IL20RB is a Prognostically Significant Gene and a Potential Biomarker in Papillary Renal Cell Carcinoma

doi:10.21203/rs.3.rs-5319680/v1

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Research Article

IL20RB is a Prognostically Significant Gene and a Potential Biomarker in Papillary Renal Cell Carcinoma

https://doi.org/10.21203/rs.3.rs-5319680/v1

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Background

Papillary renal cell carcinoma (pRCC) is the second most common type of renal cancer after clear cell renal cell carcinoma. While diagnostic and therapeutic approaches for pRCC often draw on experiences from clear cell renal cell carcinoma, specific targets for diagnosis and treatment remain elusive. IL20RB, the interleukin 20 receptor subunit β, plays a crucial role in IL20R heterodimerization. Initially associated with a variety of skin diseases, recent studies have implicated IL20RB in various malignancies, yet its role in pRCC remains unclear.

Methods

Prognostic value was determined using the TCGA database and GEPIA, with validation through the analysis of our center’s samples. Subsequent, survival analysis of pRCC patients with high and low IL20RB expression was conducted using R software (version 4.2.1). To further understand the function of IL20RB in pRCC, immune infiltration analysis as well as KEGG/GO enrichment analysis were performed using TRNA sequencing expression profiles from the TCGA database. Protein-protein interactions (PPI) between IL20RB and other proteins in human species were explored using the STRING (String-db.org) database, and drug sensitivity analysis was conducted using the GDSC database.

Results

Our results revealed specific expression of IL20RB in pRCC tumor tissues, with high expression associated with a poorer prognosis and significantly positively correlated with clinical features. Additionlly, IL20RB may function as a cytokine receptor involved in the immune-inflammatory response in the pRCC microenvironment.

Conclusions

IL20RB plays an important role in pRCC and may serve as a potential prognostic oncogene, offering a potential target for future therapeutic interventions.

Papillary renal cell carcinoma

IL20RB

Cytokine receptor

Oncogene

Prognosis

Papillary renal cell carcinoma (pRCC) is the second most common subtype of renal cell carcinoma (RCC) after clear cell renal cell carcinoma (ccRCC), accounting for approximately 15% of all RCCs, and is histologically characterized by the presence of fibrovascular cores with tumor cells arranged in a papillary configuration^[1–3]. Whilie studies in the field of prognosis-related biomarkers and targeted therapy for RCC have mainly focused on ccRCC^[4–8], there is a relative scarcity of research on pRCC, leaving numerous gaps in our understanding. In addition, there are no clear hallmark molecular changes that can diagnose PRCC, and the diagnosis of pRCC is still based on morphological diagnosis, supplemented by IHC profiles and by exclusion of outliers^[9]. Currently, the WHO/ISUP classification, although widely accepted as the main prognostic factor for PRCC, has some limitations. Some studies have yielded varying results regarding its prognostic value including its lack of significance in multivariate analysis^[10–12]. Therefore, the identification of novel molecular targets for pRCC is imperative, which will facilitate more accurate diagnosis and better prognosis and treatment of pRCC.

With the increasing research on tumors, immune cells and immune-related molecules play a pivotal role in tumor development. Interleukins and their receptors have received much attention in many tumor studies^[13–16]. RCC is an immunogenic tumor^[17], and studies have shown that interleukins and their receptor molecules play an important role in the development of RCC. For example, Masuda et al. found that serum levels of soluble interleukin-2 receptor (sIL-2R) reflected tumor progression in patients with ccRCC, and the survival of patients with high sIL-2R levels was shorter than that of patients with low sIL-2R levels^[18]. In addition, Kansler et al. also found that cytokine interleukin 15 (IL-15) in complex with IL-15 receptor α chain (IL-15Rα) can promote the expansion and cytotoxic function of intraepithelial type 1 innate lymphoid cells (ILC1s), thereby effectively inhibiting chromophobe RCC (chRCC) progression^[19].

