Combinatorial function of Foxp1/2/4 in Purkinje cell diversification and cerebellar development

Foxp1/2/4在浦肯野细胞多样化和小脑发育中的组合功能

• 批准号: 10604350
• 负责人: JAMES Y.H LI
• 金额: $ 55.62万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10604350
• 关键词: ATAC-seq; Architecture; Ataxia; Behavioral; Bilateral; Binding; Biological Assay; Cell Differentiation process; Cells; Cerebellum; Characteristics; Chromatin; Code; Cognition Disorders; Cues; DNA Binding; Data; Defect; Development; Disease; Embryo; Excision; FOXP1 gene; FOXP2 gene; Family; Functional disorder; Genes; Genetic Transcription; Genome; Goals; Heterogeneity; Histology; Human; Image; In Vitro; Individual; Intellectual functioning disability; Knowledge; Link; Mental disorders; Molecular; Morphogenesis; Movement; Mus; Mutation; Neurodevelopmental Disorder; Neurons; Pathogenesis; Pathology; Phenotype; Proteins; Psyche structure; Purkinje Cells; RNA; Research; Role; Schizophrenia; Specific qualifier value; Speech Disorders; Subgroup; Testing; Ultrasonics; autism spectrum disorder; behavioral study; cognitive function; combinatorial; dimer; insight; language impairment; multiple omics; neural circuit; neurodevelopment; prevent; programs; prospective; single-cell RNA sequencing; transcription factor; transcriptomics; vocalization

SUMMARY Abnormalities in cerebellar development, especially pathology and dysfunction of Purkinje cells, have been implicated in a wide variety of neurodevelopmental diseases, including ataxia, autism spectrum disorder, schizophrenia, and language impairment. Being one of the earliest-born cerebellar cell groups, Purkinje cells are believed instrumental in the development, function, and pathogenesis of the cerebellum. Evidence suggests the existence of Purkinje cell subtypes with distinct molecular features. However, the molecular mechanisms underlying the diversification of Purkinje cells remain poorly understood. Consequently, we lack an entry to assess the role of individual Purkinje cell subtypes. Through single-cell RNA and chromatin accessibility analyses, we uncovered at least nine molecularly distinct subtypes of Purkinje cells in the developing mouse cerebellum. These Purkinje cell subtypes contribute to different compartments in the developing cerebellum. Remarkably, the Purkinje cell subtypes display a characteristic combinatorial expression of Foxp1, Foxp2, and Foxp4, which belong to a subgroup of the forkhead-box transcription factor family. Mutations of human FOXP1 or FOXP2 are linked to speech disorders, autism spectrum disorder, and intellectual disability, indicating that these proteins coordinate the development of the neural circuits related to cognitive diseases. In vitro evidence shows that FoxP proteins form dimers or oligomers with variable transcriptional targets and actives depending on the binding partner. We hypothesize that Foxp1/2/4 form combinatorial “FoxP codes” to specify distinct Purkinje cell subtypes, which in turn control the morphogenesis of the cerebellum. Aim 1 will combine conventional expression analysis, spatial transcriptomics, and volume imaging to determine the development of PC subtypes in relation to the morphogenesis of the cerebellum. Aim 2 will delete Foxp1/2/4, individually and in combinations, from the mouse cerebellum. We will use histology, single-cell RNA-seq, and behavioral studies to evaluate the impacts of single and compound Foxp1/2/4 mutations on cerebellar development and behavioral function. Aim 3 will use a multi- omic approach to study the molecular mechanism by which combinatorial FoxP genes regulate the transcription program for Purkinje cell differentiation. At the completion of this project, we expect to have identified the individual and combinatorial roles of Foxp1/2/4 in cerebellar development. This study will have a significant positive impact, not only on the basic knowledge of cerebellar development but also on the understanding of the molecular basis of the vast number of unexplored cerebellum-related diseases.

摘要 小脑发育异常，特别是浦肯野细胞的病理和功能障碍 与多种神经发育疾病有关，包括共济失调、自闭症 精神障碍、精神分裂症和语言障碍。作为最早出生的小脑细胞群之一， 浦肯野细胞被认为在小脑的发育、功能和发病机制中起着重要作用。 有证据表明存在具有不同分子特征的浦肯野细胞亚型。然而， 浦肯野细胞多样化的分子机制仍然知之甚少。 因此，我们缺乏一个条目来评估单个浦肯野细胞亚型的作用。通过单电池 RNA和染色质可及性分析，我们发现至少九种分子上不同的亚型 发育中的小鼠小脑中的浦肯野细胞。这些浦肯野细胞亚型对不同的 发育中的小脑中的脑室。值得注意的是，浦肯野细胞亚型表现为 Foxp1、Foxp2和Foxp4的特征组合表达，它们属于 叉头盒转录因子家族。人类Foxp1或FOXP2基因突变与言语有关 障碍、自闭症谱系障碍和智力残疾，表明这些蛋白质协调 与认知疾病相关的神经回路的发展。体外证据表明，FoxP 蛋白质形成二聚体或寡聚体，具有可变的转录靶点和活性，具体取决于 有约束力的合作伙伴。我们假设Foxp1/2/4形成组合“FoxP码”以指定不同的 浦肯野细胞亚型，进而控制小脑的形态发生。目标1将结合 常规的表达分析、空间转录和体积成像来确定 PC亚型发育与小脑形态发生的关系目标2将 将Foxp1/2/4单独或组合从小鼠小脑中删除。我们将使用组织学， 单细胞RNA-seq和行为研究，以评估单细胞和 Foxp1/2/4复合突变对小脑发育和行为功能的影响AIM 3将使用多个 用组学方法研究组合FoxP基因调控的分子机制 浦肯野细胞分化的转录程序。在这个项目完成后，我们预计将有 确定了Foxp1/2/4在小脑发育中的个体和组合作用。这项研究将 不仅对小脑发育的基本知识有重大的积极影响，而且对 对大量未探索的小脑相关疾病的分子基础的了解。