Aldosterone-producing adenomas (APAs) and micronodules (APMs) are histopathological lesions that can cause a prevalent form of secondary hypertension called primary aldosteronism. Using integrated single-cell/nucleus and spatial transcriptomics, it demonstrates substantial heterogeneity between APMs and APAs, alongside an immunosuppressive tumor microenvironment in APAs. Pseudotime analyses reveal two distinct evolutionary trajectories in APAs: a direct progression from zona glomerulosa cells to APAs, observed specifically in KCNJ5 -mutated tumors, and a stepwise path from zona glomerulosa cells to APMs and then to APAs, present in both KCNJ5 -mutated APAs and APAs without KCNJ5 mutations. Within KCNJ5 -mutated APAs, we identified two distinct cellular states with different differentiation potentials: a progenitor-like state (high differentiation potential) and a mature state (low differentiation potential). This uncovered a spectrum of molecular and functional shifts during APA progression involving pathways of oxidative stress (including mechanisms leading to cell death by ferroptosis) and focal adhesion (influenced by cell density). In vitro studies in human adrenal cells demonstrate that TAZ activation regulates ferroptosis sensitivity in response to cell density changes. The findings enhance understanding of the transcriptional diversity in APMs and APAs and elucidate complex molecular alterations underlying APA pathogenesis.

Primary aldosteronism (PA) is a major cause of endocrine hypertension and contributes substantially to blood pressure (BP)-related morbidity. Although aldosterone excess directly raises BP, the extent to which genetic susceptibility to PA overlaps with the broader polygenic architecture of BP remains unclear. We aimed to characterise the shared genetic architecture between PA and BP traits and to place shared genetic signals within adrenal tissue context. We analysed genetic overlap between PA and BP using two independent PA datasets from the French COMETE Network and the US Million Veteran Program, together with genome-wide association data for systolic BP, diastolic BP and pulse pressure. Analyses included genome-wide and regional genetic correlation, cross-trait meta-analysis to identify shared genetic signals and complementary gene-prioritisation approaches. Biological context was assessed using functional annotation, tissue-specific expression analyses and integration with single-cell and spatial transcriptomic datasets from human adrenal tissue. PA showed significant positive genetic correlations with all three BP traits, with six genomic regions showing evidence of shared regional association. Cross-trait meta-analysis and co-localisation identified 143 independent shared signals associated with both PA and BP, with variant annotations indicating regulatory activity across adrenal, vascular, cardiac, adipose and neuroendocrine tissues. Gene-based analyses prioritised 43 candidate genes supported by convergent analytical approaches and implicated pathways related to vascular remodelling, extracellular matrix organisation, ion transport and endocrine regulation. Integration with adrenal single-cell and spatial transcriptomic data demonstrated that prioritised genes are expressed within defined adrenal cellular compartments, including aldosterone-producing populations. Among these candidates, SLC24A3 showed enriched expression in aldosterone-producing adenoma cells, with spatially heterogeneous expression associated with steroidogenic versus stress-adaptive transcriptional programmes. These findings support adrenal cell-state specificity of shared genetic signals and argue against a purely generic background BP-genetic explanation, without establishing causality for individual genes. PA and BP share a polygenic architecture that maps to adrenal endocrine cell states and vascular regulatory programmes within the intrinsic adrenal microenvironment. These findings link inherited BP-related variation to adrenal tissue biology and identify shared genetic signals, prioritised genes and cell states that may inform future studies of PA risk stratification, endocrine hypertension screening and cardiovascular risk prevention.

BackgroundUveitis is a complex intraocular inflammatory disease and pathology results from the continuous production of proinflammatory cytokines in the optical axis. Qinghuo Rougan Formula (QHRGF), a traditional Chinese medicine (TCM) is now used to treat uveitis with desirable effect. However, the mechanism of action is still unclear. This study aimed to explore the potential diagnostic and therapeutic biomarkers for uveitis using systems biology methods, including network pharmacology and weighted gene co-expression network analysis (WGCNA).MethodsA molecular drug-compound-target-uveitis interaction network was established using network pharmacology. Functional enrichment analyses were performed to screen potential signaling pathways. The uveitis gene expression dataset from the Gene Expression Omnibus database was subjected to WGCNA to identify gene co-expression modules related to uveitis and explore the potential hub genes. The least absolute shrinkage and selection operator (LASSO) model was used to identify the hub genes. Additionally, molecular docking was performed to verify the accuracy and stability of the model. Finally, the suppressive effects of QHRGF on uveitis were experimentally verified in vivo.ResultsNetwork pharmacology and functional enrichment analysis showed that 18 targets and immune/inflammation-related pathways were associated with the QHRGF-targeted pathway network. The yellow module contained 120 genes had a strong correlation with uveitis using WGCNA. In total, 12 putative targets of QHRGF, differentially expressed genes, and yellow module genes were determined. Six hub genes were identified using LASSO model and the receiving operating characteristic curve analysis demonstrated the model can serve as biomarkers for uveitis. The advantages of these genes were approved using molecular docking. Finally, in vivo experiments provided evidence confirming that QHRGF was identified as the key target of the anti-inflammatory effect of uveitis.ConclusionIn conclusion, this research revealed that QHRGF can be used to treat uveitis through multiple components and targets. Meanwhile, the potential anti-inflammatory action of QHRGF in the treatment of uveitis was verified by combining network pharmacology and in vivo experiments, suggesting its potential as a quite prospective agent for the therapy of uveitis.

