PKG Regulation of Sirtuin 1 as a Novel Treatment Strategy for Age-related Osteoporosis

Sirtuin 1 的 PKG 调节作为年龄相关性骨质疏松症的新型治疗策略

基本信息

批准号：
10634657
负责人：
RENATE B PILZ
金额：
$ 46.71万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-05 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10634657
关键词：
Affect Age Age-Related Osteoporosis Aging American Antioxidants Apoptosis Architecture Bone Formation Stimulation Bone Resorption Cell Aging Clinical Trials Cyclic AMP-Dependent Protein Kinases Cyclic GMP Cyclic GMP-Dependent Protein Kinases DNA Damage Deacetylase Disease Effectiveness FDA approved Feedback Foundations Fracture Fright G-Protein Signaling Pathway G-substrate Gene Expression Profile Genes Genetic Goals Guanylate Cyclase Impairment Induction of Apoptosis Insulin-Dependent Diabetes Mellitus Knock-out Knockout Mice Knowledge Lead Longevity Mediating Mesenchymal Stem Cells Messenger RNA Metabolic Minority Mitochondria Mus NOS3 gene Nitric Oxide Nitric Oxide Donors Nitric Oxide Synthase Nuclear Osteoblasts Osteoclasts Osteocytes Osteogenesis Osteoporosis Osteoporotic Oxidative Stress Oxidative Stress Induction Pain Patients Pharmaceutical Preparations Phosphorylation Physicians Proteins RNA-Binding Proteins Regulation Ribonucleoproteins Role SIRT1 gene Signal Pathway Signal Transduction Sirtuins Testing Transgenic Organisms Wild Type Mouse Woman Work age related aged analog beta catenin bone bone loss bone mass bone strength cobinamide diabetic improved interest mRNA Expression male men mitochondrial dysfunction mortality neutralizing antibody nicotinamide-beta-riboside novel novel therapeutics osteoblast differentiation osteoporosis with pathological fracture overexpression pharmacologic posttranscriptional prevent protein activation protein expression public health relevance senescence side effect skeletal treatment strategy

项目摘要

SUMMARY Age-related osteoporosis affects millions of American men and women, and is a major cause of fractures in those over age 50. Bone loss is due mainly to impaired bone formation, but FDA-approved agents with bone- anabolic effects (PTH analogs and the sclerostin-neutralizing antibody romosozumab) have serious limitations. We have found that pharmacologic or genetic stimulation of the NO/cGMP/protein kinase G (PKG) signaling pathway enhances bone formation and prevents bone loss in diabetic and aged mice; the mechanism is through increased Wnt/β-catenin signaling and reduced oxidative stress. Silent information regulator T1 (Sirtuin1, SIRT1) and its substrate NAD+ decrease with age, contributing to many age-related diseases, including osteoporosis. Osteoblast-specific Sirt1 knockout mice are osteoporotic while Sirt1 overexpressing mice have high bone mass. We found recently that PKG activation increases SIRT1 expression in osteoblasts, mesenchymal stem cells (MSCs), and bones of aged mice by stabilizing Sirt1 mRNA; the RNA binding protein hnRNPA1 is required. SIRT1 is known to increase NO synthesis, possibly leading to a positive feedback loop with PKG. Supporting positive feedback between PKG and SIRT1, we found they synergistically increase pro-survival genes in osteoblasts and prevent oxidative stress-induced apoptosis. As part of our interest in NO/cGMP/PKG signaling, we have developed nitrosyl-cobinamide (NO-Cbi), a NO donor that is also a strong anti-oxidant. Recent work shows that a 10 week course of NO-Cbi improved bone formation, architecture, and strength in 24 month-old mice. The overall goal of the proposed studies is to define the interactions between PKG and SIRT1 in bone, and devise a novel treatment strategy for age-related osteoporosis based on PKG and SIRT1 activators. In Aim 1 we will define how PKG increases Sirt1 mRNA in MSCs and osteoblasts, focusing on PKG regulation of hnRNPA1. In Aim 2, we will determine the consequences of PKG-SIRT1 crosstalk in bones of aged mice. We will determine whether PKG requires SIRT1 to improve bone formation and strength by examining the effects of PKG-activating agents on bone in conditional osteoblast-specific Sirt1 knockout mice. To determine if SIRT1- induced NO synthesis and PKG activation is necessary for the positive effects of SIRT1 activators in bone, we will examine the skeletal effects of these agents in mice with conditional deletion of Prkg2 in osteoblasts. In Aim 3, we will test whether combining PKG- and SIRT1-activating agents can synergistically increase bone formation in aged mice, compared to each treatment alone. We will use NO-Cbi and the guanylyl cyclase activator cinaciguat to activate PKG, and direct SIRT1 activators and nicotinamide riboside, a NAD+ precursor, to activate SIRT1. The proposed work will fill important knowledge gaps, defining the role of PKG-SIRT1 crosstalk in bone during aging and determining how PKG-SIRT1 reduce age-related oxidative stress in bone; importantly, the proposed work lays the foundation for developing novel bone-anabolic therapies for age-related osteoporosis.

