Revisiting airway epithelial dysfunction and mechanisms in chronic obstructive pulmonary disease: the role of mitochondrial damage.

Chronic exposure to environmental hazards causes airway epithelial dysfunction, primarily impaired physical barriers, immune dysfunction, and repair or regeneration. Impairment of airway epithelial function subsequently leads to exaggerated airway inflammation and remodeling, the main features of chronic obstructive pulmonary disease (COPD). Mitochondrial damage has been identified as one of the mechanisms of airway abnormalities in COPD, which is closely related to airway inflammation and airflow limitation. In this review, we evaluate updated evidence for airway epithelial mitochondrial damage in COPD and focus on the role of mitochondrial damage in airway epithelial dysfunction. In addition, the possible mechanism of airway epithelial dysfunction mediated by mitochondrial damage is discussed in detail, and recent strategies related to airway epithelial-targeted mitochondrial therapy are summarized. Results have shown that dysregulation of mitochondrial quality and oxidative stress may lead to airway epithelial dysfunction in COPD. This may result from mitochondrial damage as a central organelle mediating abnormalities in cellular metabolism. Mitochondrial damage mediates procellular senescence effects due to mitochondrial reactive oxygen species, which effectively exacerbate different types of programmed cell death, participate in lipid metabolism abnormalities, and ultimately promote airway epithelial dysfunction and trigger COPD airway abnormalities. These can be prevented by targeting mitochondrial damage factors and mitochondrial transfer. Thus, because mitochondrial damage is involved in COPD progression as a central factor of homeostatic imbalance in airway epithelial cells, it may be a novel target for therapeutic intervention to restore airway epithelial integrity and function in COPD.

Targeting TFH cells is a novel approach for donor-specific antibody desensitization of allograft candidates: an in vitro and in vivo study.

The presence of donor-specific antibodies (DSA) are associated with graft failure either following human leukocyte antigen (HLA)-mismatched allogeneic stem cell transplantation or after organ transplantation. Although targeting B cells and plasma cells have been used for desensitization, there have been reports of failure. T-follicular helper (Tfh) cells assist B cells in differentiating into antibody-secreting plasma cells. We used haploidentical allograft as a platform to investigate the possibility of targeting Tfh cells to desensitize DSA. The quantities of circulating Tfh (cTfh) cell subsets in allograft candidates were abnormal, and these cells, including the cTfh2 and cTfhem cell subsets, were positively related to the production of anti-HLA antibodies. Ex vivo experiments showed that the cTfh cells of anti-HLA antibody-positive allograft candidates could induce B cells to differentiate into DSA-producing plasmablasts. The immune synapse could be involved in the assistance of cTfh cells to B cells in antibody production. In vitro experiments and in vivo clinical pilot studies indicated that targeting cTfh cells with sirolimus can inhibit their auxiliary function in assisting B cells. Ex vivo and in vivo studies demonstrated the effect of sirolimus and rituximab on DSA desensitization compared with either sirolimus or rituximab alone (60%, 43.75%, and 30%, respectively). Our findings provide new insight into the role of Tfh cells in the pathogenesis of DSA production in HLA-mismatched transplant candidates. Our data also indicate that targeting Tfh cells is a novel strategy for DSA desensitization and combination of sirolimus and rituximab might be a potential therapy. The prospective cohort of this study is registered at http://www.chictr.org.cn as #ChiCTR-OPC-15006672.

Distinct impacts of radiological appearance on lymph node metastasis and prognosis based on solid size in clinical T1 non-small cell lung cancer.