BackgroundBladder urothelial carcinoma (BLCA) is associated with high mortality and recurrence. Although mRNA-based vaccines are promising treatment strategies for combating multiple solid cancers, their efficacy against BLCA remains unclear. We aimed to identify potential effective antigens of BLCA for the development of mRNA-based vaccines and screen for immune clusters to select appropriate candidates for vaccination.MethodsGene expression microarray data and clinical information were retrieved from The Cancer Genome Atlas and GSE32894, respectively. The mRNA splicing patterns were obtained from the SpliceSeq portal. The cBioPortal for Cancer Genomics was used to visualize genetic alteration profiles. Furthermore, nonsense-mediated mRNA decay (NMD) analysis, correlation analysis, consensus clustering analysis, immune cell infiltration analysis, and weighted co-expression network analysis were conducted.ResultsSix upregulated and mutated tumor antigens related to NMD, and infiltration of APCs were identified in patients with BLCA, including HP1BP3, OSBPL9, SSH3, ZCCHC8, FANCI, and EIF4A2. The patients were subdivided into two immune clusters (IC1 and IC2) with distinct clinical, cellular and molecular features. Patients in IC1 represented immunologically ‘hot’ phenotypes, whereas those in IC2 represented immunologically ‘cold’ phenotypes. Moreover, the survival rate was better in IC2 than in IC1, and the immune landscape of BLCA indicated significant inter-patient heterogeneity. Finally, CALD1, TGFB3, and ANXA6 were identified as key genes of BLCA through WGCNA analysis, and their mRNA expression levels were measured using qRT-PCR.ConclusionHP1BP3, OSBPL9, SSH3, ZCCHC8, FANCI, and EIF4A2 were identified as potential antigens for developing mRNA-based vaccines against BLCA, and patients in IC2 might benefit more from vaccination.

Kidney renal clear cell carcinoma (KIRC) is one of the most prevalent primary malignancies with high heterogeneity in the urological system. Growing evidence implies that lactate is a significant carbon source for cell metabolism and plays a vital role in tumor development, maintenance, and therapeutic response. However, the global influence of lactate-related genes (LRGs) on prognostic significance, tumor microenvironment characteristics, and therapeutic response has not been comprehensively elucidated in patients with KIRC. In the present study, we collected RNA sequencing and clinical data of KIRC from The Cancer Genome Atlas (TCGA), E-MTAB-1980, and GSE22541 cohorts. Unsupervised clustering of 17 differentially expressed LRG profiles divided the samples into three clusters with distinct immune characteristics. Three genes (FBP1, HADH, and TYMP) were then identified to construct a lactate-related prognostic signature (LRPS) using the least absolute shrinkage and selection operator (LASSO) and Cox regression analyses. The novel signature exhibited excellent robustness and predictive ability for the overall survival of patients. In addition, the constructed nomogram based on the LRPS-based risk scores and clinical factors (age, gender, tumor grade, and stage) showed a robust predictive performance. Furthermore, patients classified by risk scores had distinguishable immune status, tumor mutation burden, response to immunotherapy, and sensitivity to drugs. In conclusion, we developed an LRPS for KIRC that was closely related to the immune landscape and therapeutic response. This LRPS may guide clinicians to make more precise and personalized treatment decisions for KIRC patients.

