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Updated: Jul 16, 2025
Stereotactic Adoptive Transfer of Cytotoxic Immune Cells in Murine Models of Orthotopic Human Glioblastoma Multiforme Xenografts
Published on: September 1, 2018
NLGN4X TCR transgenic T cells to treat gliomas
Christoper Krämer1,2, Michael Kilian1,2, Yu-Chan Chih1,3
1German Cancer Consortium (DKTK) Clinical Cooperation Unit (CCU) Neuroimmunology and Brain Tumor Immunology, German Cancer Research Center (DKFZ), Heidelberg, Germany.
Background:
Neuroligin 4 X-linked (NLGN4X) harbors a human leukocyte antigen (HLA)-A*02-restricted tumor-associated antigen, overexpressed in human gliomas, that was found to induce specific cytotoxic T cell responses following multi-peptide vaccination in patients with newly diagnosed glioblastoma.
Methods:
T cell receptor (TCR) discovery was performed using droplet-based single-cell TCR sequencing of NLGN4X-tetramer-sorted T cells postvaccination. The identified TCR was delivered to Jurkat T cells and primary human T cells (NLGN4X-TCR-T). Functional profiling of NLGN4X-TCR-T was performed by flow cytometry and cytotoxicity assays. Therapeutic efficacy of intracerebroventricular NLGN4X-TCR-T was assessed in NOD scid gamma (NSG) major histocompatibility complex (MHC) I/II knockout (KO) (NSG MHC I/II KO) mice bearing NLGN4X-expressing experimental gliomas.
Results:
An HLA-A*02-restricted vaccine-induced T cell receptor specifically binding NLGN4X131-139 was applied for preclinical therapeutic use. Reactivity, cytotoxicity, and polyfunctionality of this NLGN4X-specific TCR are demonstrated in various cellular models. Intracerebroventricular administration of NLGN4X-TCR-T prolongs survival and leads to an objective response rate of 44.4% in experimental glioma-bearing NSG MHC I/II KO mice compared to 0.0% in control groups.
Conclusion:
NLGN4X-TCR-T demonstrate efficacy in a preclinical glioblastoma model. On a global scale, we provide the first evidence for the therapeutic retrieval of vaccine-induced human TCRs for the off-the-shelf treatment of glioblastoma patients.Keywords cell therapy | glioblastoma | T cell receptor | tumor antigen.
Insights
Researchers identified a T cell receptor targeting the NLGN4X tumor antigen in glioblastoma patients. This NLGN4X-specific T cell receptor therapy showed significant efficacy in preclinical models, offering a new avenue for glioblastoma treatment.
Area of Science:
- Oncology
- Immunology
- Cell Therapy
Background:
- Neuroligin 4 X-linked (NLGN4X) is a tumor-associated antigen overexpressed in human gliomas.
- NLGN4X elicits specific cytotoxic T cell responses following multi-peptide vaccination in glioblastoma patients.
Purpose of the Study:
- To discover and characterize T cell receptors (TCRs) targeting the NLGN4X antigen.
- To evaluate the therapeutic potential of NLGN4X-specific TCR-engineered T cells in glioblastoma models.
Main Methods:
- Single-cell TCR sequencing was used to identify vaccine-induced TCRs against NLGN4X.
- TCRs were engineered into Jurkat and primary human T cells (NLGN4X-TCR-T).
- Functional assays and in vivo efficacy studies in a preclinical glioma model were performed.
Main Results:
- An HLA-A*02-restricted TCR specific for NLGN4X131-139 was identified and functionally validated.
- NLGN4X-TCR-T cells demonstrated reactivity, cytotoxicity, and polyfunctionality.
- Intracerebroventricular administration of NLGN4X-TCR-T in mice significantly prolonged survival and achieved a 44.4% objective response rate.
Conclusions:
- NLGN4X-TCR-T cells exhibit therapeutic efficacy in a preclinical glioblastoma model.
- This study provides the first evidence for retrieving vaccine-induced human TCRs for off-the-shelf glioblastoma treatment.