BACKGROUND: Solid nodules (SN) had more aggressive features and a poorer prognosis than part-solid nodules (PSN). This study aimed to evaluate the specific impacts of nodule radiological appearance (SN vs. PSN) on lymph node metastasis and prognosis based on solid size in cT1 non-small cell lung cancer (NSCLC). METHODS: Patients with cT1 NSCLC who underwent anatomical resection between 2010 and 2019 were retrospectively screened. Univariable and multivariable logistic regression analyses were adopted to evaluate the associations between nodule radiological appearance and lymph node metastasis. The log-rank test and Cox regression analyses were applied for prognostic evaluation. The cumulative recurrence risk was evaluated by the competing risk model. RESULTS: There were 958 and 665 NSCLC patients with PSN and SN. Compared to the PSN group, the SN arm had a higher overall lymph node metastasis rate (21.7% vs. 2.7%, P < 0.001), including nodal metastasis at N1 stations (17.7% vs. 2.1%), N2 stations (14.0% vs. 1.6%), and skip nodal metastasis (3.9% vs. 0.6%). However, for cT1a NSCLC, no significant difference existed between SN and PSN (0 vs. 0.4%, P = 1). In addition, the impacts of nodule radiological appearance on lymph node metastasis varied between nodal stations. Solid NSCLC had an inferior prognosis than part-solid patients (5-year disease-free survival: 79.3% vs. 96.2%, P < 0.001). The survival inferiority only existed for cT1b and cT1c NSCLC, but not for cT1a. Strikingly, even for patients with nodal involvement, SN still had a poorer disease-free survival (P = 0.048) and a higher cumulative incidence of recurrence (P < 0.001) than PSN. Specifically, SN had a higher recurrence risk than PSN at each site. Nevertheless, the distribution of recurrences between SN and PSN was similar, except that N2 lymph node recurrences were more frequent in solid NSCLC (28.21% vs. 7.69%, P = 0.041). CONCLUSION: SN had higher risks of lymph node metastasis and poorer prognosis than PSN for cT1b and cT1c NSCLC, but not for cT1a. SN exhibited a greater proportion of N2 lymph node recurrence than PSN. SN and PSN needed distinct strategies for nodal evaluation and postoperative follow-up.

Aerobic exercise training engages the canonical wnt pathway to improve pulmonary function and inflammation in COPD.

BACKGROUND: We studied whether the exercise improves cigarette smoke (CS) induced chronic obstructive pulmonary disease (COPD) in mice through inhibition of inflammation mediated by Wnt/ß-catenin-peroxisome proliferator-activated receptor (PPAR) γ signaling. METHODS: Firstly, we observed the effect of exercise on pulmonary inflammation, lung function, and Wnt/ß-catenin-PPARγ. A total of 30 male C57BL/6J mice were divided into the control group (CG), smoke group (SG), low-intensity exercise group (LEG), moderate-intensity exercise group (MEG), and high-intensity exercise group (HEG). All the groups, except for CG, underwent whole-body progressive exposure to CS for 25 weeks. Then, we assessed the maximal exercise capacity of mice from the LEG, MEG, and HEG, and performed an 8-week treadmill exercise intervention. Then, we used LiCl (Wnt/ß-catenin agonist) and XAV939 (Wnt/ß-catenin antagonist) to investigate whether Wnt/ß-catenin-PPARγ pathway played a role in the improvement of COPD via exercise. Male C57BL/6J mice were randomly divided into six groups (n = 6 per group): CG, SG, LiCl group, LiCl and exercise group, XAV939 group, and XAV939 and exercise group. Mice except those in the CG were exposed to CS, and those in the exercise groups were subjected to moderate-intensity exercise training. All the mice were subjected to lung function test, lung histological assessment, and analysis of inflammatory markers in the bronchoalveolar lavage fluid, as well as detection of Wnt1, ß-catenin and PPARγ proteins in the lung tissue. RESULTS: Exercise of various intensities alleviated lung structural changes, pulmonary function and inflammation in COPD, with moderate-intensity exercise exhibiting significant and comprehensive effects on the alleviation of pulmonary inflammation and improvement of lung function. Low-, moderate-, and high-intensity exercise decreased ß-catenin levels and increased those of PPARγ significantly, and only moderate-intensity exercise reduced the level of Wnt1 protein. Moderate-intensity exercise relieved the inflammation aggravated by Wnt agonist. Wnt antagonist combined with moderate-intensity exercise increased the levels of PPARγ, which may explain the highest improvement of pulmonary function observed in this group. CONCLUSIONS: Exercise effectively decreases COPD pulmonary inflammation and improves pulmonary function. The beneficial role of exercise may be exerted through Wnt/ß-catenin-PPARγ pathway.

