Imaging and mechanistic analyses of face-brain dysmorphology in a CLP model

CLP 模型中面脑畸形的成像和机制分析

• 批准号: 8299845
• 负责人: Robert Lipinski
• 金额: $ 7.95万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8299845
• 关键词: Address; Animal Model; Animals; Apoptosis; Attenuated; Awareness; Basic Science; Behavioral; Biological Assay; Brain; Candidate Disease Gene; Caring; Cell Proliferation; Cleaved cell; Clinical; Cognitive; Collaborations; Complex; Congenital Abnormality; Congenital neurologic anomalies; Coupled; Defect; Development; Dorsal; Dysmorphology; Embryo; Embryology; Environment; Environmental Risk Factor; Erinaceidae; Exhibits; Exposure to; Face; Fetus; Fostering; Foundations; Frontonasal Prominence; Future; Gene Expression; Gene Targeting; Genes; Genetic; Goals; Growth; Health; Human; Image; In Situ Hybridization; In Situ Nick-End Labeling; Individual; Intervention; Investigation; Knowledge; Lateral; Magnetic Resonance; Magnetic Resonance Imaging; Mediating; Mediator of activation protein; Medical; Mentorship; Microarray Analysis; Microscopy; Modeling; Molecular; Molecular Genetics; Molecular Profiling; Morbidity - disease rate; Morphogenesis; Mus; Neuraxis; North Carolina; Oral; Pathogenesis; Pathway interactions; Patients; Pattern; Perinatal Exposure; Phenotype; Population; Prevalence; Prosencephalon; Research; Research Personnel; Resolution; Reverse Transcriptase Polymerase Chain Reaction; Role; Scientist; Shapes; Signal Transduction; Staining method; Stains; Structure; Study models; Surface; Testing; Time; Tissues; Toxicology; Training; Universities; Validation; Work; base; career; chemical genetics; cleft lip and palate; clinical phenotype; craniofacial; cyclopamine; design; experience; indexing; malformation; mouse model; multidisciplinary; neurodevelopment; novel; pregnant; progenitor; relating to nervous system; research study; shape analysis; smoothened signaling pathway; spatiotemporal

DESCRIPTION (provided by applicant): Clefts of the lip and palate (CLP) are the most commonly occurring craniofacial birth defect, cause significant morbidity, and require extensive medical intervention. In addition to the facial malformations and secondary complications of CLP, central nervous system (CNS) abnormalities, and cognitive and behavioral deficits are known to co-occur. Importantly, the etiological basis of CLP, as well as the cause and extent of associated CNS abnormalities remain poorly understood. This knowledge gap is best addressed through basic research. However, significant research progress has been hampered by two major limitations: a paucity of suitable animal models, and a lack of investigators with the expertise to integrate traditionally disparate conceptual and experimental approaches. An important advance in this arena has been our recent demonstration that in utero exposure to the Hedgehog (Hh) signaling antagonist, cyclopamine, induces CLP in the mouse, mimicking clinical phenotypes. Employing this model, the studies proposed herein use high-resolution magnetic resonance imaging to define CNS dysmorphology that co-occurs with CLP and to determine whether unique patterns of these abnormalities can be predicted by specific facial phenotypes (Aim 1). Mechanistic assays will test the hypothesis that cyclopamine exposure induces associated brain-face abnormalities by disrupting inductive signals between the developing forebrain and midface (Aim 2). The Hh pathway target genes which mediate the initial pathogenesis of these malformations will be identified and characterized (Aim 3), providing a set of candidate genes whose function in craniofacial development and etiological role clinical CLP will be pursued in future studies. Cumulatively, it is expected that these studie will define specific CNS abnormalities that are present in patients with CLP and provide foundational etiopathological characterization that will drive future efforts to examine the interaction of chemical and genetic perturbations to the Hh signaling pathway in the genesis of CLP in clinical populations. To augment the candidate's previous experience in toxicology and cell signaling, these research aims are coupled with didactic and hands-on training components in embryology, neurodevelopment, and clinical and molecular genetics. The opportunities outlined in this proposal will provide the candidate with a multidisciplinary conceptual background and experimental toolset ideal for elucidation of the complex etiopathology of CLP. The successful completion of these undertakings will be fostered by the exceptional research environment provided at the University of North Carolina at Chapel Hill, and mentorship by and collaboration with experts in craniofacial/neurodevelopment (Dr. Kathleen Sulik), small animal imaging (Dr. Al Johnson), 3D face-shape analysis (Dr. Peter Hammond), craniofacial genetics (Dr. Eric Everett), and clinical genetics (Dr. Arthur Aylsworth). Ultimately, the undertaking and completion of these endeavors will provide a foundation for the candidate's goal of pursuing an academic career in a setting which incorporates clinical and basic scientists to advance oral and craniofacial health. PUBLIC HEALTH RELEVANCE: The experiments in this project will use MRI and mechanistic analyses to examine the cause, extent, and co- occurrence of face and brain abnormalities in a mouse model of cleft lip and palate. The goal of this work is to increase our understanding of structural brain abnormalities that occur in individuals with CLP as well as the genetic and environmental factors that cause these common and morbid birth defects.

