Ryanodine Receptor Calcium Leak in Circulating B-Lymphocytes as a Biomarker in Heart Failure.
Ryanodine Receptor Calcium Leak in Circulating B-Lymphocytes as a Biomarker in Heart Failure.
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DOI：
10.1161/circulationaha.117.032703
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发表时间：
2018-09-11
期刊：
影响因子：
37.8
通讯作者：
Marks AR
中科院分区：
文献类型：
--
作者：
Kushnir A;Santulli G;Reiken SR;Coromilas E;Godfrey SJ;Brunjes DL;Colombo PC;Yuzefpolskaya M;Sokol SI;Kitsis RN;Marks AR
充血性心力衰竭（CHF）管理的进展取决于监测疾病进展和治疗反应的生物标志物。在心脏收缩期间，细胞内Ca 2+通过2型兰尼碱受体/Ca 2+释放通道（RyR 2）从肌浆网（SR）释放到细胞质中。在CHF中，慢性升高的循环儿茶酚胺水平引起RyR 2的病理性重构，导致舒张SR Ca 2+渗漏和心肌收缩力降低。类似地，骨骼肌收缩需要通过1型兰尼碱受体（RyR 1）释放SR Ca 2+，CHF中慢性升高的儿茶酚胺水平导致RyR 1介导的SR Ca 2+泄漏，导致肌病和虚弱。循环B淋巴细胞表达RyR 1和儿茶酚胺反应性信号级联，使其成为CHF中由于RyR通道泄漏导致的细胞内Ca 2+处理缺陷的潜在替代物。从CHF患者、CHF状态-左心室辅助装置（LVAD）后患者和对照组中采集全血。还从患有缺血性CHF、缺血性CHF + S107（特异性减少RyR通道Ca 2+渗漏的药物）和WT对照的小鼠收集血液。通道大分子复合物通过免疫染色RyR 1免疫沉淀淋巴细胞富集制剂进行评估。在不存在和存在排空内质网（ER）内RyR 1 Ca 2+储存的RyR 1激动剂的情况下，使用流式细胞术评估RyR 1 Ca 2+泄漏，以测量B淋巴细胞中的Ca 2+荧光。来自患有CHF的人和小鼠的循环B淋巴细胞表现出重构的RyR 1和减少的ER Ca 2+储存，与慢性细胞内Ca 2+渗漏一致。这种Ca 2+渗漏与循环中的儿茶酚胺水平有关。在用Rycal S107处理的小鼠中，细胞内Ca 2+渗漏显著减少。接受LVAD治疗的CHF患者表现出异质性反应。在CHF中，B淋巴细胞表现出重构的渗漏RyR 1通道和减少的ER Ca 2+储存，与慢性细胞内Ca 2+渗漏一致。使用流式细胞术评估的B淋巴细胞中RyR 1介导的Ca 2+泄漏提供了细胞内Ca 2+处理和全身交感神经负荷的替代测量，为监测对药物和机械CHF治疗的反应提供了新的生物标志物。
Advances in congestive heart failure (CHF) management depend on biomarkers for monitoring disease progression and therapeutic response. During systole, intracellular Ca2+ is released from the sarcoplasmic reticulum (SR) into the cytoplasm through type 2 ryanodine receptor/Ca2+ release channels (RyR2). In CHF, chronically elevated circulating catecholamine levels cause pathologic remodeling of RyR2 resulting in diastolic SR Ca2+ leak, and decreased myocardial contractility. Similarly, skeletal muscle contraction requires SR Ca2+ release through type-1 ryanodine receptors (RyR1), and chronically elevated catecholamine levels in CHF cause RyR1 mediated SR Ca2+ 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 Ca2+ handling due to leaky RyR channels in CHF. Whole blood was collected from patients with CHF, CHF status-post left-ventricular assist devices (LVAD), and controls. Blood was also collected from mice with ischemic CHF, ischemic CHF + S107 (a drug that specifically reduces RyR channel Ca2+ leak), and WT controls. Channel macromolecular complex was assessed by immunostaining RyR1 immunoprecipitated from lymphocyte enriched preparations. RyR1 Ca2+ leak was assessed using flow cytometry to measure Ca2+ fluorescence in B-lymphocytes, in the absence and presence of RyR1 agonists that empty RyR1 Ca2+ stores within the endoplasmic reticulum (ER). Circulating B-lymphocytes from humans and mice with CHF exhibited remodeled RyR1 and decreased ER Ca2+ stores, consistent with chronic intracellular Ca2+ leak. This Ca2+ leak correlated with circulating catecholamine levels. The intracellular Ca2+ leak was significantly reduced in mice treated with the Rycal S107. CHF patients treated with LVAD exhibited a heterogeneous response. In CHF, B-lymphocytes exhibit remodeled leaky RyR1 channels and decreased ER Ca2+ stores consistent with chronic intracellular Ca2+ leak. RyR1 mediated Ca2+ leak in B-lymphocytes assessed using flow cytometry provides a surrogate measure of intracellular Ca2+ handling and systemic sympathetic burden, presenting a novel biomarker for monitoring response to pharmacologic and mechanical CHF therapy.