Generation of tools for visualizing understudied KvS potassium channel proteins

摘要 电压门控离子通道受到许多现有药物和药物靶点的调制 治疗各种疾病的发展计划。开发离子通道药物的一个主要障碍是 识别和验证在限制副作用的同时改善疾病的分子靶点。最近， 转录组学项目已经确定了躯体感觉和KVS电压门控K通道的转录 运动神经元亚型的研究表明，研究这些KVS亚基可能会确定 改善神经病理性疼痛或运动功能障碍。KVS蛋白是电压门控的K通道亚基 除非它们与Kv2家族的电压门控K通道亚单位共同组装，否则它们是“电沉默的”。这个 然而，许多KVS亚基的功能及其在疾病中的作用在很大程度上仍未被探索，因为几乎没有 或者没有针对这些亚单位的研究工具。照亮可药物基因组计划有适当的 确定了几个KVS家族成员，包括KCNG2-4(编码Kv6.2-6.4)和KCNS1 (编码Kv9.1)。基因表达研究表明，这些KVS亚基在选定的脊髓中强势表达 脊髓和背根神经节神经元在其中参与电信号转导 影响运动、本体感觉和疼痛。然而，关于基因表达的信息很少。 由于缺乏在天然组织中检测KVS亚基的有效试剂，这些KVS亚基在蛋白质水平上存在。 该项目的目标是(I)研制出特定的单抗，并对其进行基因编码 可视化未被研究的KVS亚基的工具，以及(Ii)使用这些试剂来定义细胞和 KVS亚单位在脊髓和背根神经节神经元中的亚细胞定位。这些工具和 关于Kv2/KvS异四聚体通道在哪里表达的信息将为以下方面提供必要的平台 KVS亚基的生理作用及其在神经系统疾病和潜能中的作用的进一步研究 作为药物靶标的公用事业。这项研究可能会确定特定的KVS亚基作为选择性药物靶点 运动神经元和感觉神经元疾病的治疗，包括肌萎缩侧索硬化症的兴奋性毒性， 神经病理性疼痛的过敏性。

Abstract Voltage-gated ion channels are modulated by many existing drugs and the targets of pharmaceutical development programs to treat a diverse array of maladies. A major obstacle to developing ion channel drugs is identification and validation of molecular targets that ameliorate disorders while limiting side effects. Recently, transcriptomics projects have identified transcripts for KvS voltage-gated K+ channels in somatosensory and motor neuron subtypes, suggesting that studying these KvS subunits could potentially identify strategies to ameliorate neuropathic pain or motor dysfunctions. KvS proteins are voltage-gated K+ channel subunits which are “electrically silent” unless they coassemble with voltage-gated K+ channel subunits of the Kv2 family. The function of many KvS subunits and their role in disease remains largely unexplored, however, as there are few or no research tools for these subunits. The Illuminating the Druggable Genome program has appropriately identified several KvS family members as “understudied” including KCNG2-4 (encoding Kv6.2-6.4) and KCNS1 (encoding Kv9.1). Gene expression studies show that these KvS subunits are robustly expressed in select spinal cord and dorsal root ganglia neurons where their participation in electrical signaling would be expected to influence movement, proprioception, and pain. However, there is little information available on the expression of these KvS subunits at the protein level due to a lack of well-validated reagents for their detection in native tissue. The objectives of this project are (i) to develop specific monoclonal antibody and genetically-encoded tools to visualize understudied KvS subunits, and (ii) to use these reagents to define the cellular and subcellular localization of KvS subunits in spinal cord and dorsal root ganglia neurons. These tools and information on where Kv2/KvS heterotetrameric channels are expressed will provide an essential platform for further studies on the physiological role of KvS subunits and their role in neurological disorders and potential utility as drug targets. This research could potentially identify specific KvS subunits as selective drug targets for the treatment of motor and sensory neuron disorders, including excitotoxicity in amyotrophic lateral sclerosis, and hypersensitivity in neuropathic pain.