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Genetic Analysis of Idiopathic-Type Curvature in Model Teleosts

模型硬骨鱼特发型弯曲的遗传分析

基本信息

批准号：
7646365
负责人：
Felix J Breden
金额：
$ 11.71万
依托单位：
SIMON FRASER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7646365
关键词：
Accounting Adult Advisory Committees Affect Algorithms Animal Genetics Animal Model Animals Architecture Basic Science Biochemical Biological Biological Process Blast Cell Braces-Orthopedic appliances Canada Candidate Disease Gene Caring Characteristics Child Childhood Chromosome Mapping Clinic Collaborations Complex Congenital Abnormality Coupled Deformity Detection Disease Early Diagnosis Etiology Exhibits Expressed Sequence Tags Family Fishes Foundations Friends Generations Genes Genetic Genetic Models Genome Genomics Genotype Growth Guppy Healthcare Systems Hereditary Disease Human Idiopathic scoliosis Individual Laboratories Lead Letters Libraries Maps Measures Methodology Methods Microsatellite Repeats Modeling Molecular Nature Online Mendelian Inheritance In Man Operative Surgical Procedures Partner in relationship Pathway interactions Patients Phenotype Physicians Physiological Population Positioning Attribute Preventive Principal Investigator Procedures Quantitative Trait Loci Research Resources Scanning Screening procedure Services Single Nucleotide Polymorphism Spinal Spinal Curvatures Spinal Fusion Syndrome Synteny Testing Therapeutic Intervention Tissues Transgenic Organisms Visit Work base comparative cost density design developmental genetics gene discovery genetic analysis genetic linkage genetic pedigree genome sequencing girls human disease improved innovation insight knockout gene medical specialties mutant novel therapeutic intervention offspring programs public health relevance scoliosis spine bone structure teleost tool

项目摘要

DESCRIPTION (provided by applicant): Familial/idiopathic-type scoliosis (IS) accounts for 80% of all human spinal curvature and affects 3%-4% of the pediatric population. The deformity imposes a substantial cost on the healthcare system through physician care, bracing treatments, and spinal fusion surgeries. Efforts to identify causative factors have failed so far, due to variability within human pedigrees, and the lack of an animal genetic model with a similar phenotype. Despite many efforts, no causative genes have been discovered for this syndrome. The proposed research uses two closely related species of teleosts each having mutants with idiopathic-type curvature similar to humans. These animals are the first models to demonstrate spinal curvature that is not induced nor caused by congenital malformation of the vertebrae. It is well established that humans and fish share many genes with similar tissue and temporal expression characteristics, and comparisons between human and fish genomes have proven to be valuable for understanding the genetics of human diseases. The combined analysis in these two highly-tractable, laboratory models will identify candidate genes for idiopathic-type curvature to be tested for association with IS in human pedigrees (see letter of collaboration from Dr. Nancy Miller). The identification of genes associated with human IS could allow for effective screening, facilitating early curve detection and treatment, mitigating curve progression before surgery is necessary. Understanding physiological mechanisms can suggest novel therapeutic interventions. Objectives: (1) To identify genes involved in the idiopathic-type curvature phenotype using two closely related model teleosts, the first non-induced animal models for IS. (2) To investigate the complex genetic architecture of idiopathic-type curvature in a comprehensive pedigree of the curveback mutant. Hypothesis: Candidate genes discovered in either of these models will help elucidate the genetic causes and physiological pathways leading to the deformity in humans. Research plan: We will use guppy and medaka mapping crosses combined with a high- density genetic map and a fully-annotated, complete genome sequence for medaka, to identify candidate genes for spinal deformities in these highly syntenic, model genetic teleosts. We will use a comprehensive, multi-generation pedigree for a detailed analysis of inheritance of the curveback phenotype. The curveback guppy presents several opportunities for discovering genes that cause idiopathic-type spinal curvature: a multi-generation curveback pedigree based on sib-mating with over 1500 normal and curved individuals, most of which have been preserved for genotyping; mapping cross comprising 129 BC offspring of which an average of 48% have curves of variable magnitude, and 175 F2s (a total of 16.5% of 127 adult F2s are curved; the distribution of curve magnitude is variable); 189 expressed sequence tag (EST)-based single nucleotide polymorphisms (SNPs), and 839 random genomic SNPs, designed for high-throughput genotyping; recessive phenotype; and QTL study based on 84 BC offspring screened for 189 EST-based SNPs. The ESTs markers have been mapped to positions on the medaka genome using the BLAST algorithm. Therefore, synteny between guppy and medaka has been confirmed, and therefore the fully-annotated medaka sequence can be used to identify candidate genes for guppy curveback. The wavy medaka mutant also presents several opportunities and resources for mapping genes responsible for idiopathic-type scoliosis: the wavy lineage has been inbred for more than 60 years; a fully-annotated genome is available; and a genetic linkage map has been described, based on 232 microsatellite markers; a full range of genomic tools are available for medaka, including transgenics and gene knockout libraries. We have completed an extensive mapping cross of 437 F2 offspring from one family, 103 of which exhibit curvature. Using microsatellite markers, we have performed an initial coarse-scan of approximately one half of the medaka genome and identified a candidate locus for association with curvature. Our proposed research will use the extensive mapping cross and the genomic resources available for medaka to fine-map candidate genes for idiopathic- type spinal curvature. Public Health Relevance: Human idiopathic-type scoliosis (IS) is a debilitating disease, affecting 3%-4% of the global pediatric population. The genetic basis is unknown. The proposed research offers a highly innovative approach to mapping genes associated with IS, using the first animal models known to exhibit idiopathic-type curvature similar to humans. How the IS phenotype arises in model teleosts is a worthwhile basic science pursuit that may lead to greater understanding of the molecular and cellular mechanisms in humans. Identification of genes associated with idiopathic-type spinal curvature could allow for effective screening to facilitate early curve detection in humans and suggest new therapeutic approaches.

