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RASA2 deletion rescues immune synapse dysfunction, enhancing CAR T cell efficacy against DMGs.

Jorge Ibanez-Vega1, Robert Teis1, Jennifer K Ocasio2

  • 1Bone Marrow Transplantation and Cellular Therapy, St Jude Children's Research Hospital, Memphis, Tennessee, USA.

Journal for Immunotherapy of Cancer
|March 30, 2026
PubMed
Summary

Diffuse midline gliomas (DMGs) resist chimeric antigen receptor (CAR) T-cell therapy due to impaired immune synapse formation. Deleting RASA2 enhances CAR T-cell function and anti-tumor response against these pediatric brain tumors.

Keywords:
Central Nervous System CancerChimeric antigen receptor - CARImmunotherapy

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Area of Science:

  • Immunology
  • Oncology
  • Pediatric Neuro-oncology

Background:

  • Chimeric antigen receptor (CAR) T-cell therapy shows promise but limited efficacy in diffuse midline gliomas (DMGs).
  • Mechanisms of CAR T-cell resistance in DMGs are not well understood.
  • DMGs are aggressive pediatric brain tumors requiring novel therapeutic strategies.

Purpose of the Study:

  • To investigate the mechanisms underlying CAR T-cell resistance in DMGs.
  • To compare CAR T-cell efficacy against DMGs versus adult glioblastoma.
  • To explore RASA2 deletion as a strategy to enhance CAR T-cell function in DMGs.

Main Methods:

  • Compared B7-H3 CAR T-cell efficacy in DMG and glioblastoma cell lines in vitro and in vivo.
  • Utilized live-cell imaging and single-cell RNA sequencing to analyze immune synapse (IS) formation.
  • Genetically deleted RASA2 in CAR T-cells to assess its impact on IS formation and anti-tumor activity.

Main Results:

  • DMG cells impaired IS formation, leading to reduced CAR T-cell activation and anti-tumor response.
  • RASA2 deletion in CAR T-cells improved IS formation, calcium flux, and signaling molecule accumulation.
  • RASA2-deleted CAR T-cells demonstrated enhanced migration, in vitro functionality, and superior early in vivo anti-tumor efficacy against DMGs.

Conclusions:

  • Tumor-specific factors, particularly impaired IS formation, limit CAR T-cell efficacy in DMGs.
  • RASA2 deletion represents a viable strategy to enhance CAR T-cell function against DMGs.
  • Cytoskeleton remodeling and T-cell motility are potential therapeutic targets for next-generation CAR T-cell engineering.