Feedback responsive cell factories for biomanufacturing

Mammalian cells rely on complex and highly dynamic gene networks to maintain cellular homeostasis in response to environmental stimuli and intracellular signals. Efficient cellular reprogramming thus requires integration of orthogonal components for cell engineering with innate cellular networks through feedback control systems. We explored the use of post-translational tools for superior feedback regulation of dynamic behaviors. Specifically, we demonstrated efficient detection and manipulation of the signaling network mediating the cellular stress response for the design of high producing cell lines for protein manufacturing and for the development of cell therapies for sustained protein secretion.

Our approach is based on the use of the NanoDeg (ACS Synth Biol. 2018 Feb 16;7(2):540-552) – a bifunctional system that mediates proteasomal degradation of a target protein with high specificity and exquisite control over rate of decay. Iterations of experimental measurements and predictive modeling resulted in the design of a platform technology for quantitative, multiplexed profiling of gene expression signatures of the stress response with high sensitivity and dynamic resolution of the stimulus causing proteotoxic stress (Nature Chem Biol. 2020 May;16(5):520-528).

Current work is focused on deploying this technology for manipulating stress signaling pathways in response to feedback signals generated at different stages of protein production, with the overall goal to engineer cells for enhanced and sustained expression of recombinant proteins. This talk with describe strategies for engineering cells based on feedback signals that enhance the stress attenuation response induced in response to proteotoxic stress and delay ER-stress induced apoptosis with respect to activation of the stress attenuation response.