Kidney renal clear cell carcinoma (KIRC) has long been identified as a highly immune-infiltrated tumor. However, the underlying role of pyroptosis in the tumor microenvironment (TME) of KIRC remains poorly described. Herein, we systematically analyzed the prognostic value, role in the TME, response to ICIs, and drug sensitivity of pyroptosis-related genes (PRGs) in KIRC patients based on The Cancer Genome Atlas (TCGA) database. Cluster 2, by consensus clustering for 24 PRGs, presented a poor prognosis, likely because malignancy-related hallmarks were remarkably enriched. Additionally, we constructed a prognostic prediction model that discriminated well between high- and low-risk patients and was further confirmed in external E-MTAB-1980 cohort and HSP cohort. By further analyzing the TME based on the risk model, higher immune cell infiltration and lower tumor purity were found in the high-risk group, which presented a poor prognosis. Patients with high risk scores also exhibited higher ICI expression, indicating that these patients may be more prone to profit from ICIs. The sensitivity to anticancer drugs that correlated with model-related genes was also identified. Collectively, the pyroptosis-related prognosis risk model may improve prognostic information and provide directions for current research investigations on immunotherapeutic strategies for KIRC patients.

In this study, we investigated the prognostic significance of the expression of N6-methyladenosine (m6A) RNA methylation regulatory genes in kidney renal papillary cell carcinoma (KIRP). RNA-sequencing data analysis showed that 14 of 20 major m6A RNA methylation regulatory genes were differentially expressed in the KIRP tissues from The Cancer Genome Atlas (TCGA) database. We constructed a prognostic risk signature with three m6A RNA methylation regulatory genes, IGF2BP3, KIAA1429 and HNRNPC, based on the results from univariate and LASSO Cox regression analyses. Multivariate Cox regression analysis confirmed that the risk score based on the three-gene prognostic risk signature was an independent predictive factor in KIRP. The overall survival of high-risk KIRP patients was significantly shorter than the low-risk KIRP patients. Expression of the three prognostic risk-related genes correlated with the AJCC and TNM stages of KIRP patients from TCGA and GEPIA datasets. ROC curve analysis showed that the three-gene prognostic risk signature precisely predicted the 1-year, 3-year and 5-year survival of KIRP patients. These findings demonstrate that expression of three prognostic risk-related m6A RNA methylation regulatory genes accurately predicts survival outcomes in KIRP patients.

Kidney renal clear cell carcinoma (KIRC) is the predominant pathological subtype of renal cell carcinoma (RCC) in adults. Long non-coding RNAs (lncRNAs) are an important class of gene expression regulators and serve fundamental roles in immune regulation. The intent of this study is to develop a novel immune-related lncRNA signature to accurately predict the prognosis for KIRC patients. Here, we performed genome-wide comparative analysis of lncRNA expression profiles in 537 KIRC patients from The Cancer Genome Atlas (TCGA) database. Cox regression model-identified immune-related lncRNAs were extracted for constructing a novel five immune-related lncRNA signature (AC008105.3, LINC02084, AC243960.1, AC093278.2, and AC108449.2) with the ability to predict the prognosis of KIRC patients. Univariate and multivariate Cox regression analyses demonstrated that the signature could act as an independent prognostic predictor for overall survival (OS). With the further investigation on different clinicopathological parameters, we found that the signature could divide KIRC samples into high-risk groups with shorter OS and low-risk groups with longer OS in different subgroups. Principal component analysis suggested that the five immune-related lncRNA signature drew a clear distinction between high- and low-risk groups based on the immune-related lncRNAs. The different immune status between the two groups was observed in gene set enrichment analysis and the ESTIMATE algorithm. Except for AC093278.2, the expressions of the other four lncRNAs expression were significantly upregulated in tumor tissues. In summary, the identified immune-lncRNA signature had important clinical implications in prognosis prediction and could be exploited as underlying immune therapeutic targets for KIRC patients.

