Non-viral, specifically targeted CAR-T cells achieve high safety and efficacy in B-NHL

Abstract

Recently, chimeric antigen receptor (CAR)-T cell therapy has shown great promise in treating haematological malignancies1–7. However, CAR-T cell therapy currently has several limitations8–12. Here we successfully developed a two-in-one approach to generate non-viral, gene-specific targeted CAR-T cells through CRISPR–Cas9. Using the optimized protocol, we demonstrated feasibility in a preclinical study by inserting an anti-CD19 CAR cassette into the AAVS1 safe-harbour locus. Furthermore, an innovative type of anti-CD19 CAR-T cell with PD1 integration was developed and showed superior ability to eradicate tumour cells in xenograft models. In adoptive therapy for relapsed/refractory aggressive B cell non-Hodgkin lymphoma (ClinicalTrials.gov, NCT04213469), we observed a high rate (87.5%) of complete remission and durable responses without serious adverse events in eight patients. Notably, these enhanced CAR-T cells were effective even at a low infusion dose and with a low percentage of CAR+ cells. Single-cell analysis showed that the electroporation method resulted in a high percentage of memory T cells in infusion products, and PD1 interference enhanced anti-tumour immune functions, further validating the advantages of non-viral, PD1-integrated CAR-T cells. Collectively, our results demonstrate the high safety and efficacy of non-viral, gene-specific integrated CAR-T cells, thus providing an innovative technology for CAR-T cell therapy.

Cost Effectiveness Analysis of CAR-T Cell Therapy for Patients with Relapsed/Refractory Multiple Myeloma in China

“…Whether it is from the perspective of drug development or from the perspective of economic evaluation, it is necessary to develop a more effective CAR T-cell regimen for R/R MM, especially achieving a substantial proportion of long-term survivors. Recently, a new strategy to develop non-viral, gene-speci c targeted CAR-T cells by CRISPR-Cas9 was reported and the innovative technology for CAR-T cell therapy achieved the high safety and e cacy in clinical trial [39] . Therefore, when the CAR-T with this novel development strategy could be approved by regulatory authorities, the ICER of the CAR-T therapy would be reduced greatly.…”

Tumor buster - where will the CAR-T cell therapy ‘missile’ go?

“…Still, these approaches often hinder CAR expression, carry a high tumor risk, and are more expensive to manufacture [33][34][35][36][37]. A paper published in Nature reports that by using non-viral targeted integration, researchers have prepared CD19 CAR-T cells (AAVS1-19bbz) that effectively eradicate tumor cells in the BL cell line Raji and cell line-derived xenograft mouse models [38]. In this study, the researchers also produced CD19 CAR-T cells (PD1-19bbz) with programmed cell death 1 (PD1) knocked out by CRISPR-Cas9 technology, which showed strong eradication ability against Raji cells that were either high or low in programmed death-ligand 1 (PD-L1) expression [38].…”

“…A paper published in Nature reports that by using non-viral targeted integration, researchers have prepared CD19 CAR-T cells (AAVS1-19bbz) that effectively eradicate tumor cells in the BL cell line Raji and cell line-derived xenograft mouse models [38]. In this study, the researchers also produced CD19 CAR-T cells (PD1-19bbz) with programmed cell death 1 (PD1) knocked out by CRISPR-Cas9 technology, which showed strong eradication ability against Raji cells that were either high or low in programmed death-ligand 1 (PD-L1) expression [38]. In the phase 1 clinical trial using PD1-19bbz cells, seven out of eight relapsed/refractory B-NHL patients achieved CR and the rest PR, and no CAR-T cell-related grade 3 or higher adverse events were observed (CTCAE v5.0) [38].…”

“…In this study, the researchers also produced CD19 CAR-T cells (PD1-19bbz) with programmed cell death 1 (PD1) knocked out by CRISPR-Cas9 technology, which showed strong eradication ability against Raji cells that were either high or low in programmed death-ligand 1 (PD-L1) expression [38]. In the phase 1 clinical trial using PD1-19bbz cells, seven out of eight relapsed/refractory B-NHL patients achieved CR and the rest PR, and no CAR-T cell-related grade 3 or higher adverse events were observed (CTCAE v5.0) [38]. This demonstrates the high efficacy and safety profile of the cells.…”

A novel non-viral PDCD1 site-integrated CAR design: killing 2 birds with 1 stone

“…Recently, the teams of Mingyao Liu and He Huang (et al) developed a two-in-one approach to generate non-viral CAR-T-cells by inserting an anti-CD19 CAR cassette into the specific locus through CRISPR/Cas9. 19 First, they inserted an anti-CD19 CAR cassette containing 4-1BB and CD3ζ into the AAVS1 safe-harbor locus to demonstrate the feasibility of non-viral CAR-T-cells. Then, an innovative CAR-T-cell was developed by integrating the anti-CD19 CAR cassette into the PDCD1 locus ( Fig.…”