Department of Respiratory and Critical Care Medicine, Binzhou Medical University Hospital, Binzhou Medical University, Binzhou 256603, China.
Department of Cellular and Genetic Medicine, School of Pharmaceutical Sciences, Binzhou Medical University, Yantai 264003, China.
Aging (Albany NY). 2023 Jun 16;15(12):5514-5534. doi: 10.18632/aging.204805.
Idiopathic pulmonary fibrosis (IPF) is a fibrosing interstitial pneumonia of unknown cause. The most typical characteristic of IPF is gradual weakening of pulmonary elasticity and increase in hardness/rigidity with aging. This study aims to identify a novel treatment approach for IPF and explore mechanism of mechanical stiffness underlying human umbilical cord mesenchymal stem cells (hucMSCs) therapy. Target ability of hucMSCs was examined by labeling with cell membrane dye Dil. Anti-pulmonary fibrosis effect of hucMSCs therapy by reducing mechanical stiffness was evaluated by lung function analysis and MicroCT imaging system and atomic force microscope and . Results showed that stiff environment of fibrogenesis caused cells to establish a mechanical connection between cytoplasm and nucleus, initiating expression of related mechanical genes such as Myo1c and F-actin. HucMSCs treatment blocked force transmission and reduced mechanical force. For further exploration of mechanism, ATGGAG was mutated to CTTGCG (the binding site of miR-136-5p) in the full-length sequence of circANKRD42. Wildtype and mutant plasmids of circANKRD42 were packaged into adenovirus vectors and sprayed into lungs of mice. Mechanistic dissection revealed that hucMSCs treatment repressed circANKRD42 reverse splicing biogenesis by inhibiting hnRNP L, which in turn promoted miR-136-5p binds to 3'-Untranslated Region (3'-UTR) of YAP1 mRNA directly, thus inhibiting translation of YAP1 and reducing YAP1 protein entering nucleus. The condition repressed expression of related mechanical genes to block force transmission and reduce mechanical forces. The mechanosensing mechanism mediated directly by circANKRD42-YAP1 axis in hucMSCs treatment, which has potential general applicability in IPF treatment.
特发性肺纤维化（IPF）是一种原因不明的纤维性间质性肺炎。IPF 的最典型特征是肺弹性逐渐减弱，随着年龄的增长硬度/刚性增加。本研究旨在为 IPF 寻找一种新的治疗方法，并探讨人脐带间充质干细胞（hucMSCs）治疗中机械僵硬的潜在机制。通过细胞膜染料 Dil 标记来检查 hucMSCs 的靶向能力。通过肺功能分析、MicroCT 成像系统和原子力显微镜评估 hucMSCs 治疗通过降低机械僵硬来减少肺纤维化的效果。结果表明，纤维化形成的刚性环境导致细胞在细胞质和细胞核之间建立机械连接，启动了 Myo1c 和 F-肌动蛋白等相关机械基因的表达。hucMSCs 治疗阻断了力的传递并降低了机械力。为了进一步探讨机制，将 ATGGAG 突变为 CTTGCG（miR-136-5p 的结合位点）在 circANKRD42 的全长序列中。将 circANKRD42 的野生型和突变型质粒包装到腺病毒载体中并喷洒到小鼠肺部。机制分析表明，hucMSCs 治疗通过抑制 hnRNP L 抑制 circANKRD42 的反向剪接生物发生，从而促进 miR-136-5p 直接结合 YAP1 mRNA 的 3'-非翻译区（3'-UTR），从而抑制 YAP1 的翻译并减少 YAP1 蛋白进入细胞核。该条件抑制了相关机械基因的表达，从而阻断力的传递并降低机械力。circANKRD42-YAP1 轴介导的机械感觉机制在 hucMSCs 治疗中具有直接的潜在普遍性，可用于 IPF 的治疗。
