Identification of JUN gene and cellular microenvironment in response to PD-1 blockade treatment in lung cancer patients via single-cell RNA sequencing.

Exploring the molecular mechanisms of PD-1/PDL-1 blockade for non-small cell lung cancer (NSCLC) would facilitate understanding for tumor microenvironment (TME) and development of individualized medicine. To date, biomarkers of response to PD-1 blockade therapy were still limited. In this study, we hypothesize that cell type in the tumor microenvironment can influence the effect of PD-1 blockade immunotherapy through specific genes. Therefore, we re-analyze the single-cell RNA sequencing data and validation in tissue from lung adenocarcinoma patients. Dynamic changes of cellular subpopulation were observed after anti-PD-1 immunotherapy among TMEs between primary/metastasis or good/poor response patients. Non-exhausted CD8 T cells and dysregulated genes were observed in responsing patients from PD-1 blockade therapy. Among all changed genes, JUN, involved in PD-1 blockade immunotherapy pathway, and could be considered as a PD-1 responsing biomarker.

A Single-Arm Multi-Center Phase II Clinical Trial of Cadonilimab (anti-PD-1/CTLA-4) in Combination with or without Conventional Second-Line Treatment for Patients with Extensive Stage Small Cell Lung Cancer.

BACKGROUND: Cadonilimab (AK104) is a bispecific IgG-single-chain Fv fragment (ScFv) antibody that binds to PD-1 and CTLA-4. Cadonilimab has shown encouraging anti-tumour activity and a favourable safety profile in several tumour types. In second-line treatment, there is no defined standard of care for patients with extensive-stage small-cell lung cancer (ES-SCLC). Cadonilimab is expected to show substantial clinical efficacy. OBJECTIVE: To assess the antitumor activity and safety of cadonilimab monotherapy or combination with conventional therapy in ES-SCLC patients who failed first-line treatment. METHODS: In this multicenter, open-label, phase II study, ES-SCLC patients who had failed first-line treatment, also aged 18 years to 70 years with histologically or cytologically confirmed ES-SCLC, and an Eastern Cooperative Oncology Group performance status (ECOG-PS) of 0-2 were eligible. Patients will receive cadonilimab 10 mg/kg every three weeks (Q3 W) among 24 months until progressive disease (PD) or adverse events (AE) discovery. The primary endpoint is progression-free survival (PFS). TRIAL REGISTRATION: NCT05901584.

MAL, a potential immunotherapy target, is associated with poor prognosis in cancer patients.

The MAL gene encodes Myelin and Lymphocyte Protein, mainly expressed in T cells with immunomodulatory effects, showing the potential as a target for immunotherapy. However, the mechanism of MAL in the regulation of immune infiltration and its association with the prognosis in pan-cancer patients remain elusive. We used the TCGA, TIMER2.0, GTEx, UCSC, and TISCH databases and the R programming tool to explore the role of MAL in cancers. MAL was differently expressed in the majority of malignancies relative to the matched healthy controls. Patients with low MAL levels had adverse survival outcomes in the BRCA and LUAD cohorts. In all cancer types, MAL showed a significant correlation to specific immune-subpopulation abundance in particular T cells as well as B cells. MAL was also implicated in immunological pathways in BRCA and LUAD, suggesting the important role of MAL in cancer immune regulation. In conclusion, the pan-cancer study indicates that MAL with excellent prognostic value is a potential immunotherapy target in multiple cancers.

A novel sub-regional radiomics model to predict immunotherapy response in non-small cell lung carcinoma.

