Revolutionizing cancer treatment: enhancing CAR-T cell therapy with CRISPR/Cas9 gene editing technology

Abstract

CAR-T cell therapy, a novel immunotherapy, has made significant breakthroughs in clinical practice, particularly in treating B-cell-associated leukemia and lymphoma. However, it still faces challenges such as poor persistence, limited proliferation capacity, high manufacturing costs, and suboptimal efficacy. CRISPR/Cas system, an efficient and simple method for precise gene editing, offers new possibilities for optimizing CAR-T cells. It can increase the function of CAR-T cells and reduce manufacturing costs. The combination of CRISPR/Cas9 technology and CAR-T cell therapy may promote the development of this therapy and provide more effective and personalized treatment for cancer patients. Meanwhile, the safety issues surrounding the application of this technology in CAR-T cells require further research and evaluation. Future research should focus on improving the accuracy and safety of CRISPR/Cas9 technology to facilitate the better development and application of CAR-T cell therapy. This review focuses on the application of CRISPR/Cas9 technology in CAR-T cell therapy, including eliminating the inhibitory effect of immune checkpoints, enhancing the ability of CAR-T cells to resist exhaustion, assisting in the construction of universal CAR-T cells, reducing the manufacturing costs of CAR-T cells, and the security problems faced. The objective is to show the revolutionary role of CRISPR/Cas9 technology in CAR-T cell therapy for researchers.

Keywords: CAR-T cell; CRISPR/Cas9; cost; durability; immune checkpoints; security.

Figure 1

Structural changes to CAR constructs from first to fifth generation. The first generation CAR consists of a single-chain variable fragment (scFv), a transmembrane domain (TM) and an intracellular signaling domain (CD3ζ). The second generation CAR adds a costimulatory molecule (CD28 or 4-1BB) to the intracellular domain. The third generation CAR adds two costimulatory molecules (CD28 and 4-1BB) to the intracellular domain; The fourth generation CAR is based on the second generation CAR or the third generation CAR, and the cytokine receptor genes are added to the intracellular domain. The fifth generation CAR is to add an intermediate system in the extracellular domain, separating the scFv from the transmembrane domain in the extracellular domain; or insert an IL2 RB chain fragment between the costimulatory domain (CD28) and CD3ζ, inducing activation of the JAK-STAT pathway, thereby promoting cell proliferation. In addition, universal CAR-T cells generated using universal T cells (TCR gene and/or HLA class I locus of allogeneic T cells have been destroyed) also belong to the fifth generation CAR-T cells.