Disrupting usp14-mediated PARP1 dynamics reinstates mic-a/b-driven antigen-independent CD8⁺ t-cell killing in glioma

Antigen loss is a major mechanism of resistance to immunotherapy. MIC-A/B are stress-inducible ligands expressed by tumour cells that activate NKG2D on cytotoxic immune cells and mediate NKG2D-dependent, antigen-independent tumour cell killing, yet the mechanisms underlying their reduced expression in glioma remain unclear. Using single-cell RNA sequencing and spatial transcriptomics, we investigated ectopic MIC-A/B in mouse glioma and identified USP14 as a key regulator through deubiquitinase screening. Proteomic, coimmunoprecipitation, chromatin immunoprecipitation, immunofluorescence and ubiquitination assays characterized the interactions between USP14, PARP1 and NFIL3, while an intracranial tumour model combined USP14 inhibition and immunotherapy to evaluate effects on tumourigenesis and antitumour immunity. We found that MIC-A/B increased CD8⁺ T-cell infiltration and reversed exhaustion, and that USP14 stabilized PARP1 via K63-linked deubiquitination at lysine 653, reducing NFIL3 ..., , , # Disrupting usp14-mediated PARP1 dynamics reinstates mic-a/b-driven antigen-independent CD8⁺ t-cell killing in glioma

Dataset DOI: [10.5061/dryad.vq83bk47n](https://doi.org/10.5061/dryad.vq83bk47n)

This dataset contains processed data to reproduce key transcriptomic analyses testing the hypothesis that restoring stress-ligand signaling (MIC-A/B–NKG2D axis) and/or inhibiting the deubiquitinase USP14 (IU1) can reshape the immune microenvironment in the GL261 mouse intracranial glioma model, consistent with enhanced cytotoxic immune programs. Data are provided as per-sample normalized expression matrices to facilitate downstream reuse.

## Description of the data and file structure

**The submission comprises three coordinated modules:**

(1) CD45⁺ scRNA-seq from MICAB overexpression (MICAB-OE) and matched control tumors (per-sample normalized matrices), enabling immune-cell clustering/annotation and comparison of CD8⁺ T-cell state distributions (e.g., effector vs exhausted programs); ...