Date on Master's Thesis/Doctoral Dissertation

8-2019

Document Type

Master's Thesis

Degree Name

M. Eng.

Department

Bioengineering

Committee Chair

Kopechek, Jonathan

Committee Co-Chair (if applicable)

Roussel, Thomas

Committee Member

Roussel, Thomas

Committee Member

Yaddanapudi, Kavitha

Author's Keywords

CAR-T; t-cells; transfection; sonoporation; electroporation; acute lymphoblastic leukemia

Abstract

Leukemias are the most common form of childhood cancer making up 30% of total pediatric oncological cases, and Acute Lymphoblastic Leukemia (ALL) makes up a significant portion (12%) of the total pediatric cancer diagnoses. In 2017, the FDA approved a successful immunotherapy called CAR-T therapy for the treatment of pediatric B-cell ALL. This therapy includes a CAR (chimeric antigen receptor) that is loaded into the T-cell and expressed. Currently, the loading of the CAR utilizes viral transduction, but consistency issues lead to adverse symptoms in the patients. Better methods of transduction/transfection are being studied in order to improve these consistency concerns. In this thesis, the efficiency of sonoporation as a non-viral method of transfection was assessed. Fluorescein was loaded as a fluorescent model molecule for beginning understanding of the sonoporation efficiency. It was found that by using sonoporation over electroporation for the uptake of fluorescein, the efficiency is improved by 34%. When sonoporation was used for the transfection of GFP plasmid, the same increase was not proven. This leads to the conclusion that without further optimization, sonoporation is successful at loading small molecule such as fluorescein but not those as large as plasmids. With optimization, sonoporation could eventually be used as a non-viral method to transfect T-cells with CARs for CAR-T therapy and the treatment of ALL in both children and adults.

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