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Published on: July 20, 2019
TOB1 modulates neutrophil phenotypes to influence gastric cancer progression and immunotherapy efficacy
Jinfeng Zhang1, Yunlong Li2, Jing Chen3
1Scientific Research Center, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, China.
Introduction:
The ErbB-2.1(TOB1) signaling transducer protein is a tumor-suppressive protein that actively suppresses the malignant phenotype of gastric cancer cells. Yet, TOB1 negatively regulates the activation and growth of different immune cells. Understanding the expression and role of TOB1 in the gastric cancer immune environment is crucial to maximize its potential in targeted immunotherapy.
Methods:
This study employed multiplex immunofluorescence analysis to precisely delineate and quantify the expression of TOB1 in immune cells within gastric cancer tissue microarrays. Univariate and multivariate Cox analyses were performed to assess the influence of clinical-pathological parameters, immune cells, TOB1, and double-positive cells on the prognosis of gastric cancer patients. Subsequent experiments included co-culture assays of si-TOB1-transfected neutrophils with AGS or HGC-27 cells, along with EdU, invasion, migration assays, and bioinformatics analyses, aimed at elucidating the mechanisms through which TOB1 in neutrophils impacts the prognosis of gastric cancer patients.
Results:
We remarkably revealed that TOB1 exhibits varying expression levels in both the nucleus (nTOB1) and cytoplasm (cTOB1) of diverse immune cell populations, including CD8+ T cells, CD66b+ neutrophils, FOXP3+ Tregs, CD20+ B cells, CD4+ T cells, and CD68+ macrophages within gastric cancer and paracancerous tissues. Significantly, TOB1 was notably concentrated in CD66b+ neutrophils. Survival analysis showed that a higher density of cTOB1/nTOB1+CD66b+ neutrophils was linked to a better prognosis. Subsequent experiments revealed that, following stimulation with the supernatant of tumor tissue culture, the levels of TOB1 protein and mRNA in neutrophils decreased, accompanied by enhanced apoptosis. HL-60 cells were successfully induced to neutrophil-like cells by DMSO. Neutrophils-like cells with attenuated TOB1 gene expression by si-TOB1 demonstrated heightened apoptosis, consequently fostering a malignant phenotype in AGS and HCG-27 cells upon co-cultivation. The subsequent analysis of the datasets from TCGA and TIMER2 revealed that patients with high levels of TOB1 combined neutrophils showed better immunotherapy response.
Discussion:
This study significantly advances our comprehension of TOB1's role within the immune microenvironment of gastric cancer, offering promising therapeutic targets for immunotherapy in this context.
Insights
TOB1, a tumor-suppressive protein, is found in gastric cancer immune cells, particularly neutrophils. Higher levels of TOB1 in neutrophils correlate with better patient prognosis and immunotherapy response.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- The ErbB-2.1 (TOB1) signaling transducer protein suppresses gastric cancer malignancy but inhibits immune cell activation.
- Understanding TOB1's role in the gastric cancer immune microenvironment is vital for targeted immunotherapy.
Purpose of the Study:
- To investigate the expression and function of TOB1 in immune cells within the gastric cancer microenvironment.
- To determine the prognostic significance of TOB1 and its association with immune cells in gastric cancer patients.
- To elucidate the mechanism by which TOB1 in neutrophils influences gastric cancer prognosis and immunotherapy response.
Main Methods:
- Multiplex immunofluorescence analysis to quantify TOB1 expression in various immune cells.
- Univariate and multivariate Cox regression analyses for prognostic assessment.
- Co-culture assays with si-TOB1-transfected neutrophils and gastric cancer cells, alongside EdU, invasion, and migration assays.
- Bioinformatics analysis of TCGA and TIMER2 datasets.
Main Results:
- TOB1 expression was detected in multiple immune cell types, with notable concentration in CD66b+ neutrophils.
- Higher density of cytoplasmic and nuclear TOB1 in neutrophils correlated with improved patient prognosis.
- Reduced TOB1 in neutrophils led to increased apoptosis and promoted gastric cancer cell malignancy.
- High TOB1 and neutrophil levels predicted a better response to immunotherapy.
Conclusions:
- TOB1 expression in neutrophils is a significant prognostic biomarker in gastric cancer.
- Targeting TOB1 in neutrophils may offer a novel therapeutic strategy for enhancing immunotherapy efficacy in gastric cancer.
- This study provides critical insights into the interplay between TOB1, neutrophils, and the immune microenvironment in gastric cancer.
