Letter to the Editor Open Access
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Hepatol. Jan 27, 2023; 15(1): 116-122
Published online Jan 27, 2023. doi: 10.4254/wjh.v15.i1.116
Prognostic role of ring finger and WD repeat domain 3 and immune cell infiltration in hepatocellular carcinoma
Fang Zhang, Xin Dong, Jian Gan, Jiang-Tao Wang, Wu-Xia Quan, Yan-Dong Miao
Yan-Dong Miao, Fang Zhang, Cancer Center, Yantai Affiliated Hospital of Binzhou Medical University, Yantai 264000, Shandong Province, China
Wu-Xia Quan, Department of Electrocardiogram, Yantai Affiliated Hospital of Binzhou Medical University, Yantai 264100, Shandong Province, China
Jiang-Tao Wang, Department of Thyroid and Breast Surgery, Yantai Affiliated Hospital of Binzhou Medical University, Yantai 264000, Shandong Province, China
Jian Gan, Department of Gastroenterology, Yantai Affiliated Hospital of Binzhou Medical University, Yantai 264000, Shandong Province, China
Xin Dong, Second Department of Oncology, Yantai Affiliated Hospital of Binzhou Medical University, Yantai 264000, Shandong Province, China
ORCID number: Yan-Dong Miao (0000-0002-1429-8915); Wu-Xia Quan (0000-0002-1898-850X); Jiang-Tao Wang (0000-0002-1222-164X); Jian Gan (0000-0003-2645-6076); Xin Dong (0000-0002-4318-3228); Fang Zhang (0000-0001-5120-0991).
Author contributions: Miao YD and Zhang F designed the research; Miao YD wrote the paper; Quan WX and Wang JT performed the data analysis, Gan J and Dong X prepared the tables and figures; Zhang F and Miao YD reviewed the manuscript; Miao YD and Quan WX contributed equally to the article; All authors approved the final manuscript.
Conflict-of-interest statement: All the authors report having no relevant conflicts of interest for this article.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Fang Zhang, MD, Chief Doctor, Professor, Cancer Center, Yantai Affiliated Hospital of Binzhou Medical University, No. 717 Jinbu Street, Muping District, Yantai 264000, Shandong Province, China. zhangfang820127@163.com
Received: October 17, 2022
Peer-review started: October 17, 2022
First decision: November 15, 2022
Revised: October 29, 2022
Accepted: December 23, 2022
Article in press: December 23, 2022
Published online: January 27, 2023

Abstract

We have found that the expression of ring finger and WD repeat domain 3 (RFWD3) is significantly higher in unpaired and paired hepatocellular carcinoma (HCC) tissues than in normal tissues. Moreover, this expression has a significant correlation with the infiltration level of 14 immune cell types and when the detected RFWD3 expression levels were grouped as high and low, a prominent difference was revealed for overall survival, disease-specific survival, and progression-free interval. Through statistical analysis (univariate Cox), we were also able to identify RFWD3 as an independent prognostic element for HCC, with RFWD3 having an ability to accurately predict HCC prognosis (area under the curve of 0.863). Finally, we have generated prognostic nomograms for probabilities of 1-, 3- and 5-year overall survival in HCC via integrating the factors of age, pathologic stage, alpha-fetoprotein level, and RFWD3 expression.

Key Words: Hepatocellular carcinoma, Ring finger and WD repeat domain 3, Immune cell infiltration, Bioinformatics

Core Tip: We have discovered that ring finger and WD repeat domain 3 (RFWD3) expression is remarkably higher in tumor tissues compared to corresponding non-tumor tissues, regardless of hepatocellular carcinoma (HCC) tissue type (unpaired or paired). The RFWD3 expression also showed a significant correlation with the infiltration level of 14 immune cell types and was identified as an independent prognostic element in HCC by univariate Cox regression analysis. Our collective findings suggest that RFWD3 has the ability to accurately predict prognosis of HCC.



TO THE EDITOR

We perused the recently published paper by Liang et al[1] with much interest. The authors reported on their assessments of ring finger and WD repeat domain 3 (RFWD3) expression levels in hepatocellular carcinoma (HCC) patients. Their findings included RFWD3 effects on HCC prognosis, the processes of proliferation, invasion and metastasis, and the underlying mechanisms, specifically regulation via the Wnt/β-catenin signaling pathway. We have a particular appreciation for these authors' novel investigation into the prognostic implication of RFWD3 in HCC as we have also discovered that the expression of RFWD3 is prominently higher in both unpaired and paired HCC tissues from HCC patients than in their corresponding normal tissues (Figure 1A and B).

