Han-Mo Yang, Joo-Eun Lee, Ju-Young Kim, Jihye You, Joonoh Kim, Hak Seung Lee, Hee Min Yoo, Min Gyu Kong, Jung-Kyu Han, Hyun-Jai Cho, Kyung Woo Park, Hyun-Jae Kang, Bon-Kwon Koo, Young-Bae Park, Hyo-Soo Kim
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As compared with vascular smooth muscle cells (VSMCs) differentiated from iPSCs of normal subjects with negative provocation test, VSA patient-specific iPSCs-derived VSMCs showed very strong contraction in response to stimulants. Moreover, VSA patient-specific VSMCs exhibited a significant increase in stimulation-induced intracellular calcium efflux (Changes in the relative fluorescence unit [ΔF/F]; Control group vs. VSA group, 2.89 ± 0.34 vs. 10.32 ± 0.51, p < 0.01), and exclusively induced a secondary or tertiary peak of calcium efflux, suggesting that those findings could be diagnostic cut-off values for VSA. The observed hyperreactivity of VSA patient-specific VSMCs were caused by the upregulation of sarco/endoplasmic reticulum Ca<sup>2+</sup>-ATPase 2a (SERCA2a) due to its enhanced small ubiquitin-related modifier (SUMO)ylation. This increased activity of SERCA2a was reversed by treatment with ginkgolic acid, an inhibitor of SUMOylated E1 molecules (pi/µg protein; VSA group vs. VSA + ginkgolic acid, 52.36 ± 0.71 vs. 31.93 ± 1.13, p < 0.01).</p><p><strong>Conclusions: </strong>Our findings showed that abnormal calcium handling in sarco/endoplasmic reticulum could be induced by the enhanced SERCA2a activity in patients with VSA, leading to spasm. 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This increased activity of SERCA2a was reversed by treatment with ginkgolic acid, an inhibitor of SUMOylated E1 molecules (pi/µg protein; VSA group vs. VSA + ginkgolic acid, 52.36 ± 0.71 vs. 31.93 ± 1.13, p < 0.01).</p><p><strong>Conclusions: </strong>Our findings showed that abnormal calcium handling in sarco/endoplasmic reticulum could be induced by the enhanced SERCA2a activity in patients with VSA, leading to spasm. 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引用次数: 0
摘要
背景:虽然血管痉挛性心绞痛(VSA)已知是由冠状动脉痉挛引起的，但没有研究完全阐明其确切的潜在机制。此外，为了确认VSA，患者应进行有创冠状动脉造影和痉挛激发试验。在此，我们使用外周血源性诱导多能干细胞(iPSCs)研究VSA的病理生理，并开发了VSA的体外诊断方法。方法和结果:用10 mL VSA患者外周血制备iPSCs，并将其分化为靶细胞。与阴性激发试验的正常受试者iPSCs分化的血管平滑肌细胞(VSMCs)相比，VSA患者特异性iPSCs衍生的血管平滑肌细胞在兴奋剂作用下表现出非常强的收缩反应。此外，VSA患者特异性VSMCs表现出刺激诱导的细胞内钙外排显著增加(相对荧光单位的变化[ΔF/F];对照组与VSA组相比，p 2+- atp酶2a (SERCA2a)的差异为2.89±0.34比10.32±0.51，这是由于其小泛素相关修饰子(SUMO)的磷酸化增强。这种增加的SERCA2a活性被银杏酸处理逆转，银杏酸是SUMOylated E1分子的抑制剂(pi/µg蛋白;VSA组vs. VSA +银杏酸组，52.36±0.71 vs. 31.93±1.13,p结论:VSA患者SERCA2a活性增强可诱导sarco/内质网钙处理异常，导致痉挛。这种新的冠状动脉痉挛机制可能对VSA的药物开发和诊断有用。
Identification of cell-biologic mechanisms of coronary artery spasm and its ex vivo diagnosis using peripheral blood-derived iPSCs.
Background: Although vasospastic angina (VSA) is known to be caused by coronary artery spasm, no study has fully elucidated the exact underlying mechanism. Moreover, in order to confirm VSA, patients should undergo invasive coronary angiography with spasm provocation test. Herein, we investigated the pathophysiology of VSA using peripheral blood-derived induced pluripotent stem cells (iPSCs) and developed an ex vivo diagnostic method for VSA.
Methods and results: With 10 mL of peripheral blood from patients with VSA, we generated iPSCs and differentiated these iPSCs into target cells. As compared with vascular smooth muscle cells (VSMCs) differentiated from iPSCs of normal subjects with negative provocation test, VSA patient-specific iPSCs-derived VSMCs showed very strong contraction in response to stimulants. Moreover, VSA patient-specific VSMCs exhibited a significant increase in stimulation-induced intracellular calcium efflux (Changes in the relative fluorescence unit [ΔF/F]; Control group vs. VSA group, 2.89 ± 0.34 vs. 10.32 ± 0.51, p < 0.01), and exclusively induced a secondary or tertiary peak of calcium efflux, suggesting that those findings could be diagnostic cut-off values for VSA. The observed hyperreactivity of VSA patient-specific VSMCs were caused by the upregulation of sarco/endoplasmic reticulum Ca2+-ATPase 2a (SERCA2a) due to its enhanced small ubiquitin-related modifier (SUMO)ylation. This increased activity of SERCA2a was reversed by treatment with ginkgolic acid, an inhibitor of SUMOylated E1 molecules (pi/µg protein; VSA group vs. VSA + ginkgolic acid, 52.36 ± 0.71 vs. 31.93 ± 1.13, p < 0.01).
Conclusions: Our findings showed that abnormal calcium handling in sarco/endoplasmic reticulum could be induced by the enhanced SERCA2a activity in patients with VSA, leading to spasm. Such novel mechanisms of coronary artery spasm could be useful for drug development and diagnosis of VSA.
期刊介绍：
Biomaterials Research, the official journal of the Korean Society for Biomaterials, is an open-access interdisciplinary publication that focuses on all aspects of biomaterials research. The journal covers a wide range of topics including novel biomaterials, advanced techniques for biomaterial synthesis and fabrication, and their application in biomedical fields. Specific areas of interest include functional biomaterials, drug and gene delivery systems, tissue engineering, nanomedicine, nano/micro-biotechnology, bio-imaging, regenerative medicine, medical devices, 3D printing, and stem cell research. By exploring these research areas, Biomaterials Research aims to provide valuable insights and promote advancements in the biomaterials field.
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