IL20RB, or interleukin 20 receptor subunit β, plays a crucial role in the constitution of both type I IL20R (IL20RA and IL20RB) and type II IL20R (IL22RA1 and IL-20RB). Specifically, the IL20RA/IL20RB dimer functions as the receptor for IL19, IL20, and IL24, while the IL22RA1/IL20RB dimer serves as the receptor for IL20 and IL24^{[20, 21]}. IL20RB is the common chain for both receptor types, therefore, its role in IL20R heterodimerization is important. Although IL20RB was initially found to be associated with a variety of skin diseases, such as vitiligo and psoriasis^{[22, 23]}, recent studies have found that IL20RB is also associated with a variety of malignancies, such as nasopharyngeal carcinoma, colorectal cancer, breast cancer and pancreatic ductal adenocarcinoma^[24–27]. Recently, Guo et al. also demonstrated that IL20RB plays an important role in ccRCC and is a novel prognostic and potential therapeutic biomarker for ccRCC^[28]. However, the relationship between IL20RB and pRCC is still unclear and there is still much room for research.

In this study, we investigated specific aspects of IL20RB’s role in pRCC, with a focus on its specific expression, clinical prognostic value, immune infiltration, related functional pathways and drug treatment sensitivity analysis. Additionally, we validated the expression of IL20RB in tumor tissues and assessed its prognostic relevance in conjunction with clinical data.

2.1 Patients. We included a group of 42 patients with pRCC who underwent partial or radical nephrectomy from January 1, 2016 to December 31, 2022 at the Department of Urology, Zhongda Hospital, Southeast University. The inclusion criteria for the study population were as follows: 1. Histopathologic diagnosis of papillary renal cell carcinoma; 2. No other renal disease in the past; 3. No drug therapy prior to surgery. We collected patients' tumor tissue wax blocks to produce tissue paraffin sections, and we also collected patients' clinical data, including age, gender, pathological type, laterality, T stage, clinical stage, surgical methods (partial versus radical nephrectomy) and so on. The clinical baseline characteristics of the patients are shown in Supplementary Table 1. Our study was approved by the ethical committee of Zhongda Hospital Southeast University (2022ZDSYLL392-P01). We also included 88 patients with pRCC from the TCGA database, and the clinical baseline characteristics are shown in Supplementary Table 2.

2.2 Screening of Key Prognostic Genes in pRCC. Clinical data and transcriptome RNA-seq data of 291 pRCC patients were obtained from The Cancer Genome Atlas (TCGA). The survival package in R software (version 4.2.1) was used to perform batch fit survival regression analysis of TCGA pRCC data to obtain OS (Overall Survival) differential genes, DSS (Disease Specific Survival) differential genes, and PFI (Progression Free Interval) differential genes, respectively. The glmnet package was used to perform lasso regression analyses to re-screen for prognostic differential genes. The pROC package was used for ROC analysis and ROC testing. Venn diagrams were produced via the Bioinformatics & Evolutionary Genomics website (http://bioinformatics.psb.ugent.be/webtools/Venn/).

2.3 Specific Expression of IL20RB and IL20 Subfamily Genes in pRCC. The median expression of IL20RB and IL20 subfamily genes (IL-19, IL-20, IL-22, IL-24 and IL-26) in tumor tissues and normal tissues of pRCC patients was obtained through the GEPIA2 website (GEPIA 2 (cancer-pku.cn)). We also compared the differences in IL20RB expression between patients with different T stages and clinical stages based on TCGA pRCC transcriptome RNA-seq data using the ggplot2 (3.3.6) package, stats (4.2.1) package, and car (3.1-0) package in R (version 4.2.1).

2.4 Immunohistochemistry. The tissue was fixed in 10% buffered formalin, dehydrated in ascending order of alcohols, embedded in paraffin blocks, and cut to a thickness of 3–4 microns. We performed IL20RB immunohistochemical staining on their tissue sections (Anti-IL-20RB Rabbit Polyclonal Antibody, HuaBio, ER63751). Semi-quantitative scoring of IL20RB immunohistochemical staining intensity was as follows: 0: no staining; 1: light brown was weakly positive; 2: brown was positive; 3: dark brown was strongly positive. The pathologist observed the staining of cancerous and paracancerous tissues in each kidney cancer specimen separately and assessed the staining intensity using histochemical score. Patients whose cancerous tissue staining intensity was greater than that of the paracancerous tissue were categorized into the IL20RB high expression group, and patients whose cancerous tissue staining intensity was lower than that of the paracancerous tissue were categorized into the IL20RB low expression group. There were 33 patients in the IL20RB high expression group and 9 patients in the IL20RB low expression group.

