GENETIC BASES FOR DISEASES OF THE RANK SIGNALING PATHWAY

Rank 信号通路疾病的遗传基础

基本信息

批准号：
8249359
负责人：
STEVEN R MUMM
金额：
$ 31.22万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8249359
关键词：
Accounting Adolescent Adult Affect Binding Biological Assay Biological Models Blood capillaries Bone Diseases Bone remodeling CSF1 gene CSF1R gene Candidate Disease Gene Cell model Childhood Clinical Code DNA DNA Sequence Defect Deformity Detection Diagnosis Disease Exons Familial disease Family Foundations Founder Effect Fracture Genes Genetic Genetic Markers Genomics Goals Grant Heterogeneity Human Immature Bone In Vitro Individual Interleukin-1 Journals Lead Life Literature Maps Medicine Molecular Mutation Navajo New England Nuclear Nucleic Acid Regulatory Sequences Osteoclasts Osteolysis Osteopathic Medicine Osteopenia Osteoporosis Paget&apos s Disease Pathway interactions Patients Phenotype Polymerase Chain Reaction Population Publishing RNA RNA Splicing Research Sampling Sequence Tagged Sites Serum Signal Pathway Signal Transduction Site Southern Blotting TNFRSF11B gene TNFSF10 gene TNFSF11 gene TRAF6 gene TRANCE protein Testing Tissues Tumor necrosis factor receptor 11b base bone turnover calcification capillary cytokine disease-causing mutation early childhood effective therapy genetic analysis infancy member mutant prenatal public health relevance receptor research clinical testing research study skeletal skeletal disorder young adult

项目摘要

DESCRIPTION (provided by applicant): Juvenile Paget's disease (JPD), an autosomal recessive osteopathy, features rapidly remodeling woven bone, osteopenia, fractures, and progressive skeletal deformity that can be fatal by young adult life. Homozygous deletion of the TNFRSF11B gene, encoding osteoprotegerin (OPG), was identified as the cause of JPD in two Navajo families. Previously, a defect in the RANK (receptor activator of nuclear factor ?-B) gene was determined to be the cause of a related disorder, familial expansile osteolysis (FEO). Hence, mutations in genes involved in the OPG/RANKL/RANK/NF-?B signaling pathway are responsible for JPD and related disorders of bone turnover. The long-range goal of this study is to identify other mutations in members of the OPG/RANKL/RANK/NF-?B pathway that cause JPD and related diseases in newly-ascertained patients, and to use in vitro cell modeling systems to assess the downstream effects of these mutations, to elucidate the pathobiology of these disorders. The Specific Aims of this project are: Specific Aim 1: Ascertain and evaluate additional patients worldwide with juvenile Paget's disease (JPD), familial expansile osteolysis (FEO), and related disorders. Specific Aim 2: In new patients with JPD, FEO, and related disorders, identify the genetic defects that cause these skeletal diseases. Specific Aim 3: Characterize the downstream effects of mutant OPG, RANK, and other members of the OPG/RANKL/RANK/NF-?B signaling pathway on osteoclast formation and action. Additional patients will be ascertained, and evaluated radiographically, biochemically, and molecularly. DNA and RNA samples will be screened for mutations in TNFRSF11B, encoding OPG, TNFRSF11A, encoding RANK, and TNFSF11, encoding RANKL, using PCR and capillary DNA sequencing, and RT-PCR. We will also use a microarray-based copy number analysis to identify potential genomic defects in these patients, and identify new candidate genes. For patients without mutations in the OPG, RANK, or RANKL genes, large-scale Solexa sequencing will be used to search other candidate genes in the OPG/RANKL/RANK/NF-?B signaling pathway for disease-causing mutations, including SQSTM1, VCP, TRAF6, aPKC, NIK, and others. Modulators of RANK signaling will also be examined, including TRAIL, IL-1, MCSF, c-FMS and additional cytokines and their cognate receptors. In vitro osteoclast culture assays will be used to determine downstream effects of mutations on osteoclast formation and function. These studies will further elucidate the genetic bases for JPD, FEO, and related disorders of bone turnover, caused by defects in the OPG/RANKL/RANK/NF-?B signaling pathway, which is critical for osteoclast formation and action. PUBLIC HEALTH RELEVANCE: This project combines clinical evaluation, genetic analysis, and experimental studies for patients and their families with rare bone diseases, including juvenile Paget's disease, familial expansile osteolysis, and related disorders. By understanding the genetic causes and cellular mechanisms of these skeletal diseases, new and better treatments can be developed. The information gained from this study may also be useful for therapy in common bone diseases, such as osteoporosis.

描述（由申请人提供）：青少年佩吉特病 (JPD) 是一种常染色体隐性遗传性骨病，其特点是快速重塑编织骨、骨质减少、骨折和进行性骨骼畸形，这些畸形可能对年轻人致命。编码骨保护素 (OPG) 的 TNFRSF11B 基因的纯合性缺失被确定为两个纳瓦霍家族中 JPD 的病因。此前，RANK（核因子β-B受体激活剂）基因的缺陷被确定为相关疾病家族性膨胀性骨溶解症（FEO）的原因。因此，涉及 OPG/RANKL/RANK/NF-κB 信号通路的基因突变是 JPD 和相关骨转换疾病的原因。本研究的长期目标是在新确定的患者中鉴定导致 JPD 和相关疾病的 OPG/RANKL/RANK/NF-κB 通路成员中的其他突变，并使用体外细胞建模系统来评估这些突变的下游影响，以阐明这些疾病的病理学。该项目的具体目标是：具体目标 1：确定并评估世界范围内患有青少年佩吉特病 (JPD)、家族性扩张性骨溶解 (FEO) 和相关疾病的其他患者。具体目标 2：在患有 JPD、FEO 和相关疾病的新患者中，确定导致这些骨骼疾病的遗传缺陷。具体目标 3：表征突变型 OPG、RANK 和 OPG/RANKL/RANK/NF-κB 信号通路其他成员对破骨细胞形成和作用的下游影响。将对其他患者进行确定，并进行放射学、生化和分子学方面的评估。将使用 PCR 和毛细管 DNA 测序以及 RT-PCR 筛查 DNA 和 RNA 样本中编码 OPG 的 TNFRSF11B、编码 RANK 的 TNFRSF11A 和编码 RANKL 的 TNFSF11 中的突变。我们还将使用基于微阵列的拷贝数分析来识别这些患者潜在的基因组缺陷，并识别新的候选基因。对于OPG、RANK或RANKL基因没有突变的患者，将使用大规模Solexa测序来搜索OPG/RANKL/RANK/NF-κB信号通路中的其他候选基因是否存在致病突变，包括SQSTM1、 VCP、TRAF6、aPKC、NIK 等。还将检查 RANK 信号传导调节剂，包括 TRAIL、IL-1、MCSF、c-FMS 和其他细胞因子及其同源受体。体外破骨细胞培养测定将用于确定突变对破骨细胞形成和功能的下游影响。这些研究将进一步阐明 JPD、FEO 和骨转换相关疾病的遗传基础，这些疾病是由 OPG/RANKL/RANK/NF-κB 信号通路缺陷引起的，该信号通路对于破骨细胞的形成和作用至关重要。公共健康相关性：该项目结合了针对患有罕见骨病（包括青少年佩吉特氏病、家族性扩张性骨溶解和相关疾病）的患者及其家人的临床评估、遗传分析和实验研究。通过了解这些骨骼疾病的遗传原因和细胞机制，可以开发新的更好的治疗方法。从这项研究中获得的信息也可能有助于治疗常见的骨疾病，例如骨质疏松症。