Lung transcriptomics reveals the underlying mechanism by which aerobic training enhances pulmonary function in chronic obstructive pulmonary disease.

BACKGROUND: Aerobic training is the primary method of rehabilitation for improving respiratory function in patients with chronic obstructive pulmonary disease (COPD) in remission. However, the mechanism underlying this improvement is not yet fully understood. The use of transcriptomics in rehabilitation medicine offers a promising strategy for uncovering the ways in which exercise training improves respiratory dysfunction in COPD patients. In this study, lung tissue was analyzed using transcriptomics to investigate the relationship between exercise and lung changes. METHODS: Mice were exposed to cigarette smoke for 24 weeks, followed by nine weeks of moderate-intensity treadmill exercise, with a control group for comparison. Pulmonary function and structure were assessed at the end of the intervention and RNA sequencing was performed on the lung tissue. RESULTS: Exercise training was found to improve airway resistance and lung ventilation indices in individuals exposed to cigarette smoke. However, the effect of this treatment on damaged alveoli was weak. The pair-to-pair comparison revealed numerous differentially expressed genes, that were closely linked to inflammation and metabolism. CONCLUSIONS: Further research is necessary to confirm the cause-and-effect relationship between the identified biomarkers and the improvement in pulmonary function, as this was not examined in the present study.

The impact of delayed tracheostomy on critically ill patients receiving mechanical ventilation: a retrospective cohort study in a chinese tertiary hospital.

OBJECTIVES: The timing of tracheostomy for critically ill patients on mechanical ventilation (MV) is a topic of controversy. Our objective was to determine the most suitable timing for tracheostomy in patients undergoing MV. DESIGN: Retrospective cohort study. SETTING AND PARTICIPANTS: One thousand eight hundred eighty-four hospitalisations received tracheostomy from January 2011 to December 2020 in a Chinese tertiary hospital. METHODS: Tracheostomy timing was divided into three groups: early tracheostomy (ET), intermediate tracheostomy (IMT), and late tracheostomy (LT), based on the duration from tracheal intubation to tracheostomy. We established two criteria to classify the timing of tracheostomy for data analysis: Criteria I (ET ≤ 5 days, 5 days < IMT ≤ 10 days, LT > 10 days) and Criteria II (ET ≤ 7 days, 7 days < IMT ≤ 14 days, LT > 14 days). Parameters such as length of ICU stay, length of hospital stay, and duration of MV were used to evaluate outcomes. Additionally, the outcomes were categorized as good prognosis, poor prognosis, and death based on the manner of hospital discharge. Student's t-test, analysis of variance (ANOVA), Mann-Whitney U test, Kruskal-Wallis test, Chi-square test, and Fisher's exact test were employed as appropriate to assess differences in demographic data and individual characteristics among the ET, IMT, and LT groups. Univariate Cox regression model and multivariable Cox proportional hazards regression model were utilized to determine whether delaying tracheostomy would increase the risk of death. RESULTS: In both of two criterion, patients with delayed tracheostomies had longer hospital stays (p < 0.001), ICU stays (p < 0.001), total time receiving MV (p < 0.001), time receiving MV before tracheostomy (p < 0.001), time receiving MV after tracheostomy (p < 0.001), and sedation durations. Similar results were also found in sub-population diagnosed as trauma, neurogenic or digestive disorders. Multinomial Logistic regression identified LT was independently associated with poor prognosis, whereas ET conferred no clinical benefits compared with IMT. CONCLUSIONS: In a mixed ICU population, delayed tracheostomy prolonged ICU and hospital stays, sedation durations, and time receiving MV. Multinomial logistic regression analysis identified delayed tracheostomies as independently correlated with worse outcomes. TRIAL REGISTRATION: ChiCTR2100043905. Registered 05 March 2021. http://www.chictr.org.cn/listbycreater.aspx.

