Single-cell RNA sequencing in double-hit lymphoma: IMPDH2 induces the progression of lymphoma by activating the PI3K/AKT/mTOR signaling pathway.

BACKGROUND: IMPDH2 is the rate-limiting enzyme of the de novo GTP synthesis pathway and has a key role in tumors; however, the specific mechanism underlying IMPDH2 activity in diffuse large B cell lymphoma (DLBCL) is still undetermined. This study aims to explore the potential mechanism of IMPDH2 in DLBCL, and its possible involvement in double-hit lymphoma (DHL), i.e., cases with translocations involving MYC and BCL2 and/or BCL6. METHODS: Using single-cell sequencing and bioinformatics analysis to screen for IMPDH2. Exploring the differential expression of IMPDH2 and its correlation with prognosis through multiplexed immunofluorescence analysis. Using CCK8, EdU, clone formation assay, and animal model to analyze biological behavior changes after inhibiting IMPDH2. Explaining the potential mechanism of IMPDH2 in DLBCL by Western blot and multiplexed immunofluorescence. RESULTS: Prognostic risk model was constructed by single-cell sequencing, which identified IMPDH2 as a DHL-related gene. IMPDH2 was highly expressed in cell lines and tissues, associated with poor patient prognosis and an independent prognostic factor. In vitro and in vivo experiments showed that IMPDH2 inhibition significantly inhibited DHL cell proliferation. Flow cytometry showed apoptosis and cycle arrest. Western blot results suggested that c-Myc regulated the activation of PI3K/AKT/mTOR signaling pathway by IMPDH2 to promote tumor development in DHL. Moreover, multiplex immunofluorescence revealed decreased T-cell infiltration within the tumor microenvironment exhibiting concurrent high expression of IMPDH2 and PD-L1. CONCLUSIONS: Our results suggest that IMPDH2 functions as a tumor-promoting factor in DHL. This finding is expected to generate novel insights into the pathogenesis of these patients, thereby identifying potential therapeutic targets.

Case report: Immune modulation after PD-1 inhibitor therapy in a patient with extranodal NK/T-cell lymphoma secondary to chronic active Epstein-Barr virus disease unveiled by single-cell transcriptomics.

Chronic active Epstein-Barr virus disease (CAEBV) is a systemic lymphoproliferative disorder that is closely linked to Epstein-Barr virus (EBV) infection. The clinical course and severity of CAEBV can vary, and in some cases, it can progress to overt lymphoma, which is characterized by extranodal natural killer/T-cell lymphoma (ENKTL) and has a poor clinical outcome. Although anti-programmed cell death protein-1 (PD-1) therapy has shown effectiveness in some patients with EBV-associated disease, it has been less successful in others, and the exact mechanism of action of PD-1 inhibitor therapy in these diseases remains unclear. In this report, we describe a patient who was diagnosed with ENKTL secondary to CAEBV and experienced rapid disease progression accompanied by hyperinflammation after receiving PD-1 inhibitor therapy. Single-cell RNA sequencing revealed a significant increase in the patient's lymphocyte count, especially in natural killer cells, with increased activity following PD-1 inhibitor therapy. This case raises questions about the efficacy and safety of PD-1 inhibitor therapy in patients with EBV-associated diseases.

Association between environmental phenols and all-cause and cancer mortality.

To study the association between environmental phenols exposure, including benzophenone-3 (BP3) and triclosan, and all-cause mortality and cancer mortality in adults. A total of 8035 participants were included from the National Health and Nutrition Examination Survey (2003-2012). Urinary BP3 and triclosan were measured with liquid chromatography-tandem mass spectrometry. The associations between urinary BP3 and triclosan with cancer and all-cause mortality were explored by multivariable logistic regressions and restricted cubic splines. During an average of 7.25-year follow-up, 696 cases (8.7%) of all-cause mortality and 137 cases (1.71%) of cancer mortality occurred. The average levels of BP3 and triclosan were 12.2 and 10.3 ng/mL, respectively. Compared with the lowest quartile, the multivariable-adjusted hazard ratio of the highest quartile was 0.64 (95% confidence interval: 0.50 to 0.81; P < 0.001) for BP3 and 0.76 (0.61 to 0.94; P = 0.011) for triclosan. However, no significant associations between urinary BP3 and triclosan and cancer mortality were observed. BP3 and triclosan exposure decreased the risk of all-cause mortality while they were not associated with cancer mortality.

