Obesity-induced dysfunction of human MSC in peripheral microvascular repair

肥胖引起的人间充质干细胞在外周微血管修复中的功能障碍

• 批准号: 10516515
• 负责人: Alfonso Eirin
• 金额: $ 70.66万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10516515
• 关键词: Address; Adipose tissue; Age; Attenuated; Biological Models; Bobcat; Body Weight decreased; Bone Marrow; Cardiovascular Diseases; Cell physiology; Cells; Chronic; Clinical; Cost Savings; Cytosine; Diabetes Mellitus; Down-Regulation; Endocrine; Endothelial Cells; Enhancers; Enzyme Inhibitor Drugs; Epidemic; Epigenetic Process; Event; Exhibits; Family suidae; Fatty acid glycerol esters; Functional disorder; Gene Expression; Gene Targeting; Genes; Harvest; Histones; Human; Impairment; In Vitro; Inflammation; Injury; Kidney; Kidney Diseases; Life Style Modification; Link; Magnetic Resonance Imaging; Mediating; Mesenchymal; Metabolic; Methods; Methylation; MicroRNAs; Mitochondria; Mitochondrial DNA; Modification; Mus; Obesity; Organ; Pathogenesis; Patients; Peptide antibodies; Peptides; Peripheral; Peripheral Vascular Diseases; Peripheral arterial disease; Plasma; Predisposition; Prevalence; Process; Property; Quality of life; Regulation; Renal Artery Stenosis; Role; Sampling; Secondary to; Stromal Cells; Structure; System; Techniques; Testing; Therapeutic; Thinness; Tissues; Vascular Diseases; angiogenesis; bariatric surgery; base; cardiovascular risk factor; cell injury; citrate carrier; critical limb Ischemia; effective therapy; falls; functional disability; human subject; improved; in vivo; in vivo imaging; injured; injury recovery; microCT; mitochondrial dysfunction; mortality risk; mouse model; neutralizing antibody; novel; obese patients; paracrine; promoter; protective effect; protein expression; repair function; repaired; reparative capacity; restoration; stem; tissue repair; tool; transcriptome sequencing

Abstract Obesity triggers cellular damage and impedes tissue recovery from injury, and its escalating prevalence may promote complications of peripheral vascular disease, such as critical limb ischemia (CLI) or renal artery stenosis (RAS). Reducing complications of obesity could diminish the risk of death, improve quality of life, and produce extensive cost savings. This application is based on the scientific premise that obesity increases tissue susceptibility to injury by interfering with normal defense and repair processes associated with mesenchymal stem/stromal cells (MSCs). MSCs constitute an effective endogenous cellular repair system, but obesity may blunt their efficacy. We found that obesity-induced MSC dysfunction in pigs was associated with altered mitochondrial structure and function, but the mechanisms of mitochondrial damage in human MSC and its contribution to regulation of MSC function in human obesity remain unknown. Our central hypothesis is that human obesity engages epigenetic mechanisms that impair human MSC mitochondrial structure and function and render MSC functionally deficient. We speculate that obesity alters in MSC the epigenetic states of micro-RNA (miR) miR-181a, a key miR that targets mitochondrial DNA and negatively regulates their function. A consequent fall in levels of the mitochondrial derived peptide (MDP) MOTS-c in turn impairs function and tissue repair capacity of MSC in obesity. To test our hypothesis, we will define gene expression and epigenetic states of mitochondrial targeting miRNAs and MOTS-c in human adipose tissue-derived MSC and elucidate their functional significance for both MSCs and their mitochondria. Our Specific Aims will pursue 3 hypotheses. Aim 1: Human obesity induces MSC miR-181a expression and in turn mitochondrial and MSC structural damage and dysfunction. Using RNA-seq we will identify miR-181a as a key miR upregulated in MSCs from patients with obesity vs. healthy controls. Its role in regulating MSC and mitochondrial function and structure will be assessed in vitro and in vivo (in mice with CLI or RAS) using novel in vivo imaging and ex vivo techniques. Aim 2: Human obesity engages epigenetic mechanisms to alter miR- 181a. We will define the epigenetic landscape of miR-181a using MeDIP-seq, and its contribution to MSC repair in vitro and in vivo using an epigenetic modifier. Aim 3: A fall in MOTS-c owing to mitochondrial damage contributes to functional impairment of ‘obese MSC’. Using novel MDP-seq we will pinpoint MOTS-c as a unique MDP linking mitochondrial to cellular integrity in MSC. MSC treated with MOTS-c peptide or neutralizing antibody will be characterized, and restoration of ‘obese’ MSC function tested both in vitro and in vivo. The proposed studies, employing cutting edge techniques, may uncover novel mechanisms underlying cell damage and impaired repair in human obesity. These studies will advance understanding of the pathogenesis of cellular damage, and likely contribute towards management of patients with obesity and vascular disease.

抽象的 肥胖会触发细胞损伤并阻碍组织从损伤中恢复及其不断升高的患病率 可能促进周围血管疾病的并发症，例如临界肢体缺血（CLI）或肾动脉 狭窄（RAS）。减少肥胖并发症可能会降低死亡的风险，改善生活质量和 节省了大量成本。该应用基于肥胖增加的科学前提 通过干扰正常的防御和维修过程，组织对损伤的易感性 间充质干/基质细胞（MSC）。 MSC构成有效的内源性细胞修复系统， 但是肥胖可能会钝其效率。我们发现肥胖引起的猪中的MSC功能障碍与 线粒体结构和功能的改变，但是人类MSC中线粒体损伤的机制 它对人类肥胖症中MSC功能调节的贡献尚不清楚。 我们的中心假设是，人肥胖与损害人类MSC的表观遗传机制有关 线粒体结构和功能以及渲染MSC功能不足。我们推测肥胖症 在MSC中变化微-RNA的表观遗传态（miR）miR-181a，靶向线粒体DNA的关键miR 并负面调节其功能。随之而来的线粒体衍生肽（MDP）的水平下降 MOTS-C反过来会损害MSC在肥胖症中的功能和组织修复能力。为了检验我们的假设，我们将 定义人体中线粒体靶向moRNA和MOTS-C的基因表达和表观遗传态 脂肪组织衍生的MSC并阐明了其在MSC及其线粒体中的功能意义。 我们的具体目标将提出3个假设。目标1：人肥胖会诱导MSC miR-181a表达和 进而线粒体和MSC结构损伤和功能障碍。使用RNA-seq，我们将识别miR-181a为 肥胖症患者与健康对照患者的MSC中上调的关键miR。它在规范MSC和 线粒体功能和结构将在体外和体内评估（在具有CLI或RAS的小鼠中），并使用新颖 体内成像和离体技术。目标2：人类肥胖与表观遗传机制不同以改变mir- 181a。我们将使用MEDIP-SEQ定义miR-181a的表观遗传景观及其对MSC的贡献 使用表观遗传修饰剂在体外和体内修复体内。 AIM 3：由于线粒体损伤，MOTS-C下降 有助于“肥胖MSC”的功能障碍。使用新颖的MDP-seq，我们将把MotPoint Chim-C作为一个 独特的MDP将线粒体与MSC中的细胞完整性联系起来。 MSC用Mots-C Pepper处理或中和 抗体将被表征，并且在体外和体内测试了“肥胖” MSC功能的恢复。 提出的研究采用尖端技术可能会发现细胞下的新机制 人类肥胖症的损害和修复受损。这些研究将提高对发病机理的理解 细胞损伤，可能有助于肥胖和血管疾病患者的治疗。