Engineering a Universal CAR T Cell

UCSD Department of Bioengineering

Senior Design 2019

Group 8

Group 8 Video.mp4

Senior Design Project

The Senior Design Project is a capstone project for all senior Bioengineering undergraduate students at UCSD. It challenges students to identify a problem the can be solved through the hands-on application of skills they have learned throughout their undergraduate education.

The Problem

Many traditional cancer therapies - such as surgical removal, chemotherapy, and radiation - suffer from extensive limitations or can cause severe off-tumor toxicity.

One of the more recent cancer therapies - CAR T cell therapy - is an autologous immunotherapy which may circumvent some of these drawbacks; it demonstrates higher specificity, lower toxicity, and more long-lasting efficacy. However, existing CAR T cells therapies cannot treat different tumors types with only one construct. It is time-consuming and expensive to design new CAR constructs every time we deal with a different cancer cell.

Our Solution

The goal of our senior design is to address and improve some deficiencies of traditional CAR T cells by designing a new universal CAR T cell construct, PEbody CAR, and performing validation experiments on its potency in vitro.

The PEbody CAR is composed of a universal receptor on the T cell surface and a tumor targeting scFv molecules, which can bind to any antibody conjugated with R-phycoerythrin, or PE, a fluorescent dye. The first stage of the project is to compare the CAR-T cell expression level between a traditional CAR construct, CD19 CAR and PEbody CAR in Jurkat cells. The second step is to test both CAR constructs in primary T cells (PBMC) and compare their killing abilities of blood cancer and solid tumor cells. It is expected that PEbody CAR will have similar, or better, expression level, activation and killing efficacy as the CD19 control.

Step 1

CAR T Cell Activation

T cell activation induces downstream signaling cascade and secretes cytokines, both of which facilitate tumor eradication. To verify if our PEbody CAR T cells can function normally, we tested the activation level for PEbody CAR as compared to the commercially available CD19 CAR T cells.

Step 2

Tumor Killing Efficacy

One of the most important functions for CAR T cells is to efficiently kill cancer cells. We evaluated the killing ability of PEbody CAR system by performing luciferase killing assays. We co-cultured PEbody CAR T cells with a B cell precursor leukemia cell line, Nalm6 for 24 hours. Then we measured the percentage of alive cancer cells by registering fluorescent signals.

Step 3

Future Directions

One great setback of CAR T cell therapy is that the CAR gets quickly exhausted in vivo. Therefore, if the universal CAR gives solid results, we also want to incorporate a control element into the CAR construct so we can have spatial and temporal control over CAR T cell activation.

Contributors

Group 8

Jiayi Li

Yuan Yuan

Emma Zelus

Wang Lab

Dr. Yingxiao Peter Wang (PI)

Dr. Praopim Mint Limsakul (Mentor)

UCSD Bioengineering

Dr. Bruce Wheeler

Ali Zamat (TA)

Page designed by Jiayi Li, Yuan Yuan, and Emma Zelus.

Report abuse