Premature aging disorders, metabolites, and atherosclerosis

项目概要/摘要 电离辐射（IR）诱导的早衰触发血管疾病（PmSVD），以及 如Hutchinson-Gilford早老综合征（HGPS），其显著特征是加速过程 动脉粥样硬化（AthS）和冠状动脉疾病（CAD）。虽然PmSVD中的内皮功能障碍是好的， 已知，缺乏预防PmSVD诱导的CAD的可用治疗;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）， 在癌症复发和耐药性中起核心作用，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诱导PISP的分子机制。 内皮细胞（EC）和CAD。PmSVD通过激活PKC β显著上调TOP2β降解。的 EC TOP2β的耗竭引起PARP活化和PISP;它还加速AthS。我们展示了 mtROS在PKC β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中的关键作用已经被很好地确定。最后，通过在两个样品中进行IC-MS分析， 在IR和HGPS EC中，我们还发现以下3条代谢相关途径在IR和HGPS EC中受到调节 共同的EC：1）核苷酸糖-糖胺聚糖（GAG）和硫酸盐，2）谷氨酸盐，和3）NAD+- 硫化氢（H2S）。尽管所有3种代谢途径对CAD和衰老的贡献已经被证实， 因此，PmSVD在调节PmSVD中的确切作用和机制仍然是未知的。我们建议 PmSVD诱导的线粒体ROS激活PKC β-TOP 2 β模块，随后激活TOP 2 β的新假说 降解，并引发TL DNA损伤。TL DNA损伤促进PARP活化，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。我们将通过追踪 以下3个具体目标：在目标1中，我们将确定以下3个常见的作用和调节机制 体外PmSVD中的代谢物相关途径; 1）核苷酸糖-GAG和硫酸盐，2）谷氨酸盐，3）NAD+- 硫化氢在目的2中，我们将描述PKC β-TOP2β模块和PARP 1在PmSVD介导的代谢物中的作用。 改变和MT功能障碍。在目标3中，我们将确定PKC β-TOP2β模块的作用， 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 预期建立PKC β-TOP2β和PARP作为调节PmSVD诱导的细胞凋亡的中枢分子的作用。 代谢物变化和PISP。这种方法是创新的，因为我们将使用iPSC，离子等新技术。 色谱-质谱（IC-MS），机器学习，成像质谱细胞术，和一种新的小鼠 CAthS模型。拟议的研究应该通过导致一种新的方法来积极影响PmSVD， 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.