A novel cell-autonomous role for β-adrenergic receptor signaling in osteoclasts

破骨细胞中β-肾上腺素能受体信号传导的新型细胞自主作用

• 批准号: 10608343
• 负责人: Katherine Jean Motyl
• 金额: $ 5.04万
• 依托单位: MAINEHEALTH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10608343
• 关键词: Adrenergic Antagonists; Adrenergic beta-Agonists; Age; Age-Related Bone Loss; Aging; Agonist; Albuterol; Alleles; Atenolol; Award; Binding; Biological Assay; Bone Density; Bone Diseases; Bone Marrow; Bone Resorption; Bone remodeling; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcineurin; Calcineurin inhibitor; Cells; Clinical; Clinical Trials; Cohort Studies; Complement; Cyclic AMP-Dependent Protein Kinases; Data; Doctor of Philosophy; Dose; Engineering; Ensure; G Protein-Coupled Receptor Signaling; Genes; Genetic; Goals; Hand; Human; In Vitro; Laboratories; Lead; Ligand Binding; Maine; Marrow; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Meta-Analysis; Modeling; Mus; Myelogenous; Osteoblasts; Osteoclasts; Osteogenesis; Osteoporosis; Osteoporosis prevention; PPP3CA gene; PTH gene; Parents; Pathology; Pathway interactions; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacology; Phenotype; Physiological; Postmenopause; Preventive therapy; Production; Property; Propranolol; Receptor Signaling; Refractory; Research Project Grants; Role; Schools; Science; Serum; Signal Pathway; Signal Transduction; Sympathetic Nervous System; TRANCE protein; Testing; Universities; Woman; Work; beta-adrenergic receptor; bone; bone loss; career development; efficacy evaluation; efficacy testing; fracture risk; hormone therapy; human model; improved; in vivo; in vivo evaluation; monocyte; mouse model; novel; nuclear factors of activated T-cells; osteoclastogenesis; preclinical study; prevent; prospective; receptor; recruit; response; transcriptome sequencing

ABSTRACT The overall goal of this supplemental research project is to support the career development of Rebecca Peters, who is a PhD candidate at the University of Maine Graduate School of Biomedical Sciences and Engineering. The sympathetic nervous system (SNS) is known to promote osteoclastogenesis via osteoblast-mediated RANKL production. However, recent work from our laboratory has demonstrated that β-blockers can limit osteoclast activity directly, without influencing RANKL production, in vitro and in mice treated with anabolic parathyroid hormone (PTH) therapy. Furthermore, the efficacy of the cardio-selective β-blocker atenolol to prevent osteoporosis is currently being tested in humans in the Atenolol to Prevent Osteoporosis (APO) trial, but mechanisms of efficacy with relationship to osteoclasts have not been studied in humans or mouse models. In the parent award, we are using genetic mouse models to test the hypotheses that β1AR and β2AR signaling in osteoclasts specifically promote osteoclast differentiation and function. However, the award does not examine the pharmacological properties of β-blockers that allow them to limit osteoclast differentiation directly, nor does it test the translatability of the findings to human osteoclast models. In this supplemental award, we will study the function of β1AR and β2AR in response to β-blockers in human and murine osteoclasts. We will also test whether our in vitro findings of atenolol-mediated suppression of osteoclast differentiation and bone resorption can be recapitulated in vivo. The proposed studies will not overlap with ongoing work on the parent award, but will complement and add translational significance to the findings. Supplement Specific Aim 1: Test the function of βARs in human osteoclasts in vitro. We have found that propranolol potentiates PTH-induced Ca2+ signaling in osteoblasts, however, β-blockers may have a distinct function in osteoclasts. Specifically, we hypothesize that β1AR and β2AR promote intracellular Ca2+-induced NFATc1 activity to support osteoclast differentiation, and that this is prevented by β-blockers. In this aim, we will use primary human peripheral blood mononuclear cell (PBMC) and murine bone marrow-derived osteoclasts to test this hypothesis. Supplement Specific Aim 2: Test the efficacy of atenolol to prevent bone resorption in vivo. We have preliminary data that atenolol also prevents human osteoclast differentiation in vitro, but whether it could be a potential co-therapy for osteoporosis treatment with PTH is unknown. Furthermore, whether atenolol would limit sympathetic nervous system-mediated resorption in vivo is unknown. We hypothesize that atenolol will limit resorption in both cases, indicating it could be clinically useful as an anti- resorptive therapy, whether or not SNS activity is high. To test whether atenolol will prevent bone resorption in the context of PTH therapy and elevated SNS tone, we will treat mice with atenolol and PTH or salbutamol (β2AR agonist, to mimic high SNS activity) and examine bone microarchitecture and resorption.

摘要 这个补充研究项目的总体目标是支持丽贝卡彼得斯的职业发展， 他是缅因州大学生物医学科学与工程研究生院的博士候选人。 已知交感神经系统（SNS）通过成骨细胞介导的成骨细胞分化促进破骨细胞生成。 RANKL生产。然而，我们实验室最近的工作表明，β受体阻滞剂可以限制 在体外和用合成代谢药物处理的小鼠中，破骨细胞活性直接，不影响RANKL的产生 甲状旁腺激素（PTH）治疗。此外，心脏选择性β受体阻滞剂阿替洛尔对 预防骨质疏松症目前正在阿替洛尔预防骨质疏松症（APO）试验中进行人体试验， 但与破骨细胞相关的疗效机制尚未在人类或小鼠中进行研究 模型在父母奖中，我们使用遗传小鼠模型来检验β 1 AR和β 2 AR 破骨细胞中的信号传导特异性地促进破骨细胞分化和功能。然而，该奖项 没有检查β受体阻滞剂的药理学特性，使其限制破骨细胞分化 它也没有直接测试这些发现对人类破骨细胞模型的可转化性。本补充 因此，我们将研究β 1 AR和β 2 AR在人和小鼠对β受体阻滞剂反应中的功能 破骨细胞我们还将测试阿替洛尔介导的破骨细胞抑制的体外研究结果是否 分化和骨吸收可以在体内重演。拟议的研究不会与 目前正在进行的关于家长奖的工作，但将补充和增加翻译的意义，调查结果。 补充具体目标1：检测β AR在体外人破骨细胞中的功能。我们发现 普萘洛尔可增强PTH诱导的成骨细胞内钙离子信号传导，然而，β受体阻滞剂可能具有明显的 在破骨细胞中发挥作用。具体地说，我们假设β 1 AR和β 2 AR促进细胞内Ca2+诱导的 NFATc1活性，以支持破骨细胞分化，这是由β-阻滞剂阻止。为此，我们 将使用原代人外周血单核细胞（PBMC）和鼠骨髓来源的 破骨细胞来验证这个假设。补充具体目标2：测试阿替洛尔预防骨质疏松的功效 体内再吸收。我们有初步的数据表明阿替洛尔也能阻止人类破骨细胞的分化， 体外，但它是否可能是一个潜在的共同治疗骨质疏松症治疗与PTH是未知的。 此外，阿替洛尔是否会限制体内交感神经系统介导的吸收尚不清楚。 我们假设阿替洛尔在这两种情况下都会限制吸收，表明它可能在临床上作为一种抗- 再吸收疗法，无论SNS活性是否高。为了测试阿替洛尔是否会防止骨吸收， 在PTH治疗和SNS张力升高的背景下，我们将用阿替洛尔和PTH或沙丁胺醇治疗小鼠 （β 2AR激动剂，模拟高SNS活性），并检查骨微结构和再吸收。