Premature aging disorders, metabolites, and atherosclerosis

项目概要/摘要 电离辐射 (IR) 诱发的过早衰老引发的血管疾病 (PmSVD) 以及 哈钦森-吉尔福德早衰综合症 (HGPS) 的显着特点是加速进程 动脉粥样硬化（AthS）和冠状动脉疾病（CAD）。尽管 PmSVD 中的内皮功能障碍是很常见的 众所周知，预防 PmSVD 诱发的 CAD 的可用治疗方法很少；因此，迫切需要 填补这个空白。由 TL 功能障碍引起的持续性衰老相关分泌表型 (PISP) 发挥着重要作用 在癌症复发和耐药中发挥核心作用，但其调节机制和对 AthS 的贡献仍然存在 未知。我们的长期目标是确定 PmSVD 在中诱导 PISP 的分子机制 内皮细胞 (EC) 和 CAD。 PmSVD 通过 PKC z 激活显着上调 TOP2β 降解。的 EC TOP2β 的耗尽会引发 PARP 激活和 PISP；它还加速了 AthS。我们展示了关键的 mtROS 在 PKC z 激活中的作用，这是 Mt 核反馈回路的初始步骤之一。值得注意的是， mtROS 在 IR 和 HGPS 中的关键作用已经得到充分证实。最后，通过对两者进行 IC-MS 分析 IR和HGPS ECs，我们还发现以下3个代谢相关途径在IR和HGPS中受到调节 EC 的共同点：1) 核苷酸糖-糖胺聚糖 (GAG) 和硫酸盐，2) 谷氨酸，以及 3) NAD - 硫化氢（H2S）。尽管所有 3 种代谢物途径对 CAD 和衰老的贡献已被证实 研究表明，调节 PmSVD 的确切作用和机制仍然很大程度上未知。我们建议 新假设：PmSVD 诱导的 mtROS 激活 PKC z-TOP2β 模块，然后激活 TOP2β 降解，并引发 TL DNA 损伤。 TL DNA 损伤促进 PARP 激活，从而诱导 mt 功能障碍并形成线粒体核反馈回路，导致持续的代谢物变化，包括 核苷酸糖和 NAD -H2S 途径，导致 PISP 和 CAD。我们将通过追求 以下 3 个具体目标：在目标 1 中，我们将确定以下 3 个常见的角色和监管机制 PmSVD 体外代谢相关途径； 1) 核苷酸糖-GAG 和硫酸盐，2) 谷氨酸，3) NAD - 硫化氢。在目标 2 中，我们将描述 PKC z-TOP2β 模块和 PARP1 在 PmSVD 介导的代谢物中的作用 体外变化和 mt 功能障碍。在目标 3 中，我们将确定 PKC z-TOP2β 模块的作用和 PmSVD 诱导的体内冠状动脉 AthS (CAthS) 中随后的 PARP 激活。拟议的工作是 预计将确定 PKCζ-TOP2β 和 PARP 作为调节 PmSVD 诱导的中枢分子的作用 代谢变化和 PISP。该方法具有创新性，因为我们将使用 iPSC、ion 色谱-质谱联用仪 (IC-MS)、机器学习、成像质谱流式细胞仪和新型小鼠 CAthS 模型。拟议的研究应该通过提出一种新的方法来对 PmSVD 产生积极影响 抑制 PISP。

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.