Immune infiltration in TP53 missense mutant contributes to poor prognosis in hepatocellular carcinoma, unlike CTNNB1 mutations

Abstract

Background: Hepatocellular carcinoma (HCC) is one of the deadliest cancer over the world. In this study, we aimed to determine the most critical molecular event in HCC patients with tumor protein p53 (TP53) or catenin beta 1 (CTNNB1) mutations, and to explore how these two mutations differ in their impact on HCC prognostication.

Methods: We performed an integrated comparative analysis of exome and transcriptome data from The Cancer Genome Atlas (TCGA) for HCC patients. Patient prognosis and correlation with the immune infiltration characteristics were performed. HCC cell line based in vitro experiments were also performed to validate the mechanistic insights.

Results: The 3-year progression-free survival (PFS) analysis of HCC patients with TP53 mutations indicated a significantly poorer clinical outcome compared to those with CTNNB1 mutations. Functional annotation of the TP53 mutant cohort revealed a substantial upregulation of genes associated with immune regulation, while the CTNNB1 mutant cohort displayed a prominent activation of metabolic pathways. Further comparative analysis and in vitro experiments showed that TP53 missense mutations activated the signal transducer and activator of transcription 3 (STAT3) signaling pathway, which fostered neutrophil infiltration and enhanced the enrichment of regulatory T (Treg) cells by secreting specific inflammatory molecules in the tumor microenvironment. Notably, treatment with a an STAT3 inhibitor suppressed the expression of these inflammatory molecules, underscoring how an immunosuppressive tumor microenvironment in the TP53 mutant cohort contributes to a poor prognosis.

Conclusion: Our study provides valuable insights, revealing that HCC patients with TP53 missense mutations exhibit a distinct immune profile associated with poorer clinical outcome compared to those with CTNNB1 mutations.

Keywords

hepatocellular carcinoma / progression-free survival / gene expression / p53 / β-catenin / neutrophil / T regulatory cell / chemokine / signal transduction / The Cancer Genome Atlas

References

[1] Rumgay H , Arnold M , Ferlay J , et al. Global burden of primary liver cancer in 2020 and predictions to 2040. J Hepatol. 2022; 77 (6): 1598- 1606.

[2] Dhanasekaran R , Nault JC , Roberts LR , Zucman-Rossi J . Genomic medicine and implications for hepatocellular carcinoma prevention and therapy. Gastroenterology. 2019; 156 (2): 492- 509.

[3] Hassin O , Oren M . Drugging p53 in cancer: one protein, many targets. Nat Rev Drug Discovery. 2023; 22 (2): 127- 144.

[4] Kim S , Jeong S . Mutation hotspots in the β-catenin gene: lessons from the human cancer genome databases. Mol Cells. 2019; 42 (1): 8- 16.

[5] Namani A , Veeraiyan D , Patra T . A comparative analysis indicates SLC7A11 expression regulate the prognostic value of KEAP1-NFE2L2-CUL3 mutations in human uterine corpus endometrial carcinoma. Free Radic Biol Med. 2024; 222: 223- 228.

[6] Sherman BT , Hao M , Qiu J , et al. DAVID: a web server for functional enrichment analysis and functional annotation of gene lists (2021 update). Nucleic Acids Res. 2022; 50 (W1): W216- W221.

[7] Li T , Fu J , Zeng Z , et al. TIMER2.0 for analysis of tumor-infiltrating immune cells. Nucleic Acids Res. 2020; 48 (W1): W509- W514.

[8] Patra T , Meyer K , Ray RB , Kanda T , Ray R . Akt inhibitor augments anti-proliferative efficacy of a dual mTORC1/2 inhibitor by FOXO3a activation in p53-mutated hepatocarcinoma cells. Cell Death Dis. 2021; 12 (11): 1073.

[9] Lin L , Jou D , Wang Y , et al. STAT3 as a potential therapeutic target in ALDH⁺ and CD44⁺/CD24⁺ stem cell-like pancreatic cancer cells. Int J Oncol. 2016; 49 (6): 2265- 2274.

