Foxp-regulated signaling pathways in brain development - Diversity

雷切尔·沃尔默的研究与训练 A.基金赠款摘要 母基金（R 01 MH 126481;资助期06/01/21-05/31/2026）的标题是：“Foxp调节的信号传导 大脑发育的途径。”转录因子FOXP家族的两个成员FOXP 1和FOXP 2， 与单基因形式的智力残疾、自闭症谱系障碍和特殊语言有关 语言缺陷。FOXP 1或FOXP 2的变异体是最重要的基因之一， 自闭症谱系障碍我们之前已经证明Foxp 1和Foxp 2都对细胞凋亡有显著的贡献。 皮质和纹状体发育。我们通过对基因表达的研究将这些发育变化联系起来， 电生理学和行为学。我们进一步确定了基因表达的非细胞自主变化， 最新的单细胞RNA测序技术。基于这些数据，驱动 在父母补助金和拟议的补充工作是，Foxp 1和Foxp 2是关键的协调， 转录信号级联以细胞类型特异性的方式对神经元功能很重要， 神经发育障碍的风险，如自闭症。我们建议识别这些细胞类型特异性 通过使用啮齿动物模型，通过两个具体的目标，将利用snATAC- seq：1）确定在发育中由Foxp 1编排的细胞类型特异性基因调控程序， 2）确定Foxp 2在发育中的细胞类型特异性基因调控程序。 皮层总之，这些目标将描绘Foxp 1和Foxp 2对皮质神经元的细胞类型贡献。 发展啮齿类动物模型和细胞类型特异性基因组数据集将提供对基本的 控制正常哺乳动物大脑发育的分子机制。这些研究目标支持 NIMH的使命是了解和开发精神障碍的治疗方法。最后，培训计划， 将为Vollmer女士提供神经基因组学方面的尖端技能。 目的1和2的父母补助金是确定细胞类型特异性发育转录程序 使用单核RNA测序（snRNA-seq）在小鼠脑中由Foxp 1或Foxp 2调节， 时间点。这些实验将利用先前产生的Foxp 1和Foxp 2 cKO小鼠， 并使用Emx1.Cre发表（例如Araujo等人，Journal of Neuroscience 2017; Mr. et al.，基因& Development 2017; Co等人，大脑皮层2020）。我们的实验室最近也采用了单一的方法- 细胞核ATAC测序，以揭示基因调控和信号传导的细胞类型特异性贡献 级联（例如Berto等人，Nature Neuroscience 2021）。将这种染色质可及性方法与 在细胞类型水平的基因表达，应该提供一个新的水平的分辨率，了解如何这些 转录因子组织发育信号级联。如下所述，Rachael Vollmer将 通过使用与snRNA-seq相同的Foxp 1和Foxp 2 cKO小鼠来推进父母资助的目标; 然而，她将在相同的时间点应用snATAC-seq来理解染色质可及性。这个牢房 特异性的方法将使我们能够缩小Foxp 1和Foxp 2的直接转录靶点， 在这些模型系统中观察到的表型，如受损的皮质迁移和行为可识别性。

RESEARCH AND TRAINING PLAN FOR RACHAEL VOLLMER A. SUMMARY OF THE FUNDED GRANT The parent grant (R01MH126481; funding period 06/01/21-05/31/2026) is titled: “Foxp-regulated signaling pathways in brain development.” Two members of the FOXP family of transcription factors, FOXP1 and FOXP2, have been linked to monogenetic forms of intellectual disability, autism spectrum disorders, and specific speech and language deficits. Variants in FOXP1 or FOXP2 are among the most significant genes associated with autism spectrum disorders. We previously showed that Foxp1 and Foxp2 both have significant contributions to cortical and striatal development. We linked these developmental changes via studies of gene expression, electrophysiology, and behaviors. We further identified non-cell-autonomous changes in gene expression using newly available single-cell RNA-sequencing technology. Based on these data, the central hypothesis driving the work in the parent grant and the proposed supplement is that Foxp1 and Foxp2 are key orchestrators of transcriptional signaling cascades in a cell type-specific manner that are important for neuronal function and are at risk in neurodevelopmental disorders such as autism. We propose to identify these cell type-specific contributions in the developing cortex by using rodent models through two specific aims that will utilize snATAC- seq: 1) Determine the cell type-specific gene regulatory programs orchestrated by Foxp1 in the developing cortex; and 2) Determine the cell type-specific gene regulatory programs orchestrated by Foxp2 in the developing cortex. Together, these aims will delineate the cell type contribution of both Foxp1 and Foxp2 to cortical development. The rodent models and cell-type specific genomic datasets will provide insight into the basic molecular mechanisms governing normal mammalian brain development. These research aims support the mission of the NIMH to understand and develop treatments for mental disorders. Finally, the training plan that we have described in this proposal will provide Ms. Vollmer with a cutting-edge skill set in neurogenomics. Aims 1 and 2 of the parent grant are to identify the cell-type specific developmental transcriptional program regulated by Foxp1 or Foxp2 in the mouse brain using single-nuclei RNA-sequencing (snRNA-seq) at several time points. These experiments will utilize previously generated Foxp1 and Foxp2 cKO mice already generated and published using Emx1.Cre (e.g. Araujo et al., Journal of Neuroscience 2017; Usui et al., Genes & Development 2017; Co et al., Cerebral Cortex 2020). Our lab has also recently applied the approach of single- nuclei ATAC-sequencing to uncover the cell type specific contributions of gene regulation and signaling cascades (e.g. Berto et al., Nature Neuroscience 2021). Coupling this chromatin accessibility approach with gene expression at the cell type level, should provide a new level of resolution for understanding how these transcription factors organize developmental signaling cascades. As described below, Rachael Vollmer will advance the aims of the parent grant by using the same Foxp1 and Foxp2 cKO mice proposed for snRNA-seq; however, she will apply snATAC-seq at the same timepoints to understand chromatin accessibility. This cell- specific approach will allow us to narrow down the direct transcriptional targets of Foxp1 and Foxp2 driving the observed phenotypes in these model systems such as impaired cortical migration and behavioral inflexibility.