2.5 Survival Analysis of IL20RB. We analyzed the pRCC data in TCGA using the survminer package and the survival package in R (version 4.2.1) to obtain the Kaplan-Meier curve for IL20RB. In addition, to further validate the results of the TCGA data, we obtained Kaplan-Meier curves for IL20RB in our patients in conjunction with the data from our center.

2.6 Immune Infiltration Analysis. TRNA-sequencing expression profiles and corresponding clinical data for IL20RB were obtained from the TCGA dataset (https://portal.gdc.com). Correlations between gene expression and immune score were visualized using R with the ggstatsplot package, and multi-gene correlations were depicted using the pheatmap package. Spearman’s correlation analysis, suitable for non-normally distributed variables, was employed to illustrate relationships between quantitative variables. Statistical significance was set at P values below 0.05 (P < 0.05). All analyses were conducted using R (version 4.2.1).

2.7 Functional Analysis of IL20RB in pRCC. IL20RB's RNA-sequencing expression profiles and clinical data were acquired from TCGA (https://portal.gdc.com). Differential mRNA expression was studied using the limma package in R, defining significance as "Adjusted P < 0.05 and |Log2 (Fold Change) |> 1." To explore potential targets, functional enrichment analysis was conducted, employing Gene Ontology (GO) for molecular function (MF), biological pathways (BP), and cellular components (CC), along with Kyoto Encyclopedia of Genes and Genomes (KEGG) Enrichment Analysis. Carcinogenesis insights for mRNA were gained using the Cluster Profiler package (version: 3.18.0) in R (version 4.2.1), analyzing GO functions and enriching KEGG pathways. Heatmaps were created using the R pheatmap package. STRING (String-db.org) database was utilized to explore the protein-protein interactions (PPI) between IL20RB and other proteins in human species. The PPI network map was constructed and drawn by connecting the hub genes with specific lines, which has been verified by clinical samples, experiments, and protein crystal structure.

2.8 Drug Sensitivity Analysis of IL20RB in pRCC. IL20RB's RNA-sequencing expression profiles and clinical data were sourced from the TCGA dataset (https://portal.gdc.com). Chemotherapeutic response predictions were made using the Genomics of Drug Sensitivity in Cancer (GDSC) database (https://www.cancerrxgene.org/) via the R package "pRRophetic." Ridge regression was applied to estimate half-maximal inhibitory concentration (IC50), with default parameter settings. To address batch effects and consider tissue types, combat was utilized, and duplicate gene expression values were averaged.

3.1 IL20RB is a potential key prognosis-related gene in pRCC. We used the transcriptome RNA-seq data from the TCGA database to obtain the OS differential genes, DSS differential genes, and PFI differential genes of pRCC, respectively, and Venn diagrams were used to visualize their intersection. As shown in Fig.1a, there are 47 genes that are common intersections of the OS differential gene, the DSS differential gene, and the PFI differential gene. Lasso regression can narrow down the gene screening scope and improve the accuracy and interpretability of the analysis results through some mechanisms such as automatic feature selection and prevention of overfitting. Therefore, by lasso regression analysis, we further screened OS differential genes, DSS differential genes and PFI differential genes to obtain four key prognostic genes of pRCC, which were IGF2BP3, SLC7A11, FBXO43 and IL20RB, and visualized them in a Venn diagram (Fig.1b). As shown in Fig.1c and Fig.1d, four genes were significantly more expressed in tumor tissues than in normal tissues in both non-paired and paired samples of pRCC. To determine whether these four genes have diagnostic value for pRCC, we performed ROC correlation analysis in combination with TCGA data and visualized the results. As shown in Fig.1e, the AUC (Area Under Curve) of IGF2BP3, SLC7A11, FBXO43, and IL20RB were all greater than 0.5, but the AUC of IL20RB was significantly larger, which indicates that IL20RB has a higher diagnostic value in pRCC and is worth focusing on.

