via the caudal vein for treatment. The survival of rats was analyzed, and reverse transcription qualitative PCR (RT-qPCR) and immunohistochemistry (IHC) were employed to detect the expression of related genes. Histopathological examination was conducted using hematoxylin and eosin (HE) staining, and flow cytometry was utilized to analyze T-cell apoptosis. Proteomics and RNA-seq analyses were performed on Exos. The data were subjected to functional enrichment analysis using the R language. A protein interaction network was constructed using the STRING database, and miRWalk, TargetScan, and miRDB databases predicted the target genes, differentially expressed miRNAs, and transcription factors (TFs). Exos from BMSCs overexpressing IDO1 prolonged the survival time of rats undergoing allogeneic HTx. These Exos reduced inflammatory cell infiltration, mitigated myocardial damage, induced CD4 T-cell apoptosis, and alleviated transplantation rejection. The correlation between Exos from BMSCs overexpressing IDO1 and immune regulation was profound. Notably, 13 immune-related differential proteins (Anxa1, Anxa2, C3, Ctsb, Hp, Il1rap, Ntn1, Ptx3, Thbs1, Hspa1b, Vegfc, Dcn, and Ptpn11) and 10 significantly different miRNAs were identified. Finally, six key immune proteins related to IDO1 were identified through common enrichment pathways, including Thbs1, Dcn, Ptpn11, Hspa1b, Il1rap, and Vegfc. Thirteen TFs of IDO1-related key miRNAs were obtained, and a TF-miRNA-mRNA-proteins regulatory network was constructed. Exosome miRNA derived from BMSCs overexpressing IDO1 may influence T-cell activation and regulate HTx rejection by interacting with mRNA.

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过表达 IDO1 BMSCs 外泌体 miRNA 和蛋白质对大鼠同种异体心脏移植排斥反应的免疫调节机制探索

免疫调节和吲哚胺2,3-二氧化酶1（IDO1）在异体器官移植的排斥反应中起着关键作用。本研究旨在阐明骨髓间充质干细胞（BMSCs）过表达IDO1的外泌体（Exos）在异体心脏移植（HTx）排斥反应中与免疫相关的功能机制。建立了异体心脏移植大鼠模型。大鼠BMSCs转染oe-IDO1和oe-NC后提取外泌体。通过尾静脉注射外显子进行治疗。分析大鼠的存活率，并采用逆转录定性 PCR（RT-qPCR）和免疫组织化学（IHC）检测相关基因的表达。组织病理学检查采用苏木精和伊红（HE）染色，流式细胞术用于分析 T 细胞凋亡。对 Exos 进行了蛋白质组学和 RNA-seq 分析。使用 R 语言对数据进行了功能富集分析。使用 STRING 数据库构建了蛋白质相互作用网络，miRWalk、TargetScan 和 miRDB 数据库预测了靶基因、差异表达的 miRNA 和转录因子 (TF)。过表达 IDO1 的 BMSCs 外显子延长了接受异体 HTx 大鼠的存活时间。这些Exos减少了炎症细胞浸润，减轻了心肌损伤，诱导了CD4 T细胞凋亡，并减轻了移植排斥反应。来自过表达 IDO1 的 BMSCs 的 Exos 与免疫调节之间的关系非常密切。值得注意的是，研究发现了13种与免疫相关的差异蛋白（Anxa1、Anxa2、C3、Ctsb、Hp、Il1rap、Ntn1、Ptx3、Thbs1、Hspa1b、Vegfc、Dcn和Ptpn11）和10种显著不同的miRNA。最后，通过共同的富集途径确定了 6 个与 IDO1 相关的关键免疫蛋白，包括 Thbs1、Dcn、Ptpn11、Hspa1b、Il1rap 和 Vegfc。获得了13个与IDO1相关的关键miRNA的TFs，并构建了一个TF-miRNA-mRNA-蛋白质调控网络。从过量表达IDO1的BMSCs中提取的外泌体miRNA可能通过与mRNA相互作用影响T细胞活化并调控HTx排斥反应。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cell Transplantation 生物-细胞与组织工程

CiteScore

6.00

自引率

3.00%

发文量

审稿时长

6 months

期刊介绍： Cell Transplantation, The Regenerative Medicine Journal is an open access, peer reviewed journal that is published 12 times annually. Cell Transplantation is a multi-disciplinary forum for publication of articles on cell transplantation and its applications to human diseases. Articles focus on a myriad of topics including the physiological, medical, pre-clinical, tissue engineering, stem cell, and device-oriented aspects of the nervous, endocrine, cardiovascular, and endothelial systems, as well as genetically engineered cells. Cell Transplantation also reports on relevant technological advances, clinical studies, and regulatory considerations related to the implantation of cells into the body in order to provide complete coverage of the field.