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.

项目概要促炎细胞因子 TNFα 被认为是糖尿病中观察到的骨折愈合延迟的原因，然而，对于 TNFα 抑制对骨形成的影响尚未达成共识，这表明寻找具有除此之外的独特功能的新药物的需求尚未得到满足。我们的基因筛选发现 TNFR 是颗粒体蛋白前体 (PGRN) 的新型受体（Tang 等人，Science，2011），这是一种已被研究的软骨形成因子。 1 型糖尿病是最常见的自身免疫性疾病，其特征是慢性炎症和 TNFα 活性升高，尽管 TNFα 活性主要通过 TNFR1 介导，但我们很高兴地发现 PGRN 刺激的骨再生在很大程度上取决于。这些矛盾的发现表明再生 PGRN/TNFR2 通路在 PGRN 刺激的骨折愈合中发挥着重要作用。此外，还发现了 14-3-3ε。作为响应 PGRN 刺激的 TNFR2 通路的一个组成部分，我们还开发了一种名为 Atsttrin 的工程蛋白，它由 PGRN 的三个 TNFR 结合域组成，考虑到 TNFα 升高，Atsttrin 比 PGRN 更有效。被认为是导致糖尿病骨折愈合延迟的原因，我们研究 PGRN 和 Atsttrin 通过 a) 抑制 TNFα/TNFR1 炎症和骨吸收来刺激糖尿病骨折愈合b) 将 14-3-3ε 招募到 TNFR2，然后激活骨再生途径。具体目标是：(1) 确定 PGRN，特别是其衍生物 Atsttrin 在糖尿病骨折愈合中的作用。将使用全身性和诱导性 PGRN 敲除小鼠来确定 PGRN 敲除是否会延迟糖尿病骨折愈合，以及重组 PGRN 和 Atsttrin 是否可以逆转它(SA#1A)；骨折愈合的哪个阶段需要PGRN才能成功完成糖尿病骨折愈合(SA#1B)；以及PGRN，尤其是Atsttrin是否对治疗糖尿病骨折具有治疗功效(SA#1C)。适当的可注射水凝胶局部输送不同剂量的 PGRN 或 Atsttrin (2) 阐明 PGRN 和 Atsttrin 刺激糖尿病骨折愈合的分子机制。 PGRN、Atsttrin 和 TNFα 对糖尿病骨髓干细胞软骨形成、信号通路、TNFR 和 14-3-3ε 的相互作用和依赖性的影响 (SA#2A)；这两种 TNFR 是否对于介导 PGRN 在糖尿病骨愈合中的作用很重要； (SA#2B)；以及通过建立 PGRN 和 Atsttrin 的保护作用是否依赖于 14-3-3ε诱导性 14-3-3ε[-/-] 小鼠糖尿病骨折模型 (SA#2C) 拟议的研究不仅将增进我们对糖尿病骨折愈合分子事件的理解，而且还可能导致糖尿病骨折的新治疗干预措施。愈合和其他骨折愈合受损的情况。