3.2 Specific Expression of IL20RB and IL20 Subfamily Genes in pRCC. In order to have a visual panorama of IL20RB expression in pRCC, we used the GEPIA2 website to obtain the average expression of IL20RB in tumor tissues as well as normal tissues of pRCC. Also considering that IL20RB is a key receptor subunit of IL20, we also visualized the average expression of IL20 subfamily genes in tumor tissues as well as normal tissues of pRCC. As shown in Fig.1f, IL20RB was highly expressed in pRCC tumor tissues relative to normal tissues. In addition, except for IL24, other IL20 subfamily genes were not expressed in tumor tissues or were more prominently expressed in normal tissues. It has been demonstrated that IL24 is an immunomodulatory cytokine and is mainly characterized by the induction of tumor cell apoptosis to inhibit tumor growth in cancer^{[29, 30]}. The above suggests that IL20RB may not play a biological role in pRCC through direct binding with IL20 subfamily genes. To further understand the correlation between IL20RB expression in pRCC and clinical features, we compared IL20RB expression in pRCC patients with different T stages as well as clinical stages based on TCGA data. As illustrated in Figure 2, the expression of IL20RB was different and the difference was statistically significant at different pathologic stages as well as clinical stages. The expression of IL20RB gradually increased with the elevation of pathologic T stage, N stage, and M stage. Similarly, with the increase of clinical T stage, N stage and M stage, the expression of IL20RB also gradually increased. This suggests that IL20RB expression is positively correlated with clinical features.

3.3 Immunohistochemistry of IL20RB in pRCC. We collected a total of 42 postoperative tissue samples from patients with pRCC at Zhongda Hospital, Southeast University. All participants provided informed consent, and the study was approved by the Ethics Committee of Zhongda Hospital, Southeast University (2022ZDSYLL392-P01). Fig.3A and Fig.3B demonstrates the carcinoma tissue of pRCC, while Fig.3C and Fig.3D show paracarcinoma renal tissue of pRCC with IL20RB staining in brown. We observed the staining of carcinoma and paracancerous tissues in each pRCC specimen separately and compared the staining intensity. Staining intensity of carcinoma tissues was greater than that of paracancerous tissues implying upregulation of IL20RB expression, while staining intensity of carcinoma tissues was lower than that of paracancerous tissues implying decreased IL20RB expression. We found that IL20RB was mainly expressed in the cytoplasm of cancer cells in papillary renal carcinoma tissues, and some of it was also expressed in the cell membrane, and most of the expression was positive and strongly positive. Meanwhile, IL20RB was also widely expressed in the cytoplasm of renal tubular epithelial cells adjacent to cancer cells. Among the 42 patients with pRCC in our center, 33 patients presented high expression of IL20RB in immunohistochemically stained sections (Fig.3E and Fig.3F) and 9 patients presented low expression of IL20RB in immunohistochemically stained sections (Fig.3G and Fig.3H), which shows that IL20RB is overall upregulated in expression in tumor cells of pRCC. Meanwhile, in the IL20RB high expression group, we compared IL20RB staining results in different T stages (Fig.4A-4D). We found that IL20RB expression gradually increased with increasing T stage, which could also be reflected by the positive cell index of IL20RB (Fig.4E).

3.4 Survival Analysis of IL20RB. We obtained the Kaplan-Meier curve for IL20RB by analyzing the data of pRCC patients in TCGA. As shown in Figure 5a, pRCC patients with high expression of IL20RB had a significantly worse prognosis than pRCC patients with low expression of IL20RB. To further validate this result, we combined the data from our center and divided the patients into IL20RB high-expression and low-expression groups based on the results of immunohistochemical staining, thus obtaining the Kaplan-Meier curve of IL20RB in pRCC patients in our center (Figure 5b). We suggest that IL20RB may be an oncogene and its high expression may be associated with a worse prognosis in pRCC.

3.5 Immune Infiltration Analysis. To analyze the potential correlation between the expression of IL20RB and the immune infiltration status in the tumor microenvironment of pRCC, we utilized the Quantiscore method to explore the relationship between IL20RB and a range of major immune cells. A significant negative correlation was found between IL20RB expression and the infiltration of CD4+ T cells (Fig.6g), which indicated that IL20RB may promote the malignant progression of pRCC through downregulation of adaptive anticancer immunity. In addition, although the CD8+ T cells also exhibit positive correlation with the expression of IL20RB, the overall expressive extent is relatively low (Fig.6h). More importantly, as the major immune suppressive cells^[31], regulatory T cells were found to be significantly positively-associated with the up-regulation of IL20RB (Fig.6i), which demonstrated that IL20RB may also induce immune tolerance in pRCC through induction of polarization or recruitment of regulatory T cells. In addition to T cells, IL20RB was also found to be correlated with a range of myeloid cells, such as neutrophils (Fig.6e), M1 macrophages (Fig.6b), and monocytes (Fig.6d), highlighting its involvement in shaping the myeloid component of the pRCC immune microenvironment.

