Targeting Cellular Senescence and RAGE in Type 2 Diabetes

靶向 2 型糖尿病中的细胞衰老和 RAGE

基本信息

批准号：
10176684
负责人：
Joshua Nicholas Farr
金额：
$ 39.75万
依托单位：
MAYO CLINIC ROCHESTER
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：
2021-05-01 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10176684
关键词：
Address Advanced Glycosylation End Products Aging Apoptosis B-Lymphocytes Biology Biomechanics Bone Resorption Bone remodeling Brain CASP8 gene CDKN2A gene Cell Aging Cells Ceramides Collagen Complement Complications of Diabetes Mellitus Cre-LoxP Cytometry Dasatinib Data Deterioration Development Dominant-Negative Mutation Energy Metabolism Fatty Acids Fatty acid glycerol esters Fracture Functional disorder Genetic Genetic Transcription Histologic Hyperglycemia In Vitro Inflammation Inflammatory Intervention Intuition LOX gene Life Liver Lysine Marrow Mass Spectrum Analysis Mediating Metabolic Metabolic dysfunction Metabolic stress Molecular Mouse Strains Mus Myelogenous Myeloid Cells Non-Insulin-Dependent Diabetes Mellitus Osteoblasts Osteoclasts Osteocytes Pathway interactions Pharmaceutical Preparations Pharmacology Phenotype Physiological Processes Population Process Property Proteins Quercetin Research Resources Risk Factors Role Serum Signal Transduction Skeleton Structure of beta Cell of islet System T-Lymphocyte Technology Testing Therapeutic Tissues Transgenes Transgenic Mice Transgenic Organisms Translations Work aged bone bone cell bone quality bone turnover cellular targeting clinical application diabetic evidence base glycation high dimensionality improved in vivo innovation innovative technologies insight non-diabetic novel novel strategies novel therapeutic intervention osteoprogenitor cell pre-clinical premature prevent promoter receptor binding receptor for advanced glycation endproducts senescence skeletal sugar tool transgenic suicide gene young adult

项目摘要

PROJECT SUMMARY Type 2 diabetes (T2D), a major risk factor for poor bone quality and fractures, is associated with the premature accumulation of senescent cells and advanced glycation endproducts (AGEs; activators of the receptor for AGE [RAGE] pathway) in multiple tissues, including bone. Intuitively, senescent cells and RAGE could act independently or interact via cross-talk to contribute substantially to skeletal fragility in T2D, yet this concept has not been rigorously tested. This proposal is founded on innovative concepts, technology, and approaches to test our central hypothesis that targeting cellular senescence or RAGE can improve T2D-related skeletal fragility. To test our hypothesis, we will use novel transgenic mice and innovative technology, including mass cytometry as well as advanced histological and molecular tools. The interplay among bone, energy metabolism, and T2D has been a topic of research for years, yet few in vivo studies have rigorously interrogated the contributions of senescent cells or RAGE signaling to skeletal dysfunction in T2D. From a translational perspective, better understanding of the cross-talk between senescence and RAGE in bone will yield impactful advances and may reveal novel strategies to ameliorate accelerated skeletal aging in T2D. To this end, in Aim 1 we will identify, locate, and characterize bone-resident senescent cell populations in mice with T2D and define their T2D-specific senescence-associated secretory phenotype (SASP). In Aim 2, using mice harboring transgenes that enable the selective elimination of p16Ink4a+ or p21Cip1+ senescent cells, we will test the hypothesis that senescent cell clearance in mice with established T2D will normalize bone remodeling and quality. Thus, we will distinguish the causal roles of p16Ink4a and p21Cip1 in mediating skeletal dysfunction in T2D using our global p16- and p21-ATTAC mouse strains by comparing the effects of systemic clearance of p16Ink4a+ vs p21Cip1+ senescent cells. In addition, we will assess the relative impact of clearing senescent osteocytes, using our novel Cre-LoxP lines – p16-LOX-ATTAC and p21-LOX-ATTAC. Global and osteocyte- specific clearance of senescent cells will be compared with pharmacological elimination using “senolytics”. Finally, in Aim 3, using our novel Cre-loxP mouse that inhibits RAGE signal transduction via cell-specific cytosolic-domain deficient dominant-negative RAGE (DN-RAGE) expression, we will define the effects of inhibiting RAGE signaling in the osteoblast/osteocyte and myeloid/osteoclast lineages on skeletal fragility in mice with T2D. Collectively, these studies will rigorously test whether cellular senescence and RAGE signaling underlie T2D-related skeletal fragility. We will address these questions by leveraging our unique resources and expertise. We will build upon compelling preliminary data and innovative approaches, including novel analytical, transgenic, and pharmacological tools that we anticipate will significantly advance our understanding of the fundamental biology of skeletal dysfunction in T2D, leading to new mechanistic insights, and evidence- based therapeutic approaches to facilitate the translation of preclinical discoveries to clinical applications.

