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.

项目概要/摘要。 已知淋巴功能障碍与涉及衰老和衰老的各种病症相关。 炎症，如阿尔茨海默病，心血管衰退和关节炎进展。在整个 随着年龄的增长，淋巴管收缩性已经显示出恶化，这与减少的基因表达有关， 在淋巴肌细胞（LMC）中，调节稳态收缩和细胞骨架完整性的途径。 同样，我们先前发现，年龄依赖性淋巴功能障碍的细胞机制， 慢性炎性关节炎肿瘤坏死因子转基因（TNF-Tg）小鼠模型与 老年人关节疾病的进展和关节引流腘淋巴管（PLV）收缩的丧失 小鼠在这些老年小鼠中，PLV在超微结构水平上显示出显著的LMC萎缩，这是因为 建议消除淋巴功能并加剧疾病进展。作为临床 与此相关，我们发现活动性类风湿关节炎患者的功能性LV较少， 手表面的淋巴清除减少。因此，我们假设炎症介导了 加速与年龄相关的LMC损伤，其中发炎的LMC将表现出相当的功能障碍， LMC收缩性和细胞骨架完整性所必需的基因途径。为了验证这一假设，我们建议 评估α平滑肌肌动蛋白（αSMA）+ PLV-LMC覆盖率并研究转录变化， 单细胞RNA测序（scRNAseq）在年轻人（2个月大），老年人（8个月大）和老年人（24个月大） old）WT和TNF-Tg小鼠。为了实现这一目标，我们已经证明了αSMA+ PLV-LMC的覆盖率是 相对于WT同窝出生小鼠，在老龄（8月龄）TNF-Tg小鼠中降低。此外，我们还初步实现了 在老化LMC的scRNAseq中的成功表明这种方法的可行性。鉴于不充分的 为了表征LMC在健康和疾病中的作用，我们的scRNAseq 数据还提供了LMC是转录上不同的血管肌细胞的证据。通过 完成本拨款申请中体现的研究目标，我们有机会提供 为未来的临床前研究和LMC的靶向治疗干预提供了相当大的创新。一致 通过申请人的研究培训计划，主要研究者将主要在实验室研究中获得宝贵的经验 提供额外的机会来培养临床试验设计、医疗保健、学术领导力和 罗切斯特大学的社区外展，有着培养成功医生的宝贵历史- 科学家对于以下PA-21-049 F30资助申请，我们建议阐明LMC的机制 在衰老和炎症过程中的功能障碍，有利于淋巴细胞生物学， 了解关节炎的神秘年龄相关进展。