Differential expression of immune-regulatory proteins C5AR1, CLEC4A and NLRP3 on peripheral blood mononuclear cells in early-stage non-small cell lung cancer patients

Abstract

Changes in gene expression profiling of peripheral blood mononuclear cells (PBMC) appear to represent the host’s response to the cancer cells via paracrine signaling. We speculated that protein expression on circulating T-lymphocytes represent T-lymphocyte trafficking before infiltration into the tumor microenvironment. The possibility of using protein expression on circulating T-lymphocytes as a biomarker to discriminate early-stage non-small cell lung cancer (NSCLC) was explored. Four independent PBMC gene expression microarray datasets (GSE12771, GSE13255, GSE20189 and GSE3934) were analyzed. We selected C5AR1, CLEC4A and NLRP3 based on their significant protein expression in tumor-infiltrating lymphocytes, but not in normal lymphoid tissue. A validation study using automated flow cytometry was conducted in 141 study participants including 76 treatment-naive early-stage non-small cell lung cancer patients (NSCLC), 12 individuals with non-malignant pulmonary diseases, and 53 healthy individuals. Median ratios of C5AR1, CLEC4A and NLRP3 specific antibody staining to CD3 positive cells in early-stage NSCLC patients compared to healthy controls were 0.014 [0–0.37] vs. 0.01 [0–0.07, p = 0.13], 0.03 [0–0.87] vs. 0.02 [0–0.13, p = 0.10] and 0.19 [0–0.60] vs. 0.09 [0.02–0.31, p < 0.0001], respectively. Median fluorescence intensity (MFI) of CD3+C5AR1+, CD3+CLEC4A+ and CD3+NLRP3+ expression in early-stage NSCLC patients compared to healthy volunteers was 185 [64.2–4801] vs. 107.5 [27–229, p < 0.0001], 91.2 [42.4–2355] vs. 71.25 [46.2–103, p = 0.0005], and 1585 [478–5224] vs. 758.5 [318–1976, p < 0.0001], respectively. NLRP3:CD3 ratio, CD3+C5AR1+, CD3+CLEC4A+ and CD3+NLRP3+ MFI were significantly higher in early-stage NSCLC than healthy volunteers with an area under the ROC curve of 0.69–0.76. The CD3+NLRP3+ MFI provided the most distinguishable expression at 71.5% sensitivity and 70% specificity. Furthermore, CD3+NLRP3+ MFI potentially discriminated between early-stage NSCLC from malignant-mimic inflammation and infection pulmonary disease. Further validation in various pulmonary inflammatory disease might be warranted. Our proof-of-principle findings strengthen the hypothesis that malignancies generate distinctive protein expression fingerprints on circulating T-lymphocytes.

The high FKBP1A expression in WBCs as a potential screening biomarker for pancreatic cancer

“…Multiple studies have postulated about the ability of cancer cells to produce cytokine and chemokine-like secretion molecules which affect gene expression changes in blood circulating immune cells, including WBCs, contributing to the progression of various forms of cancer and impacting outcomes 31 – 34 . Recent works have shown that the levels of mRNA and protein expression of PBMCs exhibited changes in both hepatocellular carcinoma and non-small cell lung cancer 27 , 35 . In line with this previous work, our findings also described the mechanism of cancer cytokine signaling in molecule-regulated gene expression of immune cells in pancreatic cancer.…”

TMErisk score: A tumor microenvironment-based model for predicting prognosis and immunotherapy in patients with head and neck squamous cell carcinoma

“…Similarly, CLEC4A is also critical in tumorigenesis. In lung cancer and hepatocellular carcinoma, CLEC4 regulates the infiltration of various immune cells, making it a potential target for novel immunotherapy [ 33 , 34 ]. Recent reports have indicated that TENT5C, an unconventional poly(A) RNA polymerase known for boosting mRNA stability and gene expression, interferes with Plk4 function [ 35 ].…”

Identification of cell death-related biomarkers and immune infiltration in ischemic stroke between male and female patients

“…Except for ACSL1, a biomarker associated with ferroptosis, SLC2A3, MMP9, C5AR1, and NLRP3 are all biomarkers associated with pyroptosis. It has been shown that SLC2A3 is co-expressed with MMP9, SLC2A3 is co-expressed with C5AR1, and C5AR1 is co-expressed with NLRP3 ( 33 – 35 ), and they are all elevated genes in the male IS patient group. Therefore, pyroptosis is the primary form of cell death determining the course of IS in male patients.…”