Mechano signals regulating tendon and ligament homeostasis

机械信号调节肌腱和韧带稳态

• 批准号: 10522344
• 负责人: HIROSHI ASAHARA
• 金额: $ 39.05万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10522344
• 关键词: Address; African American population; Age; Aging; Agonist; Biomechanics; Data; Development; Erythrocytes; Exercise; Gene Expression; Gene Expression Profiling; Genes; Genetic Research; Histologic; Homeostasis; Human; Immobilization; Inherited; Injury; Ion Channel; Kinetics; Knock-in; Ligaments; Mechanical Stimulation; Mediating; Medical; Molecular; Morphology; Motor; Movement; Mus; Muscle; Muscle Contraction; Musculoskeletal Diseases; Musculoskeletal System; Mutation; Organ; Pathologic; Performance; Phenotype; Physical Performance; Physiologic Ossification; Physiological; Physiology; Piezo 1 ion channel; Play; Property; Reporting; Resistance; Role; Signal Transduction; Small Interfering RNA; Speed; Stretching; Structure; Sum; Tendon structure; Testing; Time; Tissues; Translating; Up-Regulation; Variant; Wild Type Mouse; aged; biomechanical test; bone; gain of function; improved; in vivo; innovation; insight; knock-down; ligament development; ligament injury; loss of function; mature animal; mechanical stimulus; mechanotransduction; molecular phenotype; novel; postnatal; response; sensor; small molecule; therapeutic target; transcriptome; treadmill; viscoelasticity

ABSTRACT Mechano-stimulation in the musculoskeletal system plays a major role in the homeostasis of all tissues and organs. Particularly in tendons, the proper physiologic exercise load has an anabolic effect with increased tenogenic gene expression in humans; however, the molecular mechanisms that sense mechanical stimuli remain unclear. Piezo1 is a mechano-sensitive ion channel involved in mechano-sensing in various organs and tissues; however, it has not yet been examined in tendons or ligaments. In mice, the R2482H mutation of Piezo1 is an active form mimicking a gain-of-function (GOF) PIEZO1 variant found in humans. To test the potential effects of Piezo1 in physical performance, we generated systemic GOF Piezo1 (Piezo1systemic-GOF) mice, muscle-specific GOF Piezo1 (Piezo1muscle-GOF) mice, and tendon-specific GOF Piezo1 (Piezo1tendon-GOF) mice and found that the jumping power of Piezo1tendon-GOF mice as well as Piezo1systemic-GOF mice at 12-weeks- old was ~1.6 times greater than in both wild-type mice and Piezo1muscle-GOF mice. Consistent with this, significantly altered macroscopic and histological phenotypes were observed in the tendons of Piezo1tendon-GOF mice with increased tendon-specific gene expression. We also found that upregulation of aging related ossification gene expression was significantly reduced in aged Piezo1tendon-GOF mice. This suggests that constitutively enhanced PIEZO1 activity promotes physical performance via tendons/ligaments. These preliminary data underlie our hypothesis that PIEZO1 acts as a critical mechano-sensor in tenocytes, maintaining and promoting tendon/ligament functions by orchestrating tendon/ligament-specific gene expression. To test this hypothesis, we will pursue the following Aims: Aim 1: Analyze the function of Piezo1 in human and mouse tendon homeostasis and aging. We will examine the function of Piezo1 in tenocytes for tendon homeostasis and in aging by morphological, histological and gene expression analysis of tendons from Piezo1tendon-GOF mice and tendon specific loss of function (LOF) Piezo1 (Piezo1tendon-LOF) mice at different aging points. We will also test the role of PIEZO1 in human tenocytes by transcriptome analysis with PIEZO1 siRNA knockdown or a small molecule PIEZO1 activator. Aim 2: Evaluate the role of Piezo1 in tenocytes on physiological performance and exercise in mice. We will evaluate the role of Piezo1 in tenocytes to potentiate physical performance using postnatally induced tendon specific gain of function and loss of function Piezo1 mice at different ages. Thereafter, we will examine whether this physical ability could be strengthened by physiological exercise inducing anabolic mechanical stimulation. Aim 3: Analyze the biomechanical properties of tendon that are regulated by Piezo1. We will uncover how the kinetic energy for the enhanced jumping ability of the Piezo1tendon-GOF mouse is produced by biomechanical analysis of tendon stored energy and muscle contraction power. Completion of these aims will support the concept that Piezo1 could be a therapeutic target to maintain or enhance structure and function of the musculoskeletal system.

抽象的 肌肉骨骼系统中的机械刺激在所有组织的稳态中发挥着重要作用 器官。特别是在肌腱中，适当的生理运动负荷具有合成代谢作用，可增加 人类肌腱基因表达；然而，感知机械刺激的分子机制 仍不清楚。 Piezo1 是一种机械敏感离子通道，参与多种器官和组织的机械感应 纸巾；然而，尚未在肌腱或韧带中进行检查。在小鼠中，R2482H 突变 Piezo1 是一种活性形式，模仿人类中发现的功能获得 (GOF) PIEZO1 变体。测试 Piezo1 对身体表现的潜在影响，我们生成了系统性 GOF Piezo1 (Piezo1systemic-GOF) 小鼠、肌肉特异性 GOF Piezo1 (Piezo1muscle-GOF) 小鼠和肌腱特异性 GOF Piezo1 (Piezo1tendon-GOF) 小鼠，发现 Piezo1tendon-GOF 小鼠和 Piezo1systemic-GOF 小鼠在 12 周时的跳跃能力- old 是野生型小鼠和 Piezo1muscle-GOF 小鼠的 1.6 倍。与此相一致的是， 在 Piezo1tendon-GOF 的肌腱中观察到显着改变的宏观和组织学表型 肌腱特异性基因表达增加的小鼠。我们还发现与衰老相关的上调 老年 Piezo1tendon-GOF 小鼠中骨化基因表达显着降低。这表明 持续增强的 PIEZO1 活性通过肌腱/韧带促进身体表现。这些 初步数据支持我们的假设，即 PIEZO1 在肌腱细胞中充当关键的机械传感器， 通过协调肌腱/韧带特异性基因来维持和促进肌腱/韧带功能 表达。为了检验这一假设，我们将追求以下目标： 目标 1：分析 Piezo1 在人类和小鼠肌腱稳态和衰老中的作用。我们将检查 Piezo1 的功能 通过形态学、组织学和基因表达分析，确定肌腱细胞在肌腱稳态和衰老中的作用 来自 Piezo1tendon-GOF 小鼠和肌腱特异性功能丧失 (LOF) Piezo1 (Piezo1tendon-LOF) 小鼠的肌腱 不同的老化点。我们还将通过转录组分析来测试 PIEZO1 在人类肌腱细胞中的作用 PIEZO1 siRNA 敲低或小分子 PIEZO1 激活剂。目标 2：评估 Piezo1 在 肌腱细胞对小鼠生理性能和运动的影响。我们将评估 Piezo1 在以下方面的作用： 肌腱细胞利用出生后诱导的肌腱特异性功能增强来增强身体表现， 不同年龄的 Piezo1 小鼠功能丧失。此后，我们将检验这种身体能力是否可以 通过引起合成代谢机械刺激的生理运动来加强。目标 3：分析 受 Piezo1 调节的肌腱的生物力学特性。我们将揭示动能如何 Piezo1tendon-GOF 小鼠的跳跃能力增强是通过肌腱的生物力学分析产生的 储存能量和肌肉收缩力。这些目标的完成将支持 Piezo1 的概念 可能是维持或增强肌肉骨骼系统结构和功能的治疗靶点。