miR-29a-3p orchestrates key signaling pathways for enhanced migration of human mesenchymal stem cells.

IF 8.2 2区生物学 Q1 CELL BIOLOGY

Cell Communication and Signaling Pub Date : 2024-07-17 DOI:10.1186/s12964-024-01737-0

Dayeon Kang, Taehwan Kim, Ga-Eun Choi, Arum Park, Jin Yoon, Jinho Yu, Nayoung Suh

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引用次数: 0

Abstract

Background: The homing of human mesenchymal stem cells (hMSCs) is crucial for their therapeutic efficacy and is characterized by the orchestrated regulation of multiple signaling modules. However, the principal upstream regulators that synchronize these signaling pathways and their mechanisms during cellular migration remain largely unexplored.

Methods: miR-29a-3p was exogenously expressed in either wild-type or DiGeorge syndrome critical region 8 (DGCR8) knockdown hMSCs. Multiple pathway components were analyzed using Western blotting, immunohistochemistry, and real-time quantitative PCR. hMSC migration was assessed both in vitro and in vivo through wound healing, Transwell, contraction, and in vivo migration assays. Extensive bioinformatic analyses using gene set enrichment analysis and Ingenuity pathway analysis identified enriched pathways, upstream regulators, and downstream targets.

Results: The global depletion of microRNAs (miRNAs) due to DGCR8 gene silencing, a critical component of miRNA biogenesis, significantly impaired hMSC migration. The bioinformatics analysis identified miR-29a-3p as a pivotal upstream regulator. Its overexpression in DGCR8-knockdown hMSCs markedly improved their migration capabilities. Our data demonstrate that miR-29a-3p enhances cell migration by directly inhibiting two key phosphatases: protein tyrosine phosphatase receptor type kappa (PTPRK) and phosphatase and tensin homolog (PTEN). The ectopic expression of miR-29a-3p stabilized the polarization of the Golgi apparatus and actin cytoskeleton during wound healing. It also altered actomyosin contractility and cellular traction forces by changing the distribution and phosphorylation of myosin light chain 2. Additionally, it regulated focal adhesions by modulating the levels of PTPRK and paxillin. In immunocompromised mice, the migration of hMSCs overexpressing miR-29a-3p toward a chemoattractant significantly increased.

Conclusions: Our findings identify miR-29a-3p as a key upstream regulator that governs hMSC migration. Specifically, it was found to modulate principal signaling pathways, including polarization, actin cytoskeleton, contractility, and adhesion, both in vitro and in vivo, thereby reinforcing migration regulatory circuits.

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miR-29a-3p 可协调关键信号通路，促进人类间充质干细胞的迁移。

背景：人类间充质干细胞（hMSCs）的归宿对其疗效至关重要，其特征是多种信号模块的协调调控。方法：miR-29a-3p外源表达于野生型或狄氏综合征临界区8（DGCR8）基因敲除的hMSCs。通过伤口愈合、Transwell、收缩和体内迁移试验评估了hMSC的体外和体内迁移。利用基因组富集分析和 Ingenuity 通路分析进行了广泛的生物信息学分析，确定了富集通路、上游调控因子和下游靶标：结果：DGCR8 基因沉默（miRNA 生物发生的一个关键组成部分）导致的全球微RNA（miRNA）耗竭显著影响了 hMSC 的迁移。生物信息学分析发现，miR-29a-3p 是一个关键的上游调控因子。在 DGCR8 敲除的 hMSCs 中过表达 miR-29a-3p 能明显改善它们的迁移能力。我们的数据表明，miR-29a-3p 通过直接抑制两种关键的磷酸酶：蛋白酪氨酸磷酸酶受体卡帕型（PTPRK）和磷酸酶与天丝同源物（PTEN）来增强细胞迁移。在伤口愈合过程中，miR-29a-3p 的异位表达稳定了高尔基体和肌动蛋白细胞骨架的极化。此外，它还通过调节 PTPRK 和 paxillin 的水平来调节病灶粘附。在免疫受损的小鼠体内，过表达 miR-29a-3p 的 hMSCs 向趋化吸引物的迁移显著增加：我们的研究结果表明，miR-29a-3p 是控制 hMSC 迁移的关键上游调节因子。具体而言，研究发现 miR-29a-3p 可调节体外和体内的主要信号通路，包括极化、肌动蛋白细胞骨架、收缩性和粘附性，从而加强迁移调控回路。

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来源期刊

Cell Communication and Signaling CELL BIOLOGY-

CiteScore

11.00

自引率

0.00%

发文量

180

期刊介绍： Cell Communication and Signaling (CCS) is a peer-reviewed, open-access scientific journal that focuses on cellular signaling pathways in both normal and pathological conditions. It publishes original research, reviews, and commentaries, welcoming studies that utilize molecular, morphological, biochemical, structural, and cell biology approaches. CCS also encourages interdisciplinary work and innovative models, including in silico, in vitro, and in vivo approaches, to facilitate investigations of cell signaling pathways, networks, and behavior. Starting from January 2019, CCS is proud to announce its affiliation with the International Cell Death Society. The journal now encourages submissions covering all aspects of cell death, including apoptotic and non-apoptotic mechanisms, cell death in model systems, autophagy, clearance of dying cells, and the immunological and pathological consequences of dying cells in the tissue microenvironment.