Investigations of the molecular genetics and pathogenesis of scoliosis.

脊柱侧弯的分子遗传学和发病机制的研究。

• 批准号: 10587849
• 负责人: Ryan Scott Gray
• 金额: $ 46.3万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-10 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10587849
• 关键词: Abate; Adhesions; Adolescent; Adult; Agreement; Alleles; Animal Genetics; Animal Model; Back; Biogenesis; Biological Process; Biomechanics; Cartilage; Cell physiology; Cells; Chest; Child; Childhood; Collection; Complement; Coupled; Cyclic AMP; Cyclic AMP-Responsive DNA-Binding Protein; Data; Defect; Dense Connective Tissue; Development; Distress; Elements; Engineering; Environment; Fibrocartilages; Funding; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gene Expression; Genes; Genetic; Genetic Processes; Genetic Screening; Genetic study; Goals; Heterogeneity; Homeostasis; Human; Human Genetics; INPPL1 gene; Idiopathic scoliosis; Inositol; Intervertebral disc structure; Investigation; Knockout Mice; Knowledge; Life; Link; Modeling; Molecular; Molecular Genetics; Morphogenesis; Movement; Mus; Musculoskeletal; Musculoskeletal Diseases; Mutagenesis; Mutant Strains Mice; Mutation; Natural History; Neurologic; Nucleotides; Pain; Pathogenesis; Pathway interactions; Pattern; Perinatal; Phenotype; Phosphoric Monoester Hydrolases; Polyphosphates; Predisposition; Property; Publishing; Purinoceptor; Receptor Gene; Receptor Signaling; Regulation; Research; Role; Severities; Signal Pathway; Signal Transduction; Signaling Protein; Spinal Cord; Spinal Curvatures; Spinal Diseases; Structural defect; Susceptibility Gene; Testing; Therapeutic; Tissues; Transcriptional Activation Domain; Variant; Vertebral column; Vertebrates; Work; Zebrafish; cartilage development; experimental study; genetic analysis; genetic approach; insight; intercalation; mutant; notochord; novel; programs; protein activation; psychosocial; scoliosis; screening; skeletal; skeletal dysplasia; transcription factor; tripolyphosphate; vertebra body

ABSTRACT This is a renewal application of an established program to investigate the regulation of the development and homeostasis of the spine. During the first funding period, our studies demonstrated that the gene Adgrg6, implicated in a common human spine disorder called adolescent idiopathic scoliosis, has an essential role in maintaining spine alignment in mice. We showed that the G-protein coupled rector Adgrg6 regulates gene expression and biomechanical properties of the intervertebral discs and dense connective tissue of the spine. Furthermore, we demonstrated that Adgrg6 stimulates cAMP signaling regulate factors essential for homeostasis of fibrocartilaginous tissue of the spine. Our findings suggest a new hypothesis that stimulation of cAMP signaling can decrease the onset and severity of scoliosis caused by the loss of Adgrg6 signaling. In addition, human genetics analysis of scoliosis identified a novel variant located in the transcriptional activation domain of the transcription factor SOX9. Significantly, targeted disruption of this domain of Sox9 in mice caused scoliosis and dysregulation of gene expression in fibrocartilaginous tissues of the spine. Here, we will test the hypothesis that Adgrg6 and Sox9 are functionally linked for regulation homeostasis and alignment of the spine. To add breadth to our program goals, we continued a forward genetic screen to isolate a collection of spine disorder mutant zebrafish. We recently identified two zebrafish mutants that fail to complement a novel thoracic scoliosis phenotype, suggesting a new pathway controlling spine morphogenesis. The characterization of this unique thoracic scoliosis phenotype will expand our knowledge into the cellular and molecular heterogeneity of spine disorders. Here, we will test the hypothesis that thoracic scoliosis in zebrafish is caused by a disruption of purinergic signaling leading to defects in notochord biogenesis. We will test these hypotheses via studies divided into three Specific Aims. Specific Aim 1 will deepen our mechanistic understanding of effectors of Adgrg6 signaling in the spine and test whether stimulation of cAMP can restore homeostasis to fibrocartilaginous tissues of the spine and halt the onset and progression of scoliosis. Specific Aim 2 will characterize the cellular and molecular causes of scoliosis in a novel Sox9 mutant mouse and use this model to test whether genetic interactions between Adgrg6 and Sox9 variants increase the susceptibility to scoliosis. Specific Aim 3 will characterize novel thoracic scoliosis mutant zebrafish and test a model that purinergic signaling is essential for notochord biogenesis and spine morphogenesis in zebrafish. Our results will provide new insights into the molecular genetics and biological processes necessary for the development and homeostasis of the spine. These studies may provide fundamental insights into the biological processes and pathways associated with human skeletal dysplasia and scoliosis.

摘要 这是一个既定计划的更新申请，以调查开发的监管， 脊柱的内稳态。在第一个资助期间，我们的研究表明，基因Adgrg 6， 涉及一种常见的人类脊柱疾病，称为青少年特发性脊柱侧凸，具有重要的作用， 保持小鼠脊柱对齐。我们发现，G蛋白偶联受体Adgrg6调节基因 表达和生物力学性能的椎间盘和致密结缔组织的脊柱。 此外，我们证明Adgrg 6刺激cAMP信号调节因子， 脊柱纤维软骨组织的稳态。我们的发现提出了一个新的假设， cAMP信号通路的阻断可以减少Adgrg6缺失引起的脊柱侧凸的发生和严重程度 发信号。此外，对脊柱侧弯的人类遗传学分析发现了一种位于脊柱侧弯的新变体。 转录因子SOX 9的转录激活结构域。重要的是，有针对性地破坏这一点， Sox 9结构域在小鼠中引起脊柱侧凸和纤维软骨组织中基因表达失调， 脊柱在这里，我们将测试Adgrg 6和Sox 9在调节功能上相关联的假设。 脊柱的平衡和排列。 为了扩大我们的计划目标，我们继续进行正向遗传筛选，以分离出一组 脊椎紊乱突变斑马鱼。我们最近发现了两个斑马鱼突变体， 胸椎脊柱侧凸表型，提出了一个新的途径控制脊柱形态发生。表征 这一独特的胸椎侧凸表型将扩展我们的知识到细胞和分子 脊柱疾病的异质性。在这里，我们将测试斑马鱼胸椎侧凸的假设， 由嘌呤能信号中断导致脊索生物发生缺陷引起。 我们将通过分为三个具体目标的研究来验证这些假设。具体目标1将深化 我们对脊髓中Adgrg6信号传导效应子的机制理解，并测试刺激脊髓中Adgrg6信号传导是否有效。 cAMP可以恢复脊柱纤维软骨组织的稳态，并阻止脊髓炎的发生和发展。 脊柱侧凸特异性目标2将描述一种新的Sox 9突变体中脊柱侧凸的细胞和分子原因 小鼠，并使用该模型来测试Adgrg6和Sox 9变体之间的遗传相互作用是否增加了 脊柱侧凸的易感性。具体目标3将描述新的胸椎侧凸突变斑马鱼，并测试一种新的脊柱侧凸突变体。 模型表明嘌呤能信号对斑马鱼脊索生物发生和脊椎形态发生是必不可少的。 我们的研究结果将提供新的见解分子遗传学和生物学过程所必需的 脊柱的发育和体内平衡。这些研究可能会提供生物学的基本见解， 与人类骨骼发育不良和脊柱侧凸相关的过程和途径。