Figure 1
Figure 1 Differential expression levels of ring finger and WD repeat domain 3 in hepatocellular carcinoma and normal tissues. A: Non-paired hepatocellular carcinoma (HCC) and normal samples; B: Paired HCC and normal samples. Data source: mRNA-Seq data from the Genotype-Tissue Expression project (GTEx) of The Cancer Genome Atlas processed through the Toil process in the UCSC Xena database. (https://xenabrowser.net/datapages/)[2].

According to the current literature, cancer cells, endothelial cells, stromal cells, immune cells, and cancer-associated fibroblasts cells all exist in the tumor microenvironment (TME)[3,4]. While the TME is known to play crucial roles in development, invasion and metastasis of HCC, the immune escape of HCC cells has yet to be fully understood and continues to complicate cancer treatment[5]. Due to the ongoing and well-known limitations of chemotherapy in general, immunotherapies are a hot topic of bench and clinical research. This newly emerging cancer therapy exploits immune cells both inside and outside the TME to target and attack cancer cells; its demonstrated advantages are high specificity and low side-effects[6]. The power of this therapeutic method’s potential lies in the fact that different types of immune-related cells serve diverse roles; for HCC, the research into defining and developing those immune cells that inhibit/promote tumor processes has a long way to go[7].

Upon reading the report that Liang et al[1] found RFWD3 is able to affect the prognosis of HCC, we tested a hypothesis that the expression of RFWD3 may be associated with immune cell infiltration in HCC. Detailed information is shown in Table 1. Following our initial positive data, we systematically explored the correlation between RFWD3 expression and infiltration level of 24 immune cell types, using a single-sample gene set analysis (also known as ssGSEA) algorithm and Spearman coefficient correlation analysis[8]. We found that RFWD3 expression has a remarkable correlation with the infiltration level of 14 immune cell types (Figure 2A). Among them, RFWD3 expression was positively associated with the infiltration level of T helper (Th) cells in general, Th2 cells in particular, T follicular helper (TFH) cells, T central memory (Tcm) cells, activated dendritic cells (DCs), natural killer (NK) CD56bright cells, and eosinophils ( all P < 0.05; Figure 2B-H). There were negative associations with cytotoxic cells, DCs, plasmacytoid DCs (pDCs), neutrophils, T gamma delta (Tgd) cells, T regulatory cells (Tregs), and Th17 cells (all P < 0.05; Figure 2I-O). We hope our findings will encourage further investigations into RFWD3 as an HCC immunotherapy. Detailed information on this aspect is presented in Table 2.

Figure 2
Figure 2 Correlation analysis of ring finger and WD repeat domain 3 expression and immune cell infiltration in hepatocellular carcinoma. A: Lollipop plot manifesting the correlation between ring finger and WD repeat domain 3 (RFWD3) expression and the infiltration level of 24 immune cell types; B-H: The infiltration levels of 7 immune cell types have significant positive correlation with RFWD3 expression; B: T helper (Th)2 cells; C: Th cells; D: T follicular helper (TFH) cells; E: T central memory (Tcm) cells; F: Activated dendritic cells (aDCs); G: Natural killer (NK) CD56bright cells; H: Eosinophils; I-O: The infiltration levels of 7 immune cell types have significant negative correlation with RFWD3 expression; I: Cytotoxic cells; J: Dendritic cells (DCs); K: Plasmacytoid dendritic cells (pDCs); L: Neutrophils; M: T gamma delta (Tgd) cells; N: T regulatory cells (Tregs); O: Th17 cells.
Table 1 Detailed statistical results of differential expression of ring finger and WD repeat domain 3 in hepatocellular carcinoma and normal tissues.
Gene
Group
n
Minimum
Maximum
Median
IQR
Lower quartile
Upper quartile
Mean
SD
SE
RFWD3Normal500.5041.5040.980.2510.8831.1331.0130.2080.029
Tumor3740.623.51.5440.7551.24521.6470.5590.029
Normal500.5041.5040.980.2510.8831.1331.0130.2080.029
Tumor500.7072.9391.5770.7161.2041.921.5780.5210.074
Table 2 Detailed information on the statistical correlation between ring finger and WD repeat domain 3 expression and immune cell infiltration.
Gene
Immune cell type
Pearson’s correlation coefficient
Pearson’s P value
Spearman’s correlation coefficient
Spearman’s P value
RFWD3Th2 cells0.499< 0.0010.501< 0.001
Th cells0.434< 0.0010.436< 0.001
Cytotoxic cells-0.304< 0.001-0.314< 0.001
DCs-0.281< 0.001-0.304< 0.001
pDCs-0.261< 0.001-0.261< 0.001
Neutrophils-0.210< 0.001-0.214< 0.001
TFH cells0.226< 0.0010.213< 0.001
Tcm cells0.187< 0.0010.1640.002
Tgd cells-0.1050.043-0.1420.006
Tregs-0.1550.003-0.1210.019
aDCs0.1410.0060.1140.027
NK CD56bright cells0.1280.0130.1120.031
Th17 cells-0.170< 0.001-0.1100.033
Eosinophils0.0770.1350.1060.041
Macrophages0.0960.0630.0710.171
Th1 cells0.0900.0810.0640.214
iDCs-0.0340.507-0.0610.241
Mast cells-0.0530.309-0.0600.247
CD8 T cells-0.0470.368-0.0580.260
T cells-0.0130.796-0.0330.522
Tem cells0.0850.1000.0200.704
B cells0.0330.5250.0170.744
NK cells0.0350.494-0.0120.810
NK CD56dim cells0.0200.699-0.0010.979

