Applications of a robust, vector-free intracellular delivery platform for immune cells

ABSTRACTTraditional vaccines have relied upon production of antibodies to surface exposed antigens to clear a pathogen from the body and prevent infectionyet this approach has proven ineffective against intracellular bacteriaparasitesand certain virusesOrganisms that infect a cell and replicate intracellularly are protected from antibodies and can only be cleared by cytotoxic T cellsCTLsthat target the infected cell and destroy itA CTL response has also shown to be beneficial in treating cancerThis is particularly clear in the field of immuno oncology where checkpoint inhibitors are revolutionizing the field by removing theoff switchfrom T cells and enabling them to effectively fight multiple cancer typesAlthough the need for treatments that produce a robust CTL response is clearprevious efforts have failed due to the inability to deliver the antigen to the cytosola requirement for a robust CTL responseAntigen delivery alternativesincluding electroporation and nanoparticle based systemscan often result in toxicityoff target effectslow efficiency and or accumulation of material in endosomal compartmentsFurthermorethese processes are not amenable to scalable deploymentlimiting the number of patients able to be treatedThusto develop the next generation of CTL driven therapiesthe field requires new technologies capable of versatileefficient and non toxic delivery of antigensIn Phase IISQZ Biotechnologies proposes to develop our vector free microfluidic platform to deliver antigens with high efficiency directly into the cytosol of dendritic cells that can then be used for the treatment of infectious diseases and cancerThe principle underlying this approach is temporary membrane disruption by rapid mechanical deformationorsqueezingto facilitate cytosolic delivery of antigensIn SBIR Phase Iwe demonstrated the feasibility of squeezing immune cells to deliver antigens and showed that with this processwe observe CTL responses that are more effective than current methodsThrough cytosolic antigen deliverywe can overcome key limitations related to scalability and efficacyIn Phase IIwe willAIMmeasure antigen specific CTL activation and proliferation using SQZed mouse and human dendritic cellsAIMmeasure efficacy of antigenspecific cytotoxic T cell responses generated in vivo through tumor and infectious disease models andAIMdetermine if cell lysate is a potential antigen source to elicit a CTL responseTogether this data will drive commercialization and partnerships for the clinical development of our novel squeezing platform Project Narrative Our ability to empower our immune system to respond to various viral diseases and cancer is limited by our ability to engineer our immune cellsWe propose to break this paradigm and commercialize a promising newmicrofluidics baseddelivery platform that relies on the temporary disruption of the cell membrane to facilitate antigen delivery directly into the cell cytoplasm to empower our immune systemOur Phase I data has demonstrated that the proposed technology can overcome many of the disadvantages of existing treatment methodologies and in Phase II we will further de risk our next generation cell therapy platform