[10] Patra T , Sasaki R , Meyer K , Ray RB , Ray R . Transforming growth factor β acts as a regulatory molecule for lipogenic pathways among hepatitis C virus genotype-specific infections. J Virol. 2019; 93 (18): e00811- 19.

[11] Murmu N , Ghosh P , Namani A , Patra T . Glyoxylate supplementation ameliorates colitis associated colon cancer progression. J Cell Physiol. 2024; 239 (11): e31394.

[12] Schulze K , Imbeaud S , Letouzé E , et al. Exome sequencing of hepatocellular carcinomas identifies new mutational signatures and potential therapeutic targets. Nat Genet. 2015; 47 (5): 505- 511.

[13] Lombardo D , Saitta C , Giosa D , et al. Frequency of somatic mutations in TERT promoter, TP53 and CTNNB1 genes in patients with hepatocellular carcinoma from Southern Italy. Oncol Lett. 2020; 19 (3): 2368- 2374.

[14] Koch AE , Kunkel SL , Harlow LA , et al. Epithelial neutrophil activating peptide-78: a novel chemotactic cytokine for neutrophils in arthritis. J Clin Invest. 1994; 94 (3): 1012- 1018.

[15] Jovic S , Linge HM , Shikhagaie MM , et al. The neutrophil-recruiting chemokine GCP-2/CXCL6 is expressed in cystic fibrosis airways and retains its functional properties after binding to extracellular DNA. Mucosal Immunol. 2016; 9 (1): 112- 123.

[16] Yadav SK , Stojkov D , Feigelson SW , et al. Chemokine-triggered microtubule polymerization promotes neutrophil chemotaxis and invasion but not transendothelial migration. J Leukoc Biol. 2019; 105 (4): 755- 766.

[17] Nitto T , Onodera K . The linkage between coenzyme A metabolism and inflammation: roles of pantetheinase. J Pharmacol Sci. 2013; 123 (1): 1- 8.

[18] Jungnickel C , Schmidt LH , Bittigkoffer L , et al. IL-17C mediates the recruitment of tumor-associated neutrophils and lung tumor growth. Oncogene. 2017; 36 (29): 4182- 4190.

[19] Yang L , Moses HL . Transforming growth factor β: tumor suppressor or promoter? Are host immune cells the answer? Cancer Res. 2008; 68 (22): 9107- 9111.

[20] Sippel TR , White J , Nag K , et al. Neutrophil degranulation and immunosuppression in patients with GBM: restoration of cellular immune function by targeting arginase I. Clin Cancer Res. 2011; 17 (22): 6992- 7002.

[21] Maruyama T , Kono K , Izawa S , et al. CCL17 and CCL22 chemokines within tumor microenvironment are related to infiltration of regulatory T cells in esophageal squamous cell carcinoma. Dis Esophagus. 2010; 23 (5): 422- 429.

[22] Antuamwine BB , Bosnjakovic R , Hofmann-Vega F , et al. N1 versus N2 and PMN-MDSC: a critical appraisal of current concepts on tumor-associated neutrophils and new directions for human oncology. Immunol Rev. 2023; 314 (1): 250- 279.

[23] Zhou SL , Zhou ZJ , Hu ZQ , et al. Tumor-associated neutrophils recruit macrophages and T-regulatory cells to promote progression of hepatocellular carcinoma and resistance to sorafenib. Gastroenterology. 2016; 150 (7): 1646- 1658.

[24] Zou JM , Qin J , Li YC , et al. IL-35 induces N2 phenotype of neutrophils to promote tumor growth. Oncotarget. 2017; 8 (20): 33501- 33514.

[25] Rodriguez PC , Quiceno DG , Ochoa AC . L-arginine availability regulates T-lymphocyte cell-cycle progression. Blood. 2007; 109 (4): 1568- 1573.

