Title:
A microfluidics-based paradigm for clinical lentivector gene transfer
A microfluidics-based paradigm for clinical lentivector gene transfer
| dc.contributor.advisor | Lam, Wilbur A. | |
| dc.contributor.author | Tran, Reginald | |
| dc.contributor.committeeMember | Dixon, J. Brandon | |
| dc.contributor.committeeMember | Doering, Christopher B. | |
| dc.contributor.committeeMember | Le Doux, Joseph M. | |
| dc.contributor.committeeMember | Sulchek, Todd | |
| dc.contributor.department | Biomedical Engineering (Joint GT/Emory Department) | |
| dc.date.accessioned | 2018-01-22T21:05:26Z | |
| dc.date.available | 2018-01-22T21:05:26Z | |
| dc.date.created | 2016-12 | |
| dc.date.issued | 2016-11-15 | |
| dc.date.submitted | December 2016 | |
| dc.date.updated | 2018-01-22T21:05:26Z | |
| dc.description.abstract | Ex vivo gene therapy using lentiviral vectors (LVs) is a proven approach to treat and potentially cure many hematologic disorders and cancer, but remains stymied by cumbersome, cost-prohibitive, and scale-limited production processes that cannot meet the demands of current clinical protocols for widespread implementation. In this work, we describe the development, characterization, and application of a microfluidic, mass transport-based approach that overcomes the diffusion limitations of current transduction platforms to enhance LV gene transfer kinetics and efficiency. This novel ex vivo LV transduction platform is flexible in design, easy to use, scalable, and compatible with standard cell culture transduction reagents and LV preparations as it mechanistically relies solely on physical principles. Hematopoietic cell lines, primary human T cells, and primary hematopoietic stem and progenitor cells were used to assess microfluidic transduction, which was demonstrated to occur up to 5-fold faster and required as little as ~1/20th of lentivirus needed in conventional clinical transduction protocols. In vivo application of microfluidics using hematopoietic stem and 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 can significantly be reduced with microfluidics, which can be incorporated with other transduction enhancement strategies. Moreover, when using equal amounts of vector, only hemophiliac mice transplanted with microfluidic-transduced Sca-1+ cells were able to restore plasma fVIII levels to normal, demonstrating the greater efficiency of the microfluidic transduction platform. Overall, this work highlights the utility of microfluidics to overcome diffusion limitations of standard cell culture systems to improve efficiency and utilization of LVs for clinical gene therapy. | |
| dc.description.degree | Ph.D. | |
| dc.format.mimetype | application/pdf | |
| dc.identifier.uri | http://hdl.handle.net/1853/59163 | |
| dc.language.iso | en_US | |
| dc.publisher | Georgia Institute of Technology | |
| dc.subject | Microfluidics | |
| dc.subject | Gene therapy | |
| dc.subject | Hemophilia | |
| dc.subject | Hematology | |
| dc.subject | Cell manufacturing | |
| dc.subject | Cancer immunotherapy | |
| dc.subject | Hematopoietic stem cells | |
| dc.subject | T cells | |
| dc.subject | Mass transport | |
| dc.subject | Lentivirus | |
| dc.title | A microfluidics-based paradigm for clinical lentivector gene transfer | |
| dc.type | Text | |
| dc.type.genre | Dissertation | |
| dspace.entity.type | Publication | |
| local.contributor.advisor | Lam, Wilbur A. | |
| local.contributor.corporatename | Wallace H. Coulter Department of Biomedical Engineering | |
| local.contributor.corporatename | College of Engineering | |
| relation.isAdvisorOfPublication | 1c1e6049-d691-42bf-8032-715acc6c1bfa | |
| relation.isOrgUnitOfPublication | da59be3c-3d0a-41da-91b9-ebe2ecc83b66 | |
| relation.isOrgUnitOfPublication | 7c022d60-21d5-497c-b552-95e489a06569 | |
| thesis.degree.level | Doctoral |