A 3D Bioprinted Human Cardiac Cell Platform to Model the Pathophysiology of Diabetes
A 3D Bioprinted Human Cardiac Cell Platform to Model the Pathophysiology of Diabetes
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用于模拟糖尿病病理生理学的 3D 生物打印人类心肌细胞平台
DOI：
10.1161/res.127.suppl_1.465
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发表时间：
2020
影响因子：
20.1
通讯作者：
Chattopadhyay, M
中科院分区：
文献类型：
--
作者：
Joddar, B;AnilKumar, S;Alonzo, M;Thakur, V;Chattopadhyay, M
受潜在高血糖（高葡萄糖/血糖）状况影响的II型糖尿病（T2 D）患者通常患有心脏萎缩，导致组织质量减少和心脏健康衰弱。为了了解心脏萎缩进展过程中的病理生理机制，从含有人心脏细胞（包括心肌细胞（CM）、成纤维细胞（CF）和内皮细胞（EC））的水凝胶混合物开发了3D生物打印类器官平台，以模拟体内组织的功能。使用正常血糖或高血糖条件培养类器官。确认了这些类器官中肌心蛋白（Myocd）、肌钙蛋白-I（TRP-I）、成纤维细胞蛋白-I（FSP-1）和内皮素-I（ET-1）的基本生物标志物的表达。为了评估高血糖期间的生理细胞连接，在CX-43阻断剂gap 26存在下评估连接蛋白-43（CX-43）的存在。使用表观基因组工具来同时询问通过组蛋白3赖氨酸9单甲基化（H3 K9 me 1）进行的组蛋白修饰，沿着使用正常与高血糖条件培养的心脏类器官中的炎症介质，例如高迁移率族蛋白1（HMGB 1）和Toll样受体4（TLR 4）的共调节。暴露于高糖的类器官显示H3 K9 me 1以及炎症介质HMGB 1和TLR 4的表达增加。与血糖正常条件相比，高血糖还表现出类器官中Myocd和FSP-1表达的改变。在高血糖下培养的类器官中，用gap 26处理显著影响CX-43表达，这表明与长期糖尿病相关的高葡萄糖条件可能导致CM-CF偶联受损，这对维持心脏功能至关重要。H3 K9 me 1水平升高提示Myocd表达降低，这可能导致CM变性。表观遗传修饰包括在高血糖条件下调节心肌基因和间隙连接蛋白的组蛋白甲基化的改变，可能导致心脏萎缩。我们希望建立一个实际的T2 D患者iPSC细胞衍生的心脏平台，为该领域提供新的治疗机会。
Type-II diabetes (T2D) patients affected by underlying hyperglycemic (high glucose/blood sugar) conditions often suffer from cardiac atrophy, resulting in tissue mass reduction and debilitating cardiac health. To understand pathophysiological mechanisms during progression of cardiac atrophy, a 3D bioprinted organoid platform was developed from a mixture of hydrogels containing human cardiac cells, including cardiomyocytes (CM), fibroblasts (CF) and endothelial cells (EC), to mimic the functionality of the in-vivo tissue. The organoids were cultured using normoglycemic- or hyperglycemic-conditions. The expression of essential biomarkers in these organoids, for myocardin (Myocd), troponin-I (TRP-I), fibroblast protein-1 (FSP-1) and endothelin-1 (ET-1) was confirmed. To assess the physiological cellular connections during hyperglycemia, the presence of Connexin-43 (CX-43) was assessed in the presence of a CX-43 blocker, gap26. Epigenomic tools were used to simultaneously interrogate histone-modifications by histone 3 lysine 9 mono-methylation (H3K9me1) along with the co-regulation of inflammatory mediators, such as the high mobility group box 1 (HMGB1) and toll like receptor 4 (TLR4) in the cardiac organoids cultured using normal versus hyperglycemic conditions. Organoids exposed to high glucose showed an increased expression of H3K9me1 as well as inflammatory mediators HMGB1 and TLR4. Hyperglycemia also exhibited alterations in expression of Myocd and FSP-1 in the organoids, compared to normoglycemic conditions. Treatment with gap26 affected the CX-43 expression significantly, in organoids cultured under hyperglycemia suggesting that high glucose conditions associated with prolonged diabetes may lead to compromised CM-CF coupling, essential for maintenance of cardiac functionality. Increased levels of H3K9me1 suggest decreased expression of Myocd, which may lead to CM degeneration. Epigenetic modifications including alterations in histone methylation in regulation of the myocardial genes and gap junction proteins under hyperglycemic conditions, may lead to cardiac atrophy. We expect to establish an actual T2D patient iPSC cell derived cardiac platform, to offer new therapeutic opportunities within the field.