November 23, 2022

Immunogenomic Engineering of a T Cell Display Platform for Personalized Cancer Therapy

1st Call, TPdF

Short Summary

Cytotoxic T cells are able to recognize and respond to tumor cells, leading to potent responses and at times complete remission from cancer. Tumor-specific T cells rely on surface proteins, known as T cell receptors (TCRs) that recognize tumor antigens and initiate a cytotoxic response against them. This has motivated the development of TCR gene therapies, in which patient’s T cells are genetically modified to express tumor-reactive TCRs. However, a number of challenges still remain in scaling this approach so that it can be useful for personalized medicine. One such limitation is the slow and laborious discovery of tumor-specific TCRs. Using genome editing, this project aims to develop a high-throughput platform for the rapid screening and discovery of tumor-specific TCRs.

Goals

The project is divided into three major aims: First, a T cell line will be modified using genome editing in order to generate a novel “plug-and-(dis)play” cell line. This cell line will be engineered to allow for facile TCR reprogramming and for functional screening of candidate TCRs (Aim 1). Next, tumor-infiltrating T cells directly from human cancer patients will be isolated and single-cell sequenced to determine their TCR sequences (Aim 2). Candidate tumor-specific TCRs will then be integrated into the cell line developed in Aim 1 and screened at high-throughput to discover TCRs that are specific to tumor cells (Aim 3).

Significance

We expect that once our platform is established, it will be utilized for the direct screening of patients TCRs. By enabling the efficient identification of tumor-reactive TCRs, we predict that our technology will translate into a valuable tool for use in the rapidly developing field of personalized TCR gene therapy.

Background

The development of high-throughput single-cell sorting and sequencing technologies now allow for the identification of individual tumor-reactive TCRs. Accordingly, a number of recent studies have exploited these technologies to demonstrate the feasibility of personalized TCR gene therapy. An important bottleneck in the TCR identification process is the use of primary T cells for screening and characterization purposes. This can become largely impractical, as either freshly isolated or thawed T cell samples are required for screening steps. Thus, the development of a robust TCR screening platform would be extremely beneficial for TCR gene therapy applications. More specifically, the ability to transfer TCR repertoires into a T cell line for facile screening would be highly desirable.

Publications

Rodrigo Vazquez-Lombardi, Johanna S. Jung, Florian Bieberich et al CRISPR-targeted display of functional T cell receptors enables engineering of enhanced specificity and prediction of cross-reactivity. Cell In Press