Investigating Cellular Senescence at the Single Cell Level

项目摘要/摘要 老龄化是慢性疾病发展的关键风险因素，可能会影响寿命和生活质量。蜂窝 衰老已成为延缓衰老过程的潜在靶点。衰老的细胞处于一种 增殖受阻，并与衰老和病理条件高度相关。它们累积成多个 并分泌促炎细胞因子和其他种类的分子，损害周围组织。 标记物p16和p21(细胞周期蛋白依赖性激酶抑制物)通常用于识别衰老细胞， 但新出现的证据表明，这些标记并不完全是敏感或特异的标记 衰老。目前对定义衰老细胞的确切遗传标记缺乏了解。 尤其是在单细胞水平上。研究表明，衰老的细胞清除可以缓解各种 与衰老相关的疾病对更好地了解衰老细胞标志物是至关重要的，以便 精确的药物治疗可以被设计成更有效地消除衰老细胞。在本建议书中 我们的目标是在单细胞水平上研究衰老细胞的转录组随年龄和 人体脂肪组织的感光治疗。已经证实衰老的细胞在体内积累 有衰老和慢性病的脂肪组织，以及脂肪组织中存在的大量细胞类型 为研究与衰老相关的特定细胞类型和标记物提供了一个极好的模型。目标1将 探索脂肪组织中随年龄增长的自然衰老细胞群体的转录组。 我们将使用单核RNA测序来捕捉对典型的单细胞分离敏感的细胞类型 方法比较衰老组织和年轻组织中细胞的转录组差异。我们假设 来自老年捐赠者的脂肪组织将包含更多衰老细胞，以及几个新的遗传标记 将在这些细胞中被识别。目标2将观察感光治疗对人体脂肪组织的影响 在单细胞水平上。抗衰老药物是专为消除衰老细胞而设计的药物，但我们 不知道感觉剂针对的确切细胞类型。我们将使用单核RNA测序来揭示 随着衰老细胞的消除而发生的转录组的变化，并了解哪些细胞类型被去除 抗衰老治疗。该项目将有助于推动该领域实现对衰老细胞的更好理解 脂肪组织中的种群和定义它们的标志物，这对于理解和 有可能治疗多种疾病。通过这一奖学金，我的培训将包括发展我的技能 遗传学、衰老、计算分析、通信、演示、网络、科学写作、临床 知识和指导。这项训练将在康涅狄格州大学健康中心和杰克逊夫妇的环境中进行 实验室和其他研究机构。这份工作将使我能够利用培训机会 和导师来推动我作为一名未来的内科科学家的职业生涯。

Project Summary/Abstract Aging is a key risk factor for chronic disease development which can affect lifespan and quality of life. Cellular senescence has emerged as a potential target to slow down the aging process. Senescent cells are in a state of proliferative arrest and are highly associated with aging and pathological conditions. They accumulate in multiple tissues and secrete pro-inflammatory cytokines and other kinds of molecules that damage surrounding tissues. The markers p16 and p21 (cyclin dependent kinase inhibitors) are commonly used to identify senescent cells, but emerging evidence has shown that these markers are not entirely sensitive or specific markers for senescence. There is currently a lack of understanding of the exact genetic markers that define senescent cells especially on the single cell level. With studies showing that senescent cell clearance can alleviate various diseases associated with aging it is vital to achieve a greater understanding of senescent cell markers so that precision medicine treatments can be designed to more effectively eliminate senescent cells. In this proposal we aim to examine the transcriptome of senescent cells on the single cell level both with aging and senolytic treatment in human adipose tissues. Senescent cells have been demonstrated to accumulate in adipose tissue with aging and chronic disease and with the vast array of cell types present in adipose tissue it makes an excellent model to study specific cell types and markers associated with senescence. Aim 1 will explore the transcriptome of naturally occurring senescent cell populations in adipose tissue with aging. We will use single nucleus RNA sequencing to capture cell types sensitive to typical single cell dissociation methods and compare the transcriptome differences between cells in aged vs. young tissues. We hypothesize that adipose tissue from older donors will contain more senescent cells and that several novel genetic markers will be identified in these cells. Aim 2 will look at the effects of senolytic treatment on human adipose tissue on the single cell level. Senolytics are drugs that are designed to specifically eliminate senescent cells but we do not know the precise cell types targeted by senolytics. We will use single nucleus RNA sequencing to uncover the transcriptome changes that occur with senescent cell elimination and learn what cell types are removed with senolytic treatment. This project will help advance the field achieving a greater understanding of senescent cell populations in adipose tissue and the markers that define them, which is essential to understanding and potentially treating numerous diseases. Through this fellowship my training will include developing my skills in genetics, aging, computational analysis, communication, presentation, networking, scientific writing, clinical knowledge and mentorship. This training will occur in the environment of UCONN Health as well as the Jackson Laboratory and other research institutions. This work will allow me to take advantage of training opportunities and mentorship to advance my career as a future physician-scientist.