The balance of pro-apoptotic and pro-survival proteins defines a cell’s fate. These processes are controlled through an interdependent and finely tuned protein network that enables survival or leads to apoptotic cell death. The caspase family of proteases is central to this apoptotic network, with initiator and executioner caspases, and their interaction partners, regulating and executing apoptosis. In this work, we interrogate and modulate this network by exogenously introducing specific initiator or executioner caspase proteins. Each caspase is exogenously introduced using redox-responsive polymeric nanogels. Although caspase-3 might be expected to be the most effective due to the centrality of its role in apoptosis and its heightened catalytic efficiency relative to other family members, we observed that caspase-7 and caspase-9 are the most effective at inducing apoptotic cell death. By critically analyzing the introduced activity of the delivered caspase, the pattern of substrate cleavage, as well as the ability to activate endogenous caspases, we conclude that the efficacy of each caspase correlated with the levels of pro-survival factors that both directly and indirectly impact the introduced caspase. These findings lay the groundwork for developing methods for exogenous introduction of caspases as a therapeutic option that can be tuned to the apoptotic balance in a proliferating cell.

中文翻译：

---

Initiator 和 Executioner Caspase 的外源性引入导致不同的凋亡结果

促凋亡蛋白和促存活蛋白的平衡决定了细胞的命运。这些过程是通过相互依赖和微调的蛋白质网络控制的，该网络能够存活或导致细胞凋亡。Caspase 蛋白酶家族是该细胞凋亡网络的核心，具有启动子和执行子 Caspase，以及它们的相互作用伙伴，调节和执行细胞凋亡。在这项工作中，我们通过外源性引入特定的启动子或执行者半胱天冬酶蛋白来询问和调节这个网络。每种半胱天冬酶都是使用氧化还原响应性聚合物纳米凝胶外源引入的。尽管 caspase-3 可能是最有效的，因为它在细胞凋亡中的作用居于核心地位，并且相对于其他家族成员而言具有更高的催化效率，我们观察到 caspase-7 和 caspase-9 在诱导细胞凋亡方面最有效。通过批判性地分析所传递的 caspase 的引入活性、底物裂解的模式以及激活内源性 caspase 的能力，我们得出结论，每种 caspase 的功效与直接和间接影响的促存活因子的水平相关引入的半胱天冬酶。这些发现为开发外源性引入半胱天冬酶作为治疗选择的方法奠定了基础，该方法可以调节增殖细胞的凋亡平衡。我们得出结论，每种半胱天冬酶的功效与直接和间接影响引入的半胱天冬酶的促存活因子的水平相关。这些发现为开发外源性引入半胱天冬酶作为治疗选择的方法奠定了基础，该方法可以调节增殖细胞的凋亡平衡。我们得出结论，每种半胱天冬酶的功效与直接和间接影响引入的半胱天冬酶的促存活因子的水平相关。这些发现为开发外源性引入半胱天冬酶作为治疗选择的方法奠定了基础，该方法可以调节增殖细胞的凋亡平衡。