Non-Visual Opsins & Vasoregulation: Implications for Vascular Therapy

非视觉视蛋白

基本信息

批准号：
9264005
负责人：
DAN E BERKOWITZ
金额：
$ 40.29万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9264005
关键词：
ADRBK1 gene Adenoviruses Adrenergic Agents Arrestins Arteries Blood Blood Circulation Blood Vessels Blood flow Cells Cephalic Circadian Rhythms Complement Coronary Arteriosclerosis Cutaneous Cyclic GMP-Dependent Protein Kinases Data Disease Dominant-Negative Mutation Dose Electrodes Electrophysiology (science)Endothelium Erectile dysfunction Exhibits Fingers Functional disorder G protein coupled receptor kinase G-Protein-Coupled Receptors G-substrate GTP-Binding Proteins Grant Health Human Hypertension Immunohistochemistry Impairment Injury Invertebrate Photoreceptors Invertebrates Lasers Light Measurement Mediating Membrane Potentials Microdialysis Molecular Mus Muscle Cells Myography Nitric Oxide Synthase Opsin Pain Paroxetine Pathway interactions Patients Pharmacology Phosphotransferases Physiological Potassium Channel Process Protein Isoforms Proteins Publications Rattus Raynaud Disease Recording of previous events Regulation Relaxation Retinal Ganglion Cells Reverse Transcriptase Polymerase Chain Reaction Role Scleroderma Signal Pathway Signal Transduction Signal Transduction Pathway Skin Sleep Smooth Muscle Myocytes Soluble Guanylate Cyclase Stimulus Stroke Tail Techniques Testing Tissues Universities Vascular Diseases Vascular Smooth Muscle Vasodilation Vasomotor Vertebrates Visual base beta-adrenergic receptor constriction desensitization experimental study in vivo inhibitor/antagonist insight knock-down light intensity melanopsin mouse model mutant new therapeutic target novel overexpression phosphodiesterase 6 prevent public health relevance receptor response small hairpin RNA small molecule inhibitor targeted treatment vascular bed vasoconstriction

项目摘要

DESCRIPTION (provided by applicant): Impaired vasomotor function mediated by pertubations in neurohumoral signaling and/or impaired endothelial function is the proximate cause of vascular diseases such as hypertension, coronary artery disease, stroke, erectile dysfunction, and specific vasculopathies such as Raynaud's phenomenon (RP). The identification of novel pathways that regulate vascular tone provides new targets for treatment of vascular disorders. We have recently identified a novel mechanism that modulates vascular tone. Melanospin (Opn4) are classically found in the retinal ganglion cells where they regulate circadian rhythm and sleep. We have identified a non-visual light (opsin) receptor, Opn4, in blood vessels from a number of mammalian species, and from number of vascular beds. These receptors mediated intensity-dependent vasorelaxation to light of a specific wavelength (blue 430-460nM). Preliminary data suggest that signal transduction mechanism(s) involve soluble guanylate cyclase and phosphodiesterase 6 but not protein kinase G. Light activation leads to vascular hyperpolarization a process that involves K+ channels. The receptor is likely regulated as a classic G-protein coupled receptor in that inhibition of G-protein receptor kinase prevents stimulus-dependent desensitization and significantly decrease the light intensity needed to produce a relaxation response. Finally, in vivo, blue light is able evoke a physiologic response; significantly increasing blood flow in the mouse tail artery. We hypothesize that 1) Opn4 is an important regulator of vasodilatory function and mediates a physiologic function in vivo; 2) the signal transduction mechanism in vessels mimics visual opsins of vertebrates or "simple" photoreceptors of invertebrates; 3) desensitization/regulation occurs by a GRK2/arrestin mechanism; and 4) this pathway is upregulated in diseases in which NO signaling (vasodilation) is impaired and vasocontriction is enhanced such as RP. In this proposal, we plan to: 1) Further characterize the potential physiologic role of Opn 4 in vasoregulation utilizing Opn4-/- mice and newly discovered Opn4 inhibitors, opsinamides, using myography in isolated vessels and laser doppler in cranial windows as endpoints in vivo; 2) Determine signal transduction mechanism/s from receptor and G protein to channel with sharp electrode measurement of membrane potential in isolated vessels using specific inhibitors and shRNA knockdown of pathway proteins; 3) Understand receptor regulation using novel pharmacologic inhibitor of GRK2, paroxetine, vascular smooth muscle-selective GRK2-/-, and arrestin-/- mice, as well as GRK2 and arrestin shRNA adenovirus knockdown in isolated vessels. 4) Determine the role of Opn4 receptors as a target in mouse models of RP and explore mechanisms mediating photorelaxation in vivo in the cutaneous circulation of healthy humans and those with primary RP using laser flow doppler and skin microdialysis. In this way we hope to gain insight into the function and dysfunction of this novel pathway in health and disease, and determine its potential as a novel therapeutic target.

描述（由适用提供）：血管肌舒张器功能受损是由神经肿瘤信号传导和/或内皮功能受损中的衰落介导的，是血管疾病的直接原因，例如高血压，冠状动脉疾病，中风，勃起功能障碍以及特定的血管疾病，例如raynaud的现象（rp）。调节血管张力的新途径的鉴定为治疗血管疾病的新靶标提供了新的靶标。我们最近确定了一种调节血管张力的新型机制。 Melanospin（OPN4）在残留的神经节细胞中经典发现，它们调节昼夜节律和睡眠。我们已经在许多哺乳动物物种的血管和血管床数量中确定了非视光（OPSIN）受体OPN4。这些受体介导了强度依赖性的血管长度，以使特定波长（蓝色430-460nm）的光。初步数据表明，信号翻译机制涉及固体鸟烯酸盐环化酶和磷酸二酯酶6，但不蛋白激酶G。光激活导致血管超极化导致涉及K+通道的过程。接收器可能被调节为经典的G蛋白偶联受体，因为抑制G蛋白受体激酶可防止刺激依赖性脱敏，并显着降低了产生松弛反应所需的光强度。最后，在体内，蓝光能够引起身体反应。小鼠尾动脉的血流显着增加。我们假设1）OPN4是血管舒张功能的重要调节剂，并在体内介导生理功能； 2）粘膜中的信号转导机制模仿脊椎动物的视觉视蛋白或无脊椎动物的“简单”感光体； 3）脱敏/调节是通过GRK2/阻止素机制发生的； 4）此途径在没有信号传导（血管舒张）受损的疾病中进行了更新，并增强了血管成分（例如RP）。在此提案中，我们计划：1）进一步表征OPN 4在使用OPN4 - / - 小鼠和新发现的OPN4抑制剂（Opsinamides）中的血管调节中的潜在生理作用，使用myagraghy在隔离的容器中使用myagraghy，而在颅骨窗户中的激光多普勒在颅骨窗口中作为端点的端点； 2）使用特定的抑制剂和途径蛋白的shRNA敲低，确定从受体和G蛋白到通道的信号转导机制/s从受体和G蛋白到通道，并通过尖锐的电极测量来确定膜电位的尖锐测量； 3）使用新型的GRK2，Paroxetine，血管平滑肌选择性GRK2 - / - 和阻止蛋白 - / - 小鼠以及GRK2和抑制素shRNA腺病毒敲低的新药理抑制剂了解受体调节。 4）确定OPN4受体作为RP小鼠模型中的靶标的作用，并探索在健康人类皮肤循环中介导体内介导的光疗法的机制，以及使用激光流动多普勒和皮肤微透析的主要RP的循环。通过这种方式，我们希望深入了解这种新型途径在健康和疾病中的功能和功能障碍，并确定其作为一种新型治疗靶标的潜力。