CAR T cell therapy production in FNL's Biopharmaceutical Development Program

CAR T cell production setup in the Biopharmaceutical Development Program's CAR T cell therapy manufacturing suite.

Gene editing technologies are changing the way we approach cell engineering. CRISPR-based gene editing can accurately modify genomic DNA to produce allogeneic and safer CAR T cells for cancer treatment. 

Cancer is one of the leading causes of death in the United States, taking more than half a million lives each year according to the National Cancer Institute. CAR T cell technologies have given hope for cancer treatment, as they have shown the same ability as immune checkpoint inhibitors to eradicate advanced leukemias and lymphomas and to keep the cancer at bay for many years. 

CAR T cells are genetically engineered immune cells. T cells isolated from patients or suitable donors are transduced to express a chimeric antigen receptor (CAR), an engineered molecule that can recognize specific types of cancer and trigger these T cells to enable the immune system to destroy specific cancer cells. 

Diagram of CAR T donor cell graft
Elimination of HLA I through disruption of β2M on the grafted allogeneic CAR T cells prevent host vs. graft disease. Elimination of the TCRa chain (TRAC) on the grafted allogeneic CAR T cells prevents graft vs. host disease (GvHD). Double knock out.

The CAR construct is typically transduced into T cells via viral vectors, such as lentiviruses or gamma-retroviruses. Viral vectors integrate CAR constructs at random sites in the T cell genome which could result in oncogenic transformation, gene knock out, and/or transcriptional silencing. Advanced gene engineering technologies based on CRISPR (clustered regularly interspaced short palindromic repeats) enable the targeted and thus safer insertion of the CAR. In addition, it allows for knock out of specific genes, such as the endogenous T cell receptor (TCR), to enable the cells to overcome host immune responses and respond to their redirected targets more efficiently. 

With funding from the National Cancer Institute Division of Cancer Treatment and Diagnosis, the Biopharmaceutical Development Program has successfully developed CRISPR-based technologies to knock-out the endogenous TCR and knock-in a CAR construct into the TRAC locus to generate CAR T clinical products. 

In addition to knocking-out the TCR, CRISPR is used to knock-out the endogenous human leukocyte antigen (HLA) class I, which is responsible for recognition of non-self-cells by the immune system. The Biopharmaceutical Development Program’s CRISPR-based platform can generate these double-knockouts for making engineered allogeneic (healthy donor) T cells. These engineered allogeneic cells can successfully be transfused into just about any patient without the risk of immune rejection.