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）：嘴唇和pa裂（CLP）是最常见的颅面先天缺陷，引起明显的发病率，需要进行广泛的医疗干预。除了CLP的面部畸形和次要并发症外，中枢神经系统（CNS）异常以及认知和行为缺陷也已知。重要的是，CLP的病因基础以及相关中枢神经系统异常的原因和程度仍然很少了解。通过基础研究最好解决这种知识差距。然而，两个主要局限性的研究进展受到了障碍：不足合适的动物模型，缺乏研究人员 专业知识以整合传统上不同的概念和实验方法。在该领域的一个重要进步是我们最近的证明是，在子宫内暴露于刺猬（HH）信号拮抗剂环原上，诱导小鼠中的CLP，模仿临床表型。在本文中提出的研究采用该模型，使用高分辨率的磁共振成像来定义CNS畸形学，以与CLP共同相处，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（AIM 1）。机械测定将检验以下假设，即环保胺暴露通过破坏发育中的前脑和中脑之间的电感信号来诱导相关的脑面异常（AIM 2）。将确定和表征介导这些畸形的初始发病机理的HH途径靶基因（AIM 3），提供一组候选基因，在未来的研究中将追求其在颅面发育和病因学角色中的功能的临床作用。累积地，预计这些研究将定义CLP患者中存在的特定中枢神经系统异常，并提供基本的病理学特征，这将推动未来的努力，以检查临床种群中CLP创造的HH信号传导途径的化学和遗传扰动与HH信号通路的相互作用。为了增强候选人在毒理学和细胞信号传导方面的先前经验，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动手训练成分相结合。该提案中概述的机会将为候选人提供多学科的概念背景和实验工具集，非常适合阐明CLP复杂的病理学。 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 （Arthur Aylsworth博士）。最终，这些努力的承诺和完成将为候选人的目标奠定基础，即在临床和基础科学家促进口腔和颅面健康的环境中追求学术生涯。 公共卫生相关性：该项目中的实验将使用MRI和机械分析来检查唇裂和口感小鼠模型中面部和脑异常的原因，程度和共同发生。这项工作的目的是增加我们对CLP患者以及导致这些常见和病态先天缺陷的遗传和环境因素的结构性脑异常的理解。