[26] Zhang F , Xia Y , Su J , et al. Neutrophil diversity and function in health and disease. Signal Transduct Target Ther. 2024; 9 (1): 343.

[27] Zhou SL , Dai Z , Zhou ZJ , et al. Overexpression of CXCL5 mediates neutrophil infiltration and indicates poor prognosis for hepatocellular carcinoma. Hepatology. 2012; 56 (6): 2242- 2254.

[28] Végran F , Rebucci M , Chevrier S , Cadouot M , Boidot R , Lizard-Nacol S . Only missense mutations affecting the DNA binding domain of p53 influence outcomes in patients with breast carcinoma. PLoS One. 2013; 8 (1): e55103.

[29] Zhu J , Sammons MA , Donahue G , et al. Gain-of-function p53 mutants co-opt chromatin pathways to drive cancer growth. Nature. 2015; 525 (7568): 206- 211.

[30] López I , P. Oliveira L , Tucci P , Álvarez-Valín F , A. Coudry R , Marín M . Different mutation profiles associated to P53 accumulation in colorectal cancer. Gene. 2012; 499 (1): 81- 87.

[31] Schulz-Heddergott R , Stark N , Edmunds SJ , et al. Therapeutic ablation of gain-of-function mutant p53 in colorectal cancer inhibits Stat3-mediated tumor growth and invasion. Cancer Cell. 2018; 34 (2): 298- 314.

[32] Traber KE , Hilliard KL , Allen E , et al. Induction of STAT3-dependent CXCL5 expression and neutrophil recruitment by oncostatin-M during pneumonia. Am J Respir Cell Mol Biol. 2015; 53 (4): 479- 488.

[33] Vasquez-Dunddel D , Pan F , Zeng Q , et al. STAT3 regulates arginase-I in myeloid-derived suppressor cells from cancer patients. J Clin Invest. 2013; 123 (4): 1580- 1589.

[34] Lu H , Ai J , Zheng Y , et al. IGFBP2/ITGA5 promotes gefitinib resistance via activating STAT3/CXCL1 axis in non-small cell lung cancer. Cell Death Dis. 2024; 15 (6): 447.

[35] Song M , He J , Pan QZ , et al. Cancer-associated fibroblast-mediated cellular crosstalk supports hepatocellular carcinoma progression. Hepatology. 2021; 73 (5): 1717- 1735.

[36] Wang C , Wang M , Zhang X , et al. The neutrophil-to-lymphocyte ratio is a predictive factor for the survival of patients with hepatocellular carcinoma undergoing transarterial chemoembolization. Ann Transl Med. 2020; 8 (8): 541.

[37] He Q , Huangfu L , Fan B , et al. T-cells infiltration mediates the association between neutrophil/lymphocyte ratio and survival in gastric cancer. Cancer Med. 2023; 12 (15): 15893- 15902.

[38] Yang Y , Qu Y , Li Z , Tan Z , Lei Y , Bai S . Identification of novel characteristics in TP53-mutant hepatocellular carcinoma using bioinformatics. Front Genet. 2022; 13: 874805.

[39] Siolas D , Vucic E , Kurz E , Hajdu C , Bar-Sagi D . Gain-of-function p53^R172H mutation drives accumulation of neutrophils in pancreatic tumors, promoting resistance to immunotherapy. Cell Rep. 2021; 36 (8): 109578.

[40] Ham SW , Jeon HY , Jin X , et al. TP53 gain-of-function mutation promotes inflammation in glioblastoma. Cell Death Differ. 2019; 26 (3): 409- 425.

[41] Ye Y , Luo Y , Sun Y , et al. Single-cell transcriptomic profiling reveals KRAS/TP53-driven neutrophil reprogramming in LUAD: a multi-gene prognostic model and therapeutic targeting of RHOV. Oncol Res. 2025; 33 (6): 1383- 1404.

RIGHTS & PERMISSIONS

The Author(s). Malignancy Spectrum published by John Wiley & Sons Australia, Ltd on behalf of Higher Education Press.