3.6 Functional Analysis of IL20RB in pRCC. To validate the potential function of IL20RB, we performed KEGG and GO enrichment analyses based on pRCC expression profiling data from TCGA. Figure 7a illustrates that, in KEGG enrichment analysis, differentially expressed genes (DEGs) associated with IL20RB upregulation were mainly enriched in the PI3K−Akt signaling pathway and Cytokine−cytokine receptor interaction. PI3K-Akt signaling has been found to be involved in regulating tumor cell metastasis and invasion in a variety of tumors^[32-34]. Figure 7b depicts the results of GO enrichment analysis, indicating that DEGs associated with IL20RB upregulation were primarily enriched in the T cell activation, extracellular structure organization, extracellular matrix organization and positive regulation of cell adhesion. In contrast, DEGs associated with IL20RB down-regulation were predominantly enriched in the Carbon metabolism and small molecule catabolic process (Fig.7c and Fig.7d). The heatmap of IL20RB-related differential genes in pRCC is shown in Supplementary Figure 1. Significant differential genes associated with IL20RB up-regulation include TPM2, CIR, BST2, CIS, ANGPTL4 and so on. Significant differential genes associated with IL20RB down-regulation mainly include OGDHL, MIOX, AKR7A3 and SLC13A1. Moreover, we utilized the String method to explore the protein-protein interactions and potential functional molecular partners centered on IL20RB. As is demonstrated in the PPI map (Figure.8), significant interactions are found between IL20RB and a range of crucial cytokines, such as IFN-γ, IL-6, IL-10, and TNF, etc., which indicated that IL20RB may be involved in the regulation of the inflammatory microenvironment of pRCC. Moreover, multiple key molecular adaptors and signaling pathways in oncologic processes are also found to be in interaction with IL20RB, such as JAK1 and TYK2, which indicated that IL20RB may also mediates the oncogenic pathways of pRCC through classical signaling pathways.

3.7 Drug Sensitivity Analysis of IL20RB in pRCC. By using the GDSC database, we performed drug sensitivity analysis of pRCC patients in TCGA. As shown in Figure 9, individuals with high IL20RB expression were sensitized to common targeted drugs including axitinib, pazotinib, sunitinib, tivozanib, and the chemotherapeutic drug cisplatin. Notably, tivozanib had the lowest half-maximal inhibitory concentration (Fig.5e), suggesting its potential as a highly effective therapeutic option for pRCC patients with elevated IL20RB expression. Although there are no prospective studies evaluating the efficacy of tivozanib in pRCC, a phase III clinical trial by Rini, Brian I et al. also found that tivozanib as third-line or fourth-line therapy improved progression-free survival and was better tolerated compared with sorafenib in patients with metastatic renal cell carcinoma^[35], which may provide some inspiration for future clinical trials of tivozanib in pRCC. Conversely, our analysis revealed that pRCC patients with high expression of IL20RB display low sensitivity to erlotinib (Fig.9b), indicating that erlotinib may have limited therapeutic effect in this subgroup of pRCC patients in clinical practice.

Papillary renal cell carcinoma (pRCC), which ranks as the second most common subtype of renal cell carcinoma following clear cell renal cell carcinoma (ccRCC), is distinguished by a distinct set of clinicopathological and molecular features. Unlike ccRCC, which predominantly presents with a clear cytoplasmic appearance, pRCC is characterized by a papillary growth pattern and a different spectrum of genetic alterations. These differences not only influence the tumor’s biological behavior but also have implications for diagnosis, prognosis, and therapeutic strategies, making pRCC a unique entity within the broader category of renal cell carcinomas. Understanding these distinguishing characteristics is essential for optimizing clinical management and guiding targeted research efforts. Diagnostic and therapeutic strategies for pRCC have predominantly been extrapolated from studies primarily involving ccRCC tumors. Nonetheless, the distinct features of pRCC bear implications for diagnosis and management of both localized and metastatic disease. Therefore, various research studies focusing on pRCC have attracted attention from researchers. Currently, there exists a lack of consensus regarding the optimal regimen for the management of advanced pRCC. However, ongoing trials suggest promising results for both molecular targeted therapy and immunotherapy in this subtype^[36–38]. This underscores the imperative need for further research to identify novel targets that can enhance the diagnosis and treatment of pRCC.