描述（由申请人提供）：唇腭裂 (CLP) 是最常见的颅面出生缺陷，会导致严重的发病率，并且需要广泛的医疗干预。除了面部畸形和 CLP 继发并发症外，已知中枢神经系统 (CNS) 异常以及认知和行为缺陷也会同时发生。重要的是，CLP 的病因学基础以及相关中枢神经系统异常的原因和程度仍然知之甚少。这种知识差距最好通过基础研究来解决。然而，重大研究进展受到两个主要限制的阻碍：缺乏合适的动物模型，以及缺乏具有相关研究能力的研究人员。 整合传统上不同的概念和实验方法的专业知识。该领域的一个重要进展是我们最近证明，在子宫内暴露于 Hedgehog (Hh) 信号拮抗剂环巴明可诱导小鼠 CLP，模拟临床表型。本文提出的研究利用该模型，使用高分辨率磁共振成像来定义与 CLP 共同发生的 CNS 畸形，并确定是否可以通过特定的面部表型来预测这些异常的独特模式（目标 1）。机制分析将检验这一假设，即环巴明暴露通过破坏发育中的前脑和中面部之间的感应信号来诱发相关的脑面部异常（目标 2）。将鉴定和表征介导这些畸形的初始发病机制的 Hh 途径靶基因（目标 3），提供一组候选基因，其在颅面发育中的功能和临床 CLP 的病因学作用将在未来的研究中进行探讨。总之，预计这些研究将定义 CLP 患者中存在的特定 CNS 异常，并提供基础的病因病理学特征，这将推动未来研究临床人群 CLP 发生过程中化学和遗传扰动与 Hh 信号通路的相互作用。为了增强候选人之前在毒理学和细胞信号传导方面的经验，这些研究目标与胚胎学、神经发育以及临床和分子遗传学方面的教学和实践培训相结合。本提案中概述的机会将为候选人提供多学科概念背景和实验工具集，非常适合阐明 CLP 的复杂病因病理学。这些事业的成功完成将得益于北卡罗来纳大学教堂山分校提供的卓越研究环境，以及颅面/神经发育（Kathleen Sulik 博士）、小动物成像（Al Johnson 博士）、3D 面部形状分析（Peter Hammond 博士）、颅面遗传学（Eric Everett 博士）和临床遗传学专家的指导和合作。 （阿瑟·艾尔斯沃斯博士）。最终，这些努力的承担和完成将为候选人在临床和基础科学家的环境中追求学术生涯的目标奠定基础，以促进口腔和颅面健康。 公共健康相关性：该项目中的实验将使用 MRI 和机制分析来检查唇裂和腭裂小鼠模型中面部和大脑异常的原因、程度和同时发生。这项工作的目标是加深我们对 CLP 患者发生的大脑结构异常以及导致这些常见和病态出生缺陷的遗传和环境因素的了解。