GENETIC BASES FOR DISEASES OF THE RANK SIGNALING PATHWAY

Rank 信号通路疾病的遗传基础

• 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
• 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' 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

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）：常染色体隐性骨质病（JPD）的少年paget病（JPD）具有迅速重塑的编织骨，骨质骨，骨折和渐进性骨骼畸形，这是年轻人成人生活可能是致命的。编码骨蛋白酶（OPG）的TNFRSF11b基因的纯合删除被确定为两个纳瓦霍家族的JPD的原因。以前，级别（核因子的受体激活因子？-b）基因的缺陷被确定为相关疾病的原因，家族膨胀骨溶解（FEO）。因此，涉及OPG/RANKL/RANK/NF-？B信号传导途径的基因突变是造成JPD和骨转换的相关疾病的原因。这项研究的远距离目标是鉴定OPG/RANKL/RANK/NF-？B途径的其他突变，导致新近确定的患者的JPD和相关疾病，并使用体外细胞建模系统来评估这些疾病病理生物学的体外细胞建模系统来评估这些突变的下游效应。该项目的具体目的是：特定目标1：确定和评估全球少年paget病（JPD），家族性扩张骨溶解（FEO）（FEO）和相关疾病的其他患者。 特定目标2：在新的JPD，FEO和相关疾病的新患者中，确定引起这些骨骼疾病的遗传缺陷。 具体目标3：表征突变体OPG，等级和其他成员的下游效应OPG/RANKL/RANK/NF-？B信号传导途径在破骨细胞形成和作用上。 将在射线照相，生化和分子上确定其他患者。 DNA和RNA样品将在TNFRSF11b，编码OPG，TNFRSF11A，编码等级和TNFSF11中进行筛选，并使用PCR和毛细管DNA测序和RT-PCR进行编码。我们还将使用基于微阵列的拷贝数分析来识别这些患者的潜在基因组缺陷，并确定新的候选基因。对于OPG，等级或RANKL基因中没有突变的患者，大规模的Solexa测序将用于搜索OPG/RANKL/RANK/RANK/NF-？B？B信号传导途径的其他候选基因，以搜索引起疾病的突变，包括SQSTM1，VCP，VCP，VCP，TRAF6，APKC，APKC，NIK和其他。还将检查等级信号传导的调节因子，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法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由OPG/RANKL/RANKL/RANK/NF-？B信号传导途径缺陷引起的JPD，FEO和相关疾病的遗传基础，这对于破骨细胞形成和作用至关重要。 公共卫生相关性：该项目结合了患有罕见骨骼疾病的患者及其家庭的临床评估，遗传分析和实验研究，包括少年paget病，家族性膨胀骨溶解以及相关疾病。通过了解这些骨骼疾病的遗传原因和细胞机制，可以开发出新的和更好的治疗方法。从这项研究中获得的信息也可能对常见骨疾病（例如骨质疏松症）的治疗有用。