The role of the protocadherin gene cluster in neurodevelopment and the implications for neurodevelopmental disorders

原钙粘蛋白基因簇在神经发育中的作用及其对神经发育障碍的影响

基本信息

批准号：
10808516
负责人：
Erin Flaherty
金额：
$ 13.17万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
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：
2023-09-05 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10808516
关键词：
Address Award Behavior Biological Biophysics Brain Cell Nucleus Cell membrane Cells Cellular Assay Chromatin Complex Cytoplasm Data Development Dimerization Disease Disease model Etiology Evaluation Event Faculty Feedback Foundations GTP-Binding Proteins Gene Cluster Gene Expression Genes Genetic Genetic Transcription Goals Human In Vitro Individual Institution Integral Membrane Protein Knowledge Laboratories Learning Link Lymphocyte Mammals Mediating Membrane Mentors Mentorship Microfilaments Modeling Molecular Movement Mus Needs Assessment Neurites Neurodevelopmental Disorder Neurons Nuclear Organoids Patients Phase Positioning Attribute Postdoctoral Fellow Process Protein Isoforms Proteins Research Role Schizophrenia Scientist Series Signal Pathway Signal Transduction Single Nucleotide Polymorphism Sister Structure Surface Synapses Synaptic Transmission Techniques Tertiary Protein Structure Testing Training Transcriptional Regulation Variant Vertebrates WNT Signaling Pathway Work autism spectrum disorder axon guidance gene function genetic approach in vivo in vivo Model induced pluripotent stem cell insight membrane biogenesis neural neural circuit neurodevelopment neuronal circuitry neuropsychiatric disorder notch protein novel proband protein function screening synaptogenesis

项目摘要

PROJECT SUMMARY During development, individual neurons extend highly branched neurites that innervate the surrounding territory with minimal overlap. In mammals, the clustered protocadherins (Pcdh) provide each neuron with a unique cell specific identity required for self-identification and self-avoidance in the brain. Individual PCDHa/b/g protein isoforms cis-dimerize, and engage in homophilic trans interactions to create a lattice-like structure at contacting membrane surfaces, providing the diversity required for self-recognition. Deletion of Pcdh genes in mice results in deficits in neuronal wiring and altered behaviors. Biophysical and structural data, cellular assays, and functional studies support a model in which perfect homophilic matching of PCDH isoforms leads to the assembly of an extended PCDH lattice structure on sister neurites, triggering intracellular signaling, and a series of yet to be identified steps leading to self-avoidance. Many neuronal transmembrane proteins execute neurodevelopmental processes through both nuclear and membrane associated signaling mechanisms; however, the mechanisms regulating PCDH mediated self-avoidance remain unknown. My recent work in the Maniatis lab has demonstrated that the PCDHa/g intracellular domain (ICDs) are required for interactions with protein partners at the membrane (e.g. WAVE complex and WNT signaling modulators) and in the nucleus (e.g. chromatin and transcriptional regulators). In the nucleus, the PCDH-ICD interacts with and may regulate the expression of genes involved in axon guidance, synapse formation and participate in an auto-regulatory feedback loop. During the K99 phase, I will leverage these preliminary findings to assess the need for each downstream signaling mechanism for proper self-avoidance in vitro (Aim 1) and in vivo (Aim 2). During the independent R00 phase, I will implement knowledge and techniques acquired in the mentored phase to investigate whether PCDH SNVs from ASD cases disrupt the identified downstream signaling mechanisms, proper self-avoidance and neural circuitry during development (Aim 3). Understanding the mechanisms of self-avoidance is a fundamental step toward revealing how neural circuits are formed during development in vertebrates and could have important implications for neurodevelopmental disorders. The work described in the mentored portion of this proposal will be conducted under the mentorship of Dr. Maniatis, who has a strong record of mentoring postdoctoral research scientists who have transitioned into their own laboratories. Ultimately, this award will allow me to accomplish three broad training goals including; (1) learning to address biological questions using in vivo models, (2) developing and extending my knowledge of molecular and genetic approaches to probe gene function, and (3) expanding on my background in iPSC based disease modeling by probing PCDH function in cortical organoids. Achieving these goals and training will provide me with the strong foundation required to pursue a faculty position at a research institution where I can integrate in vitro and in vivo approaches to investigate novel questions in the field of neurodevelopment.

项目概要在发育过程中，单个神经元会延伸出高度分支的神经突，支配周围的神经细胞。重叠最小的区域。在哺乳动物中，簇状原钙粘蛋白 (Pcdh) 为每个神经元提供大脑中自我识别和自我回避所需的独特细胞特异性身份。个人 PCDHa/b/g 蛋白质异构体顺式二聚化，并参与同亲反式相互作用，在接触膜表面，提供自我识别所需的多样性。删除 Pcdh 基因小鼠的神经元连接缺陷和行为改变。生物物理和结构数据，细胞分析和功能研究支持一个模型，其中 PCDH 亚型的完美同亲匹配导致在姐妹神经突上组装延伸的 PCDH 晶格结构，触发细胞内信号传导，以及一系列尚未确定的导致自我回避的步骤。许多神经元跨膜蛋白执行通过核和膜相关信号机制的神经发育过程；然而，调节 PCDH 介导的自我回避的机制仍然未知。我最近在 Maniatis 实验室的工作表明 PCDHa/g 胞内结构域 (ICD) 与膜上的蛋白质伙伴相互作用所需的（例如 WAVE 复合物和 WNT 信号传导）调节剂）和细胞核（例如染色质和转录调节剂）。在细胞核中，PCDH-ICD 与轴突引导、突触形成和相关基因相互作用并可能调节这些基因的表达参与自动调节反馈循环。在 K99 阶段，我将利用这些初步发现评估每种下游信号机制在体外（目标 1）和体内适当自我回避的需要体内（目标 2）。在独立R00阶段，我将运用在独立R00阶段获得的知识和技术指导阶段，调查 ASD 病例中的 PCDH SNV 是否会破坏已识别的下游发育过程中的信号传导机制、适当的自我回避和神经回路（目标 3）。理解自我回避机制是揭示神经回路如何形成的基本一步在脊椎动物的发育过程中，可能对神经发育障碍产生重要影响。本提案的指导部分中描述的工作将在博士的指导下进行。 Maniatis 在指导博士后研究科学家方面有着良好的记录，这些科学家已经转变为他们自己的实验室。最终，这个奖项将使我能够实现三个广泛的培训目标，包括： (1) 学习使用体内模型解决生物学问题，(2) 发展和扩展我的知识探索基因功能的分子和遗传学方法，以及 (3) 扩展我在 iPSC 方面的背景通过探测皮质类器官中的 PCDH 功能来建立基于疾病模型。实现这些目标和培训将为我提供在我所在的研究机构担任教职所需的坚实基础可以整合体外和体内方法来研究神经发育领域的新问题。