BACKGROUND: Identifying precise biomarkers of immunotherapy response for non-small cell lung carcinoma (NSCLC) before treatment is challenging. This study aimed to construct and investigate the potential performance of a sub-regional radiomics model (SRRM) as a novel tumor biomarker in predicting the response of patients with NSCLC treated with immune checkpoint inhibitors, and test whether its predictive performance is superior to that of conventional radiomics, tumor mutational burden (TMB) score and programmed death ligand-1 (PD-L1) expression. METHODS: We categorized 264 patients from retrospective databases of two centers into training (n = 159) and validation (n = 105) cohorts. Radiomic features were extracted from three sub-regions of the tumor region of interest using the K-means method. We extracted 1,896 features from each sub-region, resulting in 5688 features per sample. The least absolute shrinkage and selection operator regression method was used to select sub-regional radiomic features. The SRRM was constructed and validated using the support vector machine algorithm. We used next-generation sequencing to classify patients from the two cohorts into high TMB (≥ 10 muts/Mb) and low TMB (< 10 muts/Mb) groups; immunohistochemistry was performed to assess PD-L1 expression in formalin-fixed, paraffin-embedded tumor sections, with high expression defined as ≥ 50% of tumor cells being positive. Associations between the SRRM and progression-free survival (PFS) and variant genes were assessed. RESULTS: Eleven sub-regional radiomic features were employed to develop the SRRM. The areas under the receiver operating characteristic curve (AUCs) of the proposed SRRM were 0.90 (95% confidence interval [CI] 0.84-0.96) and 0.86 (95% CI 0.76-0.95) in the training and validation cohorts, respectively. The SRRM (low vs. high; cutoff value = 0.936) was significantly associated with PFS in the training (hazard ratio [HR] = 0.35 [0.24-0.50], P < 0.001) and validation (HR = 0.42 [0.26-0.67], P = 0.001) cohorts. A significant correlation between the SRRM and three variant genes (H3C4, PAX5, and EGFR) was observed. In the validation cohort, the SRRM demonstrated a higher AUC (0.86, P < 0.001) than that for PD-L1 expression (0.66, P = 0.034) and TMB score (0.54, P = 0.552). CONCLUSIONS: The SRRM had better predictive performance and was superior to conventional radiomics, PD-L1 expression, and TMB score. The SRRM effectively stratified the progression-free survival (PFS) risk among patients with NSCLC receiving immunotherapy.

Facile synthesis of isoquinolines and isoquinoline N-oxides via a copper-catalyzed intramolecular cyclization in water.

A highly efficient method for the facile access of isoquinolines and isoquinoline N-oxides via a Cu(i)-catalyzed intramolecular cyclization of (E)-2-alkynylaryl oxime derivatives in water has been developed. This protocol was performed under simple and mild conditions without organic solvent, additives or ligands. By switching on/off a hydroxyl protecting group of oximes, the selective N-O/O-H cleavage could be triggered, delivering a series of isoquinolines and isoquinoline N-oxides, respectively, in moderate to high yields with good functional group tolerance and high atom economy. Moreover, the practicality of this method was further demonstrated by the total synthesis of moxaverine in five steps.

Pd-Catalyzed Sequential Formation of C-C Bonds: A New Strategy for the Synthesis of (E)-α,ß-Unsaturated Carbonyl Compounds from Sulfoxonium Ylides and 1-Iodo-2-((2-methylallyl)oxy)benzene Compounds.

α,ß-Unsaturated carbonyl compounds are significant moieties in many biological molecules and have attracted considerable attention in organic synthetic chemistry. A Pd-catalyzed cascade cyclization for the synthesis of (E)-α,ß-unsaturated carbonyl compounds with the sequential formation of C-C bonds was developed. This method offers high efficiency, good functional group tolerance, and moderate to excellent yields and generally displays high stereoselectivity.

Identification and verification of HCAR3 and INSL5 as new potential therapeutic targets of colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) is one of the most common cancers of the gastrointestinal tract and ranks third in cancer-related deaths worldwide. This study was conducted to identify novel biomarkers related to the pathogenesis of CRC based upon a bioinformatics analysis, and further verify the biomarkers in clinical tumor samples and CRC cell lines. METHODS: A series of bioinformatics analyses were performed using datasets from NCBI-GEO and constructed a protein-protein interaction (PPI) network. This analysis enabled the identification of Hub genes, for which the mRNA expression and overall survival of CRC patients data distribution was explored in The Cancer Genome Atlas (TCGA) colon cancer and rectal cancer (COADREAD) database. Furthermore, the differential expression of HCAR3 and INLS5 was validated in clinical tumor samples by Real-time quantitative PCR analysis, western blotting analysis, and immunohistochemistry analysis. Finally, CRC cells over-expressing INSL5 were constructed and used for CCK8, cell cycle, and cell apoptosis validation assays in vitro. RESULTS: A total of 286 differentially expressed genes (DEGs) were screened, including 64 genes with increased expression and 143 genes with decreased expression in 2 CRC database, from which 10 key genes were identified: CXCL1, HCAR3, CXCL6, CXCL8, CXCL2, CXCL5, PPY, SST, INSL5, and NPY1R. Among these genes, HCAR3 and INSL5 had not previously been explored and were further verified in vitro. CONCLUSIONS: HCAR3 expression was higher in CRC tissues and associated with better overall survival of CRC patients. INSL5 expression in normal tissue was higher than that in tumor tissue and its high expression was associated with a better prognosis for CRC. The overexpression of INSL5 significantly inhibited the proliferation and promoted the shearing of PARP of CRC cells. This integrated bioinformatics study presented 10 key hub genes associated with CRC. HCAR3 and INSL5 were expressed in tumor tissue and these were associated with poor survival and warrant further studies as potential therapeutic targets.

