Extracellular vesicles in cellular aging

细胞外囊泡在细胞衰老中的作用

• 批准号: RGPIN-2021-03424
• 负责人: Burger, Dylan
• 金额: $ 2.62万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=760886
• 关键词: Extracellular; vesicles; cellular; aging

The progressive decline in cellular metabolic function and replicative capacity is a hallmark of aging and a central determinant of human lifespan. These changes are highly conserved across all cell types and species and modifiable: accelerated in disease, but amenable to reprogramming to delay senescence and extend lifespan. We, and others, have shown that large EVs released through outward membrane blebbing (ectosomal pathway) induce senescence. Conversely, our preliminary data suggest that small EVs arising from endosomal pathways preserve function and replicative capacity. This research program explores the role of EVs in cell-cell communications in aging with the goal of discovering novel approaches to optimize cellular aging throughout life. Over the next five years we will test the specific hypothesis that senescent cells undergo a phenotypic shift from release of endosomal-derived EVs which preserve cell function to ectosomal-derived EVs which induce secondary senescence and progressive cellular aging. We will focus on three research questions: 1. Do aging cells undergo a shift from endosomal EV secretion to ectosomal EV secretion? We will examine models of replicative senescence, stress-induced premature senescence, and fibroblasts from individuals with progeria and compare the phenotype of EVs released into the extracellular environment. We quantify EV levels and assess changes in individual EV subpopulations. We will correlate levels of specific EV subpopulations (exosomes, ARRDC1-mediated microvesicles [small ectosomes], and microvesicles [large ectosomes]) with senescent signaling in cells and assess dysregulated pathways that may be responsible for aberrant EV secretion. 2. Do EVs regulate cellular aging pathways in target cells. We will assess the effects of EV subpopulations on cellular aging pathways in target cells. We will test whether EVs from senescent populations induce secondary senescence as well as whether EVs from young/healthy cells can inhibit senescence. We will seek to identify candidate proteins responsible for EV signaling by proteomics. 3. Can aging cells be reprogrammed to endosomal EV secretion, attenuating secondary senescence? Finally we will test whether it is possible to reprogram cells to retain their ability to secrete endosomal EVs. We will test 3 approaches. 1) Induction of endosomal EV formation, 2) Inhibition of ectosomal EV formation, and 3) Redirection of EV formation from ectosomal to endosomal pathways by targeting a novel molecular switch (diacylglycerol kinase ?) recently identified in collaboration with Dr. Stephen Gee. Our proposed research program will enhance understanding of the human ageing process at the cellular level. If successful, this project may identify novel targets for therapeutic intervention to inhibit cell senescence and delay aging. This, in turn could help mitigate the effects of aging and age-related morbidities in a rapidly aging Canadian population.

细胞代谢功能和复制能力的逐渐下降是衰老的标志，也是人类寿命的核心决定因素。这些变化在所有细胞类型和物种中高度保守，并且是可改变的：在疾病中加速，但易于重新编程以延迟衰老和延长寿命。我们和其他人已经表明，通过向外膜起泡（外体途径）释放的大EV诱导衰老。相反，我们的初步数据表明，由内体途径产生的小EV保留了功能和复制能力。该研究项目探讨了EV在衰老过程中细胞间通信中的作用，旨在发现优化整个生命过程中细胞衰老的新方法。在接下来的五年里，我们将测试衰老细胞经历从释放内体衍生的EV（其保留细胞功能）到外体衍生的EV（其诱导二次衰老和进行性细胞衰老）的表型转变的特定假设。我们将重点研究三个问题：1。衰老细胞是否经历了从内体EV分泌到外体EV分泌的转变？我们将研究复制性衰老，应激诱导的早衰和成纤维细胞的模型，从个人与早衰症和比较的表型释放到细胞外环境的EV。我们将特定EV亚群（外泌体、ARRDC 1介导的微囊泡[小核外体]和微囊泡[大核外体]）的水平与细胞中的衰老信号传导相关联，并评估可能导致EV分泌异常的失调途径。2.电动汽车是否调节靶细胞的细胞衰老途径。我们将评估EV亚群对靶细胞中细胞衰老途径的影响。我们将测试来自衰老群体的EV是否诱导二次衰老以及来自年轻/健康细胞的EV是否可以抑制衰老。我们将寻求通过蛋白质组学鉴定负责EV信号传导的候选蛋白质。3.衰老细胞能否被重编程为内体EV分泌，从而减弱继发性衰老？最后，我们将测试是否有可能重新编程细胞以保留其分泌内体EV的能力。我们将测试三种方法。1)诱导内体EV形成，2）抑制外体EV形成，3）通过靶向新型分子开关（二酰甘油激酶？）最近与Stephen Gee博士合作发现的。我们提出的研究计划将在细胞水平上提高对人类衰老过程的理解。如果成功，该项目可能会确定新的治疗干预目标，以抑制细胞衰老和延缓衰老。这反过来又有助于减轻加拿大人口迅速老龄化的老龄化和与年龄有关的发病率的影响。