Using open-source parts and 3D-printed components, the Lee lab develops a robotic system for mammalian cell cultures that accurately reproduces user-defined concentration profiles for one or more stimuli, such as cytokines or drugs. The team applies the dynamic stimulation system to investigate NF-kB signaling in single cells exposed to time-varying concentrations of TNF, a molecular network that is often deregulated in autoimmunity and cancer. Cellular responses to dynamic stimuli reveals context-dependent sensitivities and new classes of single cell responses that are distinct from the canonical NF-kB response during persistent stimulation. Guided by computational modeling, the team show that new response classes can be modulated with chemicals that target rates for basal cellular processes, including transcription and translation.
Taken together, the work shows that dynamic stimuli can be used to more accurately recapitulate biological complexity, to reveal hidden capabilities of biological systems, and to provide new opportunities to rationally manipulate disease-associated signaling mechanisms.
Mokashi CS, Schipper DL, Qasaimeh MA, Lee REC. A System for Analog Control of Cell Culture Dynamics to Reveal Capabilities of Signaling Networks. (2019) iScience [Epub ahead of print]