The Multiscale Role of Piezo Channels in Obesity-Associated Cartilage Damage

压电通道在肥胖相关软骨损伤中的多尺度作用

• 批准号: 10387891
• 负责人: Erica Valentine Ely
• 金额: $ 4.68万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10387891
• 关键词: Address; Affect; American; Animal Model; Animals; Antibodies; Atomic Force Microscopy; Attenuated; Biological; Biomechanics; Blood Circulation; Body fat; Calcium; Cartilage; Cations; Cell Death; Cells; Cessation of life; Chondrocytes; Complement; Confocal Microscopy; Degenerative polyarthritis; Development; Disease; Esthesia; Exposure to; Fatty acid glycerol esters; Goals; Harvest; Health; High Fat Diet; High Prevalence; Hindlimb; Histology; Homeostasis; Hypersensitivity; Imaging Techniques; Immunohistochemistry; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Interleukin-1 alpha; Interleukin-6; Ion Channel; Ions; Joints; Knee joint; Knock-out; Length; Leptin; Link; Maintenance; Measurement; Measures; Mechanical Stress; Mechanics; Medial meniscus structure; Messenger RNA; Modeling; Monitor; Mus; Obesity; Operative Surgical Procedures; Pain; Pathogenesis; Pathologic; Pathway interactions; Pharmaceutical Preparations; Physiological; Piezo 1 ion channel; Piezo 2 ion channel; Piezo ion channels; Play; Proteins; Risk Factors; Role; Scanning Probe Microscopes; Serum; Signal Transduction; Synovial Fluid; Synovial joint; TNF gene; Testing; Tissues; Training; Transgenic Mice; Translating; Trauma; Up-Regulation; Western Blotting; adipokines; aggrecan; behavior test; bone; cell type; cytokine; disability; experience; graduate student; healthy weight; human model; in vivo; in vivo Model; inflammatory milieu; inhibitor; insight; joint injury; joint loading; knock-down; mechanical force; mechanical load; mechanotransduction; medical specialties; meniscus injury; microCT; mouse genetics; mouse model; novel therapeutic intervention; protein expression; response; sensor; systemic inflammatory response; therapeutic target

PROJECT SUMMARY Osteoarthritis (OA) is a debilitating disease of the synovial joints and is the leading cause of pain and disability worldwide. OA affects over 30 million Americans, but there are no disease modifying drugs. Obesity is a major risk factor for OA; however, it has been difficult to disentangle the role of low-level systemic inflammation from the effects of altered joint loading with increased body mass. While cartilage requires loading to maintain tissue homeostasis, previous studies have suggested that abnormal loading due to increased body mass may explain why individuals with obesity experience cartilage damage at elevated rates when compared to healthy weight individuals. However, many recent studies demonstrate that increased body mass alone does not explain OA damage in human and animal models, and that adipokines, inflammatory mediators from body fat, play an important role in OA pathogenesis. Additionally, previous studies from the Guilak lab highlight the synergistic importance of the mechanosensitive ion channels Piezo1 and Piezo2 in cartilage health and maintenance: obesity and an OA-relevant inflammatory mediator, interleukin 1 alpha (IL-1α), modulates and sensitizes Piezo channel function. As such, this proposal investigates if cytokine/adipokine signaling is the mechanism by which Piezo channels become hypersensitized to mechanical force, ultimately leading to increased chondrocyte death. The goal of this proposal is to directly investigate the interaction between obesity-associated inflammation and the mechanosensitivity of chondrocytes. This study on the role of altered mechanosensation in the pathogenesis of OA with obesity will lead to the ultimate goal of targeting these pathways to develop novel therapeutic approaches. Specific Aim 1 focuses on determining if obesity-associated inflammatory conditions alter Piezo expression in cartilage and if this increased expression translates to increased chondrocyte sensitivity to mechanical loads. This study hones in on key dysregulated adipokines with obesity: IL-1α, Leptin, tumor necrosis factor alpha (TNF-α), and interleukin 6 (IL-6). Specific Aim 2 uses transgenic mice previously developed in the lab to investigate if the loss of chondrocyte-specific Piezo1 and/or Piezo2 ion channels protects against cartilage damage in an in vivo model of obesity and joint injury. Specifically, mice will be fed a high-fat diet (60% fat) and subjected to a destabilization of the medial meniscus (DMM) surgery, known to evoke post-traumatic OA. Together, both aims strategically develop atomic force microscopy (AFM) and calcium (Ca2+) imaging techniques with in vivo assessments of cartilage integrity to complement the use of genetically-modified mice in a model of HFD superimposed with DMM injury. The results from this study will help identify the mechanisms by which obesity affects cartilage health, ultimately leading to the development of OA disease modifying drugs, that may be more broadly applied to other tissues affected by altered mechanosensation of Piezo ion channels.

项目摘要 骨关节炎（OA）是滑膜关节的一种使人衰弱的疾病，是疼痛和残疾的主要原因 全世界。 OA会影响超过3000万美国人，但没有改变疾病的药物。肥胖是主要的 OA的危险因素；但是，很难将低级系统性炎症的作用脱离 随着体重增加的关​​节负荷改变的影响。软骨需要加载才能维持组织 稳态，先前的研究表明，由于体重增加而引起的异常负荷可能解释 与健康体重相比 个人。但是，许多最近的研究表明，仅体重增加并不能解释OA 人类和动物模型的损害，以及脂肪因子，体内炎症介质的脂肪因子，发挥 在OA发病机理中的重要作用。此外，Guilak Lab的先前研究突出了协同作用 机械敏感的离子通道在软骨健康和维护中Piezo1和Piezo2的重要性： 肥胖和与OA相关的炎症介质，白介素1 alpha（IL-1α），调节和感觉压电 通道功能。因此，该建议调查了细胞因子/脂肪因子信号传导是否是一种机制 压电通道对机械力过敏，最终导致软骨细胞死亡增加。 该提案的目的是直接研究肥胖相关注射与 软骨细胞的机理敏感性。这项关于改变机制在发病机理中的作用的研究 肥胖症的OA的最终目标是针对这些途径开发新疗法的途径 方法。特定目标1的重点是确定肥胖相关的炎症条件是否改变压电 软骨中的表达，如果这种增加的表达能够转化为软骨细胞的敏感性提高 机械载荷。这项研究对肥胖症的关键失调脂肪因子进行磨练：IL-1α，瘦素，肿瘤坏死 因子α（TNF-α）和白介素6（IL-6）。特定目标2使用以前在 实验室调查软骨细胞特异性压电和/或压电2离子通道的损失是否可以防止软骨 肥胖和关节损伤的体内模型中的损害。具体而言，小鼠将得到高脂饮食（60％脂肪）和 经受了中正态（DMM）手术的不稳定，已知会引起创伤后的OA。 两者一起，两者在战略上开发原子力显微镜（AFM）和钙（CA2+）成像技术 通过体内评估软骨完整性，以完成在模型中使用一般修饰的小鼠的使用 HFD被DMM损伤叠加。这项研究的结果将有助于确定 肥胖会影响软骨健康，最终导致OA疾病修饰药物的发展，这可能 更广泛地应用于受压电离子通道机制改变的其他组织。