Pathogenic Mechanisms of Craniometaphyseal Dysplasia

颅骨干骺端发育不良的发病机制

基本信息

批准号：
10444184
负责人：
I-Ping Chen
金额：
$ 50.28万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10444184
关键词：
1 year old Acridine Orange Actins Address Age Animal Model Ankylosis Autophagocytosis Autophagosome Binding Biochemistry Biological Biological Markers Biological Models Biology Blindness Bone Diseases Bone remodeling Cell Shape Cells Clinical Research Connexin 43 Craniofacial Abnormalities Cytoskeleton Data Defect Deformity Deposition Disease Disease Progression Dysplasia Endocrinology Energy Metabolism Event Exhibits Face Facial paralysis Fasting Funding Future Genetic Glycolysis Goals Headache Human Hyperostosis Image In Vitro Investigation Investigational Therapies Knock-in Mouse Knockout Mice Knowledge Lead Life Link Lysosomes Membrane Fusion Mendelian disorder Metabolic Metabolic Bone Diseases Metabolic Pathway Methodology Methods Microfilaments Microtubules Molecular Monitor Mouse Protein Mus Mutation Neonatal Neurologic Symptoms Operative Surgical Procedures Osteoclasts Osteoporosis Pathogenesis Pathogenicity Patient Care Patients Pharmacological Treatment Phenotype Plasma Population Property Proteins Protocols documentation Proton Pump Public Health Publications Regulation Reporter Research Sampling Shapes Signal Transduction Skeletal Development Spectrin Testing Therapeutic Therapeutic Intervention Treatment Efficacy Treatment Protocols Urine Vesicle Western Blotting base bone bone cell clinical care cost craniofacial bone deafness differential expression effective therapy genome editing immunocytochemistry improved in vivo induced pluripotent stem cell inhibitor insight long bone loss of function metabolomics mouse model mutant novel novel therapeutic intervention prevent response sensor sex skeletal disorder stem cell biology targeted treatment therapeutic target tool translational study treatment response vacuolar H+-ATPase

项目摘要

Project Summary/Abstract Investigating pathogenic mechanisms for rare Mendelian disorders is important not only to identify therapeutic strategies for lifelong debilitating diseases but also to understand fundamental biological mechanisms. In this renewal application, we propose mechanistic and translational studies for craniometaphyseal dysplasia (CMD), an understudied craniotubular bone disorder characterized by lifelong progressing hyperostosis of craniofacial bones and abnormal shape of long bones. Continued bone accrual in CMD can lead to excruciating headaches, blindness, deafness, and facial palsy. Severe cases can be life-threatening. CMD patients are treated with repetitive, costly and risky surgeries when corrections of facial deformity are needed or severe neurological symptoms occur. Mutations in the progressive ankylosis protein (ANKH) and connexin 43 (Cx43) have been identified as causes for autosomal dominant and recessive CMD, respectively. To study CMD, we have generated state-of-the-art research tools, which include mouse models carrying CMD mutations, isogenic human induced pluripotent stem cells (hiPSCs) with or without CMD mutations, and bone resorbing cells (osteoclasts) derived from these hiPSCs. In the past funding period, we have discovered the rapid degradation of mutant ANKH(Human)/ANK(Mouse) protein and studied negative effects of mutant ANKH/ANK on the cytoskeleton, which determinates cell shape, size, and polarity. We also identified differentially expressed proteins in CMD osteoclasts and preferential binding partners for mutant ANK protein. However, CMD pathogenesis is not fully understood and potential therapeutics have not been explored. Our long-term goal is to utilize our research findings for identifying potential therapeutic targets to reduce or prevent the lifelong bone deposition in craniofacial bones. In the next 5 years, we will use animal models and molecular and cellular methodologies that we have developed to focus on mechanistic investigations and prepare for future clinical studies. Based on our preliminary data and previous publications we propose three specific aims. We will study the impact of CMD-mutant ANK on cellular acidification of osteoclasts (Aim 1) and on the bi-directional regulation between the cytoskeleton and an energy metabolism regulator in CMD (Aim 2). These are likely novel dominant functions of mutant ANK leading to CMD. In Aim 3 we will identify biomarkers that can be used to monitor the disease progression in patients and mouse models. We will also evaluate shifts in biomarker expression in response to experimental treatment regimen in our model systems. We expect that the proposed studies will give deeper insight into pathogenic mechanisms of CMD, knowledge needed to discover candidate targets for therapeutics. Biomarkers that correspond to disease progression or treatment efficacy will be the basis for future clinical studies.

项目概要/摘要研究罕见孟德尔疾病的致病机制不仅对于确定治疗方法很重要治疗终生衰弱性疾病的策略，同时也了解基本的生物学机制。在这个更新申请，我们提出颅骨干骺端发育不良（CMD）的机制和转化研究，一种尚未充分研究的颅管骨疾病，其特征是颅面骨终生进行性骨质增生骨骼和长骨的异常形状。 CMD 中持续的骨质增生可能会导致痛苦头痛、失明、耳聋和面瘫。严重的情况下可能会危及生命。 CMD 患者是当需要矫正面部畸形或面部畸形严重时，需要进行重复、昂贵且有风险的手术治疗出现神经症状。进行性强直蛋白 (ANKH) 和连接蛋白 43 (Cx43) 突变已被确定为常染色体显性和隐性 CMD 的分别病因。为了研究 CMD，我们已经产生了最先进的研究工具，其中包括携带 CMD 突变的小鼠模型、同基因具有或不具有 CMD 突变的人类诱导多能干细胞 (hiPSC) 以及骨吸收细胞（破骨细胞）源自这些 hiPSC。在过去的资助期间，我们发现了快速退化突变型 ANKH（人）/ANK（小鼠）蛋白，并研究了突变型 ANKH/ANK 对细胞骨架，决定细胞的形状、大小和极性。我们还鉴定了差异表达 CMD 破骨细胞中的蛋白质和突变型 ANK 蛋白质的优先结合伴侣。然而，CMD 发病机制尚不完全清楚，潜在的治疗方法尚未探索。我们的长期目标是利用我们的研究成果来确定潜在的治疗靶点，以减少或防止颅面骨终生骨沉积。未来5年，我们将利用动物模型我们开发的分子和细胞方法专注于机制研究和为未来的临床研究做好准备。根据我们的初步数据和之前的出版物，我们提出了三个具体目标。我们将研究 CMD 突变 ANK 对破骨细胞酸化的影响（目标 1）和 CMD 中细胞骨架和能量代谢调节剂之间的双向调节（目标 2）。这些可能是导致 CMD 的突变 ANK 的新显性功能。在目标 3 中，我们将识别生物标志物可用于监测患者和小鼠模型的疾病进展。我们还将评估班次在我们的模型系统中响应实验治疗方案的生物标志物表达。我们期望拟议的研究将更深入地了解 CMD 的致病机制、所需的知识发现候选治疗靶点。与疾病进展或治疗相对应的生物标志物疗效将成为未来临床研究的基础。