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

• 批准号: 8692479
• 负责人: Catalin Filipeanu
• 金额: $ 7.55万
• 依托单位: HOWARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8692479
• 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

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收缩过度，包括雷诺病（原发性或特发性）和雷诺综合征，当与另一种结缔组织疾病如红斑狼疮或硬皮病相关时。虽然经常被低估和诊断不足，但RP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，目前仍缺少一种针对RP的特殊治疗方法，治疗方法仅包括使用一般的平滑肌松弛剂。此外，尽管这种疾病在150多年前就被描述出来了，但RP的发病机制仍然不清楚，也没有生物标志物或临床研究被证实可以早期发现和治疗。临床证据表明育亨宾，一种α 2-肾上腺素能拮抗剂（A2）可有效预防RP的血管痉挛发作。根据解剖分布，A2C是最有可能导致这些作用的肾上腺素能受体亚型，这一观点得到了实验证据的支持，实验证据表明，低温暴露后，受体在功能部位的定位增强。然而，目前还没有特异性的A2C阻滞剂可用，而且这些拮抗剂可能有许多副作用，因此无法用于RP的治疗。另一种方法是靶向控制A2C合成、成熟和细胞内运输的辅助蛋白。为了实现这一想法，在过去的几年里，我们发现了与A2C相互作用并以温度敏感的方式调节其功能的特定蛋白质。首先，我们已经证明HSP90抑制剂选择性地降低A2C靶向功能位点。此外，我们比较了细胞内运输和功能