Targeting accessory proteins of alpha2C adrenergic receptor in Raynaud Phenomenon

雷诺现象中针对 α2C 肾上腺素受体的辅助蛋白

基本信息

批准号：
8584924
负责人：
Catalin Filipeanu
金额：
$ 3.17万
依托单位：
LSU HEALTH SCIENCES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2013-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8584924
关键词：
ATP phosphohydrolase Adrenergic Antagonists Adrenergic Receptor Adverse effects Affect Amino Acids Binding Biological Markers Blood Vessels Calcium Channel Catecholamines Cell Line Cell membrane Cell surface Characteristics Clinical Clinical Treatment Clinical Trials Collaborations Connective Tissue Diseases Data Dependence Dermatology Diagnosis Diagnostic Disease Disease model Early Diagnosis Early treatment Effectiveness Emotional Stress Endoplasmic Reticulum Event Exposure to Family Foundations Future G-Protein-Coupled Receptors General Population Goals Heat shock proteins Heat-Shock Proteins 90 Human Incidence Individual Investigation Left Limb structure Lupus Erythematosus Mediating Modeling Molecular Molecular Chaperones Molecular and Cellular Biology Muscle Contraction Muscle relaxants Names Norepinephrine Numbness Pain Pathogenesis Pathology Patients Pattern Peripheral Pharmaceutical Preparations Pharmacological Treatment Phase Physiological Pilot Projects Plasma Prevention Proteins Proteomics Rattus Raynaud Disease Raynaud Phenomenon Reticulum Rodent Role Scleroderma Site Skin Smooth Muscle Smooth Muscle Myocytes Syndrome System Temperature Testing Therapeutic Transfection Woman Work Yohimbine base clinically relevant cold temperature digital experience geranylgeranylacetone golgi associated, gamma adaptin homologous, ADP-ribosylation factor interacting protein inhibitor/antagonist member men novel novel strategies novel therapeutic intervention novel therapeutics nucleophosmin overexpression pre-clinical prevent public health relevance receptor response rottlerin trafficking vasoconstriction vibration

项目摘要

DESCRIPTION (provided by applicant): Raynaud Phenomenon (RP) is characterized by exaggerated vasoconstriction in response to cold, emotional stress and vibrations and comprises from Raynaud's Disease (primary or idiopathic) and Raynaud's Syndrome when is associated with another connective tissue disorder like Lupus erythematosus or scleroderma. Although often underestimated and under diagnosed, RP is a common clinical encounter affecting between 3 to 12% of men and 6 to 20 % of women. Unfortunately, a specific treatment of RP is still missing today, the therapeutic approaches consisting only in administration of general smooth muscle relaxants. Moreover, although the disease was described more than 150 years ago, the RP pathogenesis is still not understood and no biological markers or paraclinical investigations were validated for early detection and treatment. Clinical evidence indicate that yohimbine, an alpha2-adrenergic antagonist (A2) is effective in preventing the vasospastic attacks in RP. Based on the anatomical distribution, A2C is the most probable adrenergic receptor subtype responsible for these effects and this view is supported by experimental evidence indicating that the receptor localization at the functional site is enhanced after exposure to low-temperature. However, currently no specific A2C blockers are available and also these antagonists may have numerous side effects precluding their use in the RP's treatment. An alternative approach is to target the accessory proteins controlling the synthesis, maturation and intracellular trafficking of A2C. Pursuing this idea, during the last few years we identified specific proteins interacting with A2C and regulating its function in temperature-sensitive manner. First, we have shown that HSP90 inhibitors are selectively reducing A2C targeting to the functional site. Further, we compared the intracellular trafficking and functional responses of human and rat A2C and we found that even if they are over 90 % homologous, human A2C displays a much greater temperature-dependence, indicating that the exaggerated vasoconstriction observed in RP can be studied using only the human receptor. We further identified the amino acid residues responsible for this particular pattern, due to the differential interactions with RuVBL1 protein. Accordingly, changes in the RuVBL1 cellular levels induced alterations in the temperature- dependent A2C functional responses. Similarly, increases in the cellular levels of another accessory protein named nucleophosmin, greatly enhanced the temperature-dependent A2C activation. Therefore, based on these original findings, the main goal of the present investigation is to determine using human vascular smooth muscle cells from normal and RP donors if HSP90, RuVBL1 and nucleophosmin may be used as biomarkers for early diagnostic and therapy of RP. As HSP90 inhibitors are currently in second phase clinical trial for the treatment of unrelated diseases, the expected results may have an immediate clinical relevance. Also, we aim to establish a new model to test the effectiveness of current and future therapeutic approaches in RP.

描述（由申请人提供）：雷诺现象（RP）的特征是对寒冷、情绪压力和振动做出反应时血管过度收缩，包括雷诺氏病（原发性或特发性）和与另一种结缔组织疾病（如红斑狼疮）相关的雷诺氏综合症或硬皮病。虽然 RP 经常被低估和诊断不足，但它是一种常见的临床疾病，影响 3% 至 12% 的男性和 6% 至 20% 的女性。不幸的是，目前仍然缺乏针对 RP 的特异性治疗方法，治疗方法仅包括施用一般平滑肌松弛剂。此外，尽管这种疾病在 150 多年前就已被描述，但 RP 发病机制仍不清楚，也没有验证用于早期检测和治疗的生物标志物或临床旁研究。临床证据表明，育亨宾（一种 α2 肾上腺素能拮抗剂 (A2)）可有效预防 RP 的血管痉挛发作。根据解剖学分布，A2C 是最有可能导致这些效应的肾上腺素能受体亚型，并且这一观点得到了实验证据的支持，表明功能部位的受体定位在暴露于低温后得到增强。然而，目前还没有特定的 A2C 阻滞剂可用，而且这些拮抗剂可能有许多副作用，妨碍它们在 RP 治疗中的使用。另一种方法是针对控制 A2C 合成、成熟和细胞内运输的辅助蛋白。遵循这一想法，在过去几年中，我们鉴定了与 A2C 相互作用并以温度敏感方式调节其功能的特定蛋白质。首先，我们已经证明 HSP90 抑制剂选择性地减少 A2C 靶向功能位点。此外，我们比较了细胞内运输和功能我们研究了人和大鼠 A2C 的反应，发现即使它们同源性超过 90%，人类 A2C 也表现出更大的温度依赖性，这表明仅使用人类受体就可以研究 RP 中观察到的过度血管收缩。由于差异，我们进一步鉴定了导致这种特定模式的氨基酸残基与 RuVBL1 蛋白的相互作用。因此，RuVBL1 细胞水平的变化引起温度依赖性 A2C 功能反应的改变。同样，另一种称为核磷蛋白的辅助蛋白的细胞水平增加，大大增强了温度依赖性 A2C 激活。因此，基于这些原始发现，本研究的主要目标是确定使用来自正常和 RP 供体的人血管平滑肌细胞是否可以将 HSP90、RuVBL1 和核磷蛋白用作 RP 早期诊断和治疗的生物标志物。由于 HSP90 抑制剂目前正处于治疗无关疾病的第二阶段临床试验中，因此预期结果可能具有直接的临床意义。此外，我们的目标是建立一个新模型来测试当前和未来 RP 治疗方法的有效性。