Lee Hyunho, Massaro Matteo, Abdelfattah Nourhan, Baudo Gherardo, Liu Haoran, Yun Kyuson, Blanco Elvin
Department of Nanomedicine, Houston Methodist Research Institute, Houston, Texas, USA.
Department of Neurology, Houston Methodist Research Institute, Houston, Texas, USA.
Aging Cell. 2025 Apr;24(4):e14446. doi: 10.1111/acel.14446. Epub 2024 Dec 25.
Mesenchymal stem cells (MSCs) are promising candidates for regenerative therapies due to their self-renewal and differentiation capabilities. Pathological microenvironments expose MSCs to senescence-inducing factors such as reactive oxygen species (ROS), resulting in MSC functional decline and loss of stemness. Oxidative stress leads to mitochondrial dysfunction, a hallmark of senescence, and is prevalent in aging tissues characterized by elevated ROS levels. We hypothesized that overexpression of nuclear respiratory factor-1 (NRF1), a driver of mitochondrial biogenesis, could metabolically potentiate MSCs and prevent MSC senescence. Single-cell RNA sequencing (scRNA-Seq) revealed that MSCs transfected with NRF1 messenger RNA (mRNA) exhibited upregulated expression of genes associated with oxidative phosphorylation (OXPHOS), decreased glycolytic markers, and suppression of senescence-related pathways. To test whether NRF1 induction could mitigate stress-induced premature senescence, we exposed MSCs to hydrogen peroxide (HO) and validated our findings in a replicative senescence model. NRF1 mRNA transfection significantly increased mitochondrial mass and improved aberrant mitochondrial processes associated with senescence, including reduced mitochondrial and intracellular total ROS production. Mitochondrial health and dynamics were preserved, and respiratory function was restored, as evidenced by enhanced OXPHOS, reduced glycolysis, and increased ATP production. Notably, NRF1 overexpression led to decreased senescence-associated β-galactosidase (SA-β-gal) activity and reduced expression of senescence markers p53, p21, and p16. Our findings demonstrate that NRF1 induction attenuates MSC senescence by enhancing mitochondrial function, suggesting potential translational applications for MSC-based therapies and senescence-targeted interventions.
间充质干细胞（MSCs）因其自我更新和分化能力，是再生治疗中很有前景的候选细胞。病理微环境使MSCs暴露于活性氧（ROS）等衰老诱导因子中，导致MSCs功能衰退和干性丧失。氧化应激导致线粒体功能障碍，这是衰老的一个标志，并且在以ROS水平升高为特征的衰老组织中普遍存在。我们假设，作为线粒体生物发生驱动因子的核呼吸因子-1（NRF1）的过表达可以在代谢上增强MSCs并防止MSCs衰老。单细胞RNA测序（scRNA-Seq）显示，用NRF1信使核糖核酸（mRNA）转染的MSCs表现出与氧化磷酸化（OXPHOS）相关基因的表达上调、糖酵解标志物减少以及衰老相关途径的抑制。为了测试NRF1的诱导是否可以减轻应激诱导的早衰，我们将MSCs暴露于过氧化氢（HO）中，并在复制性衰老模型中验证了我们的发现。NRF1 mRNA转染显著增加了线粒体质量，并改善了与衰老相关的异常线粒体过程，包括减少线粒体和细胞内总ROS的产生。线粒体健康和动态得以维持，呼吸功能得以恢复，这通过增强的OXPHOS、减少的糖酵解和增加的ATP产生得以证明。值得注意的是，NRF1过表达导致衰老相关β-半乳糖苷酶（SA-β-gal）活性降低以及衰老标志物p53、p21和p16的表达减少。我们的研究结果表明，NRF1的诱导通过增强线粒体功能减轻了MSCs衰老，这表明基于MSCs的治疗和以衰老为靶点的干预措施具有潜在的转化应用价值。