Application of a New Type of Protective Sputum Suction Device in Clinic against Cross-Infection between Medical Staff and Patients.

Objective: To explore the clinical application of a new type of protective sputum suction device (PSSD) in patients with tracheotomy or tracheal intubation and to evaluate the protective effect of PSSD against cross-infection between medical staffs and patients. Methods: A novel PSSD was designed which can assist closed sputum suction operation without disconnecting the ventilator. 32 patients with tracheotomy were included to study the protective effect and safety of this device. Patients' vital signs including heart rate, respiratory rate, mean arterial pressure, and blood oxygen saturation were recorded to compare the influence of open suction and closed suction (performed with this novel device). To verify the antisplash effect of this device on airway secretions, bacterial samples were collected from the hands of the suction operators and the environment near the endotracheal tube orifice before and after the two suction processes. In addition, the satisfaction of the two suction methods was compared through the questionnaire of suction staff. Finally, with the assistance of this device, an attempt was made to complete the bronchoscopy without weaning of ventilator. Results: Compared with open sputum suction, closed sputum suction has a smaller decrease in patients' blood oxygen saturation (P < 0.05), and no significant differences in other vital signs. Compared with open sputum suction, bacteria from the hands of suction staffs and the surrounding environment of the endotracheal tube were barely detected in closed suction. A questionnaire survey of sputum suction nurses suggested that the satisfaction with use and protective effect of the closed suction were better than open suction. In addition, bronchoscopy can be successfully completed with the assistance of this device, which is not possible for other breathing tubes. Conclusion: This closed sputum suction device has little effect on the oxygen saturation of patients but has excellent protective effects for medical staff against cross-infection. It has a unique advantage that can assist in completing the fiberoptic bronchoscopy with continuous ventilator-assisted breathing.

Primary Osteosarcoma of the Breast: A Rare Case Report and Literature Review.

Background: Primary osteosarcoma of the breast (POB) is an extremely aggressive and heterogeneous neoplasm that originates from nonepithelial elements of the mammary gland and accounts for fewer than 1% of breast cancers and fewer than 5% of all sarcomas. Case Presentation: An 83-year-old Chinese woman went to our hospital because of a palpable mass she had had for 8 months in the left breast accompanied by persistent dull pain for 10 days. This mass was initially misdiagnosed as a degenerating fibroadenoma and was graded as probably benign (BI-RADS category 3) by ultrasonography (US) and computed tomography (CT) plain scan and contrast enhancement of chest. Eight months later, it was presumed to be highly malignant and graded as BI-RADS category 4C because of its rapid growth and more calcifications by US and CT. 99mTc-MDP whole-body bone imaging showed that there was a mass-like abnormal radioactive concentration of Tc-99m outside the bone of the left chest. The lumpectomy of the left breast was indicated, and the pathological findings were POB. She succumbed to respiratory failure caused by multiple lung metastases 4 months after the operation. Conclusion: POB is rare, and US and CT cannot reliably distinguish the causes of calcified breast masses between benign and malignant tumors. It can be diagnosed by pathology when metaplastic carcinoma, malignant phyllodes tumor, or carcinosarcoma containing osteoid and bone is excluded. This case could help clinicians to improve the prognosis and treatment of this disease.

Identification of molecular subtypes and a novel prognostic model of diffuse large B-cell lymphoma based on a metabolism-associated gene signature.

