From in vivo to in vitro heterochronic parabiosis to identify geronic factors

项目摘要 异时联体共生模型的令人兴奋的结果表明， 使衰老的细胞和组织恢复年轻的特征。鲁棒抗衰老模型的识别 介导返老还童效应的因素是将异时共生的潜力转化为 临床应用。比较无偏倚蛋白质组学分析，随后进行血清蛋白质印迹验证 样品使我们鉴定出在小鼠和人类中随年龄下降的候选“保守”抗衰老因子， 包括PEDF（SERPINF 1），已知其在干细胞自我更新、维持和增殖中起关键作用。 小鼠的生存。该项目的总体目标是识别和验证循环抗衰老因子 在我们的最保守的候选因子中，通过利用体内和体外异时共生， 实验系统这项工作将组织解决三个关键的研究问题：1）做 候选抗衰老因子在体外和体内恢复衰老细胞和组织的年轻特性？2)做 候选抗衰老因子介导异时共生对衰老组织的有益作用， 老年干细胞异时移植;和3）候选抗衰老因子是否改变与年龄相关的 体外和体内的细胞表观基因组图谱？为了实现我们的目标：我们将优先考虑保守的候选人 抗衰老因子，通过测量它们在体外复制年轻血清的能力（目的1）;我们将测试 抗衰老因子介导异时共生对体内衰老组织有益作用的必要性 (Aim 2）;我们将讨论候选抗老年因素是否在目标1中优先考虑并在目标1中验证 2的作用，以改变年龄相关的细胞表观基因组谱在体外和体内，如所确定的全基因组 分析（目标3）。我们的长期目标是评估抗衰老因素，单独或联合使用，是否能延缓或 逆转与年龄相关的细胞和组织功能下降，从而揭示潜在的目标， 治疗干预

PROJECT SUMMARY Exciting results from the heterochronic parabiosis model suggest the presence of rejuvenation factors in the circulation that restore youthful characteristics to aged cells and tissues. Identification of robust anti-geronic factors that mediate rejuvenation effects is a key to translating the potential of heterochronic parabiosis into clinical applications. Comparative un-biased proteomic analyses followed by western blot validation of serum samples led us to identify candidate “conserved” anti-geronic factors that decline with age in mice and humans, including PEDF (SERPINF1) which is known to play a critical role in stem cell self-renewal, maintenance and survival in mice. The overall goal of the proposed project is to identify and validate circulating anti-geronic factors among our top conserved candidate factors by utilizing both in vivo and in vitro heterochronic parabiosis experimental systems. This work will be organized to address the three critical research questions: 1) Do candidate anti-geronic factors restore youthful properties to aged cells and tissues in vitro and in vivo?; 2) Do candidate anti-geronic factors mediate the beneficial effects of heterochronic parabiosis on aged tissues and of heterochronic transplantation on aged stem cells?; and 3) Do candidate anti-geronic factors alter age-related cellular epigenomic profiles in vitro and in vivo? To achieve our goals: We will prioritize conserved candidate anti-geronic factors by measuring their ability to phenocopy young serum in vitro (Aim 1); We will test for necessity of anti-geronic factors to mediate beneficial effects of heterochronic parabiosis on aged tissues in vivo (Aim 2); and We will address whether the candidate anti-geronic factors prioritized in Aim 1 and validated in Aim 2 act to alter age-related cellular epigenomic profiles both in vitro and in vivo as determined by genome-wide analyses (Aim 3). Our long-term goal is to assess whether anti-geronic factors, alone or in combination, delay or reverse the age-associated functional decline of cells and tissues, thereby revealing potential targets for therapeutic intervention.