Molecular, Cellular and Behavioral Impact of the R203W PACS1 Syndrome Mutation

R203W PACS1 综合征突变的分子、细胞和行为影响

• 批准号: 10612914
• 负责人: ANGELA M. GRONENBORN
• 金额: $ 65.49万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612914
• 关键词: Acetylation; Affect; Animals; Antisense Oligonucleotides; Behavior; Behavioral; Binding; Biochemical; Biochemistry; Biophysics; Catalytic Domain; Cellular biology; Centrosome; Cilia; Client; Complex; Contacting Client; Coupled; Data; Deacetylase; Deacetylation; Dendrites; Development; Disease; Dynein ATPase; Electroencephalography; Electrophysiology (science); Endosomes; Enzyme Inhibitor Drugs; Enzymes; Epilepsy; Goals; Golgi Apparatus; HDAC6 gene; Impairment; Intellectual functioning disability; Intervention; Maintenance; Membrane Protein Traffic; Methodology; Microtubules; Missense Mutation; Mission; Modeling; Molecular; Motor; Mus; Mutation; Neurodevelopmental Disorder; Neuronal Dysfunction; Neurons; Organelles; PACS1 Syndrome; Pathway interactions; Patients; Phenotype; Physiological; Positioning Attribute; Protein Sortings; Proteins; Public Health; Recurrence; Regulation; Research; Seizures; Signal Pathway; Signaling Molecule; Slice; Structure; Synapses; Synaptic Transmission; Testing; Therapeutic; United States National Institutes of Health; Whole Organism; alpha Tubulin; autism spectrum disorder; behavior test; biophysical analysis; curative treatments; effective therapy; flexibility; fundamental research; genome integrity; hippocampal pyramidal neuron; induced pluripotent stem cell; inhibitor therapy; innovation; insight; live cell imaging; mouse model; neurodevelopment; neurogenesis; neuron development; neurotransmission; novel; pharmacologic; postsynaptic; premature; protein complex; protein transport; therapeutic RNA; tool; trafficking; trans-Golgi Network; translational framework

Project Summary PACS1 Syndrome is a recently identified neurodevelopmental disorder caused by a recurrent de novo missense mutation in PACS1 (p.Arg203Trp). Patients carrying this missense mutation share several developmental deficits, including intellectual disability, seizures and autism. The mechanism by which PACS1R203W causes PACS1 Syndrome is unknown and no curative treatment is available. PACS1 is a multifunctional sorting protein that facilitates retrograde trafficking from endosomes to the trans-Golgi network, for delivery of proteins to the primary cilium and for genome integrity. This multifunctionality depends on a small segment of PACS1 called the furin- binding region (FBR), which binds a broad range of client proteins and signaling molecules. The R203W mutation is located in the FBR, and our biophysical studies reveal a change in the FBR dynamics when the R203W substitution is present, suggesting the possibility of an altered interaction between PACS1 and one or more of its client proteins in PACS1 Syndrome. Our preliminary studies strongly suggest PACS1R203W increases binding to the deacetylase HDAC6 to profoundly disturb membrane traffic and impair neuron development. Consequently, PACS1R203W reduces acetylation of known HDAC6 substrates, including α-tubulin, disrupting centrosome positioning and leading to Golgi fragmentation and increased dendritic arborization in pyramidal neurons. This dendritic overbranching is coupled to reduced inhibitory currents in L2/3 cortical neurons resulting in an increased excitatory:inhibitory (E:I) ratio, similar to that found in other neurodevelopmental disorders, suggesting that PACS1R203W severely affects neuronal function and behavior. Our long-term goal is to understand how PACS1R203W causes disease and to use this information to develop effective therapies. The objective of this particular application is to determine how PACS1R203W and HDAC6 combine to dysregulate neuronal arborization and synaptic transmission. We hypothesize that the aberrant interaction between PACS1R203W and HDAC6 alters organellar positioning, which contributes to excessive dendrite arborization and dysregulated synaptic activity. Guided by strong preliminary data, we will test our hypothesis by pursuing three specific aims: 1) Determine how the R203W mutation alters PACS1 structure and dynamics for influencing client protein interactions, 2) Determine how PACS1R203W and HDAC6 combine to dysregulate Golgi positioning and dendrite arborization, and 3) Determine how PACS1R203W alters synaptic activity and behavior. The approach is innovative because we will characterize, from the atomic structure to the whole-organism, the mechanism by which the recurrent R203W substitution causes neuronal dysfunction. This research is significant because it may identify new targets and therapeutic approaches to treat this debilitating disorder.

