Modulators of CaV1.3 Ca2+ regulation

CaV1.3 Ca2 调节的调节剂

• 批准号: 8408867
• 负责人: DAVID T YUE
• 金额: $ 4.05万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-10 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8408867
• 关键词: Adverse effects; Affect; Agonist; Atrial Fibrillation; Attenuated; Awareness; Basic Science; Behavior; Binding; Biological Assay; Blood Vessels; Brain; Breathing; Calcium Channel; Calmodulin; Cardiac; Cardiovascular system; Cell Nucleus; Cells; Chemicals; Circadian Rhythms; Coupled; Dihydropyridines; Doctor of Philosophy; Dose; Electrophysiology (science); Elements; Feedback; Fluorescence Resonance Energy Transfer; Functional disorder; Goals; Hair Cells; Hand; Heart; Individual; Knowledge; L-Type Calcium Channels; Laboratories; Lead; Libraries; Life; Locomotion; Mediating; Microscope; Molecular; Motor; Muscle Contraction; Neurodegenerative Disorders; Outcome; Parkinson Disease; Pathology; Periodicity; Pharmacologic Substance; Physiological; Physiology; Protocols documentation; Published Comment; Reader; Recombinants; Regulation; Role; Signal Transduction; Skeletal Muscle; Speed; Substantia nigra structure; System; Testing; Therapeutic; Toxic effect; Ursidae Family; base; counterscreen; dihydropyridine; fluorophore; improved; inhibitor/antagonist; neurotransmitter release; novel; novel strategies; patch clamp; positive mood; response; ribbon synapse; small molecule; stable cell line; tool; voltage

DESCRIPTION (provided by applicant): CaV1.3 channels are low-threshold, dihydropyridine-sensitive L-type Ca2+ channels which mediate low-voltage signaling and rhythmicity throughout the body. They are essential for neurotransmitter release at ribbon synapses such as found in cochlear hair cells; they mediate pacemaking in the heart; and they modulate oscillatory behavior throughout the brain, such as the repetitive bursting in supra-chiasmatic (circadian pacemaking circuitry) and substantia nigra (locus of primary damage in Parkinson's) nuclei. As such, overactivity of these channels may predispose for Ca2+ overload precipitating Parkinson's, and downward modulation of these channels may enhance positive mood and affect. Clearly, small-molecule compounds that selectively inhibit or enhance CaV1.3 channels, rather than the other CaV1 L-type channels would be of enormous utility for basic studies of CaV1.3 roles, and for potentially amerliorating a number of CaV1.3-related pathologies. However, though excellent L-type channels antagonists and agonists have been discovered, none can truly select among the L-type channel subtypes. Here, in the search for selective modulators, we will exploit a unique molecular interaction between ICDI and IQ domains of CaV1.3 channels, where this interaction modulates the strength Ca2+ feedback inhibition (CDI) of these channels. This promising screen will be prosecuted according to three specific aims. 1) To perform a primary screen for small molecules that disrupt or enhance a functionally critical interaction between IQ and ICDI domains of CaV1.3 channels, using the MLSMR library of 350,000-500,00 compounds. 2) To confirm and identify candidate hits from Aim 1 using a microscope-based FRET analysis of single living cells. 3) To test candidate compounds for modulation of CaV1.3 Ca2+ regulation, using patch-clamp electrophysiology. Overall, this project promises lead candidates for selective modulators of CaV1.3 versus other CaV1 L-type calcium channels.

描述（由申请人提供）：CAV1.3通道是低阈值，二氢吡啶敏感的L型Ca2+通道，可介导整个身体的低压信号传导和节奏性。它们对于在耳蜗细胞中发现的色带突触中的神经递质释放至关重要。他们介导心脏中的起搏。它们调节整个大脑中的振荡行为，例如上chiasmmantic（昼夜节律起搏回路）和底底nigra（帕金森氏症中主要损害的基因座）的重复爆发。因此，这些通道的过度活动性可能会使Ca2+过载帕金森氏症的过载易感性，并且对这些通道的下降调节可能会增强积极的情绪和影响。显然，对于有选择地抑制或增强CAV1.3通道的小分子化合物，而不是其他CAV1 L型通道对于Cav1.3角色的基本研究以及可能会进行一些社会化的CAV1.3相关病理学，将具有巨大的实用性。但是，尽管已经发现了出色的L型拮抗剂和激动剂，但没有人能真正在L型通道亚型中进行选择。在这里，在寻找选择性调节器时，我们将利用CAV1.3通道的ICDI和IQ域之间的独特分子相互作用，其中这种相互作用调节了这些通道的强度Ca2+反馈抑制（CDI）。这个有希望的屏幕将根据三个特定目标起诉。 1）使用350,000-500,00化合物的MLSMR库进行破坏或增强CAV1.3通道智商和ICDI域之间功能关键相互作用的小分子。 2）使用基于显微镜的单个活细胞分析AIM 1的候选命中。 3）使用Patch-Clamp电生理学测试候选化合物，以调节CAV1.3 Ca2+调节。总体而言，该项目有望为CAV1.3选择性调节剂与其他CAV1 L型钙通道的选择性调节剂。