用于研究心肌细胞增殖的细胞周期可视化工具——实时观察
Cell cycle visualization tools to study cardiomyocyte proliferation in real-time
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影响因子:3.6
分区:生物学2区 / 生化与分子生物学3区
发表日期:2024 Oct
作者:
Rustem Salmenov, Christine Mummery, Menno Ter Huurne
DOI:
10.1098/rsob.240167
摘要
成人人类心脏中的心肌细胞处于静止状态，心脏损伤后丢失的细胞不能通过增殖存活细胞来替代。为理解心肌细胞细胞周期的退出和重新进入机制，科学界已做出大量努力，旨在发现能够刺激心肌细胞增殖和心脏再生的治疗药物。随着大型化合物库和机器人液体处理平台的出现，单次实验中可以筛选数千种条件，但这些筛选的成功依赖于模型的适用性和质量。传统的抗体染色方法成本高、技术难度大且不能区分心肌细胞的有丝分裂与核分裂或胞质分裂的失败，限制了高通量检测的效率。活细胞成像提供了替代方案，便于高通量筛查，但也存在一些局限性。本文将（i）回顾心肌细胞的细胞周期特征；（ii）讨论各种细胞周期荧光报告系统；（iii）推测如何提升其在心肌细胞增殖预测中的价值。最后，探讨如何结合最先进的三维人类心脏类器官平台，识别可能刺激人类心脏再生的促增殖信号通路。
Abstract
Cardiomyocytes in the adult human heart are quiescent and those lost following heart injury are not replaced by proliferating survivors. Considerable effort has been made to understand the mechanisms underlying cardiomyocyte cell cycle exit and re-entry, with view to discovering therapeutics that could stimulate cardiomyocyte proliferation and heart regeneration. The advent of large compound libraries and robotic liquid handling platforms has enabled the screening of thousands of conditions in a single experiment but success of these screens depends on the appropriateness and quality of the model used. Quantification of (human) cardiomyocyte proliferation in high throughput has remained problematic because conventional antibody-based staining is costly, technically challenging and does not discriminate between cardiomyocyte division and failure in karyokinesis or cytokinesis. Live cell imaging has provided alternatives that facilitate high-throughput screening but these have other limitations. Here, we (i) review the cell cycle features of cardiomyocytes, (ii) discuss various cell cycle fluorescent reporter systems, and (iii) speculate on what could improve their predictive value in the context of cardiomyocyte proliferation. Finally, we consider how these new methods can be used in combination with state-of-the-art three-dimensional human cardiac organoid platforms to identify pro-proliferative signalling pathways that could stimulate regeneration of the human heart.