Discovery Pipeline for Genetic Defects in Hypothalamic-pituitary Development Using International Mouse Phenotyping Consortium Mice

利用国际小鼠表型联盟小鼠发现下丘脑-垂体发育遗传缺陷的管道

• 批准号: 10656660
• 负责人: Sally A. Camper
• 金额: $ 69.29万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-23 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656660
• 关键词: Abnormal Cell; Affect; Antibodies; Bioinformatics; Biological Assay; Birth; Brain; Candidate Disease Gene; Caring; Categories; Cell Culture Techniques; Cells; Central Nervous System; Cessation of life; Cilia; Complex; Congenital Abnormality; Cre driver; Cryopreservation; Data; Databases; Defect; Development; Disease; Dysmorphology; Embryo; Epigenetic Process; Equilibrium; Etiology; Face; Fertility; Functional disorder; Future; Gene Expression; Gene Expression Profiling; Genes; Genetic Diseases; Genetic Variation; Genotype; Goals; Gonadal structure; Growth; Histologic; Histological Techniques; Histology; Holoprosencephaly; Homeostasis; Human; Human Genetics; Hypopituitarism; Hypothalamic structure; In Situ Hybridization; Intellectual functioning disability; International; Investments; Knockout Mice; Knowledge; Laboratories; Lethal Genes; Mammals; Meta-Analysis; Metabolism; Methods; Molecular Diagnosis; Morbidity - disease rate; Morphology; Mouse Strains; Mus; Mutation; Neurons; Organogenesis; Partner in relationship; Pathogenesis; Pathway interactions; Patients; Penetrance; Persons; Phenotype; Pituitary Gland; Pituitary Hormones; Post-Translational Protein Processing; Pregnancy; Proliferating; Quantitative Reverse Transcriptase PCR; Reagent; Reporting; Research Personnel; Resources; Risk; Septo-Optic Dysplasia; Site; Specific qualifier value; Stains; Structural defect; Structure; Syndrome; Testing; Time; Tissues; United States National Institutes of Health; Vascularization; WNT Signaling Pathway; Water; Wild Type Mouse; Work; biological adaptation to stress; causal variant; data dissemination; gene discovery; gene function; hormone deficiency; human disease; insight; malformation; man; molecular phenotype; mortality; mutant; novel; pituitary gland development; single-cell RNA sequencing; transcription factor; treatment planning; web site

Abstract Congenital hypopituitarism (CH) is a common birth defect frequently associated with syndromic abnormalities in the central nervous system, ocular structures, face, and gonads. The most severe disorders have midline developmental anomalies and include holoprosencephaly, which is usually embryonic lethal, or septo-optic dysplasia. Less severe birth defects disrupt only hypothalamic or pituitary development, causing hormone deficiencies that affect viability, growth, fertility, metabolism, and the stress response, and may require life-long care. Over 60 genes are known to cause CH, many of which were first discovered in mice. Nonetheless, 81% of CH patients still lack a molecular diagnosis, which would be invaluable for planning treatment and predicting future risk. We propose detailed phenotyping of existing embryonic lethal knockout mice with known genetic defects that result in hypothalamic and/or pituitary malformations. This will expand the molecular diagnoses for CH and associated midline deficiencies in humans and increase our understanding of organogenesis of these critical tissues. We identified a prioritized set of 18 embryonic lethal or sub-viable mouse lines with obvious malformations in the hypothalamus and/or pituitary gland. This set of genes are grouped based on function including epigenetic regulators, components of cilia, protein modification, and other novel functional categories. We assembled a team of investigators with expertise in hypothalamic-pituitary development, mouse phenotyping, and bioinformatics. We will conduct deep and thorough phenotyping of the hypothalamus and pituitary gland in the selected IMPC mice. Our pipeline has three steps with appropriate re-prioritization at each stage. First, temporal and spatial expression of the selected genes will be determined in wild type mice, the IMPC strains will be imported, and the dysmorphology will be characterized histologically from mid- gestation to birth. Second, the strains will be assessed for defects in vascularization and specification of hypothalamic neurons and pituitary hormone producing cells using a combination of scRNA-seq and histological methods. Third, a mechanistic understanding of the pathophysiology of the developmental defects will be determined using histology, cell culture assays and/or established tissue-specific cre drivers to test hypotheses developed from scRNA-seq and meta-analyses. This work will establish genotype/phenotype relationships for novel genes that are candidates for the undiagnosed CH patients. Data dissemination will be timely and open, building on our existing mouse pituitary analysis database (mousePAD) and blog, and the IMPC website. Future expansion of our pipeline will add additional lethal or sub-viable IMPC strains with mutations in genes implicated in hypothalamic-pituitary development, but not yet studied. We are poised to make a significant impact on the discovery of genetic defects that affect hypothalamic-pituitary development in mouse and man.

抽象的 先天性下降症（CH）是经常与综合症有关的常见出生缺陷 中枢神经系统，眼部结构，面部和性腺的异常。最严重的疾病 具有中线发育异常，包括通常是胚胎致死或 九月发育不良。不太严重的先天缺陷仅破坏下丘脑或垂体发育，导致 影响生存能力，生长，生育能力，代谢和压力反应的激素缺乏，可能 需要终身照顾。已知超过60个基因引起CH，其中许多是在小鼠中首次发现的。 尽管如此，有81％的CH患者仍然缺乏分子诊断，这对于计划是无价的 治疗并预测未来的风险。我们提出了现有胚胎致命敲除的详细表型 患有已知遗传缺陷的小鼠导致下丘脑和/或垂体畸形。这将扩大 CH和相关的中线缺陷的分子诊断，并增加了我们的 了解这些关键组织的器官发生。我们确定了一组优先的18个胚胎 下丘脑和/或垂体中具有明显畸形的致命或亚下小鼠系。这 基因集基于功能进行分组，包括表观遗传调节剂，纤毛的成分，蛋白质 修改和其他新型功能类别。我们组建了一个具有专业知识的调查员团队 下丘脑 - 垂体发育，小鼠表型和生物信息学。我们将深入 在选定的IMPC小鼠中的下丘脑和垂体的彻底表型。我们的管道有 在每个阶段都有适当重新定位的三个步骤。首先，时间和空间表达 选定的基因将在野生型小鼠中确定，IMPC菌株将进口，并将 从妊娠到出生的组织学表征畸形学。其次，压力将是 评估了下丘脑神经元和垂体激素的血管形成和规格的缺陷 使用SCRNA-SEQ和组织学方法的组合产生细胞。第三，一种机械 了解发育缺陷的病理生理学将使用组织学，细胞确定 培养测定法和/或已建立的组织特异性CRE驱动因素，以测试从SCRNA-SEQ发展的假设 和荟萃分析。这项工作将建立新基因的基因型/表型关系 未诊断的CH患者的候选人。数据传播将及时开放，以我们的为基础 现有的鼠标垂体分析数据库（Mousepad）和博客以及IMPC网站。未来的扩展 我们的管道中将增加其他致命或可追溯的IMPC菌株，并在与之相关的基因中发生突变 下丘脑 - 垂体发展，但尚未研究。我们准备对 发现影响小鼠和人类下丘脑垂体发育的遗传缺陷。