Cytosolic DNA sensing instructs resident macrophage vitality and organismal longevity

胞质 DNA 传感指示常驻巨噬细胞活力和生物体寿命

基本信息

批准号：
10901044
负责人：
Shruti Sharma
金额：
$ 40.86万
依托单位：
TUFTS UNIVERSITY BOSTON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-20 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10901044
关键词：
Address Age Aging Autoimmune Diseases Brain Cell Aging Cell Death Cell Survival Cell physiology Cells Cellular Indexing of Transcriptomes and Epitopes by Sequencing Chromatin Chronic Chronic Disease Chronology Communicable Diseases Cyclic GMP DNA DNA Damage Decision Making Development Disease Drug or chemical Tissue Distribution Elements Embryo Endothelial Cells Epithelial Cells Equilibrium Etoposide Fibroblasts Functional disorder Gene Activation Genetic Health Heart Homeostasis Immune Inflammation Inflammatory Kidney Ligands Link Liver Longevity LoxP-flanked allele Lung Macrophage Malignant Neoplasms Metabolic Diseases Morbidity - disease rate Mus Nature Normal tissue morphology Organ Oxidative Stress Pathologic Pathology Pathway interactions Periodicity Peripheral Phenotype Physiological Play Population Population Decreases Population Sizes Proteomics Reporting Resolution Retroelements Role Shapes Signal Transduction Space Perception Stimulator of Interferon Genes Stimulus Stromal Cells Techniques Testing Therapeutic Tissues Work aged cell type cytokine ds-DNA expectation in vivo innovation insight male mortality mouse model multimodality multiple omics novel novel therapeutics preservation protective effect public health relevance repaired senescence sensor single-cell RNA sequencing therapeutic target tool transcriptomics

项目摘要

Project Summary Age-associated chronic inflammation is thought to be directly linked to the cellular processes termed senescence and senescence associated secretory phenotype (SASP). Recent studies suggest that the sensing of cytosolic chromatin or damaged DNA in senescent cells by the cyclic-di-GMP-AMP-synthase (cGAS) and Stimulator of Interferon Genes (STING) regulate the SASP. Based on these reports, it is presumed that cGAS/STING signaling drives age-associated pathology. However, these studies primarily focused on understanding the role of the cGAS/STING dependent senescence in fibroblasts. As fibroblasts represent only a small sliver of the cells found in tissues, we sought to extend these studies by exploring the contribution of cGAS/STING-signaling to changes in multiple tissues during chronological and physiological aging. Using single-cell RNA-sequencing, we note predominant signatures of senescence in all tissues tested (kidneys, lungs, heart, liver and brain). These signatures extend to fibroblasts and non-fibroblast stromal cells like endothelial and epithelial cells, but also to tissue-resident macrophages (TRMs). TRMs are critical for optimal organ function and repair. The loss of cGAS or STING impacts TRM viability in vivo and ex vivo, predisposing them to cell-death. Furthermore, we show that endogenous retroelements (EREs), a proven ligand for cGAS/STING activation, are expressed in TRMs and critical for normal tissue-centric functions of TRMs. Finally, contrary to expectations based on previous studies, the loss of cGAS or STING leads to significantly shorter lifespans and greater peripheral inflammatory signature. Based on these paradigm shifting findings, we propose that cGAS/STING signaling balances the protective effects of senescence with its pathological effects through cell-specific roles. Our broad objective is thus, to understand the cell-specific contribution of cGAS/STING signaling to organismal aging. Our specific aims will methodically tackle this objective by focusing on TRMs. Given the critical role DNA-sensing plays in discriminating endpoints like senescence induction or cell-survival, the unique nature of ligands may play a role in cell-fate decisions. Thus, we will first ascertain the role EREs play in shaping TRM survival and function and how their engagement of the cGAS/STING pathway factors into their role (Aim1). We then aim to clarify how cGAS/STING deficiency impacts the vitality of TRMs, predisposing them to cell-death (Aim2). Finally, we will determine if a TRM-intrinsic deficiency of cGAS/STING signaling can recapitulate the tissue dysfunction and compromised lifespans seen in mice with global cGAS/STING deficiency (Aim3). These aims will utilize cutting-edge and innovative tools like single-cell spatial multi-omics (a spatially oriented simultaneous profiling of proteomics and transcriptomics in whole tissues), multi-modal CITE-seq (phenotypic profiling and transcriptomics with single-cell resolution) and powerful genetic tools. Given the massive therapeutic push to target this pathway in cancer, several autoimmune diseases and now aging, our proposed studies are urgently needed to fully comprehend the impact of modifying this pathway long-term.

项目概要与年龄相关的慢性炎症被认为与称为“细胞过程”的细胞过程直接相关。衰老和衰老相关分泌表型（SASP）。最近的研究表明通过环二 GMP-AMP 合酶感知衰老细胞中的胞质染色质或受损 DNA (cGAS) 和干扰素基因刺激物 (STING) 调节 SASP。根据这些报告，推测 cGAS/STING 信号传导驱动与年龄相关的病理。然而，这些研究主要专注于了解 cGAS/STING 依赖性衰老在成纤维细胞中的作用。作为成纤维细胞仅代表组织中发现的一小部分细胞，我们试图通过探索 cGAS/STING 信号传导对时间和生理过程中多个组织变化的贡献老化。使用单细胞 RNA 测序，我们注意到所有测试组织中衰老的主要特征（肾、肺、心、肝和脑）。这些特征延伸至成纤维细胞和非成纤维细胞基质细胞例如内皮细胞和上皮细胞，还包括组织驻留巨噬细胞 (TRM)。 TRM 至关重要最佳的器官功能和修复。 cGAS 或 STING 的丢失会影响 TRM 在体内和离体的活力，使它们易于细胞死亡。此外，我们还发现内源性逆转录因子（ERE）是一种经过验证的 cGAS/STING 激活的配体，在 TRM 中表达，对于正常的以组织为中心的功能至关重要 TRM。最后，与之前研究的预期相反，cGAS 或 STING 的丢失导致寿命显着缩短，外周炎症特征增强。基于这些范式改变研究结果，我们提出 cGAS/STING 信号传导平衡衰老的保护作用与它通过细胞特异性作用产生病理效应。因此，我们的总体目标是了解细胞特异性 cGAS/STING 信号传导对机体衰老的贡献。我们的具体目标将有条不紊地解决这个问题通过关注 TRM 来实现目标。鉴于 DNA 传感在区分终点（例如在衰老诱导或细胞存活中，配体的独特性质可能在细胞命运的决定中发挥作用。因此，我们将首先确定 ERE 在塑造 TRM 生存和功能中所发挥的作用，以及它们如何 cGAS/STING 通路因子参与其作用（目标 1）。然后我们的目标是澄清如何 cGAS/STING 缺乏会影响 TRM 的活力，使其容易发生细胞死亡（目标 2）。最后，我们将确定 cGAS/STING 信号传导的 TRM 内在缺陷是否可以重现组织功能障碍，整体 cGAS/STING 缺陷 (Aim3) 的小鼠寿命缩短。这些目标将利用尖端和创新工具，如单细胞空间多组学（空间定向同步整个组织中的蛋白质组学和转录组学分析）、多模式 CITE-seq（表型分析和具有单细胞分辨率的转录组学）和强大的遗传工具。鉴于大规模的治疗推动针对癌症、几种自身免疫性疾病和现在衰老的这一途径，我们提出的研究是迫切需要充分理解改变这一途径的长期影响。