Mechanisms and Modulation of Opsin 3 Mediated Airway Smooth Muscle Relaxation

视蛋白 3 介导的气道平滑肌松弛的机制和调节

• 批准号: 10541204
• 负责人: Peter D Yim
• 金额: $ 16.54万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10541204
• 关键词: 11 cis Retinal; 9-cis-retinal; Agonist; Asthma; Binding; Blood Vessels; Carotene; Cell Line; Cells; Chest; Co-Immunoprecipitations; Coupled; Cyclic AMP; Cyclic GMP; Cytoskeleton; Disease; Evaluation; Exposure to; Eye; Family; Functional disorder; G-Protein Signaling Pathway; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Genomics; Human; Intestines; Ionones; Knockout Mice; Laboratories; Ligands; Light; Link; Lung; Measures; Mediating; Mediator; Methods; Modeling; Molecular Conformation; Mus; Muscle; Muscle relaxation phase; Mutation; Myosin Light Chains; Nebulizer; Opsin; Organ; Pathway interactions; Penetration; Peripheral; Phosphotransferases; Photons; Phytochemical; Proteins; Pulmonary function tests; Receptor Activation; Receptor Signaling; Relaxation; Resistance; Retina; Schiff Bases; Second Messenger Systems; Signal Pathway; Signal Transduction; Signaling Protein; Smooth Muscle; Smooth Muscle Myocytes; Specificity; Techniques; Therapeutic; Tissues; Trachea; Transfection; Uterus; Variant; Vascular Smooth Muscle; Vasodilation; Visual; chromophore; genetic regulatory protein; in vivo; light intensity; molecular subtypes; novel; novel strategies; overexpression; photoactivation; protein expression; protein protein interaction; receptor; receptor binding; respiratory smooth muscle

Project Summary/ Abstract Light-mediated relaxation of smooth muscle has been recently demonstrated. Non-visual opsins expressed on smooth muscle have been implicated as the key mediator of relaxation via light within the visible spectrum. However the mechanism of relaxation has yet to be fully understood. Our group has demonstrated that this phenomenon is also present in airway smooth muscle with light-mediated relaxation modulated by opsin receptors coupled to G protein signaling pathways. In preliminary studies, we demonstrate that airway smooth muscle photorelaxation is wavelength- and intensity-dependent and that light induces an interaction between the opsin 3 receptor and Gαs demonstrated by co-immunoprecipitation. This was associated with an increase in intracellular cyclic AMP, a known key second messenger of airway smooth muscle relaxation. We propose to identify the opsin receptors subtype(s), the G protein(s) and the cellular signaling pathways (e.g. cAMP, cGMP, and cytoskeletal regulatory proteins (RhoA, CPI-17 and myosin light chain-20) that modulate airway smooth muscle photorelaxation. We will also explore the mechanism of activation by modulating the light wavelength sensitivity of airway smooth muscle. Normally opsin receptors bind 11-cis-retinal (a chromophore and co- activator with photons of light) to form a Schiff base. The 11-cis-retinal allows for photoisomerization that changes the conformation of the opsin receptor, which induces downstream cellular signaling. Derivatives of retinal have been shown to shift the sensitivity of opsins to longer wavelengths. In our proposal we demonstrate that the use of 11-cis 2,3 didehydroretinal shifts airway smooth muscle sensitivity to longer wavelengths. We propose to evaluate additional retinal derivatives in cellular, in vivo and ex vivo models to identify chromophores that can shift the wavelength sensitivity of endogenous opsins to respond to longer wavelengths of light that may penetrate the body. In the eye, the sensitivity to different wavelengths of light is not only a function of the specific ligand but also a function of the subtype of opsin receptor expressed. Transfection of variants of OPN1 will allow evaluation of specific ligands at varying wavelengths of light in airway smooth muscle that activate cellular signaling congruent with relaxation. We will explore the direct activation of opsin receptors by carotene metabolites in the absence of light. We will demonstrate that the endogenous metabolite of carotenes, β-ionone, is a phytochemical that can directly activate opsin 3 in the absence of light. We will show that the same pro- relaxant cellular signaling occurs in airway smooth muscle when the opsin 3 receptor is directly activated by - ionine as compared to activation by chromophores/light. These proposed studies will allow for a better understanding of the activation and signaling of a previously uncharacterized receptor expressed in airway smooth muscle that modulates pro-relaxant signaling pathways.

项目总结/摘要 光介导的平滑肌松弛最近已被证明。非视觉视蛋白 在平滑肌上表达的蛋白质被认为是通过可见光内的光松弛的关键介质。 江西篇章然而，松弛的机制尚未完全理解。我们的团队已经证明了 这种现象也存在于具有由视蛋白调节光介导的松弛的气道平滑肌中 受体偶联G蛋白信号通路。在初步研究中，我们证明了气道平滑 肌肉光松弛是波长和强度依赖性的，并且光诱导肌肉光松弛之间的相互作用。 免疫共沉淀法检测视蛋白3受体和Gαs的表达。这与增加 细胞内环腺苷酸，一种已知的呼吸道平滑肌松弛的关键第二信使。我们建议 鉴定视蛋白受体亚型、G蛋白和细胞信号传导途径（例如cAMP，cGMP， 和调节气道平滑的细胞骨架调节蛋白（RhoA、CPI-17和肌球蛋白轻链-20） 肌肉光松弛我们还将通过调制光波长来探索激活机制 气道平滑肌的敏感性。正常情况下，视蛋白受体结合11-顺式-视黄醛（发色团和共- 活化剂与光的光子）以形成席夫碱。11-顺式-视黄醛允许光异构化， 视蛋白受体的构象，其诱导下游细胞信号传导。视黄醛的衍生物具有 已经被证明可以将视蛋白的敏感性转移到更长的波长。在我们的建议中，我们证明了使用 11-顺式2，3-二脱氢视网膜改变气道平滑肌对更长波长的敏感性。我们建议 在细胞、体内和离体模型中评价另外的视网膜衍生物，以鉴定可以 改变内源性视蛋白的波长敏感性以响应较长波长的光， 穿透身体在眼睛中，对不同波长的光的敏感性不仅是特定波长的函数， 配体，而且也是视蛋白受体亚型表达的功能。0 PN 1变体的转染将允许 气道平滑肌中不同波长光下激活细胞的特异性配体的评价 与放松一致的信号。我们将探讨胡萝卜素直接激活视蛋白受体 代谢物在没有光的情况下。我们将证明胡萝卜素的内源性代谢产物β-紫罗兰酮， 是一种植物化学物质，可以直接激活视蛋白3在没有光。我们将证明同样的亲- 当视蛋白3受体被β-D受体直接激活时， 与发色团/光的激活相比，这些拟议的研究将允许更好地 了解气道中表达的先前未表征的受体的激活和信号传导 调节促松弛信号通路的平滑肌。