2. Academic Validation
  3. A Tonic Signaling Code Predicts CAR-T Cell Efficacy in Diffuse Midline Glioma

A Tonic Signaling Code Predicts CAR-T Cell Efficacy in Diffuse Midline Glioma

  • bioRxiv. 2025 Oct 1:2025.09.29.679095. doi: 10.1101/2025.09.29.679095.
Emily B Deng 1,  Xiaowen Zhong 2,  Dazhuan Xin 2,  Upendra K Soni 2,  Wenkun Ma 2,  Xiao Huang 2,  Mingjun Cai 2,  Po-Yu Liang 3,  Jun Bai 3,  Qian Qin 4,  Shreya Mishra 5,  Ming Hu 5,  Arman E Bayat 2,  Jiajie Diao 1,  Mei Xin 2,  Natasha Pillay-Smiley 6,  Trent R Hummel 6,  Charles B Stevenson 7,  Jessica B Foster 8,  Peter de Blank 6,  Scott Raskin 6,  Carl Koschmann 9,  Jose A Cancelas 10,  Yi Zheng 2,  Q Richard Lu 2
Affiliations

Affiliations

  • 1 Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA.
  • 2 Department of Pediatrics, The Cure Starts Now Brain Tumor Center, Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
  • 3 Department of Computer Science, University of Cincinnati, Cincinnati, Ohio, USA.
  • 4 Broad Institute of MIT and Harvard, Cambridge, MA, USA.
  • 5 Department of Quantitative Health Sciences, Lerner Research Institute, Cleveland Clinic Foundation, Case Western Reserve University School of Medicine, Cleveland, OH, USA.
  • 6 Department of Pediatrics, The Cure Starts Now Brain Tumor Center, Division of Oncology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.
  • 7 Division of Pediatric Neurosurgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
  • 8 Division of Oncology, Children's Hospital of Philadelphia, Philadelphia, PA.
  • 9 Department of Pediatrics, Michigan Medicine, Ann Arbor, MI, USA; Rogel Cancer Center, University of Michigan, Ann Arbor, MI, USA.
  • 10 Reilly and O'Connell Families Cell Manipulation Core Facility and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA.
PMID: 41256549 DOI: 10.1101/2025.09.29.679095
Abstract

Diffuse midline glioma (DIPG/DMG) is a uniformly fatal pediatric brain tumor with no effective cure. Although CAR T-cell therapy shows promise, clinical outcomes remain inconsistent due to limited persistence and premature exhaustion. Reliable predictive biomarkers are lacking, and proposed exhaustion or stemness markers provide limited utility. Here, we systematically compare multiple CAR-T constructs targeting clinically-relevant antigen B7-H3 and identify antigen-independent CAR activation, or tonic signaling, as a key determinant of therapeutic performance. We find that B7-H3 CAR-T cells with restrained tonic signaling display superior tumor killing, persistence, and resistance to exhaustion, along with reduced CAR membrane clustering, in patient-derived DIPG models. Integrated multi-omics and single-cell profiling further reveal a CAR-T tonic signaling-associated gene signature that outperforms conventional exhaustion or stemness markers in predicting therapeutic efficacy across multiple clinical trials, including DIPG and Other tumor types. Together, these findings define a mechanistic and predictive framework to guide CAR design and improve clinical outcomes.

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