Small GTPase signaling in dendrites and spines

树突和棘中的小 GTP 酶信号传导

• 批准号: 10173123
• 负责人: Peter Penzes
• 金额: $ 78.21万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10173123
• 关键词: Affinity; Architecture; Behavior; Behavioral; Binding; Biological; Biological Assay; Biological Models; Biological Process; Biological Testing; Biology; Bipolar Disorder; Brain; Catalytic Domain; Chemicals; Cognitive; Data; Dendrites; Dendritic Spines; Development; Disease; Drug Targeting; Family; Functional disorder; Funding; Future; Glutamates; Grant; Guanine Nucleotide Exchange Factors; Human; In Vitro; Intervention; Ion Channel; Measurable; Modeling; Monomeric GTP-Binding Proteins; Morphology; Mus; Neurons; Pathology; Pathway interactions; Pharmacology; Phosphotransferases; Play; Pre-Clinical Model; Proteins; Publishing; Regulation; Risk Factors; Rodent; Rodent Model; Role; Schizophrenia; Series; Signal Transduction; Signaling Molecule; Structure-Activity Relationship; Synapses; Testing; Therapeutic; Validation; Vertebral column; autism spectrum disorder; base; cell type; drug discovery; experimental study; follow-up; genetic regulatory protein; high throughput screening; in silico; induced pluripotent stem cell; inhibitor/antagonist; innovation; mouse model; neuropsychiatric disorder; novel; novel drug class; postnatal development; receptor; rho; screening; small molecule; stem cell model; synaptic function; therapeutic target; three dimensional structure; tool; transmission process

ABSTRACT Dendritic and spine plasticity plays key roles in brain development, function, behavior, and disease. Indeed, spine and dendrite pathology is a common feature of many neuropsychiatric disorders (NPDs), including autism spectrum disorder (ASD), schizophrenia (SZ), and bipolar disorder (BPD). Rho-like small GTPases, including Rac1, are a family of regulatory proteins with central roles in dendrite and spine plasticity. Their extensive implication in NPDs suggests that these pathways can serve as therapeutic targets in NPDs. The activity of small GTPases is enhanced by guanine-nucleotide-exchange factors (GEFs), among which the Rac1- GEF kalirin is highly enriched in spines, and is perhaps the best-characterized GEF in the brain. Kalirin is a central regulator of dendrite arborization, spine plasticity, glutamatergic transmission, neuronal connectivity, and cognitive behavior. While small-molecule pharmacological modulators have been invaluable tools for studying the biological functions of kinases, receptors, or ion channels, no such tools exist for Rho-GEFs, including kalirin. Kalirin is an optimal drug target for several reasons: its expression is largely restricted to the CNS, it is highly enriched in spines, it is a signaling hub in a synaptic network including many NPD risk factors, its enzymatic activity can be modulated, and the 3D structure of its GEF domain has been determined. Here we outline a novel and innovative hit validation cascade that will allow us to develop small-molecule tools to investigate a previously unapproachable target relevant to NPDs. The brain-specific expression of kalirin and its highly compartmentalized subcellular localization at synapses suggests that regulation of Rac1 signaling, through pharmacological interventions targeting kalirin, may allow neuron- and synapse-specific effects. This is key to developing tools that produce cell type-specific and context-dependent Rac1 modulation. Using a combination of high-throughput screening (HTS) and in silico screening against the target proteins kalirin/Rac1, we produced a hit list of potential regulators of kalirin activity suitable for follow-up analysis in well-characterized and optimized assays. We hypothesize that small-molecule compounds isolated in HTS and in silico screens modulate kalirin's GEF and biological activity in rodent and human iPSC-derived neuron models, and reverse neuroarchitectural abnormalities in models of NPDs. We will test this hypothesis in the following aims: 1) Hit validation and in vitro characterization, selection, and prioritization. 2) Prioritization of mouse and human neuronal model systems for testing validated hits. 3) Characterization of validated hit compounds in mouse and iPSC models.