项目概要 PACS1 综合征是最近发现的一种神经发育障碍，由反复出现的新错义引起 PACS1 (p.Arg203Trp) 突变。携带这种错义突变的患者有一些发育缺陷， 包括智力障碍、癫痫和自闭症 PACS1R203W 导致 PACS1 的机制。 PACS1 是一种多功能分选蛋白，其症状尚不清楚，也没有可用的治疗方法。 促进从内体到跨高尔基体网络的逆行运输，将蛋白质递送到初级 纤毛和基因组完整性取决于 PACS1 的一小部分，称为弗林-。 结合区（FBR），结合广泛的客户蛋白和信号分子 R203W 突变。 位于 FBR 中，我们的生物物理研究揭示了当 R203W 存在替代，表明 PACS1 与一种或多种之间的相互作用可能发生改变 我们的初步研究强烈表明 PACS1R203W 会增加结合。 去乙酰化酶 HDAC6 会严重干扰膜交通并损害神经元发育， PACS1R203W 减少已知 HDAC6 底物的乙酰化，包括 α-微管蛋白，破坏中心体 定位并导致高尔基体破碎和锥体神经元中树突状树枝化的增加。 树突过度分支与 L2/3 皮质神经元抑制电流减少相结合，导致 兴奋性：抑制性 (E:I) 比率，类似于其他神经发育障碍中发现的比率，表明 PACS1R203W 严重影响神经功能和行为 我们的长期目标是了解如何影响神经功能和行为。 PACS1R203W 会导致疾病，并利用这些信息来开发有效的疗法。 特定应用是确定 PACS1R203W 和 HDAC6 如何结合来调节神经树枝化 我们勇敢地承认 PACS1R203W 和 HDAC6 之间的异常相互作用会改变。 细胞器定位，导致树突过度分枝和突触活动失调。 在强有力的初步数据的指导下，我们将通过追求三个具体目标来检验我们的假设：1）确定如何 R203W 突变改变 PACS1 结构和动力学，从而影响客户蛋白相互作用，2) 确定 PACS1R203W 和 HDAC6 如何结合来失调高尔基体定位和树突分枝， 3) 确定 PACS1R203W 如何改变突触活动和行为 该方法具有创新性，因为。 我们将从原子结构到整个有机体，描述周期性的机制 R203W 替代会导致神经功能障碍，这项研究意义重大，因为它可能会识别出新的靶点。 以及治疗这种使人衰弱的疾病的治疗方法。

项目成果

Genome-wide kinase-MAM interactome screening reveals the role of CK2A1 in MAM Ca2+ dynamics linked to DEE66.

全基因组激酶-MAM 相互作用组筛选揭示了 CK2A1 在与 DEE66 相关的 MAM Ca2 动力学中的作用。

• 发表时间: 2023-08-08
• 影响因子: 11.1
• 作者: Thi My Nhung, Truong;Phuoc Long, Nguyen;Diem Nghi, Tran;Suh, Yeongjun;Hoang Anh, Nguyen;Jung, Cheol Woon;Minh Triet, Hong;Jung, Minkyo;Woo, Youngsik;Yoo, Jinyeong;Noh, Sujin;Kim, Soo Jeong;Lee, Su Been;Park, Seongoh;Thomas, Gary;Simmen, Tho
• 通讯作者: Simmen, Tho

Do PACS1 variants impeding adaptor protein binding predispose to syndromic intellectual disability?

阻碍接头蛋白结合的 PACS1 变异是否容易导致综合征性智力障碍？

• DOI: 10.1002/ajmg.a.63232
• 发表时间: 2023-05-04
• 期刊: American Journal of Medical Genetics Part A
• 影响因子: 2
• 作者: Ashley Moller;Duha Hejla;Hyun Kyung Lee;J. Lyles;Yunhan Yang;Kun Chen;W. L. Li;G. Thomas;C. Boerkoel
• 通讯作者: C. Boerkoel