Background: Kidney renal clear cell carcinoma (KIRC) is the most common and lethal renal cell carcinoma (RCC) histological subtype. Ferroptosis is a newly discovered programmed cell death and serves an essential role in tumor occurrence and development. The purpose of this study is to analyze ferroptosis-related gene (FRG) expression profiles and to construct a multi-gene signature for predicting the prognosis of KIRC patients. Methods: RNA-sequencing data and clinicopathological data of KIRC patients were downloaded from The Cancer Genome Atlas (TCGA). Differentially expressed FRGs between KIRC and normal tissues were identified using ‘limma’ package in R. GO and KEGG enrichment analyses were conducted to elucidate the biological functions and pathways of differentially expressed FRGs. Consensus clustering was used to investigate the relationship between the expression of FRGs and clinical phenotypes. Univariate and the least absolute shrinkage and selection operator (LASSO) Cox regression analysis were used to screen genes related to prognosis and construct the optimal signature. Then, a nomogram was established to predict individual survival probability by combining clinical features and prognostic signature. Results: A total of 19 differentially expressed FRGs were identified. Consensus clustering identified two clusters of KIRC patients with distinguished prognostic. Functional analysis revealed that metabolism-related pathways were enriched, especially lipid metabolism. A 7-gene ferroptosis-related prognostic signature was constructed to stratify the TCGA training cohort into high- and low-risk groups where the prognosis was significantly worse in the high-risk group. The signature was identified as an independent prognostic indicator for KIRC. These findings were validated in the testing cohort, the entire cohort, and the International Cancer Genome Consortium (ICGC) cohort. We further demonstrated that the signature-based risk score was highly associated with the KIRC progression. Further stratified survival analysis showed that the high-risk group had a significantly lower overall survival (OS) rate than those in the low-risk group. Moreover, we constructed a nomogram that had a strong ability to forecast the OS of the KIRC patients. Conclusion: We constructed a ferroptosis-related prognostic signature, which might provide a reliable prognosis assessment tool for clinician to guide clinical decision-making and outcomes research.

In this study, we analyzed the prediction accuracy of an autophagy-related long non-coding RNA (lncRNA) prognostic signature using bladder urothelial carcinoma (BLCA) patient data from The Cancer Genome Atlas (TCGA) database. Univariate and multivariate Cox regression analyses showed significant correlations between five autophagy-related lncRNAs, LINC02178, AC108449.2, Z83843.1, FAM13A-AS1 and USP30-AS1, and overall survival (OS) among BCLA patients. The risk scores based on the autophagy-related lncRNA prognostic signature accurately distinguished high- and low-risk BCLA patients that were stratified according to age; gender; grade; and AJCC, T, and N stages. The autophagy-related lncRNA signature was an independent prognostic predictor with an AUC value of 0.710. The clinical nomogram with the autophagy-related lncRNA prognostic signature showed a high concordance index of 0.73 and accurately predicted 1-, 3-, and 5-year survival times among BCLA patients in the high- and low-risk groups. The lncRNA-mRNA co-expression network contained 77 lncRNA-mRNA links among 5 lncRNAs and 49 related mRNAs. Gene set enrichment analysis showed that cancer- and autophagy-related pathways were significantly enriched in the high-risk group, and immunoregulatory pathways were enriched in the low-risk group. These findings demonstrate that an autophagy-related lncRNA signature accurately predicts the prognosis of BCLA patients.

BACKGROUND:Qinghuo Rougan Formula (QHRGF) is a traditional Chinese medicine (TCM) that has been widely apllied to treat uveitis for several decades. However, the inhibitory mechanism of QHRGF in uveitis has remained to be an enigma.METHODS:The Chinese herbal medicine pharmacology data and analysis platform wereused to search and screen for the effective components of the QHRGF compound injection and to analyse possible therapeutic targets based on network topology. In addition, various known disease target databases were enraolled, the therapeutic target proteins in uveitis were screened, and a protein-protein interaction (PPI) network was constructed. Enrichment analysis was performed on key nodes. Finally, the inhibitory effect of QHRGF on uveitis was verified by experiments.RESULTS:We identified 259 major candidate targets of QHRGF and successfully constructed a 'QHRGF-compound-target-uveitis' network. Above-mentioned targets revealed by Gene enrichment analysis have played an significant role in the cell cycle, autoimmune disease, apoptosis and related signal pathways. We demonstrated that QHRGF attenuates local inflammation in experimental autoimmune uveoretinitis (EAU) rats by regulating natural killer T (NKT) cells and inhibiting MAPK signal pathways.CONCLUSION:QHRGF may regulate the local immune response and inflammatory factors mainly through the MAPK signal pathway. For autoimmune uveitis, QHRGF may be a promising, long-lasting treatment strategy.

