Cell autonomous and non-autonomous mechanisms bv which endogenous DNA damage pro

该项目的长期目标是确定损伤，特别是由于内源性过程而自发产生的核基因组损伤促进与衰老有关的退行性变化的机制。目前的模型是，随机损伤通过细胞自主机制促进衰老，即，通过驱动细胞衰老或细胞死亡。我们试图通过仔细研究自发DNA损伤如何在DNA修复缺陷引起的人类早老综合征小鼠模型中驱动衰老来挑战这一模型。该方法将是联合收割机结合独特的遗传和质谱分析工具，以实现以下目标： 1)使用高灵敏度LC-MS/MS/MS测定来测量多个年龄的早老性和野生型小鼠组织中的内源性氧化DNA损伤，并确定损伤水平是否可预测这些组织中衰老相关病理的程度。 2)一次在一种组织或细胞类型中从基因上耗尽DNA修复，并确定对这些细胞、邻近细胞和远处组织的影响，以确定损伤是否通过细胞自主或非自主机制驱动衰老。 3)利用正常小鼠和早衰小鼠之间的联体共生来确定循环因子是否可以克服由于过多DNA损伤而导致的加速衰老。 4)使用无偏见和有针对性的蛋白质组学方法，以确定系统信号激活的遗传毒性压力和老年。 这些目标的成功完成预计将产生关于衰老过程的新信息，以及是否存在多种衰老相关疾病的共同潜在机制。蛋白质组学工作可能会产生新的生物年龄生物标志物。此外，我们的初步工作通过这个项目的扩展，预计将产生合理的策略，以延长老年人的健康寿命，通过减少随机损伤和/或抑制损伤反应，促进衰老。

The long-term objective of this project is to identify the mechanism by which damage, in particular damage to the nuclear genome that arises spontaneously as a consequence of endogenous processes, promotes aging-related degenerative changes. The current model is that stochastic damage promotes aging via a cell autonomous mechanism, i.e., by driving cell senescence or cell death. We seek to challenge this model by carefully examining how spontaneous DNA damage drives aging in a mouse model of a human progeriod syndrome caused by a defect in DNA repair. The approach will be to combine unique genetic and mass spectrometry tools to accomplish the following aims: 1) To use highly sensitive LC-MS/MS/MS assays to measure endogenous oxidative DNA lesions in tissues of progeroid and wild-type mice at multiple ages and determine if the level of damage predicts the extent of aging-related pathology in each of these tissues. 2) To genetically deplete DNA repair in one tissue or cell type at a time and determine the impact on those cells, neighboring cells and distant tissues, to determine if damage drives aging via a cell-autonomous or non-autonomous mechanism. 3) To use parabiosis between normal and progeroid mice to determine if circulating factors can overcome accelerated aging due to too much DNA damage. 4) To use unbiased and targeted proteomics approaches to identify systemic signals activated in response to genotoxic stress and old age. Successful completion of these aims is anticipated to yield novel information about the aging process and if there are common underlying mechanisms for multiple aging-related diseases. The proteomics work may yield new biomarkers of biologic age. In addition, extension of our preliminary work via this project is anticipated to yield rational strategies to extend healthspan ofthe elderly by reducing stochastic damage and/or inhibiting the damage responses that promote aging.