Barrios Daniela, Bachhav Bhagyashree, Carlos-Alcalde Wendolyn, Llanos Carlos D, Zhou Wenchang, Segatori Laura
Department of Bioengineering, Rice University, Houston, TX, USA.
Department of Chemical and Biomolecular Engineering, Rice University, Houston, TX, USA.
Nat Commun. 2025 May 2;16(1):4106. doi: 10.1038/s41467-025-58994-x.
Engineering cell factories that support the production of large quantities of protein therapeutics remains a significant biomanufacturing challenge. The overexpression of secretory proteins causes proteotoxic stress, affecting cell viability and protein productivity. Proteotoxic stress leads to the activation of the Unfolded Protein Response (UPR), a series of signal transduction pathways regulating protein quality control mechanisms aimed at restoring homeostasis. Sustained UPR activation culminates with the induction of apoptosis. Current strategies for enhancing the production of therapeutic proteins have focused on the deregulated modulation of key components of the UPR. These strategies have resulted in limited and often protein-specific improvements as they may lead to adaptation and cell toxicity and do not account for natural population heterogeneities. We report here feedback-responsive cell factories that sense proteotoxic stress and, in response, modulate the UPR to enhance stress attenuation and delay cell death, addressing the limitations of current strategies. We demonstrate that our cell engineering approach enables dynamic UPR modulation upon proteotoxic stress. The sense-and-respond systems that mediate dynamic UPR modulation enhance the production of the therapeutic enzyme tissue plasminogen activator and the bispecific antibody blinatumomab. Our feedback-responsive cell factories provide an innovative strategy for dynamically adjusting the innate cellular stress response and enhancing therapeutic protein manufacturing.
构建能够支持大量生产蛋白质疗法药物的细胞工厂仍然是生物制造领域的一项重大挑战。分泌蛋白的过表达会导致蛋白毒性应激，影响细胞活力和蛋白质生产力。蛋白毒性应激会导致未折叠蛋白反应（UPR）的激活，这是一系列调节蛋白质质量控制机制以恢复体内平衡的信号转导途径。持续的UPR激活最终会诱导细胞凋亡。目前提高治疗性蛋白质产量的策略主要集中在对UPR关键组分的失控调节上。这些策略导致的改善有限且往往具有蛋白质特异性，因为它们可能导致细胞适应性和毒性，并且没有考虑到天然群体的异质性。我们在此报告了一种反馈响应型细胞工厂，它能够感知蛋白毒性应激，并相应地调节UPR，以增强应激缓解并延迟细胞死亡，解决了当前策略的局限性。我们证明，我们的细胞工程方法能够在蛋白毒性应激时实现动态UPR调节。介导动态UPR调节的传感与响应系统提高了治疗性酶组织型纤溶酶原激活剂和双特异性抗体博纳吐单抗的产量。我们的反馈响应型细胞工厂为动态调节先天性细胞应激反应和增强治疗性蛋白质制造提供了一种创新策略。
