Recent advances in clinical gene therapy highlight the need for improved lentiviral vector (LV) and adeno- associated viral (AAV) vector manufacture and utilization efficiencies for enhanced therapeutics outcomes. Expression Therapeutics in collaboration with Emory University are developing a microfluidic (µFluidic), technology-based solution that overcomes the mass-transport and diffusion limitations of current transduction platforms to enhance ex vivo vector gene transfer kinetics and efficiency. This novel ex vivo transduction platform is flexible in design, scalable, and compatible with standard cell culture transduction reagents and vector preparations as it mechanistically relies solely on physical principles. Using hematopoietic cell lines, primary human T-cells, and primary hematopoietic stem/progenitor cells, we have demonstrated that μFluidic transduction occurs up to 5-fold faster and requires ~1/20th of LV needed in conventional clinical transduction protocols. In vivo application of μFluidics using hematopoietic stem/progenitor cells of C57BL/6J hemophilia A mice transduced with factor VIII-encoding LV and transplanted into hemophilic donors demonstrated that LV usage and transduction time is significantly reduced with our platform, without loss of hematopoietic stem cell engraftment potential. The milestones proposed for the current phase I SBIR application are to 1) design and test microfluidic devices scaled-up to process > 108 target cell for genetically-modified cell product manufacture validation at clinically-relevant scales, 2) validate these devices using clinically-relevant primary human cell targets (CD34+ cells and CD3+ T cells), and 3) explore the opportunity for application of the microfluidic transduction devices to ex vivo AAV gene transfer, which is being pursued clinically for ex vivo gene-editing and transient gene expression applications.
Public Health Relevance Statement: PROJECT NARRATIVE Gene therapy products are currently demonstrating tremendous potential in clinical trials for many genetic disorders and malignancies, but commercialization progress is hampered by manufacturing limitations and costs. The objective of the current project is to commercialize a novel microfluidic transduction device that will dramatically improve gene therapy modalities by reducing costs and time required for the generation of genetically-modified cell therapy products. In the current phase I SBIR project, Expression Therapeutics will focus on continued development of our microfluidic transduction device to achieve proof of concept data at clinically-relevant scales as well as demonstrate expanded utility of the device for ex vivo gene therapy and gene editing-based application.
NIH Spending Category: Bioengineering; Biotechnology; Gene Therapy; Genetics; Hematology; Immunotherapy; Orphan Drug; Rare Diseases; Regenerative Medicine; Stem Cell Research; Stem Cell Research - Nonembryonic - Human; Stem Cell Research - Nonembryonic - Non-Human; Transplantation
Project Terms: adeno-associated viral vector; Affect; base; Blood Coagulation Disorders; cancer immunotherapy; CD3 Antigens; CD34 gene; Cell Culture Techniques; Cell Line; Cell Therapy; cell type; Cells; cellular transduction; Clinical; Clinical Trials; clinically relevant; Collaborations; commercialization; cost; CRISPR/Cas technology; Data; design; Development; Devices; Diffusion; DNA; engineered T cells; Engineering; Engraftment; Evaluation; Factor VIII; flexibility; Foundations; Future; Gene Expression; gene therapy; Gene Transfer; gene transfer vector; Generations; Genes; Genetic Diseases; genetically modified cells; Goals; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hemophilia A; Human; Immunotherapy; improved; in vivo; in vivo evaluation; Kinetics; Label; Laboratories; Lead; lead candidate; Lentivirus Vector; Malignant Neoplasms; Microfluidic Microchips; microfluidic technology; Microfluidics; Modality; Mus; novel; nuclease; particle; Patients; Persons; Phase; preclinical development; Preparation; Process; Production; Program Development; Protocols documentation; prototype; Publications; Quality Control; Reagent; Research; Running; Safety; scale up; Site; Small Business Innovation Research Grant; System; T-Lymphocyte; Testing; Therapeutic; therapeutic gene; therapy outcome; Time; transduction efficiency; Translating; Transplantation; Universities; Validation; vector; Viral; Viral Genes; Viral Vect