摘要 树突状细胞和脊椎的可塑性在大脑发育、功能、行为和疾病中起着关键作用。的确， 棘突和树突病理是许多神经精神障碍(NPD)的共同特征，包括 自闭症谱系障碍(ASD)、精神分裂症(SZ)和双相情感障碍(BPD)。Rho样的小GTP酶， 包括rac1在内，是一个调节蛋白家族，在树突和棘突的可塑性中起核心作用。他们的 对NPD的广泛研究表明，这些通路可以作为NPD的治疗靶点。这个 鸟嘌呤核苷酸交换因子(GEF)可增强小分子GTP酶的活性，其中，Rac1-GTP酶活性最高。 全球环境基金卡利林的脊椎高度丰富，可能是大脑中最具特征的全球环境基金。加里林是一名 树突分枝、脊髓可塑性、谷氨酸能传递、神经元连接的中枢调节因子， 和认知行为。虽然小分子药物调节剂一直是治疗 在研究激酶、受体或离子通道的生物学功能时，Rho-GEF还没有这样的工具， 包括加里林。Kalirin是一个理想的药物靶点有几个原因：它的表达在很大程度上限于 中枢神经系统，它的脊椎高度丰富，是突触网络中的信号枢纽，包括许多NPD风险因子， 它的酶活性是可以调节的，并确定了它的环境蛋白结构域的三维结构。在这里我们 概述一个新颖和创新的HIT验证级联，它将使我们能够开发小分子工具来 调查以前无法接近的与NPD相关的目标。Kalirin和Kalirin的脑特异性表达 它在突触的高度划分的亚细胞定位表明，对rac1信号的调节， 通过针对Kalirin的药物干预，可能允许神经元和突触特异的效应。这 是开发产生特定细胞类型和上下文相关的rac1调制的工具的关键。使用 针对靶蛋白的高通量筛选(HTS)和电子筛选相结合 Kalirin/rac1，我们制作了一份适用于后续分析的Kalirin活性潜在调节因子的命中列表 具有良好特性和优化的分析方法。我们假设在高温超导中分离出的小分子化合物 在电子屏幕上调节Kalirin在啮齿动物和人IPSC来源神经元中的全环和生物活性 模型，并逆转NPD模型的神经结构异常。我们将在 以下目标：1)HIT验证和体外表征、选择和优先排序。2)确定优先顺序 用于测试有效命中的小鼠和人类神经元模型系统。3)验证的HIT的特征 小鼠和ipsc模型中的化合物。

项目成果

PAK1 protein expression in the auditory cortex of schizophrenia subjects.

• DOI: 10.1371/journal.pone.0059458
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Deo AJ;Goldszer IM;Li S;DiBitetto JV;Henteleff R;Sampson A;Lewis DA;Penzes P;Sweet RA
• 通讯作者: Sweet RA

A Kalirin missense mutation enhances dendritic RhoA signaling and leads to regression of cortical dendritic arbors across development.

• DOI: 10.1073/pnas.2022546118
• 发表时间: 2021-12-07
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Grubisha MJ;Sun T;Eisenman L;Erickson SL;Chou S;Helmer CD;Trudgen MT;Ding Y;Homanics GE;Penzes P;Wills ZP;Sweet RA
• 通讯作者: Sweet RA

Disrupted-in-Schizophrenia 1 (DISC1) regulates spines of the glutamate synapse via Rac1.

• DOI: 10.1038/nn.2487
• 发表时间: 2010-03
• 期刊: NATURE NEUROSCIENCE
• 影响因子: 25
• 作者: Hayashi-Takagi, Akiko;Takaki, Manabu;Graziane, Nick;Seshadri, Saurav;Murdoch, Hannah;Dunlop, Allan J.;Makino, Yuichi;Seshadri, Anupamaa J.;Ishizuka, Koko;Srivastava, Deepak P.;Xie, Zhong;Baraban, Jay M.;Houslay, Miles D.;Tomoda, Toshifumi;Brandon, Nicholas J.;Kamiya, Atsushi;Yan, Zhen;Penzes, Peter;Sawa, Akira
• 通讯作者: Sawa, Akira

Epac2-mediated dendritic spine remodeling: implications for disease.

EPAC2介导的树突状脊柱重塑：对疾病的影响。

• DOI: 10.1016/j.mcn.2010.11.008
• 发表时间: 2011-02
• 期刊: MOLECULAR AND CELLULAR NEUROSCIENCE
• 影响因子: 3.5
• 作者: Penzes, Peter;Woolfrey, Kevin M.;Srivastava, Deepak P.
• 通讯作者: Srivastava, Deepak P.

Developmental Trajectories of Auditory Cortex Synaptic Structures and Gap-Prepulse Inhibition of Acoustic Startle Between Early Adolescence and Young Adulthood in Mice.

• DOI: 10.1093/cercor/bhv040
• 发表时间: 2016-05
• 期刊: Cerebral cortex
• 影响因子: 3.7
• 作者: Caitlin E. Moyer;S. Erickson;K. Fish;E. Thiels;P. Penzes;R. Sweet