Du Fang-Ying, Zhou Feng, Zhao Na, Bao Lei, Hu Cheng-Biao, Lei Jing, Liu An-Qi, Gao Ying-Feng, Bao Li-Hui, Ni Hua, Yu Xiao-Rui, Chen Ji, Sui Bing-Dong
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, China.
State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi International Joint Research Center for Oral Diseases, Center for Tissue Engineering, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, China.
Theranostics. 2025 Jan 6;15(5):1930-1948. doi: 10.7150/thno.103314. eCollection 2025.
Mesenchymal stem cells (MSCs) possess potent immunomodulatory capability, but occasionally, clinical application of MSCs is hindered by compromised cell functionality and insufficient therapeutic efficacy. Here, well-established mouse models of dextran sulfate sodium (DSS)-induced colitis and streptozotocin (STZ)-induced type 1 diabetes (T1D) were used to evaluate therapeutic immunomodulatory effects of human umbilical cord-derived MSCs. MSCs were examined at the fifth (P5) and the fifteenth (P15) passages, and three-dimensional (3D) culture was conducted by Matrigel incorporation. A series of biochemical, histopathological and cellular assays were performed to investigate the MSC function and therapeutic performance, and immunoregulation was evaluated by co-culture with T cells and analyses of T-cell infiltration into target tissues. RNA sequencing (RNA-seq) analysis followed by immunofluorescence staining, gene expression analyses and chemical regulation were used to investigate the molecular targets. MSCs lose therapeutic immunomodulatory effects after extensive expansion to P15 when cell senescence occurs. Intriguingly, 3D preconditioning of MSCs in Matrigel promotes diminished immunoregulatory capability despite extensive passages, which benefits function of P15-MSCs to modulate T-cell subsets in co-culture, suppress infiltration of pro-inflammatory T cells in the colon and pancreas tissues after infusion, ameliorate systemic inflammation, and alleviate colitis and T1D in mice. Mechanistically, 3D culture provokes transcriptomic reprogramming of MSCs toward a Yes-associated protein 1 (YAP1)-marked, Hippo signaling pathway-upregulated state with promoted release of the anti-inflammatory cytokine, transforming growth factor-beta1 (TGF-β1). Moreover, chemical regulation of YAP1 by clinically relevant drugs, verteporfin (VP) and prostaglandin E2 (PGE2), affects TGF-β1 expression and the immunomodulatory capability of MSCs during dimensional culture. Taken together, these findings unravel YAP1-based dimensional and chemical coordination of expanded MSC immunoregulation, which will shed light on precisely controlled translational application.
间充质干细胞（MSCs）具有强大的免疫调节能力，但在临床上，MSCs的应用偶尔会受到细胞功能受损和治疗效果不足的阻碍。在这里，利用已建立的硫酸葡聚糖钠（DSS）诱导的结肠炎和链脲佐菌素（STZ）诱导的1型糖尿病（T1D）小鼠模型来评估人脐带间充质干细胞的治疗性免疫调节作用。在第5代（P5）和第15代（P15）传代时对间充质干细胞进行检测，并通过掺入基质胶进行三维（3D）培养。进行了一系列生化、组织病理学和细胞分析以研究间充质干细胞的功能和治疗性能，并通过与T细胞共培养以及分析T细胞浸润到靶组织中来评估免疫调节。采用RNA测序（RNA-seq）分析，随后进行免疫荧光染色、基因表达分析和化学调控来研究分子靶点。当细胞衰老发生时，间充质干细胞在广泛传代至P15后失去治疗性免疫调节作用。有趣的是，在基质胶中对间充质干细胞进行3D预处理，尽管传代广泛，但仍能促进免疫调节能力减弱，这有利于P15间充质干细胞在共培养中调节T细胞亚群，抑制输注后结肠和胰腺组织中促炎性T细胞的浸润，改善全身炎症，并减轻小鼠的结肠炎和1型糖尿病。从机制上讲，3D培养促使间充质干细胞的转录组重编程为以Yes相关蛋白1（YAP1）为标志、Hippo信号通路上调的状态，并促进抗炎细胞因子转化生长因子-β1（TGF-β1）的释放。此外，临床相关药物维替泊芬（VP）和前列腺素E2（PGE2）对YAP1的化学调控影响3D培养期间间充质干细胞的TGF-β1表达和免疫调节能力。综上所述，这些发现揭示了基于YAP1的扩展间充质干细胞免疫调节的维度和化学协调，这将为精确控制的转化应用提供启示。