概括与年龄有关的骨质疏松症影响数百万的美国男性和女性，是骨折的主要原因 50岁以上的人。骨质流失主要是由于骨骼形成受损，但是经FDA批准的药物具有骨 - 合成代谢作用（PTH类似物和硬化素中和抗体romosozumab）具有严重的局限性。我们发现NO/CGMP/蛋白激酶G（PKG）信号的药理或遗传刺激途径可以增强骨骼形成，并防止糖尿病和老年小鼠的骨质流失；该机制是通过 Wnt/β-catenin信号传导增加并减少氧化应激。无声信息调节器T1（SIRTUIN1，SIRT1）其底物NAD+随着年龄的增长而减少，导致许多与年龄有关的疾病，包括骨质疏松症。成骨细胞特异性的SIRT1基因敲除小鼠是骨质疏松性的，而SIRT1过表达的小鼠的骨骼质量很高。我们最近发现，PKG激活增加了成骨细胞，间充质干细胞中的SIRT1表达（MSC）和老年小鼠的骨头通过稳定SIRT1 mRNA；需要RNA结合蛋白HNRNPA1。众所周知，SIRT1不会增加合成，可能导致与PKG的正反馈回路。支持 PKG和SIRT1之间的积极反馈，我们发现它们合成增加了生存基因成骨细胞并预防氧化应激诱导的细胞凋亡。作为我们对NO/CGMP/PKG信号的兴趣的一部分，我们已经开发了硝基辛酰胺（NO-CBI），这是一种也是强抗氧化剂的供体。最近的工作表明10周的无CBI课程改善了24个月大的骨骼形成，建筑和力量老鼠。拟议研究的总体目标是定义骨骼中PKG和SIRT1之间的相互作用，并设计了一种基于PKG和SIRT1激活剂的年龄相关骨质疏松症的新型治疗策略。目标 1我们将定义PKG如何增加MSC和成骨细胞中的SIRT1 mRNA，重点是PKG调节 HNRNPA1。在AIM 2中，我们将确定PKG-SIRT1串扰在老年小鼠的骨骼中的后果。我们将确定PKG是否要求SIRT1通过检查 PKG激活剂在有条件成骨细胞特异性SIRT1基因敲除小鼠中。确定SIRT1-- 诱导的无合成和PKG激活对于骨骼中SIRT1激活剂的积极作用是必要的，我们将检查这些药物在成骨细胞中有条件缺失的小鼠中这些药物的骨骼作用。目标 3，我们将测试合并PKG和SIRT1激活剂是否可以协同增加骨形成在老年小鼠中，与单独的每种治疗相比。我们将使用No-CBI和Guanylyl Cyclese激活剂 cinaciguat激活PKG，直接SIRT1激活剂和NAD+前体烟酰胺核苷激活 SIRT1。拟议的工作将填补重要的知识空白，从而定义了PKG-SIRT1串扰在骨骼在衰老和确定PKG-SIRT1如何减少骨骼中与年龄相关的氧化应激；重要的是，拟议的工作为开发与年龄相关的骨质疏松症开发新型骨食性疗法的基础。