Importantly, we agree with the finding of Liang et al[1] that indicates higher RFWD3 expression is related to worse overall survival (OS) in HCC. We have found that OS, disease-free survival, and progression-free interval were prominently shorter in HCC patient tissues with high RFWD3 expression than in those with low RFWD3 expression (all P < 0.05; Figure 3A-C). Our further statistical analysis via univariate Cox regression identified RFWD3 as an independent prognostic element for HCC (Table 3). Generation of the receiver operating characteristic curve showed that RFWD3 has the ability to accurately predict prognosis in HCC (area under the curve of 0.863). Finally, we generated prognostic nomograms for probabilities of 1-, 3- and 5-year OS in HCC via integrating the factors of age, pathologic stage, alpha-fetoprotein level, and RFWD3 expression; each element was assigned a score according to its contribution to survival (Figure 3E).

Figure 3
Figure 3 Survival analysis of ring finger and WD repeat domain 3 in hepatocellular carcinoma and the nomogram for prognosis. A-C: Ring finger and WD repeat domain 3 (RFWD3) expression is related to overall survival, disease-specific survival (B) and progression-free interval (C)[9] in the The Cancer Genome Atlas-liver hepatocellular carcinoma (HCC) data; D: Receiver operating characteristic curves for the RFWD3 gene’s prognosis predictive ability. The nomogram can predict 1-, 3- and 5-year overall survival of HCC based on clinicopathological features and the expression of RFWD3.
Table 3 Univariate Cox regression analysis in hepatocellular carcinoma.
Characteristics
Total, n
Univariate analysis
Hazard ratio (95%CI)
P value
Pathologic stage349
I173
II861.417 (0.868-2.312)0.164
III852.734 (1.792-4.172)< 0.001
IV55.597 (1.726-18.148)0.004
Child-Pugh grade240
A218
B211.595 (0.757-3.361)0.219
C12.138 (0.294-15.544)0.453
Fibrosis Ishak score214
075
1/2310.935 (0.437-2.002)0.864
3/4280.698 (0.288-1.695)0.428
5/6800.737 (0.410-1.325)0.308
Histologic grade368
G155
G21781.162 (0.686-1.969)0.576
G31231.185 (0.683-2.057)0.545
G4121.681 (0.621-4.549)0.307
RFWD33731.557 (1.148-2.110)0.004

Ultimately, our new findings highlight that the research of Liang et al[1] is worthy of attention and that subsequent efforts to build upon it, such as our related discoveries, may promote the next generation of effective and safe therapeutics, such as immunotherapies.

Statistical analysis

R statistical software (version 4.1.2; R Foundation for Statistical Computing, Vienna, Austria; https://www.R-project.org/) was used for all statistical analyses. Wilcoxon rank-sum test was used to perform the differential expression analysis of RFWD3 between HCC samples and corresponding normal samples, with results demonstrated by the “ggplot2” R package[10]. Survival analysis was carried out by log-rank test and univariate Cox regression. The association between RFWD3 expression and immune cell infiltration were performed by Spearman and Pearson analysis. Positive values of correlation coefficient indicate positive correlation, negative values indicate negative correlation.

Footnotes

Provenance and peer review: Unsolicited article; Externally peer reviewed.

Peer-review model: Single blind

Specialty type: Oncology

Country/Territory of origin: China

Peer-review report’s scientific quality classification

Grade A (Excellent): A

Grade B (Very good): 0

Grade C (Good): 0

Grade D (Fair): D

Grade E (Poor): 0

P-Reviewer: Ding Y, China; Papadopoulos N, Greece S-Editor: Xing YX L-Editor: Filipodia P-Editor: Xing YX

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