Three-Dimensional Guided Cone-Shaped Segmentectomy Versus Lobectomy for Small-sized Non-Small Cell Lung Cancer in the Middle Third of the Lung Field.

BACKGROUND: Segmentectomy has been recommended for peripheral small-sized non-small cell lung cancer (NSCLC). This study aimed to evaluate whether three dimensionally (3D) guided cone-shaped segmentectomy can achieve long-term outcomes comparable with lobectomy for small-sized NSCLC in the middle third of the lung parenchyma. METHODS: This study retrospectively screened patients with small NSCLC (≤2 cm) who underwent segmentectomy or lobectomy between January 2012 and June 2019. Tumor location was determined by 3D multiplanar reconstruction. The cone-shaped segmentectomy was performed with the guidance of 3D computed tomographic bronchography and angiography. The log-rank test, Cox hazard proportional regression, and propensity score-matching analyses were adopted for prognostic evaluation. RESULTS: After screening, 278 patients with segmentectomy and 174 subjects undergoing lobectomy were selected. All the patients had R0 resection, and no 30- or 90-day mortality was observed. The median follow-up time was 47.3 months. The 5-year overall survival (OS) was 99.6 %, and the disease-free survival (DFS) was 97.5 % for the patients undergoing segmentectomy. After propensity score-matching, the patients with segmentectomy (n = 112) had an OS (P = 0.530) and a DFS (P = 0.390) similar to those of the patients who underwent lobectomy (n = 112). The multivariable Cox regression analysis indicated no significant survival differences between segmentectomy and lobectomy [DFS: hazard ratio, 0.56 (95 % confidence interval (CI) 0.16-1.97, P = 0.369); OS: HR, 0.35 (95 % CI 0.06-2.06, P = 0.245)] after adjustment for other factors. Further analysis showed that segmentectomy achieved comparable OS (P = 0.540) and DFS (P = 0.930) for NSCLC in the middle-third and peripheral lung parenchyma (n = 454). CONCLUSIONS: For selected NSCLCs size 2 cm or smaller in the middle third of the lung field, 3D-guided cone-shaped segmentectomy was able to achieve long-term outcomes comparable with lobectomy.

Difficulty of Video-Assisted Thoracoscopic Surgery Segmentectomy: Proposal for a New Classification.

BACKGROUND: Segmentectomy has classically been distinguished as "simple" or "complex" based on the number of intersegmental planes (ISPs) dissected. However, with the increasing variety and complexity of segmentectomies, it is clear that a classification based on the number of ISPs alone is inadequate. This study aimed to develop a new classification to predict the surgical difficulty of video-assisted thoracoscopic surgery (VATS) segmentectomy. METHODS: The study retrospectively reviewed 1868 patients who underwent VATS segmentectomy between January 2014 and December 2019. Uni- and multivariate analyses were performed to identify predictors associated with prolonged operative time (>140 min), and a scoring system was constructed to classify the surgical difficulty of VATS segmentectomy. RESULTS: Altogether, 1868 VATS segmentectomies were divided into three groups: group 1 (low difficulty, including segmentectomy with only one intersegmental plane [ISP] dissection), group 2 (intermediate difficulty, including a single segmentectomy with more than one ISP dissection and a single subsegmentectomy), group 3 (high difficulty level, including combined resection with more than one ISP dissection). This classification effectively differentiated the three groups in terms of operative time, estimated blood loss, major complications, and overall complications (all p < 0.001). For receiver operating characteristic analysis, the new classification showed significantly better differentiation performance in terms of operative time (p < 0.001), estimated blood loss (p = 0.004), major complications (p = 0.002), and overall complications (p = 0.012) than the simple/complex classification. CONCLUSIONS: This new three-level classification accurately predicted the surgical difficulty of VATS segmentectomy.

