CRMP2, Nav1.7 sodium channel, and chronic pain

CRMP2、Nav1.7 钠通道和慢性疼痛

• 批准号: 10113570
• 负责人: Rajesh Khanna
• 金额: $ 38.29万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10113570
• 关键词: Accounting; Action Potentials; Acute Pain; Address; Afferent Neurons; Alanine; American; Analgesics; Arizona; Axon; Behavior; Binding; Budgets; Calcium Channel; Cell membrane; Cells; Clathrin; Clinical; Collaborations; Core Facility; Department of Defense; Disease; Endocytosis; Excision; Foundations; Generations; Genes; Genetic; Genetically Engineered Mouse; High Prevalence; Human; Injury; Institute of Medicine (U.S.); Knock-in Mouse; Link; Locomotion; Maintenance; Measures; Mechanics; Mediating; Membrane; Memory; Modeling; Modification; Mus; Mutation; Neurons; Nociception; Pain; Pain Research; Pain Threshold; Pathogenesis; Peptides; Peripheral nerve injury; Pharmaceutical Preparations; Plasmids; Play; Post-Translational Modification Site; Post-Translational Protein Processing; Principal Investigator; Proteins; Rattus; Recycling; Regulation; Research; Research Personnel; Role; Sedation procedure; Site; Smell Perception; Societies; Sodium Channel; Solid; Specificity; Spinal Cord; Spinal Ganglia; Stimulus; Sumoylation Pathway; Surface; Syndrome; Testing; Therapeutic; Thermal Hyperalgesias; Transfection; Transgenic Organisms; Ubiquitin; Universities; Work; chronic pain; chronic painful condition; collapsin response mediator protein-2; cost; density; design; effective therapy; gabapentin; gain of function mutation; improved; in vivo; inflammatory pain; mechanical allodynia; mouse model; mutant; neuronal excitability; novel; novel therapeutic intervention; overexpression; pain behavior; pain model; pain signal; painful neuropathy; pre-clinical; pregabalin; protein protein interaction; protein transport; recruit; response; side effect; success; therapeutic target; trafficking; voltage

Abstract Chronic pain conditions cause an immense burden on society due to their astonishingly high prevalence and lack of effective treatments. This application addresses how indirect modulation of the excitability of neurons in pain conditions can be achieved by altering the expression and function of the Nav1.7 sodium channel. The scientific premise is that because direct blockade of Nav1.7 channels has been unsuccessful, targeting regulators of Nav1.7 may offer therapeutic advantages allowing for a graded analgesic response. Dr. Rajesh Khanna, Principal Investigator on this project, first discovered that expression of Nav1.7 at the surface is regulated by a protein, axonal collapsin response mediator protein 2 (CRMP2), and that a mutant of CRMP2 lacking the small ubiquitin-like modifier (SUMO) post-translational modification (deSUMOylation) reduces Nav1.7 surface expression and currents. Importantly, the related Nav1.1, Nav1.3, Nav1.5, Nav1.6, Nav1.8, and Nav1.9 channels are unaffected. In preliminary studies, we demonstrate that loss of CRMP2 SUMOylation increases binding to endocytic proteins, potentially accounting for removal of Nav1.7 from the surface. The fraction of SUMOylated CRMP2 increases significantly following peripheral nerve injury. Excitingly, in vivo transfection of a CRMP2-K374A SUMO-null plasmid or a peptide mimicking the CRMP2 SUMOylation motif, into the spinal cord reversed mechanical allodynia in a model of neuropathic pain. Together, these findings strongly support the hypothesis that loss of CRMP2 SUMOylation reduces Nav1.7 localization at the plasma membrane, thereby decreasing nociceptive neuron excitability and thresholds to thermal and mechanical stimuli in acute and chronic pain. We will test this hypothesis in three specific aims. In Aim 1, we will test the general role of CRMP2 SUMOylation on Nav1.7 currents and neuronal excitability using a recently created new transgenic K374A Crmp2 knock-in mouse model where the SUMOylation site (K374) of CRMP2 has been replaced with an alanine mutation; this mouse was made by Dr. Thomas Doetschman, a co-Investigator on this project and Director of the Genetically Engineered Mouse Models Core facility at the University of Arizona. The mechanism by which Nav1.7 surface trafficking and internalization occur is unknown and will be examined in Aim 2 of this proposal. Aim 3 will evaluate the contribution of the CRMP2 SUMOylation state to acute pain thresholds as well as after experimentally induced pain thresholds using models in which Nav1.7 levels are increased; these studies will be performed in collaboration with Dr. Todd Vanderah, a co-Investigator on this project with deep expertise in preclinical pain modeling. Finally, in these mice, we will also measure CRMP2- dependent off-target effects on memory, locomotion/sedation, as well as behaviors linked to Nav1.7, including smell. The proposed study will considerably improve our understanding of how intracellular trafficking proteins can be modified in diseases/injuries, lay a solid foundation for unraveling mechanisms of the modification and trafficking of Nav1.7 in chronic pain, and offer novel and selective therapeutic targets for pain research.