BACKGROUND: Diffuse large B cell lymphoma (DLBCL) is the most common lymphoma in adults. Metabolic reprogramming in tumors is closely related to the immune microenvironment. This study aimed to explore the interactions between metabolism-associated genes (MAGs) and DLBCL prognosis and their potential associations with the immune microenvironment. METHODS: Gene expression and clinical data on DLBCL patients were obtained from the GEO database. Metabolism-associated molecular subtypes were identified by consensus clustering. A prognostic risk model containing 14 MAGs was established using Lasso-Cox regression in the GEO training cohort. It was then validated in the GEO internal testing cohort and TCGA external validation cohort. GO, KEGG and GSVA were used to explore the differences in enriched pathways between high- and low-risk groups. ESTIMATE, CIBERSORT, and ssGSEA analyses were used to assess the immune microenvironment. Finally, WGCNA analysis was used to identify two hub genes among the 14 model MAGs, and they were preliminarily verified in our tissue microarray (TMA) using multiple fluorescence immunohistochemistry (mIHC). RESULTS: Consensus clustering divided DLBCL patients into two metabolic subtypes with significant differences in prognosis and the immune microenvironment. Poor prognosis was associated with an immunosuppressive microenvironment. A prognostic risk model was constructed based on 14 MAGs and it was used to classify the patients into two risk groups; the high-risk group had poorer prognosis and an immunosuppressive microenvironment characterized by low immune score, low immune status, high abundance of immunosuppressive cells, and high expression of immune checkpoints. Cox regression, ROC curve analysis, and a nomogram indicated that the risk model was an independent prognostic factor and had a better prognostic value than the International Prognostic Index (IPI) score. The risk model underwent multiple validations and the verification of the two hub genes in TMA indicated consistent results with the bioinformatics analyses. CONCLUSIONS: The molecular subtypes and a risk model based on MAGs proposed in our study are both promising prognostic classifications in DLBCL, which may provide novel insights for developing accurate targeted cancer therapies.

Holistic View of ALK TKI Resistance in ALK-Positive Anaplastic Large Cell Lymphoma.

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase expressed at early stages of normal development and in various cancers including ALK-positive anaplastic large cell lymphoma (ALK+ ALCL), in which it is the main therapeutic target. ALK tyrosine kinase inhibitors (ALK TKIs) have greatly improved the prognosis of ALK+ALCL patients, but the emergence of drug resistance is inevitable and limits the applicability of these drugs. Although various mechanisms of resistance have been elucidated, the problem persists and there have been relatively few relevant clinical studies. This review describes research progress on ALK+ ALCL including the application and development of new therapies, especially in relation to drug resistance. We also propose potential treatment strategies based on current knowledge to inform the design of future clinical trials.

Block of Proliferation 1 Promotes Proliferation, Invasion and Epithelial Mesenchymal Transformation in Gastric Cancer.

BACKGROUND: Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide nowadays. Block of proliferation 1 (BOP1), a nucleolar protein involved in rRNA processing and ribosome assembly, is associated with tumor development in certain cancers of digestive system. Therefore, we hypothesized that BOP1 might play an important role in gastric cancer development. METHODS: Gene Expression Omnibus (GEO) database and The Cancer Genome Atlas (TCGA) were used to identify the differentially expressed genes and their clinical relevance. qPCR and western blot were performed further to examine the levels of BOP1 mRNA and protein, respectively. Cell viability, apoptosis, migration and invasion were investigated in gastric cancer cell lines with BOP1 silencing or overexpression. The epithelial mesenchymal transition (EMT) associated proteins, including E-cadherin and N-cadherin, were measured using immunoblotting. Finally, the downstream pathway of BOP1 were explored using bioinformatic analysis and qPCR. RESULTS: BOP1 was found up-regulated in gastric tumor tissues compared with paired normal tissues (P < 0.0001). Its expression was associated with more advanced pathological grades (P = 0.0006) and tumor location (P = 0.002), as well as a poor survival (HR 1.27, P = 0.015). BOP1 expression was increased in 4 kind of tumor cell lines compared with the normal group. The overexpression of BOP1 promoted cell proliferation and inhibit cell apoptosis, while silencing BOP1 showed a reversed trend. Immunoblotting results suggested that BOP enhanced N-cadherin, a mesenchymal marker, while reduced E-cadherin, an epithelial marker. Finally, bioinformatic prediction showed that the cell cycle could be a downstream pathway of BOP1. CONCLUSIONS: The present study demonstrated that BOP1 contributed to the development of gastric cancer by promoting proliferation, invasion and epithelial mesenchymal transformation, which could be a biomarker or therapeutic target in GC.

Successful PD-1 inhibitor treatment in a patient with refractory transformed follicular lymphoma who failed to respond to CAR-T cell therapy: a case report and literature review.

