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Generation of CAR T Cells for Adoptive Therapy in the Context of Glioblastoma Standard of Care
Published on: February 16, 2015
Multifunctional mRNA-Based CAR T Cells Display Promising Antitumor Activity Against Glioblastoma
Hanna Meister1, Thomas Look1, Patrick Roth1
1Department of Neurology, Clinical Neuroscience Center, University Hospital Zurich and University of Zurich, Zurich, Switzerland.
Purpose:
Most chimeric antigen receptor (CAR) T-cell strategies against glioblastoma have demonstrated only modest therapeutic activity and are based on persistent gene modification strategies that have limited transgene capacity, long manufacturing processes, and the risk for uncontrollable off-tumor toxicities. mRNA-based T-cell modifications are an emerging safe, rapid, and cost-effective alternative to overcome these challenges, but are underexplored against glioblastoma.
Experimental Design:
We generated mouse and human mRNA-based multifunctional T cells coexpressing a multitargeting CAR based on the natural killer group 2D (NKG2D) receptor and the proinflammatory cytokines IL12 and IFNα2 and assessed their antiglioma activity in vitro and in vivo.
Results:
Compared with T cells that either expressed the CAR or cytokines alone, multifunctional CAR T cells demonstrated increased antiglioma activity in vitro and in vivo in three orthotopic immunocompetent mouse glioma models without signs of toxicity. Mechanistically, the coexpression of IL12 and IFNα2 in addition to the CAR promoted a proinflammatory tumor microenvironment and reduced T-cell exhaustion as demonstrated by ex vivo immune phenotyping, cytokine profiling, and RNA sequencing. The translational potential was demonstrated by image-based single-cell analyses of mRNA-modified T cells in patient glioblastoma samples with a complex cellular microenvironment. This revealed strong antiglioma activity of human mRNA-based multifunctional NKG2D CAR T cells coexpressing IL12 and IFNα2 whereas T cells that expressed either the CAR or cytokines alone did not demonstrate comparable antiglioma activity.
Conclusions:
These data provide a robust rationale for future clinical studies with mRNA-based multifunctional CAR T cells to treat malignant brain tumors.
Insights
New mRNA-based CAR T-cells, engineered with NKG2D receptor and cytokines, show potent anti-glioblastoma activity. This approach offers a safer, faster alternative to traditional gene modification for brain tumor treatment.
Area of Science:
- Oncology
- Immunotherapy
- Molecular Biology
Background:
- Current chimeric antigen receptor (CAR) T-cell therapies for glioblastoma have limited efficacy due to persistent gene modification challenges.
- These limitations include restricted transgene capacity, lengthy manufacturing, and potential for severe off-tumor toxicities.
- Messenger RNA (mRNA)-based T-cell modification presents a promising, rapid, and cost-effective alternative but remains underexplored for glioblastoma.
Purpose of the Study:
- To evaluate the efficacy of mRNA-based multifunctional T cells engineered with a multitargeting natural killer group 2D (NKG2D) receptor and the cytokines IL12 and IFNα2 against glioblastoma.
- To compare the anti-glioma activity of these multifunctional T cells against T cells expressing only the CAR or cytokines alone.
- To investigate the underlying mechanisms and translational potential in patient-derived glioblastoma models.
Main Methods:
- Generation of mouse and human mRNA-based T cells coexpressing a multitargeting NKG2D CAR, IL12, and IFNα2.
- In vitro and in vivo assessment of anti-glioma activity in orthotopic immunocompetent mouse glioma models.
- Ex vivo immune phenotyping, cytokine profiling, and RNA sequencing to analyze the tumor microenvironment and T-cell exhaustion.
- Image-based single-cell analyses in patient glioblastoma samples to evaluate translational potential.
Main Results:
- Multifunctional CAR T cells demonstrated significantly enhanced anti-glioma activity compared to CAR T cells or cytokine-only T cells, both in vitro and in vivo.
- The coexpression of IL12 and IFNα2 alongside the CAR promoted a proinflammatory tumor microenvironment and reduced T-cell exhaustion.
- Human mRNA-based multifunctional NKG2D CAR T cells showed strong anti-glioma activity in complex patient glioblastoma microenvironments, unlike T cells with single modifications.
Conclusions:
- mRNA-based multifunctional CAR T cells coexpressing NKG2D CAR, IL12, and IFNα2 represent a potent therapeutic strategy against glioblastoma.
- This approach overcomes limitations of traditional CAR T-cell therapies, offering improved safety and efficiency.
- The findings provide a strong rationale for advancing mRNA-based multifunctional CAR T cells into clinical trials for malignant brain tumors.
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