Effective T cell priming is crucial to induce anti-tumor CD8+ T cell responses and requires the efficient presentation of antigen on major histocompatibility complex class I (MHC-I) by antigen presenting cells (APCs). Previous efforts using dendritic cells to prime CD8+ T cell responses have proven difficult due to limited cell availability in the blood and challenges delivering antigen to the APC cytosol, a necessary step for MHC-I presentation and CD8+ T cell activation. To overcome this limitation, we deliver antigen directly to the cytosol of target APCs using the microfluidics-based SQZ platform. SQZ uniquely facilitates antigen loading into both professional and unconventional APCs, including B cells, T cells, and heterogenous populations of cells, which can be easily obtained directly from the blood. Protein and peptide antigens are delivered using SQZ to each of these APCs effectively, leading to efficient presentation of immunogenic epitopes on MHC-I. Here, we demonstrate that murine SQZ-APCs can stimulate antigen-specific CD8+ T cell responses in vitro and in vivo as measured by expansion of antigen-specific T cells and production of IFNγ. In the TC-1 tumor model for HPV-associated cancers, antigen-loaded SQZ-APCs have strong anti-tumor effects both prophylactically and therapeutically. Following therapeutic immunization, the anti-tumor responses correlate with an increase in antigen-specific CD8+ tumor infiltrating lymphocytes compared to untreated mice. In addition, compared to a traditional subcutaneous peptide vaccine, SQZ-APCs elicit a five-fold greater intratumoral CD8+ T cell response and drive significantly more tumor growth inhibition. Importantly, this SQZ-enabled cancer cell therapy translates to human B cells, T cells, and heterogenous populations of cells engineered to function as APCs. When a peptide is delivered to the cytosol using SQZ, all of these primary human cells activate antigen-specific CD8+ T cell responses in vitro by stimulation of IFNγ from antigen-specific CD8+ T cell responders. In comparison to cells incubated in the presence of peptide antigen, SQZ-APCs stimulate a 10-fold increase in IFNγ production from antigen-specific CD8+ responder T cells (n=13 donors). Finally, the SQZ process has been scaled to engineer human SQZ-APCs in preparation for clinical trials with a throughput of greater than 4 billion cells SQZ’d per minute. Collectively, these findings highlight the significant clinical potential of the SQZ platform to engineer potent APCs for a new generation of cancer cell therapies.
Citation Format: Kelan A. Hlavaty, Matthew G. Booty, Scott Loughhead, Katarina Blagovic, Alfonso Vicente-Suarez, Defne Yarar, Howard Bernstein, Armon Sharei. Engineering a new generation of cell therapies for solid tumor oncology using the SQZ platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3187.