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Published on: October 16, 2018
Simultaneous targeting of Tim3 and A2a receptors modulates MSLN-CAR T cell antitumor function in a human cervical
Tahereh Soltantoyeh1, Behnia Akbari1, Zahra Shahosseini2,3
1Department of Medical Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Introduction:
Chimeric antigen receptor (CAR) T cell therapy has transformed the treatment of hematological malignancies. However, its efficacy in solid tumors is limited by the immunosuppressive tumor microenvironment that compromises CAR T cell antitumor function in clinical settings. To overcome this challenge, researchers have investigated the potential of inhibiting specific immune checkpoint receptors, including A2aR (Adenosine A2 Receptor) and Tim3 (T cell immunoglobulin and mucin domain-containing protein 3), to enhance CAR T cell function. In this study, we evaluated the impact of genetic targeting of Tim3 and A2a receptors on the antitumor function of human mesothelin-specific CAR T cells (MSLN-CAR) in vitro and in vivo.
Methods:
Second-generation anti-mesothelin CAR T cells were produced using standard cellular and molecular techniques. A2aR-knockdown and/or Tim3- knockdown anti-mesothelin-CAR T cells were generated using shRNA-mediated gene silencing. The antitumor function of CAR T cells was evaluated by measuring cytokine production, proliferation, and cytotoxicity in vitro through coculture with cervical cancer cells (HeLa cell line). To evaluate in vivo antitumor efficacy of manufactured CAR T cells, tumor growth and mouse survival were monitored in a human cervical cancer xenograft model.
Results:
In vitro experiments demonstrated that knockdown of A2aR alone or in combination with Tim3 significantly improved CAR T cell proliferation, cytokine production, and cytotoxicity in presence of tumor cells in an antigen-specific manner. Furthermore, in the humanized xenograft model, both double knockdown CAR T cells and control CAR T cells could effectively control tumor growth. However, single knockdown CAR T cells were associated with reduced survival in mice.
Conclusion:
These findings highlight the potential of concomitant genetic targeting of Tim3 and A2a receptors to augment the efficacy of CAR T cell therapy in solid tumors. Nevertheless, caution should be exercised in light of our observation of decreased survival in mice treated with single knockdown MSLN-CAR T cells, emphasizing the need for careful efficacy considerations.
Insights
Targeting Adenosine A2 Receptor (A2aR) and T cell immunoglobulin and mucin domain-containing protein 3 (Tim3) enhances chimeric antigen receptor (CAR) T cell therapy for solid tumors. Combined A2aR and Tim3 knockdown improved CAR T cell function, but single knockdown reduced mouse survival.
Area of Science:
- Immunotherapy
- Oncology
- Cellular Biology
Background:
- Chimeric antigen receptor (CAR) T cell therapy shows promise for hematological malignancies but faces challenges in solid tumors due to the immunosuppressive tumor microenvironment.
- Immune checkpoint receptors like Adenosine A2 Receptor (A2aR) and T cell immunoglobulin and mucin domain-containing protein 3 (Tim3) are investigated as targets to enhance CAR T cell function.
Purpose of the Study:
- To evaluate the impact of genetically targeting A2aR and Tim3 on the antitumor function of human mesothelin-specific CAR T cells (MSLN-CAR).
- To assess the in vitro and in vivo efficacy of MSLN-CAR T cells with A2aR and/or Tim3 knockdown in a solid tumor model.
Main Methods:
- Generated second-generation MSLN-CAR T cells using standard techniques.
- Created A2aR-knockdown and/or Tim3-knockdown MSLN-CAR T cells via shRNA-mediated gene silencing.
- Assessed in vitro antitumor function (cytokine production, proliferation, cytotoxicity) and in vivo efficacy (tumor growth, mouse survival) in a human cervical cancer xenograft model.
Main Results:
- In vitro, A2aR knockdown, alone or combined with Tim3 knockdown, significantly enhanced MSLN-CAR T cell proliferation, cytokine production, and cytotoxicity against tumor cells.
- In vivo, both dual knockdown and control MSLN-CAR T cells effectively controlled tumor growth in a xenograft model.
- Mice treated with single knockdown MSLN-CAR T cells exhibited reduced survival compared to controls.
Conclusions:
- Concomitant genetic targeting of Tim3 and A2a receptors can potentially augment CAR T cell therapy efficacy in solid tumors.
- Caution is warranted due to observed decreased survival with single knockdown MSLN-CAR T cells, highlighting the need for careful efficacy considerations.
