Mechanical Immunoengineering of T Cells for Enhanced Cancer Immunotherapy

Cancer is one of the leading causes of death globally. Immunotherapy represents one of the most promising cancer treatments inducing complete and durable responses in a fraction of cancer patients. Despite the unprecedented clinical success achieved by T cell-based cancer immunotherapies, there are tremendous outstanding challenges, including low response rate and severe toxicities, which substantially limit the clinical benefits in a majority of cancer patients. At present, enormous efforts have been spent on modulating biochemical interactions for improving the efficacy and safety of T cell-based cancer immunotherapy. Besides biochemical exchanges, mechanical interactions are universal in every step of T cell immunity and are crucial in regulating T cell functions. Thus, immunoengineering approaches that modulate mechanical interactions in T cell immunity hold great promise to improve current cancer immunotherapy. This thesis aims to develop novel mechanical immunoengineering approaches for improving cancer immunotherapy. Specifically, it includes the following parts:Developing a spiky artificial antigen-presenting cell (aAPC) to enhance ex vivo T cell expansion. Ex vivo expansion of T cells is critical in adoptive T cell therapy (ACT) to ensure an effective therapeutic dosage for cancer treatment. By mimicking the dendritic morphology of dendritic cells, I developed a spiky TiO2 microparticle presenting T cell stimulatory ligands as an aAPC to enhance ex vivo T cell expansion. The spiky aAPC outperformed the smooth counterpart, as well as Dynabead®, a standard aAPC used in clinical manufacturing of ACT therapy, for T cell activation and expansion depending on F-actin polymerization in T cells. Therefore, this finding identifies the morphology of aAPCs as a critical parameter to improve ex vivo T cell manufacturing for ACT therapy.Boosting T cell-mediated cytotoxicity by stiffening cancer cells. I discovered that cancer cell softening derived from cholesterol enrichment in the plasma membrane led to resistance to T cell-mediated cytotoxicity. Stiffening cancer cells via membrane cholesterol depletion substantially enhanced T cell-mediated cytotoxicity against cancer cells through augmenting T cell mechanical force but not any known biochemical pathways of T cell-mediated cytotoxicity. Cancer cell stiffening intervention in vivo enhanced the efficacy of ACT therapy in multiple preclinical solid tumor models. Cancer cell softness thus represents a new immune checkpoint of mechanical basis, and therapeutically targeting this mechanical immune checkpoint has the potential to improve the clinical response of T cell-based cancer immunotherapy.Utilizing T cell force as a trigger to achieve specific release of anti-cancer drugs. Specific delivery of T cell supporting drugs and/or anti-cancer drugs into tumors has the potential to increase the response rate of ACT therapy without causing systemic toxicity. To this end, I developed a T cell force-responsive drug release system based on a mesoporous silica microparticle with DNA force sensors as gatekeepers. T cell force as a highly specific signal upon recognizing cancer cells presenting cognate antigens was shown to trigger specific drug release when T cell receptor signaling was activated. Further, the T cell force-responsive drug release system could deliver anti-cancer drugs in vitro and in vivo in a T cell force-dependent manner and holds promise to enhance cancer immunotherapy.