Through the lasso regression analysis of prognostic differential genes, we initially screened four key prognostic genes, namely IGF2BP3, SLC7A11, FBXO43 and IL20RB, which were associated with OS, DSS and PFI in pRCC. As shown in Fig. 1c and Fig. 1d, although the expression of the four genes in tumor tissues was significantly higher than that of the normal tissues, we wished to obtain a more statistically diagnostic value gene for the study. Based on the TCGA data we performed ROC correlation analysis (Fig. 1e), and we found that the AUC of IL20RB was the largest, which indicated that IL20RB had a better statistical classification effect in pRCC, and was statistically valuable for subsequent grouping studies. So our team chose to further study IL20RB instead of the other three genes found during the screening process. Although IL20RB has been found to be associated with the biological behavior of a variety of tumors^[39–43], its specific role in pRCC remains elusive. As, a receptor subunit, IL20RB serves as the common chain for two types of IL20 receptor molecules, enabling it bind with diverse IL20 subfamily members^{[44, 45]}. Given whether IL20RB may play a biological role in pRCC by binding to IL20 subfamily members, we utilized GEPIA2 to obtain the average expression of IL20RB and IL20 subfamily members in pRCC. The results indicated a significant upregulation of IL20RB in tumor tissues within the pRCC tumor microenvironment. Additionally, IL24 expression exhibited an upregulated in tumors compared to normal tissues, while other IL20 subfamily members did not show significant expression in tumor tissues. This implies that the biological role of IL20RB in pRCC may not be strongly associated with the IL20 subfamily, raising the possiblility of existence e of other functionally relevant ligands.

In recent years, scholars have found that IL20RB plays an important role in some malignant diseases. For example, He et al. found that IL-20RB-expressing lung cancer tumor cells could mediate the response to osteoclasts and thus directly promote bone metastasis of lung cancer^[39]. A study on pancreatic cancer by Li et al. also showed that IL20RB plays a crucial role in promoting stemness in pancreatic cancer^[46]. IL20RB has also been recognized as a potential biomarker in ccRCC^[28]. To further investigate the correlation between IL20RB expression and clinical features of pRCC, we verified that IL20RB expression was positively correlated with pathological staging and clinical staging indicators in the TCGA cohort. To verify the credibility of the results, we included a group of 42 pRCC patients who underwent partial or radical nephrectomy in our center. Immunohistochemical staining for IL20RB revealed predominant expression in the cytoplasm and membrance of cancer cells in carcinoma tissues, the majority of which exhibited positive and strongly positive staining. In addition, based on the IL20RB-positive cell index, we reiterated a positive correlation between IL20RB expression and pathologic T stage. Kaplan-Meier analysis of both the TCGA cohort and our center patient cohort validated that high expression of IL20RB correlates with a worse prognosis for pRCC patients. These results imply that IL20RB may act as a potential tumor driver gene in pRCC.

To reveal the relationship between IL20RB and immune infiltrating cells, we utilized the Quantiscore method to explore the association between IL20RB and a range of major immune cell types. The results suggested a significant negative correlation between IL20RB expression and the infiltration of CD4 + T cells, coupled with a significant positive correlation with regulatory T cell infiltration. CD4 + T cells are well-known for their association with adaptive anti-tumor immunity^[47], while regulatory T cells are associated with harmful autoimmunity in tumors^[48]. This suggests that IL20RB might contribute to malignant progression of pRCC by suppressing adaptive anti-cancer immunity, or it might induce immune tolerance of pRCC by inducing polarization or recruitment of regulatory T cells. Thus, novel combination therapies targeting IL20RB and CD4 + T cells/regulatory T cells may be promising.

At present, although there are some IL20RB-related studies based on renal cancer, the possible functional pathway of IL20RB in pRCC has not been clarified. We performed KEGG and GO enrichment analyses as well as String analysis of IL20RB based on the TCGA cohort to reveal the potential function of IL20RB in pRCC. Enrichment analysis indicates that IL20RB is more likely to act as a cytokine receptor, and tumor samples with high IL20RB expression are mainly enriched in the immune inflammation-related pathway such as PI3K − Akt signaling pathway and Cytokine − cytokine receptor interaction. We utilized the String method to explore the protein-protein interactions and potential functional molecular partners centered on IL20RB. The analysis revealed significant interactions between IL20RB and crucial cytokines, including IFN-γ, IL-6, IL-10, and TNF, all established to be associated with inflammatory responses in tumors^[49–52]. This suggests that IL20RB may be involved in the regulation of the pRCC inflammatory microenvironment. In addition, several key molecular junctions and signaling pathways in the tumor process were found to interact with IL20RB, such as JAK1 and TYK2. Both the JAK1-associated signaling pathway and the TYK2-associated signaling pathway are well-studied classical pro-carcinogenic pathways^{[53, 54]}, indicating that IL20RB may also mediate the oncogenic pathway of pRCC through classical signaling pathways.

