NPRL2 Gene Therapy Induces Effective Antitumor Immunity in KRAS/STK11 Mutant Anti-PD1 Resistant Metastatic Non-Small Cell Lung Cancer (NSCLC) in a Humanized Mouse Model

Authors

Ismail M Meraz
Mourad Majidi
Renduo Song
Feng Meng
Lihui Gao
Qi Wang
Jing Wang
Elizabeth J Shpall
Jack A Roth

Language

English

Publication Date

2-11-2025

Journal

eLife

DOI

10.7554/eLife.98258

PMID

39932765

PMCID

PMC11813225

PubMedCentral® Posted Date

2-11-2025

PubMedCentral® Full Text Version

Post-print

Abstract

Expression of NPRL2/TUSC4, a tumor-suppressor gene, is reduced in many cancers including NSCLC. Restoration of NPRL2 induces DNA damage, apoptosis, and cell-cycle arrest. We investigated NPRL2 antitumor immune responses in aPD1R/KRAS/STK11^mt NSCLC in humanized-mice. Humanized-mice were generated by transplanting fresh human cord blood-derived CD34 stem cells into sub-lethally irradiated NSG mice. Lung-metastases were developed from KRAS/STK11^mt/aPD1R A549 cells and treated with NPRL2 w/wo pembrolizumab. NPRL2-treatment reduced lung metastases significantly, whereas pembrolizumab was ineffective. Antitumor effect was greater in humanized than non-humanized-mice. NPRL2 + pembrolizumab was not synergistic in KRAS/STK11^mt/aPD1R tumors but was synergistic in KRAS^wt/aPD1S H1299. NPRL2 also showed a significant antitumor effect on KRAS^mt/aPD1R LLC2 syngeneic-tumors. The antitumor effect was correlated with increased infiltration of human cytotoxic-T, HLA-DR+DC, CD11c+DC, and downregulation of myeloid and regulatory-T cells in TME. Antitumor effect was abolished upon in-vivo depletion of CD8-T, macrophages, and CD4-T cells whereas remained unaffected upon NK-cell depletion. A distinctive protein-expression profile was found after NPRL2 treatment. IFNγ, CD8b, and TBX21 associated with T-cell functions were significantly increased, whereas FOXP3, TGFB1/B2, and IL-10RA were strongly inhibited by NPRL2. A list of T-cell co-inhibitory molecules was also downregulated. Restoration of NPRL2 exhibited significantly slower tumor growth in humanized-mice, which was associated with increased presence of human cytotoxic-T, and DC and decreased percentage of Treg, MDSC, and TAM in TME. NPRL2-stable cells showed a substantial increase in colony-formation inhibition and heightened sensitivity to carboplatin. Stable-expression of NPRL2 resulted in the downregulation of MAPK and AKT-mTOR signaling. Taken-together, NPRL2 gene-therapy induces antitumor activity on KRAS/STK11^mt/aPD1R tumors through DC-mediated antigen-presentation and cytotoxic immune-cell activation.

Keywords

Animals, Mice, Humans, Lung Neoplasms, Carcinoma, Non-Small-Cell Lung, Proto-Oncogene Proteins p21(ras), Genetic Therapy, Tumor Suppressor Proteins, Disease Models, Animal, Antibodies, Monoclonal, Humanized, Cell Line, Tumor, Drug Resistance, Neoplasm, Mutation

Published Open-Access

yes

Recommended Citation

Meraz, Ismail M; Majidi, Mourad; Song, Renduo; et al., "NPRL2 Gene Therapy Induces Effective Antitumor Immunity in KRAS/STK11 Mutant Anti-PD1 Resistant Metastatic Non-Small Cell Lung Cancer (NSCLC) in a Humanized Mouse Model" (2025). Faculty, Staff and Student Publications. 6631.
https://digitalcommons.library.tmc.edu/uthgsbs_docs/6631