描述(申请人提供)：家族性/特发性脊柱侧凸(IS)占所有人类脊柱弯曲的80%，影响3%-4%的儿科人口。这种畸形通过医生护理、支撑治疗和脊柱融合手术给医疗系统带来了巨大的成本。到目前为止，识别致病因素的努力失败了，原因是人类谱系内的变异性，以及缺乏具有类似表型的动物遗传模型。尽管做了很多努力，但还没有发现这种综合征的致病基因。这项拟议的研究使用了两种关系密切的硬骨鱼，每种都具有与人类相似的特发性弯曲型突变。这些动物是第一个展示脊柱弯曲的模型，这种弯曲既不是由椎骨先天性畸形引起的，也不是由先天性椎骨畸形引起的。众所周知，人类和鱼类拥有许多相似的组织和时间表达特征的基因，人类和鱼类基因组的比较已被证明对理解人类疾病的遗传学有价值。在这两个高度易处理的实验室模型中的联合分析将确定特发性曲度的候选基因，以便在人类家系中进行与IS的关联测试(参见南希·米勒博士的合作信函)。识别与人类IS相关的基因可以进行有效的筛查，促进早期曲线检测和治疗，在必要的手术前减缓曲线进展。了解生理机制可以提出新的治疗干预措施。目的：(1)利用两种亲缘关系密切的硬骨鱼模型，首次建立IS的非诱导动物模型，寻找与特发性曲度表型相关的基因。(2)研究特发性曲度突变家系的复杂遗传结构。假设：在这两个模型中发现的候选基因将有助于阐明导致人类畸形的遗传原因和生理途径。研究计划：我们将使用孔雀鱼和青竹杂交作图，结合高密度遗传图谱和青竹的完全注释的完整基因组序列，在这些高度共生的模型遗传硬骨鱼中识别脊柱畸形的候选基因。我们将使用一个综合的、多世代的家系来详细分析曲折表型的遗传。曲背孔雀鱼为发现导致特发性脊柱弯曲的基因提供了几个机会：基于与1500多个正常和弯曲个体的同胞交配的多代曲折家系，其中大多数已被保存用于基因分型；由129个BC后代组成的杂交图谱，其中平均48%具有可变大小的曲线，175个F2(127个成年F2中总共16.5%是弯曲的；曲线大小分布可变)；189个基于表达序列标签(EST)的单核苷酸多态(SNPs)，和839个随机基因组SNPs，设计用于高通量基因分型；隐性表型；以及基于84个BC后代的QTL研究，筛选出189个基于EST的SNPs。已使用BLAST算法将EST标记映射到青竹基因组上的位置。因此，孔雀鱼和青竹之间的同步性已经得到证实，因此完全注释的青竹序列可以用于识别孔雀鱼弯曲病的候选基因。波状青竹突变体还为定位与特发性脊柱侧弯有关的基因提供了几个机会和资源：波状血统已被近亲繁殖60多年；已有完全注释的基因组；基于232个微卫星标记描述了遗传连锁图谱；青竹有一系列基因组工具可用，包括转基因和基因敲除库。我们已经完成了来自一个家系的437个F2后代的广泛的杂交作图，其中103个表现出曲率。使用微卫星标记，我们已经对青竹基因组的大约一半进行了初步的粗略扫描，并确定了与曲率相关的候选基因座。我们建议的研究将使用广泛的定位交叉和青竹可用的基因组资源来精细定位特发性脊柱弯曲型的候选基因。公共卫生相关性：人类特发性脊柱侧凸(IS)是一种使人衰弱的疾病，影响全球3%-4%的儿科人口。其遗传基础尚不清楚。这项拟议的研究提供了一种高度创新的方法来定位与IS相关的基因，使用了第一个已知的类似于人类的特发性曲度的动物模型。IS表型是如何在模型硬骨鱼中出现的，这是一项有价值的基础科学研究，可能会导致对人类分子和细胞机制的更好理解。识别与特发性脊柱曲度相关的基因可以进行有效的筛查，以促进人类早期曲度的检测，并提出新的治疗方法。