Targeting TNF Receptors to Inhibit Inflammation and to Prompt Bone Regeneration in Type 1 Diabetes - Resubmission - 1

靶向 TNF 受体抑制 1 型糖尿病炎症并促进骨再生 - 重新提交 - 1

基本信息

批准号：
10453563
负责人：
Chuanju Liu
金额：
$ 40.08万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10453563
关键词：
3D Print Affinity Anti-Inflammatory Agents Autoimmune Autoimmune Diseases Binding Biochemical Bone Marrow Stem Cell Bone Regeneration Bone Resorption CRISPR/Cas technology Chondrogenesis Chronic Complex Consensus Data Dependence Development Diabetes Mellitus Disease Effectiveness Event Exhibits Force of Gravity Fracture Gene set enrichment analysis Genes Genetic Genetic Screening Growth Factor Hydrogels Impaired healing Impairment In Vitro Inflammation Inflammatory Inflammatory Arthritis Inflammatory Response Injectable Injections Insulin-Dependent Diabetes Mellitus Knock-out Knockout Mice Laboratories Lead Ligands Maleimides Mass Spectrum Analysis Mediating Modeling Molecular Mus Names Open Fractures Osteogenesis PGRN gene Pathway interactions Pilot Projects Play Polyethylene Glycols Protein Engineering Recombinants Regenerative pathway Risk Role Science Signal Pathway Signal Transduction Signaling Molecule Site Streptozocin TNF gene TNFRSF1A gene TNFRSF1B gene Testing Therapeutic Treatment Efficacy Tumor Necrosis Factor Receptor bone fracture repair bone healing cytokine diabetic diabetic patient dosage in vivo inflammatory bone resorption inhibitor interest new therapeutic target novel novel therapeutic intervention osteoinductive factor prevent protective effect receptor recruit regenerative response scaffold treatment effect

项目摘要

Project Summary Pro-inflammatory cytokine TNFα is believed to be responsible for the delayed fracture healing observed in diabetes. However, there is no consensus on the effect of TNFα inhibition on the bone formation, indicating the unmet need in searching for new regents with unique features other than pure TNF inhibitors for diabetic fracture healing. Our genetic screen led to the identification of TNFR as the novel receptor of progranulin (PGRN) (Tang, et al, Science, 2011), a chondrogenic factor that has been shown to be therapeutic against autoimmune inflammatory arthritis. Type 1 diabetes is the most common autoimmune disease, characterized by chronic inflammation and elevated TNFα activity. Although TNFα activity is mediated primarily through TNFR1, we were excited to find that PGRN-stimulated bone regeneration largely depends on TNFR2. These paradoxical findings suggest that the regenerative PGRN/TNFR2 pathway plays a major role in PGRN-stimulated fracture healing. In addition, 14-3-3ε was identified as a component of TNFR2 pathway in response to PGRN stimulation. Further, we have developed an engineered protein named Atsttrin which is composed of three TNFR-binding domains of PGRN, and Atsttrin is more effective than PGRN in inflammatory arthritis. Given that elevated TNFα is believed to be responsible for delayed diabetic fracture healing, we hypothesize that PGRN and Atsttrin stimulate diabetic fracture healing through a) inhibition of TNFα/TNFR1 inflammatory and bone resorption pathway; and primarily b) recruitment of 14-3-3ε to TNFR2, followed by activation of bone regeneration pathway. The Specific Aims are: (1) To determine the role of PGRN, especially its derivative Atsttrin, in diabetic fracture healing. We will use both systemic and inducible PGRN knockout mice to determine whether knockout of PGRN delays diabetic fracture healing, and whether recombinant PGRN and Atsttrin can reverse it (SA#1A); which stage of fracture healing requires PGRN for successful completion of diabetic fracture healing (SA#1B); and whether PGRN, especially Atsttrin, has therapeutic efficacy in treating diabetic fracture (SA#1C). We will use an appropriate injectable hydrogel to locally deliver various dosages of PGRN or Atsttrin. (2) To elucidate the molecular mechanisms by which PGRN and Atsttrin stimulate diabetic fracture healing. We will determine the effects of PGRN, Atsttrin, and TNFα on chondrogenesis of diabetic bone marrow stem cells, signaling pathways, interplays and dependence on TNFR and 14-3-3ε (SA#2A); whether both TNFRs are important for mediating PGRN's role in diabetic bone healing (SA#2B); and whether the protective effects of PGRN and Atsttrin depend on 14-3-3ε by establishing diabetic fracture models with inducible 14-3-3ε[-/-] mice (SA#2C). Proposed studies will not only advance our understanding of the molecular events underlying diabetic fracture healing, but could also lead to novel therapeutic interventions for diabetic fracture healing and other conditions in which fracture healing is impaired.

项目摘要促炎细胞因子TNFα被认为是糖尿病中观察到的延迟骨折愈合的原因。但是，关于TNFα抑制对骨形成的影响尚无共识，这表明在寻找新的关注点的需求中，除了纯TNF抑制剂以外的糖尿病裂缝愈合以外的其他特征。我们的遗传筛查导致TNFR鉴定为易生蛋白（PGRN）的新型受体（Tang等，Science，2011），这是一种软骨，已被证明对自身免疫性炎症性关节炎具有治疗性。 1型糖尿病是最常见的自身免疫性疾病，其特征是慢性感染和TNFα活性升高。尽管TNFα活性是通过TNFR1介导的，但我们很高兴发现PGRN刺激的骨再生在很大程度上取决于TNFR2。这些矛盾的发现表明，再生PGRN/TNFR2途径在PGRN刺激的断裂愈合中起主要作用。另外，响应PGRN刺激，将14-3-3ε鉴定为TNFR2途径的组成部分。此外，我们开发了一种名为Atstrin的工程蛋白质，该蛋白质由PGRN的三个TNFR结合结构域组成，而Atsttrin在炎症性关节炎中比PGRN更有效。鉴于TNFα升高被认为是造成糖尿病骨折愈合延迟的原因，我们假设PGRN和ATSTTRIN通过a）抑制TNFα/TNFR1炎症和骨骼分辨率刺激了糖尿病骨折愈合。 b）将14-3-3ε募集到TNFR2，然后激活骨再生途径。具体目的是：（1）确定PGRN在糖尿病骨折愈合中的作用，尤其是其衍生物atstrin。我们将同时使用全身和诱导的PGRN敲除小鼠来确定PGRN的敲除症状延迟糖尿病骨折愈合，以及重组PGRN和ATSTTRIN是否可以将其逆转（SA＃1A）；哪个骨折愈合的阶段需要PGRN成功完成糖尿病骨折愈合（SA＃1B）； PGRN，尤其是Atstrin是否具有治疗糖尿病骨折（SA＃1C）的治疗效率。我们将使用适当的注射水凝胶在局部提供各种剂量的PGRN或Atstrin。（2）阐明PGRN和Atstrin刺激糖尿病骨折愈合的分子机制。我们将确定PGRN，atsttrin和TNFα对糖尿病骨髓干细胞的软骨发生，信号通路，相互作用和对TNFR和14-3-3ε的依赖性的影响（SA＃2A）；这两个TNFR是否对于介导PGRN在糖尿病骨骼愈合中的作用很重要（SA＃2B）； PGRN和Atstrin的受保护作用是否通过建立具有诱导14-3-3ε[ - / - ]小鼠（SA＃2C）的糖尿病断裂模型来依赖14-3-3ε。拟议的研究不仅会提高我们对糖尿病骨折愈合基础的分子事件的理解，而且还可能导致新的糖尿病骨折愈合的热干预措施以及其他损伤骨折愈合的疾病。