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CRISPR/Cas9 mediated deletion of the adenosine A2A receptor enhances CAR T cell efficacy
Lauren Giuffrida1,2, Kevin Sek1,2, Melissa A Henderson1,2
1Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Vic, Australia.
Abstract:
Adenosine is an immunosuppressive factor that limits anti-tumor immunity through the suppression of multiple immune subsets including T cells via activation of the adenosine A2A receptor (A2AR). Using both murine and human chimeric antigen receptor (CAR) T cells, here we show that targeting A2AR with a clinically relevant CRISPR/Cas9 strategy significantly enhances their in vivo efficacy, leading to improved survival of mice. Effects evoked by CRISPR/Cas9 mediated gene deletion of A2AR are superior to shRNA mediated knockdown or pharmacological blockade of A2AR. Mechanistically, human A2AR-edited CAR T cells are significantly resistant to adenosine-mediated transcriptional changes, resulting in enhanced production of cytokines including IFNγ and TNF, and increased expression of JAK-STAT signaling pathway associated genes. A2AR deficient CAR T cells are well tolerated and do not induce overt pathologies in mice, supporting the use of CRISPR/Cas9 to target A2AR for the improvement of CAR T cell function in the clinic.
Insights
Targeting the adenosine A2A receptor (A2AAR) with CRISPR/Cas9 gene editing enhances chimeric antigen receptor (CAR) T cell efficacy against tumors. This approach improves anti-tumor immunity and mouse survival, offering a promising strategy for cancer immunotherapy.
Area of Science:
- Immunology
- Cancer Biology
- Gene Editing
Background:
- Adenosine acts as an immunosuppressive factor, inhibiting anti-tumor immunity by suppressing T cells through adenosine A2A receptor (A2AAR) activation.
- Chimeric antigen receptor (CAR) T cells are a promising cancer immunotherapy, but their efficacy can be limited by the tumor microenvironment.
Purpose of the Study:
- To investigate the efficacy of targeting the adenosine A2A receptor (A2AAR) using CRISPR/Cas9 gene editing to enhance CAR T cell anti-tumor activity.
- To compare CRISPR/Cas9-mediated A2AAR deletion with other methods like shRNA knockdown or pharmacological blockade.
Main Methods:
- Utilized murine and human chimeric antigen receptor (CAR) T cells.
- Employed a CRISPR/Cas9 gene editing strategy to delete the adenosine A2A receptor (A2AAR).
- Assessed in vivo efficacy, survival rates, cytokine production (IFNγ, TNF), and JAK-STAT pathway gene expression.
Main Results:
- CRISPR/Cas9-mediated A2AAR deletion significantly enhanced CAR T cell in vivo efficacy and improved mouse survival compared to control groups.
- A2AAR-edited CAR T cells demonstrated superior resistance to adenosine-mediated immunosuppression.
- Gene-edited CAR T cells showed enhanced production of key cytokines and increased expression of JAK-STAT pathway genes.
- CRISPR/Cas9 mediated deletion of A2AAR proved more effective than shRNA knockdown or pharmacological blockade.
Conclusions:
- CRISPR/Cas9 targeting of the adenosine A2A receptor (A2AAR) is a potent strategy to enhance CAR T cell function and anti-tumor immunity.
- A2AAR-deficient CAR T cells are well-tolerated in vivo, suggesting clinical translatability.
- This approach holds significant potential for improving CAR T cell-based cancer therapies.