Exosomal circ_0007385 enhances non-small cell lung cancer cell proliferation and stemness via regulating miR-1253/FAM83A axis.

Exosomes are critical mediators of intercellular communication in the tumor microenvironment. Exosomal circular RNAs (circRNAs) can act as biomarkers and play crucial roles in many cancers, including non-small cell lung cancer (NSCLC). The aim of this study was to explore the functions and regulatory mechanism of exosomal circ_0007385 in NSCLC. The expression levels of circ_0007385, microRNA-1253 (miR-1253), family with sequence similarity 83, member A (FAM83A) mRNA were determined by quantitative real-time PCR (qRT-PCR). Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (Edu), and colony formation assays were utilized to determine cell proliferation ability. Sphere formation efficiency was determined by sphere formation assay. All protein levels were detected by western blot assay. Exosomes were detected using transmission electron microscopy analysis. Size distribution of exosomes was analyzed by nanoparticle tracking analysis. The interaction between miR-1253 and circ_0007385 or FAM83A was confirmed by dual-luciferase reporter, RNA immunoprecipitation (RIP) and RNA pull-down assays. Mice xenograft model was established to verify the function of circ_0007385 in vivo. Circ_0007385 was upregulated in NSCLC tissues and cells. Knockdown of circ_0007385 inhibited NSCLC cell proliferation and stemness, while exosomal circ_0007385 facilitated NSCLC cell proliferation and stemness. In addition, miR-1253 was a direct target of circ_0007385, and miR-1253 reversed the inhibitory effects of circ_0007385 on cell proliferation and stemness in NSCLC cells. Moreover, FAM83A was a direct target of miR-1253, and miR-1253 suppressed NSCLC cell proliferation and stemness by targeting FAM83A. Furthermore, circ_0007385 knockdown inhibited tumor growth in vivo. Exosomal circ_0007385 promoted NSCLC cell proliferation and stemness by regulating miR-1253/FAM83A axis.

LncRNA PCBP1-AS1 correlated with the functional states of cancer cells and inhibited lung adenocarcinoma metastasis by suppressing the EMT progression.

The development of single-cell RNA sequencing (scRNA-seq) provided us an unprecedented chance to identify novel oncogenes or tumor suppressors at single-cell resolution. Long non-coding RNAs (lncRNAs) related to the functional states of cancer cells might play vital roles in the progression of lung adenocarcinoma (LUAD). In this study, lncRNAs that were associated with the functional states of LUAD cells identified in scRNA-seq studies were screened based on the CancerSEA database. Differential gene expression analysis and survival analysis were performed in TCGA, GEO and our JSPH databases. Finally, transwell and tail vein metastasis assays were used to reveal the functions of our identified novel prognostic lncRNAs. A total of 849 lncRNAs were initially identified. Among them, 11 lncRNAs were found significantly associated with LUAD prognosis in the TCGA database. Two of them (PCBP1-AS1 and ZSCAN16-AS1) were further validated in independent GEO datasets. ScRNA-seq analysis showed that PCBP1-AS1 and ZSCAN16-AS1 were significantly negatively correlated with most of the functional states of LUAD cells, especially with metastasis. Functionally, PCBP1-AS1 was aberrantly downregulated in LUAD cells and tumor tissues. Knockdown of PCBP1-AS1 significantly promoted the migration and invasion of LUAD cells. Consistently, PCBP1-AS1 overexpression suppressed the metastasis of LUAD in vitro and in vivo. Besides, PCBP1-AS1 inhibition induced decreased E-cadherin expression and increased N-cadherin, Vimentin and Snail expression. In conclusion, PCBP1-AS1 could suppress the metastasis of LUAD by targeting the epithelial-mesenchymal transition pathway and might serve as a prognostic biomarker and a potential therapeutic target of LUAD.

Association Between Intercostal Nerve Block and Postoperative Glycemic Control in Patients With Diabetes Undergoing Video-Assisted Thoracoscopic Pulmonary Resection: A Retrospective Study.

