Premature aging disorders, metabolites, and atherosclerosis

项目摘要/摘要 电离辐射(IR)引起的早衰触发的血管疾病(PmSVD) 作为Hutchinson-Gilford早衰症(HGPS)，其显著特征是过程加速 动脉粥样硬化(THs)和冠状动脉疾病(CAD)。尽管PmSVDS的内皮功能障碍很好 众所周知，预防PmSVD引起的CAD的可用治疗方法很少；因此，迫切需要 填补这一空白。TL功能障碍引起的持续性衰老相关分泌表型(PISP) 在癌症复发和耐药中发挥核心作用，但其调节机制和对通路的贡献仍然存在 未知。我们的长期目标是确定PmSVD诱导PISP的分子机制 内皮细胞(ECs)和冠心病。PmSVD通过激活蛋白激酶Cβ显著上调TOP2ζ的降解。这个 去除ECTOP2β可激活PARP和PISP，也可加速血管紧张素转换酶的活性。我们展示了关键的 线粒体ROS在蛋白激酶Cζ激活中的作用，这是线粒体核反馈环的初始步骤之一。值得注意的是， MtROS在IR和HGPS中的关键作用已得到很好的证实。最后，通过对两个样品进行IC-MS分析 IR和HGPS ECs中，我们还发现以下3条代谢产物相关途径在IR和HGPS中受到调控 共同的ECS：1)核苷酸糖-糖胺多聚糖(GAG)和硫酸盐，2)谷氨酸，3)NAD- 硫化氢(H2S)。尽管所有3种代谢物途径对冠心病和衰老的贡献 尽管如此，调控PmSVD的确切作用和机械洞察力在很大程度上仍不清楚。我们建议 一种新的假说，即PmSVD诱导的线粒体ROS激活PKCζ-TOP2β模块，随后激活TOP2β 降解，并引发TL DNA损伤。TL DNA损伤促进PARP激活，从而诱导mt 功能障碍并形成mt核反馈环，导致持续性代谢物变化，包括 核苷糖和NAD-H_2S途径，导致PISP和CAD。我们将通过以下方式测试我们的假设： 以下三个具体目标：在目标1中，我们将确定以下三个共同的作用和监管机制 PmSVD的体外代谢产物相关途径：1)核苷酸-糖和硫酸盐，2)谷氨酸，3)NAD- 硫化氢。在目标2中，我们将表征PKCζ-top2β模块和PARP1在PmSVD介导的代谢物中的作用 在体外的变化和线粒体功能障碍。在目标3中，我们将确定PKCζ-TOP2β模块和 PmSVD诱导的活体冠状动脉损伤(CATH)后PARP的激活。建议的工作是 有望确定PKC、ζ-TOP2、β和PARP作为调节PmSVD诱导的中枢分子的作用 代谢物改变和PISP。这种方法是创新的，因为我们将使用IPSC、ION的新技术 色质联用(IC-MS)、机器学习、成像质谱仪和一种新的小鼠 凯斯模型。拟议的研究应该会对PmSVD产生积极影响，因为它导致了一种新的方法 抑制PISP值。

Project Summary/Abstract Premature senescence-triggered vascular diseases (PmSVD) induced by ionizing radiation (IR), as well as Hutchinson-Gilford progeria syndrome (HGPS), are notably characterized by accelerating processes of atherosclerosis (AthS) and coronary artery disease (CAD). Although endothelial dysfunction in PmSVDs is well known, there is a paucity of available treatments to prevent PmSVD-induced CAD; hence, there is an urgent need to fill this gap. Persistent senescence-associated secretory phenotype (PISP), provoked by TL dysfunction, plays a central role in cancer recurrence and resistance, but its regulatory mechanisms and contribution to AthS remain unknown. Our long-term goal is to determine the molecular mechanisms by which PmSVD induces PISP in endothelial cells (ECs) and CAD. PmSVD significantly up-regulated TOP2β degradation via PKCζ activation. The depletion of EC TOP2β instigated PARP activation and PISP; it also accelerated AthS. We showed the critical role of mtROS in PKCζ activation, which is one of the initial steps for the Mt-nucleus feedback loop. Of note, the crucial role of mtROS in both IR and HGPS has been well established. Lastly, by performing IC-MS analysis in both IR and HGPS ECs, we also found that the following 3 metabolite-related pathways were regulated in IR and HGPS ECs in common: 1) nucleotide sugars-glycosaminoglycans (GAGs) and sulfate, 2) glutamate, and 3) NAD+- hydrogen sulfide (H2S). Although the contribution of all 3 metabolites pathways to CAD and aging has been suggested, the exact role and mechanical insights in regulating PmSVD remain largely unknown. We propose the novel hypothesis that PmSVD-induced mtROS activates the PKCζ-TOP2β module, followed by TOP2β degradation, and instigates TL DNA damage. TL DNA damage promotes PARP activation, which induces mt dysfunction and forms an mt-nucleus feedback loop, resulting in persistent metabolites changes, including nucleotide sugars and NAD+-H2S pathways, causing PISP and CAD. We will test our hypothesis by pursuing the following 3 specific aims: In Aim 1, we will determine the role and regulatory mechanisms of the following 3 common metabolites-related pathways in PmSVD in vitro; 1) nucleotide sugars-GAGs and sulfate, 2) glutamate, 3) NAD+- H2S. in Aim 2, we will characterize the role of PKCζ-TOP2β module and PARP1 in PmSVD-mediated metabolites changes and mt dysfunction in vitro. In Aim 3, we will determine the role of the PKCζ-TOP2β module and subsequent PARP activation in PmSVD-induced coronary AthS (CAthS) in vivo. The proposed work is expected to establish the roles of PKCζ-TOP2β and PARP as the hub molecules in regulating PmSVD-induced metabolite changes and PISP. The approach is innovative because we will use the new technologies of iPSC, ion chromatography-mass spectrometry (IC-MS), machine learning, imaging mass cytometry, and a novel mouse CAthS model. The proposed research should positively impact PmSVD by leading to a novel approach to inhibiting PISP.