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 先天性垂体功能减退症 (CH) 是一种常见的出生缺陷，通常与综合征相关 中枢神经系统、眼部结构、面部和性腺异常。最严重的疾病 有中线发育异常，包括前脑无裂畸形，这通常是胚胎致死的，或 视隔发育不良。不太严重的出生缺陷只会破坏下丘脑或垂体的发育，导致 激素缺乏会影响生存能力、生长、生育能力、新陈代谢和应激反应，并可能 需要终身照顾。已知超过 60 个基因可导致 CH，其中许多基因首次在小鼠中发现。 尽管如此，81% 的 CH 患者仍然缺乏分子诊断，这对于制定计划非常有价值 治疗和预测未来风险。我们建议对现有胚胎致死性敲除进行详细的表型分析 具有导致下丘脑和/或垂体畸形的已知遗传缺陷的小鼠。这将扩大 人类 CH 和相关中线缺陷的分子诊断，并提高我们的 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/或垂体有明显畸形的致死或亚存活小鼠系。这 一组基因根据功能进行分组，包括表观遗传调节因子、纤毛成分、蛋白质 修饰等新颖的功能类别。我们组建了一支具有以下专业知识的调查人员团队 下丘脑-垂体发育、小鼠表型和生物信息学。我们将深入开展 对选定的 IMPC 小鼠的下丘脑和垂体进行彻底的表型分析。我们的管道有 三个步骤，每个阶段都适当重新确定优先级。首先，时间和空间的表达 将在野生型小鼠中确定选定的基因，导入 IMPC 品系，并 畸形将从妊娠中期到出生进行组织学表征。其次，应变将是 评估下丘脑神经元和垂体激素的血管化和规格缺陷 结合 scRNA-seq 和组织学方法生产细胞。三、机械化 对发育缺陷病理生理学的理解将通过组织学、细胞学来确定 培养测定和/或建立的组织特异性 cre 驱动程序来测试 scRNA-seq 提出的假设 和荟萃分析。这项工作将为新基因建立基因型/表型关系，这些基因是 未确诊 CH 患者的候选者。数据传播将基于我们的及时和开放 现有的小鼠垂体分析数据库（mousePAD）和博客，以及IMPC网站。未来扩展 我们的管道将添加额外的致死或亚存活 IMPC 菌株，其基因突变涉及 下丘脑-垂体发育，但尚未研究。我们准备对世界产生重大影响 发现影响小鼠和人类下丘脑-垂体发育的遗传缺陷。