Elucidating the Mechanisms of Lymphatic Muscle Cell Dysfunction in Aging and Inflammation

阐明衰老和炎症中淋巴肌细胞功能障碍的机制

• 批准号: 10382881
• 负责人: Howard Mark Kenney
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-17 至 2024-10-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10382881
• 关键词: Actins; Address; Age; Aging; Alzheimer' s Disease; Ankle; Applications Grants; Area; Arthritis; Atrophic; Biology; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Caring; Cell Lineage; Cells; Cellular biology; Characteristics; Chronic; Clinical; Clinical Trials Design; Community Outreach; Contracts; Cytoskeletal Proteins; Data; Disease; Disease Progression; Down-Regulation; Elderly; Exhibits; Failure; Fluorescence-Activated Cell Sorting; Fostering; Foundations; Frequencies; Functional disorder; Future; Gene Expression; Genes; Genetic Transcription; Glaucoma; Goals; Grant; Hand; Health; Histology; Homeostasis; Image; Immune response; Indocyanine Green; Inflammation; Inflammatory Arthritis; Inflammatory Bowel Diseases; Joints; Leadership; Lymph; Lymphatic; Lymphatic System; Lymphatic clearance; Lymphatic function; Lymphedema; Lymphoid Tissue; Measures; Mediating; Medical; Metabolic Diseases; Modeling; Mus; Muscle Cells; Muscle Contraction; Near-infrared optical imaging; Neoplasm Metastasis; Pathogenesis; Pathology; Pathway interactions; Patients; Physicians; Publishing; Recording of previous events; Research; Research Training; Rest; Rheumatoid Arthritis; Role; Scientific Inquiry; Scientist; Smooth Muscle Actin Staining Method; Smooth Muscle Myocytes; Surface; TNF gene; Tamoxifen; Targeted Research; Testing; Therapeutic Intervention; Tissues; Transgenic Mice; Transgenic Organisms; Universities; Validation; Vascular Smooth Muscle; age related; aged; arthropathies; cell type; cohort; differential expression; experience; frontier; genetic signature; human old age (65+); in vivo; innovation; lymphatic development; lymphatic dysfunction; lymphatic pump; lymphatic vessel; mortality; mouse model; novel; pre-clinical research; protein expression; single-cell RNA sequencing; success; targeted treatment; therapeutic target; transcriptomics; tumor

Project Summary/Abstract. Lymphatic dysfunction is known to be associated with various disorders involved in both aging and inflammation, such as Alzheimer’s disease, cardiovascular decline, and arthritic progression. Throughout aging, lymphatic contractility has been shown to deteriorate related to reduced gene expression of essential pathways that mediate homeostatic contractions and cytoskeletal integrity in lymphatic muscle cells (LMCs). Similarly, we previously discovered that age-dependent cellular mechanisms of lymphatic dysfunction in the tumor necrosis factor transgenic (TNF-Tg) mouse model of chronic inflammatory arthritis are associated with progression of joint disease and the loss of joint-draining popliteal lymphatic vessel (PLV) contractions in aged mice. In these aged mice, the PLV demonstrates significant LMC atrophy at an ultrastructural level, which is proposed to drive the elimination of lymphatic function and exacerbate disease progression. As a clinical correlate, we have found that patients with active rheumatoid arthritis have fewer functional LVs and diminished lymphatic clearance on the surface of the hands. Thus, we hypothesize that inflammation mediates accelerated age-related damage to LMCs where inflamed LMCs will exhibit a comparable dysfunction in the gene pathways necessary for LMC contractility and cytoskeletal integrity. To test this hypothesis, we propose to assess alpha smooth muscle actin (αSMA)+ PLV-LMC coverage and investigate transcriptional changes by single cell RNA sequencing (scRNAseq) in young (2-month-old), aged (8-month-old), and elderly (24-month- old) WT and TNF-Tg mice. Towards this goal, we have demonstrated that αSMA+ PLV-LMC coverage is reduced in aged (8-month-old) TNF-Tg mice relative to WT littermates. Additionally, we achieved preliminary success in scRNAseq of aged LMCs to indicate the feasibility of this approach. Given the inadequate characterization of LMCs necessary for targeted research into their role in health and disease, our scRNAseq data also provides evidence that LMCs are transcriptionally distinct vascular muscle cells. Through the completion of the research aims embodied in this grant proposal, we have the opportunity to provide considerable innovation for future pre-clinical research and targeted therapeutic interventions of LMCs. In line with the applicant’s Research Training Plan, the PI will gain valuable experience primarily in bench research with additional opportunities to foster abilities in clinical trial design, medical care, academic leadership, and community outreach at the University of Rochester with a cherished history of producing successful physician- scientists. For the following PA-21-049 F30 grant submission, we propose to elucidate the mechanisms of LMC dysfunction during both aging and inflammation for the benefit of lymphatic cellular biology and our understanding of the enigmatic age-related progression of arthritis.

项目摘要/摘要。 已知淋巴功能障碍与衰老和涉及的各种疾病有关 炎症，例如阿尔茨海默氏病，心血管下降和艺术进展。自始至终 衰老，淋巴收缩力已显示可检测与必需基因表达降低有关 介导淋巴肌肉细胞中稳态收缩和细胞骨架完整性的途径（LMC）。 同样，我们先前发现了淋巴功能障碍的年龄依赖性细胞机制 慢性炎性关节炎的肿瘤坏死因子转基因（TNF-TG）小鼠模型与 关节疾病的进展和衰老的白杨淋巴管（PLV）损失 老鼠。在这些老年小鼠中，PLV在超微结构水平上表现出明显的LMC萎缩，这是 提议推动消除淋巴功能和加剧疾病进展。作为临床 相关，我们发现活跃类风湿关节炎患者的功能LVS较少，并且 手表面上的淋巴间隙降低。那我们假设炎症媒体 与年龄相关的LMC加速损害在发炎的LMC中会存在可比的功能障碍 LMC收缩性和细胞骨架完整性所需的基因途径。为了检验这一假设，我们提出了 评估α平滑肌肌动蛋白（αSMA）+ PLV-LMC覆盖范围，并通过 年轻（2个月大），年龄（8个月大）的单细胞RNA测序（SCRNASEQ），较早（24个月 - 旧）WT和TNF-TG小鼠。为了实现这一目标，我们已经证明了αSMA+ PLV-LMC覆盖范围 相对于wt同窝仔的老年（8个月大）TNF-TG小鼠的老化（8个月大）。此外，我们获得了初步 在老年LMC的SCRNASEQ方面取得了成功，以表明这种方法的可行性。考虑到不足 我们的SCRNASEQ的针对健康和疾病中的作用所必需的LMC的表征 数据还提供了证据表明LMC是转录不同的血管肌肉细胞。通过 该赠款提案中体现的研究目的的完成，我们有机会提供 对未来的临床前研究和针对性LMC的治疗干预措施的巨大创新。排队 借助申请人的研究培训计划，PI将获得宝贵的基准研究经验 还有其他机会促进临床试验设计，医疗保健，学术领导和 罗切斯特大学的社区宣传，拥有成功的物理历史 科学家。对于以下PA-21-049 F30赠款提交，我们建议阐明LMC的机制 淋巴细胞生物学的益处和炎症期间的功能障碍 了解关节炎的神秘年龄相关的进展。