LINC00202 attenuates the progression of gastric cancer via suppressing expression level of KLF2.

PURPOSE: We aimed to investigate the function of LINC00202 in influencing the malignant progression of gastric cancer (GC). METHODS: The relative level of LINC00202 in GC and adjacent normal tissues was examined by quantitative real-time polymerase chain reaction (qRT-PCR). The difference in LINC00202 level among GC patients with different TNM stages was compared. Subsequently, regulatory effects of LINC00202 on proliferative capacity of AGS and SGC-7901 cells were evaluated. Subcellular distribution of LINC00202 was analyzed. The interaction and correlation between LINC00202 and Krüppel-like factor 2 (KLF2) were analyzed by RNA immunoprecipitation (RIP), Chromatin immunoprecipitation (ChIP) and linear regression test. Finally, the involvement of KLF2 in LINC00202-mediated proliferative ability of GC cells was clarified. LINC00202 was upregulated in GC tissues compared with that in adjacent normal ones. Its level remained higher in GC patients with stage III-IV than those with stage I-II. Silencing LINC00202 markedly attenuated the proliferation of GC cells. RESULTS: LINC00202 was mainly enriched in nucleus. KLF2 level was negatively correlated to and interacted with LINC00202. Transfection of LINC00202 decreased the recruitment ability of EZH2 on KLF2. Importantly, KLF2 partially reversed the regulatory effect of LINC00202 on the proliferative ability of GC cells. CONCLUSIONS: LINC00202 enhances the proliferative ability of GC cells via negatively regulating KLF2, thus aggravating the progression of GC.

Segmental artery clamping versus main renal artery clamping in nephron-sparing surgery: updated meta-analysis.

OBJECTIVES: Ischemia-reperfusion injury is harmful in partial nephrectomy (PN) in renal cell carcinoma. Choosing an appropriate surgical method is important to reduce ischemia-reperfusion injury. This study aimed to compare the effect of segmental artery clamping (SAC) and main renal artery clamping (MAC) on patients who underwent PN. METHODS: Studies from January 2008 to November 2019 were identified by an electronic search of English and Chinese databases, including PubMed, Excerpt Medica Database, Cochrane Library, Wanfang, VIP, and Chinese National Knowledge Internet, without language restriction. Two reviewers were involved in the trial. The effects on operation time (OT), warm ischemia time (WIT), length of hospital stay (LOS), blood transfusion rate, postoperative complication rate, Clavien classification (≥ 3), and positive surgery margin (PSM) were evaluated using Stata software. Standardized mean difference (SMD, for continuous data) and pooled odds ratios (for count data) with 95% confidence interval (CI) were used as effect indicators. RESULTS: Thirty-two studies were included. SAC decreased the 1-week (SMD = - 0.973; 95% CI = - 1.414, - 0.532; P = 0.000), 1-month (SMD = - 0.411; 95% CI = - 0.769, - 0.053; P = 0.025), and 3-month (affected kidney: SMD = - 0.914; 95% CI = - 1.662, - 0.617; P = 0.000) percentages of postoperative changes in renal function (estimated glomerular filtration rate) between the SAC and MAC groups. Sub-group analysis showed that the SAC group had longer OT (SMD = 0.562; 95% CI = 0.252, 0.871; P = 0.000) than the MAC group. However, no differences were observed in the OT, WIT, LOS, blood transfusion rate, postoperative complication rate, Clavien classification (≥ 3), and PSM between the two groups. CONCLUSIONS: SAC is superior to MAC in terms of short-term postoperative renal function recovery. The use of SAC or MAC depends on tumor size, location, surgical modality, and surgeon's judgments.

Heterogeneity of T3 stage esophageal squamous cell carcinoma in different parts based on enhanced CT radiomics.

