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(申请人提供)：唇腭裂(CLP)是最常见的头面部出生缺陷，会导致严重的发病率，需要广泛的医疗干预。除了面部畸形和CLP的继发性并发症外，中枢神经系统(CNS)异常以及认知和行为障碍也是已知的共同发生。重要的是，CLP的病因学基础以及相关的中枢神经系统异常的原因和程度仍然知之甚少。这一知识鸿沟最好通过基础研究来解决。然而，重大的研究进展受到两大限制的阻碍：缺乏合适的动物模型，以及缺乏研究人员。 整合传统上不同的概念和实验方法的专业知识。这一领域的一个重要进展是我们最近证明，在子宫内暴露于刺猬(Hedgehog，HH)信号拮抗剂环多巴胺，可以模拟临床表型，在小鼠中诱导CLP。利用这一模型，本文提出的研究使用高分辨率磁共振成像来定义与CLP共同出现的中枢神经系统畸形，并确定这些异常的独特模式是否可以通过特定的面部表型来预测(目标1)。机械分析将测试环多巴胺暴露通过破坏发育中的前脑和面部中部之间的感应信号而导致相关的脑-面部异常的假设(目标2)。HH通路的靶基因将被鉴定和鉴定(目标3)，提供一组候选基因，其在头面部发育中的功能和临床CLP的病因作用将在未来的研究中进行。总而言之，预计这些研究将确定CLP患者中存在的特定中枢神经系统异常，并提供基础的病因学特征，以推动未来研究化学和遗传扰动对临床人群CLP发生中HH信号通路的相互作用。为了增强候选人以前在毒理学和细胞信号方面的经验，这些研究目标与胚胎学、神经发育、临床和分子遗传学的教学和实践培训部分相结合。这项建议所概述的机会将为候选人提供一个多学科的概念背景和实验工具集，理想地用于阐明CLP的复杂病因。这些项目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(凯瑟琳·苏利克博士)、小动物成像(Al Johnson博士)、三维面部形状分析(Peter Hammond博士)、颅面遗传学(Eric Everett博士)和临床遗传学(Arthur Aylsworth博士)专家的指导和合作。最终，这些努力的开展和完成将为候选人在临床和基础科学家结合以促进口腔和颅面健康的环境中追求学术生涯的目标奠定基础。 公共卫生相关性：该项目中的实验将使用核磁共振成像和机械分析来检查唇腭裂小鼠模型中面部和大脑异常的原因、程度和共存情况。这项工作的目的是增加我们对CLP患者大脑结构异常的了解，以及导致这些常见和病态出生缺陷的遗传和环境因素。