抽象的 慢性疼痛由于其患病率高得惊人并且给社会造成了巨大的负担。 缺乏有效的治疗方法。该应用解决了如何间接调节神经元的兴奋性 疼痛状况可以通过改变 Nav1.7 钠通道的表达和功能来实现。这 科学前提是，由于直接封锁Nav1.7频道不成功，针对 Nav1.7 的调节剂可能提供治疗优势，允许分级镇痛反应。拉杰什博士 该项目的首席研究员 Khanna 首先发现 Nav1.7 在地表的表达是 受轴突塌陷反应介导蛋白 2 (CRMP2) 蛋白以及 CRMP2 突变体的调节 缺乏小泛素样修饰剂 (SUMO) 翻译后修饰 (deSUMOylation) 会降低 Nav1.7 表面表达和电流。重要的是，相关的 Nav1.1、Nav1.3、Nav1.5、Nav1.6、Nav1.8 和 Nav1.9 频道不受影响。在初步研究中，我们证明 CRMP2 SUMOylation 的丧失 增加与内吞蛋白的结合，可能是从表面去除 Nav1.7 的原因。这 周围神经损伤后，SUMO 化 CRMP2 的比例显着增加。令人兴奋的是，在体内 转染CRMP2-K374A SUMO无效质粒或模拟CRMP2 SUMO化基序的肽， 进入脊髓可逆转神经性疼痛模型中的机械性异常性疼痛。综合起来，这些发现 强烈支持这样的假设：CRMP2 SUMOylation 的缺失会降低 Nav1.7 在血浆中的定位 膜，从而降低伤害性神经元的兴奋性以及热和机械的阈值 急性和慢性疼痛的刺激。我们将在三个具体目标上检验这一假设。在目标 1 中，我们将测试 使用最近创建的新方法，CRMP2 SUMOylation 对 Nav1.7 电流和神经元兴奋性的一般作用 转基因 K374A Crmp2 敲入小鼠模型，其中 CRMP2 的 SUMO 化位点 (K374) 已被 被丙氨酸突变取代；这只老鼠是由该项目的联合研究员 Thomas Doetschman 博士制作的 亚利桑那大学基因工程小鼠模型核心设施项目主任和主任。这 Nav1.7 表面贩运和内化发生的机制尚不清楚，将在 本提案的目标 2。目标 3 将评估 CRMP2 SUMO 化状态对急性疼痛的影响 阈值以及使用 Nav1.7 水平的模型进行实验诱导的疼痛阈值后 增加；这些研究将与联合研究员 Todd Vanderah 博士合作进行 项目在临床前疼痛建模方面拥有深厚的专业知识。最后，在这些小鼠中，我们还将测量 CRMP2- 对记忆、运动/镇静以及与 Nav1.7 相关的行为的依赖性脱靶效应，包括 闻。拟议的研究将大大提高我们对细胞内运输蛋白质如何进行的理解 可以在疾病/损伤中进行修饰，为揭示修饰机制奠定坚实的基础 Nav1.7 在慢性疼痛中的运输，并为疼痛研究提供新颖的选择性治疗靶点。

项目成果

In silico identification and validation of inhibitors of the interaction between neuropilin receptor 1 and SARS-CoV-2 Spike protein.

神经毡蛋白受体 1 与 SARS-CoV-2 刺突蛋白之间相互作用的抑制剂的计算机识别和验证。

• 发表时间: 2020-09-23
• 作者: Perez;Patek, Marcel;Moutal, Aubin;Cabel, Carly R;Thorne, Curtis A;Campos, Samuel K;Khanna, Rajesh
• 通讯作者: Khanna, Rajesh

Targeting the vascular endothelial growth factor A/neuropilin 1 axis for relief of neuropathic pain.

靶向血管内皮生长因子 A/神经毡蛋白 1 轴以缓解神经性疼痛。

• 发表时间: 2023-07-01
• 影响因子: 7.4
• 作者: Stratton, Harrison J;Boinon, Lisa;Gomez, Kimberly;Martin, Laurent;Duran, Paz;Ran, Dongzhi;Zhou, Yuan;Luo, Shizhen;Perez;Patek, Marcel;Ibrahim, Mohab M;Patwardhan, Amol;Moutal, Aubin;Khanna, Rajesh
• 通讯作者: Khanna, Rajesh

Dissecting the role of the CRMP2-neurofibromin complex on pain behaviors.

剖析 CRMP2-神经纤维蛋白复合物对疼痛行为的作用。

• 发表时间: 2017-11
• 影响因子: 7.4
• 作者: Moutal, Aubin;Wang, Yue;Yang, Xiaofang;Ji, Yingshi;Luo, Shizhen;Dorame, Angie;Bellampalli, Shreya S;Chew, Lindsey A;Cai, Song;Dustrude, Erik T;Keener, James E;Marty, Michael T;Vanderah, Todd W;Khanna, Rajesh
• 通讯作者: Khanna, Rajesh

Discovery of Quinoline Analogues as Potent Antivirals against Enterovirus D68 (EV-D68).

发现喹啉类似物作为肠道病毒 D68 (EV-D68) 的有效抗病毒药物。

• DOI: 10.1021/acs.jmedchem.9b00115
• 发表时间: 2019-04-25
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Musharrafieh R;Zhang J;Tuohy P;Kitamura N;Bellampalli SS;Hu Y;Khanna R;Wang J
• 通讯作者: Wang J

The role of cyclin-dependent kinase 5 in neuropathic pain.

细胞周期蛋白依赖性激酶 5 在神经性疼痛中的作用。

• 发表时间: 2020
• 影响因子: 7.4
• 作者: Gomez, Kimberly;Vallecillo, Tissiana G M;Moutal, Aubin;Perez;Delgado;Felix, Ricardo;Khanna, Rajesh
• 通讯作者: Khanna, Rajesh