Esophageal cancer is a common malignant tumor of the digestive system with a high incidence and a poor prognosis. At the present, CT-based radiomics is providing more and more valuable information. However, the heterogeneity of the study and the poor repeatability of the texture feature parameters have limited its wider clinical application. In the present study, we focused on comparing the differences in the texture features of T3 stage esophageal squamous cell carcinoma at different locations and normal esophageal wall, aiming to provide some pieces of useful information for future research on esophageal squamous cell carcinoma.Fifty seven cases with throat CT imaging, including esophageal cancer contrast enhanced CT and conventional CT of healthy control group. The texture characteristics in control group and tumor group among different parts were compared. Using Univariable analysis, we compared the difference and conducted receiver-operator curve analysis to evaluate the performance of tumor grade diagnosis model.53 radiomic features were significantly different in control group and so as 93 features for tumor group. The upper section was the mostly different from the other 2 sections. Run-length matrix (RLM) features in tumor group accounted for the highest proportion, only Surface Volume Ratio was different.There are differences in the texture features of the tube wall in different parts of the esophagus of healthy adults, and this difference is more obvious in pT3 stage esophageal squamous cell carcinoma. In the future radiomics study of esophageal squamous cell carcinoma, we need to pay attention to this to avoid affecting the accuracy of the results.

Copper-Catalyzed Cascade Cyclization of Arylsulfonylhydrazones Derived from ortho-Alkynyl Arylketones: Regioselective Synthesis of Functionalized Cinnolines.

A novel copper-catalyzed cascade reaction of arylsulfonylhydrazones derived from ortho-alkynyl arylketones was accomplished. This reaction provides concise access to diversified cinnolines in good yields. The mechanistic investigations have disclosed involvement of the key alkynyl amination, 1,4-aryl migration, desulfonylation, and diazo radical cyclization cascade in the transformation.

Direct Synthesis of Benzo[ f]indazoles from Sulfonyl Hydrazines and 1,3-Enynes by Copper-Catalyzed Annulation.

A novel and efficient strategy for the direct synthesis of benzo[ f]indazoles via copper-catalyzed cascade reaction of sulfonyl hydrazides with 1,3-enynes under mild conditions has been developed. This method achieves the formation of two C-N bonds and one C-C bond in one pot, providing a series of benzo[ f]indazoles in moderate to good yields with good functional group tolerance and remarkable regioselectivity.

lncRNA H19/miR-675 axis regulates cardiomyocyte apoptosis by targeting VDAC1 in diabetic cardiomyopathy.

We previously established a rat model of diabetic cardiomyopathy (DCM) and found that the expression of lncRNA H19 was significantly downregulated. The present study was designed to investigate the pathogenic role of H19 in the development of DCM. Overexpression of H19 in diabetic rats attenuated oxidative stress, inflammation and apoptosis, and consequently improved left ventricular function. High glucose was associated with reduced H19 expression and increased cardiomyocyte apoptosis. To explore the molecular mechanisms involved, we performed in vitro experiments using cultured neonatal rat cardiomyocytes. Our results showed that miR-675 expression was decreased in cardiomyocytes transfected with H19 siRNA. The 3'UTR of VDAC1 was cloned downstream of a luciferase reporter construct and cotransfected into HEK293 cells with miR-675 mimic. The results of luciferase assay indicated that VDAC1 might be a direct target of miR-675. The expression of VDAC1 was upregulated in cardiomyocytes transfected with miR-675 antagomir, which consequently promotes cellular apoptosis. Moreover, enforced expression of H19 was found to reduce VDAC1 expression and inhibit apoptosis in cardiomyocytes exposed to high glucose. In conclusion, our study demonstrates that H19/miR-675 axis is involved in the regulation of high glucose-induced apoptosis by targeting VDAC1, which may provide a novel therapeutic strategy for the treatment of DCM.

The pterocellins, novel bioactive alkaloids from the marine bryozoan Pterocella vesiculosa.

Two new alkaloids, pterocellins A and B, have been isolated from the New Zealand marine bryozoan Pterocella vesiculosa. Structural elucidation was achieved through NMR and mass spectral analysis in conjunction with a single-crystal X-ray diffraction study of pterocellin A. The pterocellins possess a novel heterocyclic skeleton and exhibit potent antitumor activity and antimicrobial activity in vitro but only modest activity in the in vivo hollow fiber assay at the National Cancer Institute.