Investigating sex differences in DDX3X mouse models

研究 DDX3X 小鼠模型的性别差异

基本信息

批准号：
10782849
负责人：
Stephen Nicholas Floor
金额：
$ 6.05万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2025-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10782849
关键词：
Brain Cell Cycle Cell Differentiation process Cell physiology Cells Collaborations Communities Compensation Complement Critical Thinking Data Development Developmental Delay Disorders Diagnostic Embryo Embryology Epilepsy Etiology Family Female Foundations Funding Future Goals Grant Heterozygote Histology Impairment Intellectual functioning disability Knock-out Link Medical Mentors Mentorship Messenger RNA Microcephaly Microscopy Missense Mutation Molecular Mus Mutation Nature Neurodevelopmental Disorder Neurosciences Nonsense Mutation Pathology Phenocopy Phenotype Physicians Positioning Attribute Productivity Proteins RNA Helicase RNA-Binding Proteins Research Research Personnel Role Scientist Sex Differences Syndrome Technical Expertise Techniques Technology Testing Therapeutic Therapeutic Intervention Training Translational Regulation Translations Writing autism spectrum disorder brain malformation career career development cell fate specification de novo mutation design developmental neurobiology differential expression experimental study graduate school impaired brain development insight knock-down loss of function male mouse genetics mouse model nerve stem cell nervous system disorder neural neurogenesis paralogous gene post-doctoral training sex sexual dimorphism single cell analysis single-cell RNA sequencing skills stem cell proliferation stem cells transcriptome transcriptomics virtual

项目摘要

Abstract De novo mutations in the RNA binding protein DDX3X result in DDX3X syndrome. DDX3X mutations are also associated with autism spectrum disorder (ASD) and may account for 1-3% of unexplained developmental delay (DD) in females. Yet, our understanding of DDX3X function in the developing brain is limited. Our funded grant aims to understand the mechanisms by which DDX3X loss of function and missense mutations perturb cellular function. This supplement will focus on sexually dimorphic phenotypes linked to DDX3X. Most de novo mutations are in DDX3X females though increasing numbers of cases in males have been observed. We have discovered that complete loss of Ddx3x in females causes striking microcephaly. In contrast, complete knockout in males phenocopies haploinsufficiency in females. Our data suggest that these differences between males and females may be due to altered expression of the paralog, Ddx3y. In this supplement, we will test the hypothesis that phenotypic differences in males and females associated with Ddx3x loss are due to compensation by the paralog, Ddx3y. To test this, we will use single cell transcriptomics to quantify cell composition and transcriptome changes associated with Ddx3x loss in males and female embryonic brains. Additionally, we will use knockdown and rescue experiments to test the functional redundancy of Ddx3y and Ddx3x for embryonic brain development. These experiments will give valuable new insights to understand roles for Ddx3x in neurogenesis at a cellular and molecular level. Importantly, this proposal is designed to give fundamental experimental training in mouse genetics, embryology, sequencing, histology and microscopy. This supplement also includes a comprehensive mentoring plan for career development.

抽象的 RNA 结合蛋白 DDX3X 的从头突变会导致 DDX3X 综合征。 DDX3X 突变也与自闭症谱系障碍 (ASD) 相关，可能占 1-3% 女性不明原因的发育迟缓（DD）。然而，我们对 DDX3X 功能的理解大脑的发育是有限的。我们资助的赠款旨在了解 DDX3X 功能丧失和错义突变会扰乱细胞功能。本补充将重点关注与 DDX3X 相关的性二态表型。大多数从头突变发生在 DDX3X 中尽管观察到男性病例数量不断增加，但女性病例较多。我们发现女性 Ddx3x 的完全缺失会导致显着的小头畸形。相比之下，完整雄性表型敲除，雌性单倍体不足。我们的数据表明这些男性和女性之间的差异可能是由于旁系同源物 Ddx3y 的表达改变所致。在本补充材料中，我们将检验以下假设：男性和女性的表型差异与 Ddx3x 损失相关的原因是旁系同源 Ddx3y 的补偿。为了测试这一点，我们将使用单细胞转录组学来量化细胞组成和相关转录组变化男性和女性胚胎大脑中 Ddx3x 缺失。此外，我们将使用击倒和拯救实验测试 Ddx3y 和 Ddx3x 对胚胎大脑的功能冗余发展。这些实验将为理解 Ddx3x 在以下领域中的作用提供有价值的新见解：细胞和分子水平上的神经发生。重要的是，该提案旨在提供小鼠遗传学、胚胎学、测序、组织学和显微镜。该补充材料还包括全面的职业指导计划发展。