循环 B 淋巴细胞兰尼碱受体钙漏作为心力衰竭的生物标志物。
Ryanodine Receptor Calcium Leak in Circulating B-Lymphocytes as a Biomarker in Heart Failure.
机构信息
Department of Physiology and Cellular Biophysics, Clyde and Helen Wu Center for Molecular Cardiology, Columbia University College of Physicians and Surgeons, New York (A.K., G.S., S.R.R., A.R.M.).
Department of Medicine, Division of Cardiology, Columbia University Medical Center, New York (A.K., E.C., S.J.G., D.L.B., P.C.C., M.Y., A.R.M.).
出版信息
Circulation. 2018 Sep 11;138(11):1144-1154. doi: 10.1161/CIRCULATIONAHA.117.032703.
BACKGROUND
Advances in congestive heart failure (CHF) management depend on biomarkers for monitoring disease progression and therapeutic response. During systole, intracellular Ca is released from the sarcoplasmic reticulum into the cytoplasm through type-2 ryanodine receptor/Ca release channels. In CHF, chronically elevated circulating catecholamine levels cause pathological remodeling of type-2 ryanodine receptor/Ca release channels resulting in diastolic sarcoplasmic reticulum Ca leak and decreased myocardial contractility. Similarly, skeletal muscle contraction requires sarcoplasmic reticulum Ca release through type-1 ryanodine receptors (RyR1), and chronically elevated catecholamine levels in CHF cause RyR1-mediated sarcoplasmic reticulum Ca leak, contributing to myopathy and weakness. Circulating B-lymphocytes express RyR1 and catecholamine-responsive signaling cascades, making them a potential surrogate for defects in intracellular Ca handling because of leaky RyR channels in CHF.
METHODS
Whole blood was collected from patients with CHF, CHF following left-ventricular assist device implant, and controls. Blood was also collected from mice with ischemic CHF, ischemic CHF+S107 (a drug that specifically reduces RyR channel Ca leak), and wild-type controls. Channel macromolecular complex was assessed by immunostaining RyR1 immunoprecipitated from lymphocyte-enriched preparations. RyR1 Ca leak was assessed using flow cytometry to measure Ca fluorescence in B-lymphocytes in the absence and presence of RyR1 agonists that empty RyR1 Ca stores within the endoplasmic reticulum.
RESULTS
Circulating B-lymphocytes from humans and mice with CHF exhibited remodeled RyR1 and decreased endoplasmic reticulum Ca stores, consistent with chronic intracellular Ca leak. This Ca leak correlated with circulating catecholamine levels. The intracellular Ca leak was significantly reduced in mice treated with the Rycal S107. Patients with CHF treated with left-ventricular assist devices exhibited a heterogeneous response.
CONCLUSIONS
In CHF, B-lymphocytes exhibit remodeled leaky RyR1 channels and decreased endoplasmic reticulum Ca stores consistent with chronic intracellular Ca leak. RyR1-mediated Ca leak in B-lymphocytes assessed using flow cytometry provides a surrogate measure of intracellular Ca handling and systemic sympathetic burden, presenting a novel biomarker for monitoring response to pharmacological and mechanical CHF therapy.
背景
充血性心力衰竭 (CHF) 管理的进展依赖于生物标志物来监测疾病进展和治疗反应。在收缩期，细胞内 Ca 通过 2 型兰尼碱受体/Ca 释放通道从肌浆网释放到细胞质中。在 CHF 中，慢性升高的循环儿茶酚胺水平导致 2 型兰尼碱受体/Ca 释放通道发生病理性重塑，导致舒张期肌浆网 Ca 渗漏和心肌收缩力降低。同样，骨骼肌收缩需要通过 1 型兰尼碱受体 (RyR1) 释放肌浆网 Ca，而 CHF 中慢性升高的儿茶酚胺水平导致 RyR1 介导的肌浆网 Ca 渗漏，导致肌肉病和无力。循环 B 淋巴细胞表达 RyR1 和儿茶酚胺反应性信号级联，使它们成为细胞内 Ca 处理缺陷的潜在替代物，因为 CHF 中的漏 RyR 通道。
方法
从 CHF 患者、左心室辅助装置植入后的 CHF 患者和对照者采集全血。还从缺血性 CHF 小鼠、缺血性 CHF+S107（一种专门减少 RyR 通道 Ca 渗漏的药物）和野生型对照小鼠中采集血液。通过免疫染色从富含淋巴细胞的制剂中免疫沉淀 RyR1 来评估通道大分子复合物。使用流式细胞术评估 RyR1 Ca 渗漏，以测量无 RyR1 激动剂和有 RyR1 激动剂时 B 淋巴细胞内的 Ca 荧光，RyR1 激动剂可排空内质网内的 RyR1 Ca 储存。
结果
来自 CHF 患者和小鼠的循环 B 淋巴细胞表现出重塑的 RyR1 和减少的内质网 Ca 储存，这与慢性细胞内 Ca 渗漏一致。这种 Ca 渗漏与循环儿茶酚胺水平相关。用 Rycal S107 治疗的小鼠的细胞内 Ca 渗漏明显减少。接受左心室辅助装置治疗的 CHF 患者表现出异质性反应。
结论
在 CHF 中，B 淋巴细胞表现出重塑的渗漏性 RyR1 通道和减少的内质网 Ca 储存，这与慢性细胞内 Ca 渗漏一致。使用流式细胞术评估的 B 淋巴细胞中的 RyR1 介导的 Ca 渗漏提供了细胞内 Ca 处理和全身交感神经负荷的替代测量，为监测药理学和机械性 CHF 治疗的反应提供了一种新型生物标志物。
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