Follicular lymphoma (FL) accounts for approximately 35% of all non-Hodgkin lymphomas and can progress to diffuse large B cell lymphoma (DLBCL) at a rate of 2% per year. Here, we present a 56-year-old female patient who was diagnosed with grade 3a FL. Further pathological investigation revealed that the lymphoma had transformed into DLBCL following six courses of R-CHOP regimen, and further disease progression was observed after two courses of R2-GemOx. We ultimately failed to collect hematopoietic stem cells after two courses of R2-ICE. CD-22 CAR-T cell therapy salvaged the patient; however, a new soft tissue mass of 4.8 × 4.1 cm rapidly emerged in the patient's right lung. Following the observation of strong tissue staining of PD-L1 (90%), the patient was administered PD-1 inhibitor and 26 Gy of radiotherapy and has maintained progression-free survival at more than 15 months of follow-up. Transformed FL with strong PD-L1 expression showed a poor response to standard immunochemotherapy. Our findings support the potential benefit of PD-1 inhibitor and combination therapies in this type of transformed FL.

Colon cancer recurrence­associated genes revealed by WGCNA co­expression network analysis.

The present study aimed to identify the recurrence­associated genes in colon cancer, which may provide theoretical evidence for the development of novel methods to prevent tumor recurrence. Colon cancer and matched normal samples microarray data (E­GEOD­39582) were downloaded from ArrayExpress. Genes with significant variation were identified, followed by the screening of differentially expressed genes (DEGs). Subsequently, the co­expression network of DEGs was constructed using the weighted correlation network analysis (WGCNA) method, which was verified using the validation dataset. The significant modules associated with recurrence in the network were subsequently screened and verified in another independent dataset E­GEOD­33113. Function and pathway enrichment analyses were also conducted to determine the roles of selected genes. Survival analysis was performed to identify the association between these genes and survival. A total of 434 DEGs were identified in the colon samples, and stress­associated endoplasmic reticulum protein family member 2 (SERP2) and long non­coding RNA­0219 (LINC0219) were determined to be the vital DEGs between all the three sub­type groups with different clinical features. The brown module was identified to be the most significant module in the co­expression network associated with the recurrence of colon cancer, which was verified in the E­GEOD­33113 dataset. Top 10 genes in the brown module, including EGF containing fibulin like extracellular matrix protein 2 (EFEMP2), fibrillin 1 (FBN1) and secreted protein acidic and cysteine rich (SPARC) were also associated with survival time of colon cancer patients. Further analysis revealed that the function of cell adhesion, biological adhesion, extracellular matrix (ECM) organization, pathways of ECM­receptor interaction and focal adhesion were the significantly changed terms in colon cancer. In conclusion, SERP2, EFEMP2, FBN1, SPARC, and LINC0219 were revealed to be the recurrence­associated molecular and prognostic indicators in colon cancer by WGCNA co­expression network analysis.

Classifier of cross talk genes predicts the prognosis of hepatocellular carcinoma.

The present study aimed to establish a prediction model for hepatocellular carcinoma (HCC) based on the cross talk genes from important biological pathways involved in HCC. Differentially expressed genes (DEGs) for HCC were identified from mRNA profiles of GSE36376, which were mapped to protein­protein interaction (PPI) networks from BioGrid and the human protein reference database. Then critical genes based on the deviation score and the degree of node were selected from the novel PPI network. Cross talk genes were screened from the network established based on the associations of gene­gene, gene­pathway and pathway­pathway. A classifier based on specific cross talk genes was constructed for prediction of HCC using the random forest algorithm. Finally, the diagnostic performance of this prediction model was verified by predicting survival time of patients with HCC from the genome cancer atlas (TCGA) and other independent gene expression omnibus (GEO) databases. From the novel PPI network, a total of 200 critical genes were screened out and they were significantly enriched in 23 pathways, which have been reported to be significantly associated with the development of HCC. Based on these identified pathways, cross talk genes were identified including AKT1, SOS1, EGF, MYC, IGF1, ERBB2, CDKN1B, SHC2, VEGFA and INS. The prediction model has a relative average classification accuracy of 0.94 for HCC, which has a stable predicting efficacy for survival time of HCC patients validated in the TCGA database and two other independent GEO datasets. In conclusion, a total of 39 cross talk genes in HCC were identified and a classifier based on the cross talk genes was constructed, which indicates a high prognosis prediction efficacy in several independent datasets. The results provide a novel perspective to develop a multiple gene diagnostic tool for HCC prognosis, which also provided potential biomarkers or therapeutic targets for HCC.