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: 8828670
• 负责人: Robert Lipinski
• 金额: $ 24.14万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8828670
• 关键词: Address; Animal Model; 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 Profiling; 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; Medical Genetics; 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; animal imaging; base; career; chemical genetics; cleft lip and palate; clinical phenotype; craniofacial; craniofacial development; cyclopamine; design; differential expression; experience; indexing; malformation; mouse model; multidisciplinary; neurodevelopment; novel; pregnant; progenitor; public health relevance; relating to nervous system; research study; shape analysis; smoothened signaling pathway; spatiotemporal

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 studies 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.

唇腭裂（CLP）是最常见的颅面部出生缺陷，可导致严重的出生缺陷。 发病率高，需要广泛的医疗干预。除了面部畸形和继发性 CLP 的并发症、中枢神经系统 (CNS) 异常以及认知和行为缺陷 已知同时发生。重要的是，CLP 的病因学基础，以及相关 CNS 的病因和程度 异常现象仍知之甚少。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，以及缺乏具有专业知识的研究人员来整合传统上不同的概念和 实验方法。我们最近证明，在子宫内 暴露于 Hedgehog (Hh) 信号拮抗剂环杷明可在小鼠中诱导 CLP，模拟 临床表型。采用该模型，本文提出的研究使用高分辨率磁 共振成像来定义与 CLP 共同发生的 CNS 畸形并确定是否是独特的 这些异常的模式可以通过特定的面部表型来预测（目标 1）。机械分析将 检验环杷明暴露通过扰乱诱发相关脑面部异常的假设 发育中的前脑和中面部之间的感应信号（目标 2）。 Hh 途径的靶基因 将鉴定和表征介导这些畸形的初始发病机制（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 的病因学作用将是 在未来的学习中继续追求。总的来说，预计这些研究将定义特定的中枢神经系统异常 存在于 CLP 患者中并提供基本的病因病理学特征，这将推动 未来努力检查化学和遗传扰动与 Hh 信号通路的相互作用 临床人群中 CLP 的起源。增强候选人之前在毒理学方面的经验 细胞信号传导，这些研究目标与胚胎学的教学和实践培训相结合， 神经发育、临床和分子遗传学。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。这些事业的顺利完成将 得益于北卡罗来纳大学教堂山分校提供的卓越研究环境， 颅面/神经发育专家（Kathleen Sulik 博士）的指导和合作， 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（Arthur Aylsworth 博士）。最终，承担并完成 这些努力将为候选人在特定环境中追求学术生涯的目标奠定基础 它汇集了临床和基础科学家来促进口腔和颅面健康。

项目成果

A Simple and Reliable Method for Early Pregnancy Detection in Inbred Mice.

一种简单可靠的近交小鼠早期妊娠检测方法。

• 发表时间: 2015
• 期刊: Journal of the American Association for Laboratory Animal Science : JAALAS
• 作者: Heyne,GalenW;Plisch,ErinH;Melberg,CalG;Sandgren,EricP;Peter,JodyA;Lipinski,RobertJ
• 通讯作者: Lipinski,RobertJ

Definition of critical periods for Hedgehog pathway antagonist-induced holoprosencephaly, cleft lip, and cleft palate.

• DOI: 10.1371/journal.pone.0120517
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Heyne GW;Melberg CG;Doroodchi P;Parins KF;Kietzman HW;Everson JL;Ansen-Wilson LJ;Lipinski RJ
• 通讯作者: Lipinski RJ