Mutating PD-1 to Avoid PD-L1 From Binding Therefore Regulates NSCLC

Abstract

Lung cancer is one of the most common types of cancer in human clinical cases. It has been a worldwide problem for hundreds of years. Currently, there are more research done on blocking the pathway between PD-1 and PD-L1 using drugs or antibodies; however, there are not that many papers that write about other ways to block the pathway such as knocking down ASN 45 in the PD-1 and PD-L1 binding domain using CRISPR to change PD-1’s structure, therefore, regulating the binding. This study investigates the effect of genetic engineered T cells on NSCLC xenograft mice. CRISPR to mutate PD1 amino acids(ASN 45) in the PDL1 binding domain in effector T cells and then inject the cells into NSCLC xenograft mice and see if tumor shrinks more than Wild Type PD1 control injections. Also measure the activation of the t cells in the tumor by isolating them by FACS and using FACS for CD69, pSrc, pErk to detect activation. Negative control is Wild Type PD1, positive control is Keytruda treatments with Wild Type PD1. The result of this study will provide essential information for the future research of PD-1 mutation. By mutating certain amino acid that is on the binding domain of PD-1 would change the structure of the glycoprotein and therefore avoid the combination of PD-1 and PD-L1, which would then forbid cancerous cells from evading immune detection.