Objective:To study the molecular mechanism of LongdanXiegan Tang for treating uveitis.Methods:This was a experimntal study.The Chinese herbal medicine pharmacology data and analysis platform were used to search and to screen the effective components of a compound injection of LongdanXiegan Tang,and to analyze its possible therapeutic targets based on the network topology.By searching a variety of known disease target databases,we can screen the therapeutic target of uveitis disease and construct the active ingredient-disease treatment target network with the target of effective component treatment and to screen out the key nodes.Using topological analysis of the key nodes involved in the signal pathways and enrichment analysis,and screening out the possible Chinese medicine compound Radix LongdanXiegan Tang for the treatment of uveitis,may help to identify the signal pathways.Results:After the main active ingredient for screening and target prediction of LongdanXiegan Tang were completed,the therapeutic target of uveitis was screened,and an enrichment analysis of the signal pathways was completed.LongdanXiegan Tang can reduce the production of inflammatory cytokines and infiltration and affect the immune status,regulating the occurrence and development of uveitis by mainly regulating the MAPK signaling pathway and promoting both the GnRH and estrogen signaling pathways.Conclusions:LongdanXiegan Tang can treat uveitis primarily through the MAPK,GnRH and estrogen signaling pathways,regulating the inflammatory cells and inflammatory cytokines and autoimmunity.

Objective: To investigate the effects of total glucosides of paeony (TGP) on natural killer T (NKT) cell gene expression in liver of experimental autoimmune uveitis (EAU) rats. Methods: Experimental study. Forty-five Lewis rats were randomly divided into three groups: normal controls, EAU-stimulated rats (model group), and EAU-stimulated rats treated with TGP (TGP intervention group). Control animals received an emulsion of phosphate-buffered saline (PBS) and CFA, which was injected subcutaneously. The other two groups were injected an equal volume of chylomicron containing interphotoreceptor retinoid-binding protein polypeptide (IRBP). Inflammation was observed by daily slit-lamp microscopy of the anterior segments after immunization. The incidence of inflammation and the inflammation scores were recorded. At 12 days after immunization, we extracted the eyes for pathologic examination, and CD4+ T cells were collected from the livers. The expression levels of NKT cells were detected by flow cytometry. Reverse transcription polymerase change reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to measure the gene and protein expression levels of interleukin (IL)-4 and interferon (IFN)-γ. Data were analyzed by one-way ANOVA. Results: Twelve days after immunization, the inflammatory score and pathological grade of the TGP group were significantly lower than in the EAU group, but higher than in the normal group. Flow cytometry showed that compared with normal control group, the number of liver NKT cells in the EAU group and in the TGP intervention group were significantly increased. Further, the increased in the number of NKT expression in the TGP intervention group was higher than in the EAU group (P<0.05) during the inflammatory phase. RT-PCR showed that on day 12 the expression of IFN-γ mRNA in the liver of the TGP intervention group was significantly higher than in the control group (P<0.001), but lower than in the EAU group (P<0.001). At the same time, the expression of liver IL-4 mRNA in the TGP group was lower than that of the normal group (P<0.001), but significantly higher than the EAU group (P<0.001). ELISA showed that the liver IFN-γ levels in the TGP group were significantly lower in the EAU group after 12 days (P=0.01). The concentration of liver IL-4 protein was higher in the TGP group than in the EAU group (P<0.001). Conclusions: TGP can treat uveitis in EAU rats by regulating the expression of liver IL-4 and IFN-γ genes and increasing the number of NKT cells.

RATIONALE:Both inflammatory myofibroblastic tumor (IMT) and retroperitoneal fibrosis are rare lesions, but kidney involvement is more rare. It is the first study about IMT of the kidney in a patient with perinephric and periureteric fibrosis and we hold that fibroblast proliferation may be an intermediate status in oncogenesis of IMT. But further investigation is necessary in order to better clarify the relationship between fibroblast proliferation and IMT.PATIENT CONCERNS:A 54-year-old female presented no positive signs except dull back pain after overwork.DIAGNOSES:On the basis of the urinary ultrasonography and computed tomography (CT) scan, we strongly suspected a renal cell carcinoma.INTERVENTIONS:Considering the little remaining function of the right kidney and the possibility of malignancy, we performed a laparoscopic right radical nephrectomy.OUTCOMES:According to the analysis of the postoperative paraffin section and immunohistochemistry assay, a final diagnosis of IMT and retroperitoneal fibrosis nodules was made.LESSONS:Both IMTs are rare lesions and its etiology and pathogeny are unclear. It is the first study about IMT of the kidney in a patient with perinephric and periureteric fibrosis. This report suggested that fibroblast proliferation may be an intermediate status in oncogenesis of IMT, but further investigation is necessary in order to better clarify the relationship between fibroblast proliferation and IMT. The preoperative diagnosis of renal IMT remains difficult. Preoperative fine-needle aspiration or percutaneous biopsy and intraoperative frozen section were applied to confirm the diagnosis to avoid unnecessary nephrectomy, especially in patients with renal insufficiency, bilateral masses, or a solitary kidney.