Some studies have shown that cytokines play a multifaceted important role in targeted therapy of RCC, such as synergistic enhancement, increased sensitivity, immune regulation and regulation of angiogenesis^[55–57]. Given the recent development of several clinical trials for pRCC, we attempted to perform a drug sensitivity analysis of pRCC patients within the TCGA using the GDSC database. Our findings revealed that pRCC patients exhibiting high IL20RB expression demonstrated increased sensitivity to commonly targeted drugs, including axitinib, pazotinib, sunitinib, tivozanib, and the chemotherapeutic drug cisplatin. It's important to note that, due to the limited number of drugs included in the GDSC database, the sensitivity analysis for some cutting-edge drugs could not be covered. Nevertheless, this underscores the substantial research space in the realm of drug therapy for pRCC. The quest for more sensitive and effective targeted drugs remains an imperative task in the ongoing landscape of clinical trials.

As is known, numerous genes, such as PTTG1, TPX2, claudin 6, CENPA, P4HB and others, have been identified as potential biomarkers in pRCC^[58–62]. Despite the focus on different genes in these studies, all identified genes hold promise for future pRCC investigations, contributing to the deepening of our understanding of this condition. In this study, we focused on the role of the prognostically critical gene IL20RB in pRCC. Our emphasis encompassed the expression, clinical relevance, immune infiltration and functional analysis of IL20RB in pRCC, as well as drug sensitivity analysis of commonly used targeted agents. We are pleased to contribute our own efforts to the study of pRCC. Overall, there are still some shortcomings in our study, for example, since pRCC is a rare renal cancer, we included a relatively small sample size from our center, and we need to combine multicenter data in the next study. In addition, we need more in-depth studies to elucidate the mechanism of IL20RB in pRCC. However, we believe that our research will become meaningful if our exploration can benefit patients in the future.

PRCC, the second most common histologic subtype of renal cancer, has been relatively limited studied. Our investigation revealed the specific expression of IL20RB in pRCC tumor tissues, and its elevated expression associates with a worse prognosis, suggesting its potential role as a prognostic relevant oncogene. Furthermore, IL20RB appears to play a role in the immune-inflammatory response in the pRCC tumor microenvironment, positioning it as a potential target for future targeted drugs.

Ethics approval and consent to participate

Our study was approved by the ethical committee of Zhongda Hospital Southeast University (2022ZDSYLL392-P01). We obtained previous postoperative pathology tissue from clinical patients at our center and did not perform interventional experiments on the participants. Disease information for all participants was obtained from their verbal consent at follow-up.

Consent for publication

Not applicable.

Availability of data and materials

The results presented in this paper rely, in part or in whole, on data from the TCGA Research Network (https://www.cancer.gov/tcga), Expression Atlas (https://www.ebi.ac.uk/gxa), and the Cancer Cell Line Encyclopedia (https://sites.broadinstitute.org/ccle).

Competing interests

The authors declare no conflicts of interest regarding the publication of this paper.

Funding

This study was funded by National Natural Science Foundation of China (82070773, 82100732); Natural Science Foundation of Jiangsu Province (BK20201271, BK 20200360);

Authors’ contributions

Tiancheng Jiang conducted the experiments and data analysis, and drafted the paper. Lei Zhang, Weipu Mao and Yongkun Zhu worked together on sample collection. Ganlin Wang refined the details of the article. Guoqing Wang was responsible for the analysis of the pathology slides as a pathologist. Shuqiu Chen and Zhenglin Zhang provided expert advice and support throughout. All authors reviewed and revised the manuscript and approved the final version.

Acknowledgment

We are very grateful for the technical support from the Department of Pathology, School of Medicine, Southeast University.

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IL20RB is a Prognostically Significant Gene and a Potential Biomarker in Papillary Renal Cell Carcinoma

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