PhD Proposal by Tong Yu

Event Details
  • Date/Time:
    • Wednesday December 1, 2021
      10:00 am - 12:00 pm
  • Location: Atlanta, GA; REMOTE
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  • URL: Bluejeans
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Summaries

Summary Sentence: A biomechanics-based delivery strategy to primary immune cells for generating allogeneic cell therapy with multiple gene knockout

Full Summary: No summary paragraph submitted.

Tong Yu
BME PhD Proposal Presentation

Date:2021-12-01
Time: 10:00 am
Location / Meeting Link: https://bluejeans.com/709126474/9128

Committee Members:
Advisor: Prof. Todd Sulchek (School of Mechanical Engineering, Georgia Institute of Technology) Committee: Prof. Sunil Raikar (School of Medicine, Winship Cancer Institute, Emory University)) Prof. James Dahlman (School of Biomedical Engineering, Georgia Institute of Technology) Prof. Wilbur Lam (School of Biomedical Engineering, Georgia Institute of Technology) Prof. Gabriel Kwong (School of Biomedical Engineering, Georgia Institute of Technology)


Title: A biomechanics-based delivery strategy to primary immune cells for generating allogeneic cell therapy with multiple gene knockout

Abstract: Genetically engineered immune cells, such as those used in Chimeric Antigen Receptor-modified (CAR) T cell therapy and T Cell Receptor-modified (TCR-T) T cell therapy, have transformed the treatment of oncological diseases. Yet manufacturing of cell therapies faces challenges, including low scalability, inefficient workflow, and high production cost. Allogeneic T cell products, in which cells are sourced from healthy donors, genetically modified, and supplied to multiple patients, will greatly reduce the cost of manufacturing and shorten the treatment regimen. Manufacturing process for allogenic CAR-T and TCR-T cells requires genetic knockout of multiple genes related to foreign antigen presentation and recognition to improve safety and persistency of infused cells. Additionally, gene editing has been applied in negative regulator (e.g., PD1) knockout to improve CAR-T function against solid tumor, and in shared target antigen knockout to reduce CAR-T cell fratricide. These applications demand a more efficient and safer strategy for gene reprogramming, which is unmet by current methods. CRISPR/Cas9 system is a powerful choice of gene reprogramming system. Because Cas9 protein induces DNA double stranded breaks (DSB) at targeted loci, multiplexed cas9/gRNA delivery in one batch increases the possibility of chromosomal deletion and translocation, leading to genetic instability. A TCR-T cells carrying triple edits had 1-4% of cells harboring chromosomal translocation. These cells pose a direct safety concern to patients and lead to low in vivo fitness and persistence that results in low long-term potency. Therefore, a new gene editing workflow is needed to reduce incidence of multiple DSB. The goal of this research project is to test a microfluidic cell transfection technology with the potential to permit multiple CRISPR edits with high transfection efficiency and viability, and minimal negative impact on genome stability and therapeutic potency. To achieve this goal, the proposed study will apply a microfluidic Volume Exchange for Convective Transfection (VECT) platform for biomechanical transfections. This study pursues 3 aims: Aim 1: Optimize VECT device for efficient delivery to primary T cells. We will test various device design and fabrication method to achieve optimal transfection and cell viability. Aim 2: Demonstrate new editing workflows consisting of sequential triple gene editing and test impact on genome stability and CAR-T cell potency. We will evaluate VECT’s ability to enable consecutive single gene knockout to avoid generating multiple DSB. Aim 3: Relate biomechanical features of T cell to transfection outcome. We will test the hypothesis that VECT preferentially deliver to cells with certain mechanical features, and altering mechanics can improve transfection.

Additional Information

In Campus Calendar
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Graduate Studies

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Faculty/Staff, Public, Undergraduate students
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Keywords
Phd proposal
Status
  • Created By: Tatianna Richardson
  • Workflow Status: Published
  • Created On: Nov 18, 2021 - 9:19am
  • Last Updated: Nov 18, 2021 - 9:19am