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）：Raynaud现象（RP）的特征是响应寒冷，情绪压力和振动而夸张的血管收缩，并由Raynaud的疾病（原发性或特发性）和Raynaud's综合征组成，何时与其他结缔组织疾病（如其他结缔组织疾病）相关。尽管经常被低估和被诊断出诊断，但RP是一种常见的临床相遇，影响了3至12％的男性和6％至20％的女性。不幸的是，今天仍缺少对RP的特定治疗方法，这种治疗方法仅包括一般平滑肌松弛剂。此外，尽管150年前描述了该疾病，但仍未了解RP发病机理，也没有验证生物学标记或层状研究以进行早期检测和治疗。临床证据表明，α2-肾上腺素能拮抗剂Yohimbine有效防止RP的血管疗法攻击。基于解剖学分布，A2C是负责这些作用的最可能的肾上腺素能受体亚型，并且通过实验证据支持这种观点，表明在暴露于低温后，功能部位的受体定位会增强。但是，目前尚无特定的A2C阻滞剂可用，而且这些拮抗剂可能会产生许多副作用，无法在RP的治疗中使用。另一种方法是靶向控制A2C的合成，成熟和细胞内运输的辅助蛋白。追求这一想法，在过去的几年中，我们确定了与A2C相互作用的特定蛋白质，并以温度敏感的方式调节其功能。首先，我们已经表明HSP90抑制剂有选择地减少对功能部位的A2C靶向。此外，我们比较了细胞内贩运和功能人和大鼠A2C的反应，我们发现，即使它们超过90％同源，人A2C也会显示出更大的温度依赖性，这表明只能使用人体受体研究RP中观察到的夸张的血管收缩。由于差异，我们进一步识别了该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的依赖性A2C功能响应的变化。同样，另一种名为核磷脂的辅助蛋白的细胞水平增加，大大增强了温度依赖性的A2C激活。因此，基于这些原始发现，本研究的主要目标是确定如果HSP90，RUVBL1和核素蛋白可以用作RP的早期诊断和治疗的生物标志物，则使用正常和RP供体的人血管平滑肌细胞。由于HSP90抑制剂目前正在第二阶段临床试验中治疗无关疾病，因此预期的结果可能具有直接的临床相关性。此外，我们旨在建立一个新模型，以测试RP中当前和未来治疗方法的有效性。