项目概要 2 型糖尿病 (T2D) 是骨质量差和骨折的主要危险因素，与早产有关。衰老细胞和晚期糖基化终产物（AGEs；受体激活剂）的积累包括骨骼在内的多个组织中的 AGE [RAGE] 通路）直观地说，衰老细胞和 RAGE 可以发挥作用。独立或通过串扰相互作用，极大地导致 T2D 骨骼脆弱，但这个概念该提案尚未经过严格测试。测试我们的中心假设，即针对细胞衰老或 RAGE 可以改善与 T2D 相关的骨骼为了检验我们的假设，我们将使用新型转基因小鼠和创新技术，包括质量。细胞计数以及先进的组织学和分子工具骨骼、能量之间的相互作用。代谢，T2D 多年来一直是研究课题，但很少有体内研究严格证实 From a 探讨了衰老细胞或 RAGE 信号传导对 T2D 骨骼功能障碍的影响。转化视角，更好地理解骨骼衰老和RAGE之间的相互作用取得了有影响力的进展，并可能揭示改善 T2D 骨骼加速老化的新策略。为此，在目标 1 中，我们将识别、定位和表征小鼠骨中的衰老细胞群在目标 2 中，使用 T2D 并定义其 T2D 特异性衰老相关分泌表型 (SASP)。携带能够选择性消除 p16Ink4a+ 或 p21Cip1+ 衰老细胞的转基因的小鼠，我们将检验以下假设：患有 T2D 的小鼠中的衰老细胞清除将使骨重塑正常化因此，我们将区分 p16Ink4a 和 p21Cip1 在介导骨骼功能障碍中的因果作用。 T2D 使用我们的全球 p16- 和 p21-ATTAC 小鼠品系，通过比较全身清除的影响 p16Ink4a+ 与 p21Cip1+ 衰老细胞此外，我们将评估清除衰老细胞的相对影响。骨细胞，使用我们的新型 Cre-LoxP 系 - p16-LOX-ATTAC 和 p21-LOX-ATTAC Global 和骨细胞-。衰老细胞的特异性清除将与使用“senolytics”的药物消除进行比较。最后，在目标 3 中，使用我们的新型 Cre-loxP 小鼠，通过细胞特异性抑制 RAGE 信号转导胞浆结构域缺陷显性失活 RAGE (DN-RAGE) 表达，我们将定义抑制成骨细胞/骨细胞和骨髓/破骨细胞谱系中的 RAGE 信号传导对骨骼脆性的影响总的来说，这些研究将严格测试细胞衰老和 RAGE 信号传导是否相关。我们将利用我们独特的资源和技术来解决这些问题。我们将建立在令人信服的初步数据和创新方法的基础上，包括新颖的方法。我们预计分析、转基因和药理学工具将显着增进我们的理解 T2D 骨骼功能障碍的基础生物学研究，带来新的机制见解和证据- 基于治疗方法，促进临床前发现转化为临床应用。