Three-dimensional (3D) human induced pluripotent stem cell-derived engineered cardiac tissues (hiPSC-ECTs) have emerged as a promising alternative to two-dimensional hiPSC-cardiomyocyte monolayer systems because hiPSC-ECTs are a closer representation of endogenous cardiac tissues and more faithfully reflect the relevant cardiac pathophysiology. The ability to perform functional and molecular assessments using the same hiPSC-ECT construct would allow for more reliable correlation between observed functional performance and underlying molecular events, and thus is critically needed. Herein, for the first time, we have established an integrated method that permits sequential assessment of functional properties and top-down proteomics from the same single hiPSC-ECT construct. We quantitatively determined the differences in isometric twitch force and the sarcomeric proteoforms between two groups of hiPSC-ECTs that differed in the duration of time of 3D-ECT culture. Importantly, by using this integrated method we discovered a new and strong correlation between the measured contractile parameters and the phosphorylation levels of alpha-tropomyosin between the two groups of hiPSC-ECTs. The integration of functional assessments together with molecular characterization by top-down proteomics in the same hiPSC-ECT construct enables a holistic analysis of hiPSC-ECTs to accelerate their applications in disease modeling, cardiotoxicity, and drug discovery. Data are available via ProteomeXchange with identifier PXD022814.

中文翻译：

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用于人类诱导多能干细胞衍生的工程心脏组织标准化评估的功能集成自上而下蛋白质组学

三维 (3D) 人类诱导多能干细胞衍生的工程心脏组织 (hiPSC-ECT) 已成为二维 hiPSC-心肌细胞单层系统的有希望的替代品，因为 hiPSC-ECT 更接近内源性心脏组织等忠实反映了相关的心脏病理生理。使用相同的 hiPSC-ECT 结构进行功能和分子评估的能力将允许观察到的功能性能和潜在的分子事件之间更可靠的相关性，因此是迫切需要的。在这里，我们第一次建立了一种集成方法，该方法允许对同一单个 hiPSC-ECT 构建体的功能特性和自上而下的蛋白质组学进行顺序评估。我们定量确定了两组 hiPSC-ECT 之间等长抽搐力和肌节蛋白形式的差异，这些差异在 3D-ECT 培养的持续时间上有所不同。重要的是，通过使用这种综合方法，我们发现测量的收缩参数与两组 hiPSC-ECT 之间的 α-原肌球蛋白磷酸化水平之间存在新的强相关性。在同一个 hiPSC-ECT 结构中通过自上而下的蛋白质组学将功能评估与分子表征相结合，可以对 hiPSC-ECT 进行整体分析，以加速其在疾病建模、心脏毒性和药物发现中的应用。数据可通过 ProteomeXchange 获得，标识符为 PXD022814。