Human Genetics and Clinical Translation

人类遗传学和临床转化

基本信息

批准号：
10458400
负责人：
CAROL A WISE
金额：
$ 35.7万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10458400
关键词：
ATAC-seq Adolescent Adult Affect Alleles Award Back Pain Biological Biological Response Modifier Therapy CRISPR/Cas technology Candidate Disease Gene Cartilage Cells Child Childhood Collagen Type XI Connective Tissue Coupled Databases Deformity Development Diagnosis Early Diagnosis Embryo Engineering Epiphysial cartilage Ethylnitrosourea Etiology Extended Family Extracellular Matrix Family Female Functional disorder Genes Genetic Recombination Genetic Screening Genetic Translation Genetically Engineered Mouse Genomics Goals Growth Health Histologic Human Human Genetics Idiopathic scoliosis Image Immunohistochemistry Intervention Intervertebral disc structure Knock-in Knock-in Mouse Knockout Mice LacZ Genes Life MMP14 gene Matrix Metalloproteinases Mechanics Medical Meta-Analysis Modeling Molecular Monitor Morphology Mus Musculoskeletal Diseases Mutation Operative Surgical Procedures Pathogenesis Pathway interactions Patients Phenotype Physiological Prevention Preventive Resistance Reverse engineering Risk Risk Factors Role School-Age Population Sex Bias Spinal Susceptibility Gene Testing Time Tissues Untranslated RNA Variant Vertebral column Zebrafish biobank bone quality boys clinical translation cohort cost experimental study gene discovery genetic architecture genome sequencing genome wide association study girls high risk multi-ethnic mutant novel offspring postnatal programs risk variant scoliosis sex sexual dimorphism single-cell RNA sequencing skeletal skeletal disorder spatiotemporal tool transcriptomics whole genome

项目摘要

Project Summary Adolescent idiopathic scoliosis (AIS) is a twisting condition of the spine and is the most common pediatric musculoskeletal disorder, affecting 3% of children worldwide. Children with AIS risk severe disfigurement, back pain, and physiologic dysfunction later in life, and are treated symptomatically rather than preventively because the underlying etiology is unknown. Girls requiring treatment for AIS outnumber boys by more than five-fold. Our overall purpose is to understand the biologic causes of AIS as a means to early diagnosis, prevention and non- invasive biologic treatment. With support from this P01 in the previous award period we substantially increased the number of validatedAIS susceptibility loci by large-scale multi-ethnic GWAS meta-analysis. Together with the other projects in the program we also established that the cartilage extracellular matrix (ECM) is a functional tissue in AIS. We subsequently discovered that nonsynonymous variants in genes encoding ECM components COL11A1 and MMP14 are associated with AIS. Together with genomics Project 3 we uncovered evidence of sex-biased PAX1-COL11A1 expression. In parallel, whole genome sequencing in families identified candidate mutations in AIS families that zebrafish Project 2 is engineering into orthologous zebrafish genes. Here we propose to drive these discoveries forward to develop mechanistic understanding of AIS pathogenesis. In one aim of the project, we will evaluate the consequences of Col11a1 loss from chondrogenic lineages by generating Col11a1fl/fl /ATC Cre lines, inducing cre recombination at embryologic and at early postnatal timepoints. Mutant offspring and their littermates will be evaluated phenotypically and morphologically, by imaging, quantitative immunohistochemistry and mechanical strength testing of growth plate and intervertebral disc (IVD) cartilages. With Project 3-Genomics we will also perform single cell RNAseq coupled with ATAC-seq in IVD to define the role of Col11a1 at cellular levels. To characterize the role of Mmp14 in mouse spine, we will characterize its spatio-temporal expression, and its cell-specific role in spine development using MT1-MMPlacZ/+ knockin lines. In a second aim, we will continue reverse engineering together with Project 2-Zebrafish and Project 3-Genomics to characterize the molecular and functional consequences of spine deformity-associated mutations in vertebrate models. In a reciprocal fashion we will cross-reference new scoliosis candidate alleles identified in Project 2-Zebrafish or Project 3-Genomics with those identified in our human cohorts. In a third aim we will discover novel high-risk scoliosis-associated variants by genome sequencing extended families and a unique cohort of AIS treatment non-responders. We will also identify new spine deformity mutants from our highly productive ENU-induced genetic screen in mice. Building on the momentum of our previous discoveries, we will synergize with Zebrafish Project 2 and Genomics Project 3 in further defining pathways and developmental mechanisms of spine development and deformity. Our collaborative will also continue to define the genetic architecture of AIS and yield tools that will enable better diagnosis, interventions, and preventions.

项目摘要青春期特发性脊柱侧弯（AIS）是脊柱的扭曲状况，是最常见的小儿肌肉骨骼疾病，影响全球3％的儿童。 AIS风险严重毁容的孩子，返回疼痛和生理功能障碍以后生活，并受到症状的治疗，而不是预防性治疗潜在的病因是未知的。女孩需要对AIS的男孩的治疗超过五倍。我们的总体目的是了解AIS的生物学原因是早期诊断，预防和非 - 侵入性生物治疗。在上一个奖项期间的P01的支持下，我们大大提高了通过大规模多种族GWAS荟萃分析的验证易感基因座数量。与其他项目一起在该程序中，我们还确定软骨外基质（ECM）是AIS中的功能组织。我们随后发现编码ECM组件COL11A1和MMP14的基因中的非同义变体为与AIS相关。与基因组学项目3一起，我们发现了性别偏见的PAX1-COL11A1的证据表达。同时，家族中的整个基因组测序鉴定了斑马鱼的AIS家族中的候选突变项目2正在工程直系同源的斑马鱼基因。在这里，我们建议推动这些发现向前发展以发展 AIS发病机理的机械理解。以该项目的一个目的，我们将评估 COL11A1通过产生COL11A1FL /FL /ATC CRE系导致CRE重组而导致软骨谱系损失在胚胎学和产后时间点。将评估突变的后代及其同窝室通过成像，定量免疫组织化学和机械强度在表型上和形态上生长板和椎间盘（IVD）软骨的测试。通过项目3基因组学，我们还将执行单细胞RNASEQ与IVD中的ATAC-SEQ结合，以定义Col11a1在细胞水平上的作用。表征 MMP14在小鼠脊柱中的作用，我们将表征其时空表达及其细胞特异性作用在使用MT1-MMPLACZ/+敲击线的脊柱发育中。在第二个目标中，我们将继续反向工程与项目2-泽布拉夫和第三基因组技术一起表征分子和功能脊椎畸形相关突变在脊椎动物模型中的后果。以相互的方式我们将跨参考的新脊柱侧弯候选等位基因在项目2- Zebrafish或Project 3基因组中确定的等位基因那些在我们的人类人群中确定的。在第三个目标中，我们将发现新型的高风险脊柱侧弯相关变体通过基因组测序扩展家庭和独特的AIS治疗无反应者。我们也会从我们高生产力的ENU诱导的小鼠遗传筛选中确定新的脊柱畸形突变体。建筑在我们以前发现的势头上，我们将与斑马鱼项目2和基因组学协同作用项目3在进一步定义脊柱发育和畸形的途径和发育机制中。我们的协作还将继续定义AIS的遗传体系结构，并产生工具，这将使更好诊断，干预和预防。