Characterizing the mechanistic basis for sex-dimorphic responses to retinoic acid signaling in the developing brain

表征发育中大脑对视黄酸信号的性别二态性反应的机制基础

基本信息

批准号：
10607935
负责人：
Kelsey Hennick
金额：
$ 4.36万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-30 至 2025-06-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10607935
关键词：
Address Anatomy Anterior Basic Science Biological Assay Biomedical Research Brain CRISPR interference Central Nervous System Data Data Set Development Disease Disparate Enhancers Enzymes Epigenetic Process Etiology Exhibits Exposure to FOXP1 gene Female Fragile X Syndrome Gene Dosage Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genomics Glutamates Gonadal Steroid Hormones Histones Human Immunohistochemistry In Vitro Incidence KDM5B gene Lead Link Lysine Mediating Methylation Modeling Molecular Mutation Neurodevelopmental Disorder Neurons Organoids Pattern Penetrance Phenotype Prefrontal Cortex RORA gene Regulator Genes Retinoic Acid Receptor Rett Syndrome Sampling Sex Bias Sex Chromosomes Sex Differences Signal Pathway Signal Transduction Site Structure Testing Therapeutic Intervention Tissues Transcription Initiation Transcriptional Regulation Tretinoin Up-Regulation Vitamin A Work X Chromosome X Inactivation autism spectrum disorder cell type demethylation differential expression dosage experimental study genome-wide gray matter histone modification human female insight knock-down male morphogens nervous system development nervous system disorder neurodevelopment programs receptor function response retinoic acid receptor alpha risk variant sex sexual dimorphism single-cell RNA sequencing stem cell differentiation stem cells targeted treatment therapeutic candidate transcription factor

项目摘要

Project Summary/Abstract Sex hormones and sex chromosome genes instruct development of sex-specific features during neurodevelopment, including differences in somatic gene expression and gray matter volume. Together, these developmental programs lead to fundamental differences at the molecular, cellular, and tissue level between males and females. While some neurological disorders have clear links to X chromosome genes, such as Rett Syndrome and Fragile X Syndrome, other disorders including Autism show sex-biased penetrance with no clear genetic mechanism. This proposal aims to address the mechanistic basis of sex-dimorphic transcriptional responses to retinoic acid signaling in the developing brain. Retinoic acid signaling is indispensable for modulating gene regulatory programs that orchestrate proper nervous system development, and recent work has shown that the Autism risk gene FOXP1 is upregulated in response to retinoic acid signaling. Preliminary studies have shown specific upregulation of FOXP1 in response to retinoic acid signaling in female cortical organoids, and not in males. This project will similarly leverage stem cell-derived cortical organoids to model neurodevelopment in vitro to better understand the molecular basis of sex-dimorphic phenotypes upon exposure to retinoic acid. Aim 1 will characterize sex-dimorphic genome wide expression changes in response to retinoic acid. Male and female organoids will be treated with vitamin A, the precursor to retinoic acid, and used for scRNA-seq. This dataset will uncover genes upregulated specifically in female organoids in response to retinoic acid. Additionally, I will identify cell types in both male and female organoids that exhibit the greatest gene expression changes in response to retinoic acid, lending insight into cell type-specific sensitives to retinoic acid during neurodevelopment. In Aim 2, the relationship between dosage of X-linked lysine demethylase KDM5C and retinoic acid-induced sex-specific gene regulatory programs will be characterized. Knockdown of KDM5C in female stem cell-derived organoids and subsequent genome-wide changes in H3K4 methylation, which is demethylated by KDM5C, will be determined by CUT&Tag. A putative enhancer at the FOXP1 locus coincides with H3K4me3 marks, and KDM5C-dependent methylation at this genomic site will be of particular importance. The impact of KDM5C knockdown on retinoic acid-induced FOXP1 expression will then be interrogated by immunohistochemistry. Together, these experiments will further our understanding of the intersection of epigenetics, gene expression, and cell signaling pathways during neurodevelopment, providing an an important mechanistic basis for sex-dimorphic developmental programs. This will further our understanding of the etiology of neurodevelopmental disorders with sex-biased penetrance, while uncovering potential candidates for therapeutic interventions.

项目概要/摘要性激素和性染色体基因指导性别特异性特征的发育神经发育，包括体细胞基因表达和灰质体积的差异。在一起，这些发育程序导致分子、细胞和组织水平上的根本差异男性和女性。虽然一些神经系统疾病与 X 染色体基因有明显的联系，例如 Rett 综合症和脆性 X 综合症以及自闭症等其他疾病的外显率存在性别偏见，但没有明确的遗传机制。该提案旨在解决性别二态性转录的机制基础发育中的大脑对视黄酸信号的反应。视黄酸信号传导对于调节基因调控程序，协调适当的神经系统发育，以及最近的工作研究表明，自闭症风险基因 FOXP1 响应视黄酸信号而上调。初步的研究表明，女性皮质中 FOXP1 响应视黄酸信号而特异性上调类器官，而不是男性。该项目将类似地利用干细胞衍生的皮质类器官来建模体外神经发育，以更好地了解性别二态性表型的分子基础接触视黄酸。目标 1 将表征响应中性别二态性全基因组表达变化至视黄酸。男性和女性类器官将使用维生素 A（视黄酸的前体）进行处理，并且用于 scRNA 测序。该数据集将揭示女性类器官中特异性上调的基因至视黄酸。此外，我将确定男性和女性类器官中表现出最大的细胞类型基因表达因视黄酸而变化，有助于深入了解细胞类型特异性的敏感性神经发育过程中的视黄酸。在目标2中，X-连锁赖氨酸的剂量之间的关系去甲基化酶 KDM5C 和视黄酸诱导的性别特异性基因调控程序将得到表征。女性干细胞衍生类器官中 KDM5C 的敲低以及随后 H3K4 的全基因组变化由 KDM5C 去甲基化的甲基化将通过 CUT&Tag 测定。假定的增强剂 FOXP1 基因座与 H3K4me3 标记重合，并且该基因组位点处的 KDM5C 依赖性甲基化将被特别重要。 KDM5C 敲低对视黄酸诱导的 FOXP1 表达的影响将然后通过免疫组织化学进行询问。总之，这些实验将进一步加深我们对神经发育过程中表观遗传学、基因表达和细胞信号通路的交叉，为性别二态性发育计划提供重要的机制基础。这将进一步推动我们了解具有性别偏见外显率的神经发育障碍的病因，同时揭示治疗干预的潜在候选者。