Poliovirus receptor (PVR) mediates carboplatin-induced PD-L1 expression in non-small-cell lung cancer cells.

Programmed death-ligand-1 (PD-L1) is an immune suppressor that inhibits T cell based immunity. Anti-PD-L1/PD-1 immunotherapy benefits those patients receiving platinum-based combinational chemotherapy. However, the underlying mechanism is still largely unknown. In this study, we found that carboplatin could induce PD-L1 expression in NSCLC H292, A549 and H1299 cells in a dose-dependent manner. mRNA sequencing and the subsequent validation assays found that carboplatin significantly induced PVR expression, which is considered as an immuno-adhesion molecule. Mechanistically, PVR knockdown significantly abrogated carboplatin-induced PD-L1 expression. Functionally, knockdown of PVR significantly reversed the CD3+ T cells proliferation inhibition caused by carboplatin increased PD-L1. Moreover, the carboplatin-induced PVR and subsequent up-regulation of PD-L1 might be mediated via the EGFR, PI3K/AKT, and ERK signaling pathways. Immunohistochemical staining results showed that the PD-L1 expression was positively associated with PVR expression in clinical NSCLC samples. Our study reveals a novel regulatory mechanism of PD-L1 expression, provides evidence that carboplatin inhibits tumor immune response by up-regulating PD-L1 expression and explains the rationale for combining platinum-based chemotherapy with PD-L1/PD-1 inhibitors.

TRPV1 is a risk factor for sleep disturbance in patients with gastro-oesophageal reflux disease: a case control study.

BACKGROUND/AIMS: Gastro-oesophageal reflux disease (GORD) is a chronic high-morbidity disease with a bidirectional relationship with sleep disturbance (SD) that may occur via the transient receptor potential vanilloid type 1 receptor (TRPV1) in the oesophageal mucosa. Yet the related mechanism was still unclear, the aim of this study is to investigate whether TRPV1 is associated with the presence of SD in GORD patients. METHODS: A case-control study was performed. After the screening, A total of 88 subjects were assigned to GORD without sleep disturbance (GORD + NOSD, n = 28), GORD comorbid sleep disturbance (GORD + SD, n = 30) and matched healthy controls (n = 30). Mucosal tissue was obtained from the participants by digestive endoscopy, the levels of TRPV1 expressed in the oesophageal mucosa were detected via RT-qPCR and western blot in different groups, and the correlation between GORD and SD were also analysed. RESULTS: In this study, we found that the Gastroesophageal Reflux Disease Diagnostic Questionnaire (GerdQ) scores was positively correlated with Pittsburgh Sleep Quality Index (PSQI) scores but negatively correlated with total sleep time (TST). We also found that the level of TRPV1 expressed in the oesophageal mucosa of GORD + SD was significantly higher than GORD + NOSD patients, and they were all higher than healthy controls. CONCLUSION: The current study suggested a closer link exists between GORD and sleep disturbance, and TRPV1 in oesophageal mucosa may be a crucial factor affecting sleep in GORD patients.

REM Sleep Fragmentation in Patients With Short-Term Insomnia Is Associated With Higher BDI Scores.

Objective: To observe the changes in sleep characteristics and BDI scores in patients with short-term insomnia disorder (SID) using a longitudinal observational study. Methods: Fifty-four patients who met the criteria for SID of the International Classification of Sleep Disorders, third edition, were recruited. Depression levels were assessed using the Beck depression inventory (BDI) at enrollment and after 3 months of follow-up, respectively. Sleep characteristics were assessed by polysomnography. Results: After 3 months of follow-up, the group was divided into SID with increased BDI score (BDI >15) and SID with normal BDI score (BDI ≤ 15) according to the total BDI score of the second assessment. The differences in rapid eye movement (REM) sleep latency, REM sleep arousal index, and NREM sleep arousal index between the two groups were statistically significant. The total BDI score was positively correlated with REM and NREM sleep arousal index and negatively correlated with REM sleep latency, which were analyzed by Pearson correlation coefficient. Multiple linear regression was used to construct a regression model to predict the risk of depression in which the prediction accuracy reached 83.7%. Conclusion: REM sleep fragmentation is closely associated with future depressive status in patients with SID and is expected to become an index of estimating depression risk.

STAT3-induced SMYD3 transcription enhances chronic lymphocytic leukemia cell growth in vitro and in vivo.

OBJECTIVE AND DESIGN: The purpose of this study was to investigate the roles of SMYD3 and STAT3 in chronic lymphocytic leukemia (CLL) and the possible underlying mechanisms. MATERIALS: Blood samples were collected from 20 patients with CLL and 20 hematologically normal donors. Human cell lines K562, HL-60, MEG-1, and BALL-1 were performed in vitro and BALB/c nude mouse was used in subcutaneous tumor experiments. TREATMENT: WP1066 (30 mg/kg) was also injected intratumorally two days after the first lentivirus treatment and then every four days for a total of four injections and 3 µM WP1066 was carried out for 48 h to downregulate STAT3 phosphorylation. METHODS: We performed studies using the human CLL cell line MEG-1 in vitro and nude mouse subcutaneous tumor experiments in vivo. Differential expression of RNAs was determined using qRT-PCR. The CCK-8 assay and colony formation assay were conducted to evaluate cell proliferation. Flow cytometry was performed to assess cell apoptosis. The relative protein levels were detected using western blotting. Chromatin immunoprecipitation (ChIP) assays, luciferase reporter assays and WP1066, a STAT3 inhibitor, were used to explore the regulatory mechanisms of proteases and transcription factors. A subcutaneous tumor model was constructed to verify the results in vivo. RESULTS: SMYD3 and STAT3 expressions positively correlated with the progression of CLL. Upregulation of SMYD3 significantly promoted the proliferation and inhibited the expression of apoptosis-related genes. The results of the ChIP assays and luciferase reporter assays suggested that STAT3 targeted the promoter region of SMYD3 and, thus, promoted SMYD3 transcription. Downregulation of the phosphorylation of STAT3 by WP1066 notably inhibited the binding of STAT3 to the SMYD3 promoter, and subsequently downregulated SMYD3 transcription. The STAT3 inhibitor inhibited CLL cell growth in vivo, and overexpression of SMYD3 promoted CLL cell growth. Furthermore, overexpression of SMYD3 reversed the inhibitory effects of the STAT3 inhibitor on CLL cell growth. CONCLUSIONS: The STAT3-mediated transcription of SMYD3 plays a role in promoting the progression of chronic lymphocytic leukemia.