Single-Cell Sequencing of Peripheral Mononuclear Cells Reveals Distinct Immune Response Landscapes of COVID-19 and Influenza Patients.

The coronavirus disease 2019 (COVID-19) pandemic poses a current world-wide public health threat. However, little is known about its hallmarks compared to other infectious diseases. Here, we report the single-cell transcriptional landscape of longitudinally collected peripheral blood mononuclear cells (PBMCs) in both COVID-19- and influenza A virus (IAV)-infected patients. We observed increase of plasma cells in both COVID-19 and IAV patients and XIAP associated factor 1 (XAF1)-, tumor necrosis factor (TNF)-, and FAS-induced T cell apoptosis in COVID-19 patients. Further analyses revealed distinct signaling pathways activated in COVID-19 (STAT1 and IRF3) versus IAV (STAT3 and NFκB) patients and substantial differences in the expression of key factors. These factors include relatively increase of interleukin (IL)6R and IL6ST expression in COVID-19 patients but similarly increased IL-6 concentrations compared to IAV patients, supporting the clinical observations of increased proinflammatory cytokines in COVID-19 patients. Thus, we provide the landscape of PBMCs and unveil distinct immune response pathways in COVID-19 and IAV patients.

MicroRNA-155 downregulates long noncoding RNA prostate cancer-associated transcript 29 in hepatocellular carcinoma to suppress cancer cell invasion and migration.

Long noncoding RNA (lncRNA) prostate cancer-associated transcript 29 (PCAT29) is known to suppress several cancers, but its participation in hepatocellular carcinoma (HCC) remains elusive. This study tried to explore PCAT29 function in HCC. In this study, a total of 62 HCC patients were enrolled. Tissue samples were collected from all 62 patients to isolate RNA samples. Quantitative reverse transcription polymerase chain reaction was applied for the expression analysis of PCAT29 and microRNA-155 (miR-155) in these tissue samples. The 62 HCC patients were followed up for 5 years to explore the prognostic value of PCAT29 for HCC. Correlations were analyzed using linear regression. IntaRNA 2.0 was used to predict the interaction between PCAT29 and miR-155. The role of PCAT29 and miR-155 in regulating HCC cell invasion and migration was evaluated by Transwell assay. We found that PCAT29 expression was downregulated in HCC and miR-155 expression was upregulated in HCC compared to nontumor samples (p < 0.001). Downregulation of PCAT29 was found to be closely associated with poor survival of HCC patients. MiR-155 was inversely correlated with PCAT29. It was predicted that miR-155 could target PCAT29. In HCC cells, miR-155 overexpression resulted in reduced PCAT29 expression (p < 0.05). MiR-155 counteracted the inhibitory effects of PCAT29 overexpression on HCC cell migration and invasion. These results suggest that PCAT29 may be a potential prognostic biomarker and a novel therapeutic target for treating HCC.

Construction and Validation of a Novel Nomogram Predicting Recurrence in Alpha-Fetoprotein-Negative Hepatocellular Carcinoma Post-Surgery Using an Innovative Liver Function-Nutrition-Inflammation-Immune (LFNII) Score: A Bicentric Investigation.

Purpose: We developed a nomogram based on the liver function, nutrition, inflammation, and immunity (LFNII) score to predict recurrence-free survival (RFS) post-resection in patients with hepatocellular carcinoma (HCC) exhibiting alpha-fetoprotein (AFP) negativity (AFP ≤20 ng/mL). Patients and Methods: Clinical data of 661 patients diagnosed with alpha-fetoprotein-negative hepatocellular carcinoma (AFP-NHCC) who underwent surgical resection at two medical centers between 2012 and 2021 were collected. A total of 462 and 199 patients served as the training and validation sets, respectively. Pre-operative blood markers were collected and analyzed for LFNII. The LFNII score was formulated using the least absolute shrinkage and selection operator Cox regression model. A nomogram model was developed using the training set to incorporate other relevant clinicopathological indicators and predict postoperative recurrence. Model discrimination was assessed using the receiver operating characteristic curve, calibration was evaluated using a calibration curve, and clinical applicability was assessed using clinical decision curve analysis. A comparison with liver cancer staging was performed using the nomogram model. Finally, a cohort study was conducted to validate our findings. Results: We derived the LFNII scores from nine indicators. Elevated LFNII scores correlated with unfavorable clinicopathological features. The LFNII score area under the curve revealed superior predictive efficacy at 1-, 2-, and 5-year RFS intervals, with values of 0.675, 0.658, and 0.633, respectively. Multivariate Cox analysis revealed that a high LFNII score independently increased RFS risk in patients with AFP-NHCC. The C-index of the LFNII-nomogram model was 0.686 (95% confidence interval [CI], 0.651-0.721). The nomogram model's clinical application value surpassed that of standard HCC staging systems. Conclusion: The LFNII score-derived nomogram effectively predicted the RFS of patients with AFP-NHCC after curative resection.

Tissue factor promotes HCC carcinogenesis by inhibiting BCL2-dependent autophagy.

