Synthetic Compartments to Rewire Inflammatory Signaling and Augment Immune Cell Function

University Of Pennsylvania, Philadelphia PA

Investigators

Abstract

Engineered immunotherapies harness the power of the immune system to combat diseases and to attack specified targets, such as cancer cells. While their applications have been transformative, a major challenge toward widespread use is the presence of sustained stimuli in a tissue or tumor microenvironment that strongly inhibit immune cell function. Chronic inflammatory signaling via interferons causes phenomena such as T cell exhaustion and macrophage phenotypic switching. New platforms that enable cells to robustly maintain their effector functions, despite sustained inflammatory hyperstimulation from interferons, are urgently required to drive the field forward. To address this critical unmet need, we propose development of synthetic compartments capable of insulating and sequestering signaling components. This strategy can accelerate reaction rates and rewire signaling, altering flux through the Type I and Type II interferon pathways to restore immune cell competence. We have developed a modular intracellular microcompartment platform based on the self-assembly of specific intrinsically disordered proteins (IDPs). Using this system, along with tagged enzymes, we have demonstrated selective and robust control of component sequestration and release, enabling on demand control of cell behaviors. In preliminary data, we have demonstrated the feasibility of targeting janus kinases (JAKs) to synthetic compartments, feasibility of efficiently sequestering TYK2, feasibility of expressing condensates in human T cells, and feasibility of generating knockouts of TYK2 with reduced Type I signaling. In Aim 1, we propose to further develop this platform to ‘insulate’ and shut down key components of JAK STAT signaling in Type I interferon pathway, which is known to result in T cell exhaustion. We will target and functionally insulate the kinase TYK2 via inducible expression of our synthetic compartments. We will also measure immune-related pathways as validation that our approach is feasible without major alteration of T cell physiology. In Aim 2, we propose to redirect repressive Type I signals to Type II cell stimulatory outputs, effectively reprogramming intracellular inflammatory signaling. In Aim 3, we will use these strategies in an in vitro co-culture model of mouse melanoma and T cells to restore immune cell function under conditions that cause exhaustion. We will test the extent to which transient sequestration of TYK2 shuts down interferon Type I signaling, enabling T cell cells to block tumor cell growth. The future of personalized medicine depends on engineering syngeneic cells that target disease tissues and are resistant to deactivating signals. By harnessing the power of compartmentalization and controlled release, our platform provides a new and versatile toolkit to program immune cell responsiveness for next-generation cell-regenerative therapies.