Genetic Analysis of Idiopathic-Type Curvature in Model Teleosts

模型硬骨鱼特发型弯曲的遗传分析

• 批准号: 7532864
• 负责人: Felix J Breden
• 金额: $ 14.33万
• 依托单位: SIMON FRASER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7532864
• 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; Condition; Congenital Abnormality; Coupled; Decompression Sickness; 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; Human Genetics; 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; Personal Satisfaction; Phenotype; Physicians; Physiological; Population; Positioning Attribute; Preventive; Principal Investigator; Procedures; Public Health; Quantitative Trait Loci; Range; 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; genetic analysis; genetic pedigree; genome sequencing; girls; human disease; improved; innovation; insight; knockout gene; medical specialties; mutant; novel therapeutics; programs; scoliosis; spine bone structure; teleost; tool

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（由申请人提供）：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%的儿科人群。这种畸形通过医生护理、支具治疗和脊柱融合手术给医疗保健系统带来了巨大的成本。迄今为止，由于人类谱系内的变异性以及缺乏具有相似表型的动物遗传模型，鉴定致病因素的努力失败了。尽管做了许多努力，但尚未发现这种综合征的致病基因。这项拟议中的研究使用了两种密切相关的硬骨鱼，每种硬骨鱼都有与人类相似的特发性弯曲突变体。这些动物是第一个证明脊柱弯曲不是由先天性椎骨畸形引起的模型。众所周知，人类和鱼类共享许多具有相似组织和时间表达特征的基因，并且人类和鱼类基因组之间的比较已被证明对于理解人类疾病的遗传学是有价值的。在这两个高度易处理的实验室模型中进行的联合分析将鉴定出特发性弯曲的候选基因，以在人类谱系中检测与IS的相关性（见Nancy米勒博士的合作信）。与人类IS相关的基因的鉴定可以允许有效的筛查，促进早期弯曲检测和治疗，在手术前减轻弯曲进展是必要的。了解生理机制可以提出新的治疗干预措施。目的：（1）利用两种近缘的硬骨鱼模型（第一个非诱导的IS动物模型）鉴定与特发性弯曲表型相关的基因。(2)在一个综合性的弯曲突变家系中研究特发性弯曲的复杂遗传结构。假设：在这些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鳉杂交作图，结合高密度遗传图谱和青鳉完全注释的完整基因组序列，来鉴定这些高度共线的模型遗传硬骨鱼中脊柱畸形的候选基因。我们将使用一个全面的，多代系谱的详细分析遗传的弯曲表型。驼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：一个基于超过1500个正常和弯曲个体的同胞交配的多代驼背谱系，其中大部分已被保存用于基因分型;包括129个BC后代和175个F2的作图杂交，其中平均48%具有可变大小的曲线（127个成年F2中总共有16.5%是弯曲的;曲线幅度的分布是可变的）; 189个基于表达序列标签（EST）的单核苷酸多态性（SNP），和839个随机基因组SNP，设计用于高通量基因分型;隐性表型;对84个BC后代进行QTL分析，共筛选出189个EST SNP位点。EST标记已被映射到青鳉基因组上的位置使用BLAST算法。因此，孔雀鱼和青鳉之间的同线性已被证实，因此，完全注释的青鳉序列可用于识别孔雀鱼回弯的候选基因。波浪形青鳉突变体也提供了几个机会和资源，以映射基因负责特发性脊柱侧凸：波浪谱系已近交超过60年;一个完整的注释基因组是可用的;和遗传连锁图已被描述，基于232个微卫星标记;全方位的基因组工具，青鳉，包括转基因和基因敲除库。我们已经完成了一个广泛的映射杂交437 F2后代从一个家庭，其中103个表现出曲率。使用微卫星标记，我们已经进行了初步的粗扫描约一半的青鳉基因组，并确定了一个候选位点与曲率。我们的研究计划将利用广泛的作图杂交和青鳉的基因组资源来精细定位特发性脊柱弯曲的候选基因。 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%的儿科人群。遗传基础尚不清楚。拟议的研究提供了一种高度创新的方法来绘制与IS相关的基因，使用已知的第一个表现出与人类相似的特发性曲率的动物模型。IS表型如何在硬骨鱼模型中出现是一个值得探索的基础科学，它可能会导致对人类分子和细胞机制的更深入理解。识别与特发性脊柱弯曲相关的基因可以进行有效的筛选，以促进人类早期弯曲检测，并提出新的治疗方法。