INTRODUCTION: Tissue factor (TF) is an important predictor for poor prognosis of Hepatocellular carcinoma (HCC). TF can also upregulate the expression of BCL2, which is a key inhibitor of autophagy responses. This study aims to explore the role of BCL2-dependent autophagy in TF-regulated HCC carcinogenesis. METHODS: In this study, we explored the roles of TF in HCC using gene overexpression and silencing assays. Besides, we further identified the significance of BCL2-Beclin1-autophagy signaling on TF-regulated HCC tumorigenesis by combining TF silencing with pharmacological autophagy inhibitor (3-MA). RESULTS: The experimental data showed that the overexpressed TF promoted BCL2 protein expression and inhibited the autophagy activity (shown as LC3 conversion rate, p62 expression and autophagosomes) while maintaining the survival in HCC cells. In contrast, the silent TF showed the completely opposite results. Furthermore, TF knockdown promoted the dissociation of Beclin1 from BCL2-Beclin1 complex. In addition, the enhanced autophagy and inhibited survival by TF knockdown could be reversed by autophagy inhibition with 3-MA or spautin-1 (Beclin1 specific inhibitor) in HCC cells. Xenografts assays also showed that TF-silencing HCC cells had stronger tumorigenicity in vivo, which was recovered by spautin-1 administration. CONCLUSIONS: TF inhibits autophagy-related death by enhancing BCL2 expression, whereby promoting HCC tumorigenesis.

A Novel Prognostic Model for Identifying the Risk of Hepatocellular Carcinoma Based on Angiogenesis Factors.

Hepatocellular carcinoma (HCC) is the most common primary liver cancer with poor prognosis. An optimized stratification of HCC patients to discriminate clinical benefit regarding different degrees of malignancy is urgently needed because of no effective and reliable prognostic biomarkers currently. HCC is typically characterized by rich vascular. The dysregulated vascular endothelial growth factor was proved a pivotal regulator of the development of HCC. Therefore, we investigated the capability of angiogenic factors (AFs) in stratifying patients and constructed a prognostic risk model. A total of 6 prognostic correlated AFs (GRM8, SPC25, FSD1L, SLC386A, FAM72A and SLC39A10) were screened via LASSO Cox regression, which provided the basis for developing a novel prognostic risk model. Based on the risk model, HCC patients were subdivided into high-risk and low-risk groups. Kaplan-Meier curve indicated that patients in the high-risk group have a lower survival rate compared with those in the low-risk group. The prognostic model showed good predictive efficacy, with AUCs reaching 0.802 at 1 year, 0.694 at 2 years, and 0.672 at 3 years. Univariate and multivariate cox regression analysis demonstrated that the risk score had significant prognostic value and was an independent prognostic factor for HCC. Moreover, this model also showed a good diagnostic positive rate in the ICGC-LIRI-JP and GSE144269. Finally, we demonstrated the efficacy of the AF-risk model in HCC patients following sorafenib adjuvant chemotherapy. And revealed the underlying molecular features involving tumor stemness, immune regulation, and genomic alterations associated with the risk score. Based on a large population, we established a novel prognostic model based on 6 AFs to help identify HCC patients with a greater risk of death. The model may provide a reference for better clinical management of HCC patients in the era of cancer precision medicine.

Somatic mutation profiling of liver and biliary cancer by targeted next generation sequencing.

Liver and biliary cancers are highly lethal cancer types lacking effective treatments. The somatic mutations, particularly those with low mutant allele frequencies, in Chinese patients with liver and biliary cancer have not been profiled, and the frequency of patients benefiting from targeted therapy has not been studied. The present study evaluated the tumor tissues of 45 Chinese patients with hepatocellular carcinoma (HCC) and 12 Chinese patients with biliary tract cancer (BTC) by targeted next generation sequencing, with an average coverage of 639×, to identify alterations in 372 cancer-related genes. A total of 263 variants were identified in 139 genes, with 85.6% of these variants not previously reported in the Catalogue Of Somatic Mutations In Cancer database, and the mutation profile was different from the current datasets, including The Cancer Genome Atlas dataset and the National Cancer Center Japan (NCC_JP) dataset. Patients with hepatitis B virus (HBV) infection harbored more mutations than those without HBV infection, and the mutations in HBV carriers occurred preferentially in genes involved in vascular endothelial growth factor signaling pathways. Mutations in fibroblast growth factor and RAS signaling pathways were enriched in patients with cirrhosis, and alterations in interleukin and transforming growth factor signaling pathways were more frequently identified in individuals with abnormal bilirubin expression. Of all the patients, 7% exhibited variants in the target of sorafenib, and 42% harbored variants in the targets of drugs that have been approved to treat other types of cancer. These findings indicate diverse HCC/BTC variants patterns in different populations, and that the mutation load and patterns are correlated with clinical features. Further clinical studies are now warranted to evaluate the efficacies of other targeted drugs besides sorafenib in the treatment of patients with liver and biliary cancer.