OBJECTIVES: The present study was performed to investigate the possible association between intercostal nerve block (INB) and postoperative glycemic control in patients with diabetes undergoing video-assisted thoracoscopic pulmonary resection. DESIGN: A retrospective study. SETTING: Single-center tertiary academic hospital. PARTICIPANTS: Patients with diabetes, ages 18 to 79 years, who had undergone elective video-assisted thoracoscopic pulmonary resection (segmentectomy or lobectomy) from January 1, 2015, to December 31, 2018. INTERVENTIONS: Postoperative blood glucose levels and insulin dosage were extracted from the record. MEASUREMENTS AND MAIN RESULTS: Patients with diabetes who received INB before closure of surgical incisions were compared with those who did not receive INB. The primary outcome was the daily blood glucose (BG) level. Univariate analyses and multivariate regression analysis were performed to explore risk factors of hyperglycemia within 48 hours after the surgery. Baseline characteristics were comparable between the two groups. Patients who received INB had a lower maximum BG level and amplitude of glycemic excursion from zero-to-24 hours after surgery (p = 0.007 and p = 0.041, respectively) and lower maximum and minimum BG levels from 24-to-48 hours after surgery (p = 0.023 and p = 0.006, respectively). Meanwhile, the daily insulin dose increment during zero-to-24 hours and 24-to-48 hours after surgery decreased (p = 0.010 and p = 0.003, respectively), the white blood cell counts within 48 hours after surgery were lower (p = 0.021), and the length of postoperative stay decreased in the INB group (p = 0.044). Multivariate regression analysis further confirmed that INB was an independent protective factor of postoperative hyperglycemia (Nagelkerke R2 value 0.229; odds ratio 0.298; 95% confidence interval 0.099-0.901; p = 0.032). CONCLUSION: INB, performed before closure of surgical incisions, was associated with improved glycemic control in patients with diabetes within 48 hours after video-assisted thoracoscopic pulmonary resection.

The rat model of COPD skeletal muscle dysfunction induced by progressive cigarette smoke exposure: a pilot study.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) skeletal muscle dysfunction is a prevalent malady that significantly affects patients' prognosis and quality of life. Although the study of this disease has attracted considerable attention, a definite animal model is yet to be established. This study investigates whether smoke exposure could lead to the development of a COPD skeletal muscle dysfunction model in rats. METHODS: Sprague Dawley rats were randomly divided into model (MG, n = 8) and control groups (CG, n = 6). The MG was exposed to cigarette smoke for 16 weeks while the CG was not. The body weight and forelimb grip strength of rats were monitored monthly. The pulmonary function and the strength of tibialis anterior muscle were assessed in vitro and compared after establishing the model. The histological changes in lung and gastrocnemius muscles were observed. The expressions of interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α were detected by ELISA, while the expressions of Atrogin-1 and MuRF1 in the gastrocnemius muscle were determined by Western blotting. RESULTS: Smoke exposure slowly increases the body weight and forelimb grip strength of MG rats, compared to CG rats. However, it significantly decreases the pulmonary ventilation function and the skeletal muscle contractility of the MG in vitro. Histologically, the lung tissues of MG show typical pathological manifestations of emphysema, while the skeletal muscles present muscular atrophy. The expressions of IL-6, IL-8, and TNF-α in MG rats are significantly higher than those measured in CG rats. Increased levels of Atrogin-1 and MuRF1 were also detected in the gastrocnemius muscle tissue of MG. CONCLUSION: Progressive smoking exposure decreases the contractile function of skeletal muscles, leading to muscular atrophy. It also increases the expressions of inflammatory and muscle protein degradation factors in COPD rats. This indicates that smoke exposure could be used to establish a COPD skeletal muscle dysfunction model in rats.

Effects of resistance training on exercise capacity in elderly patients with chronic obstructive pulmonary disease: a meta-analysis and systematic review.

OBJECTIVE: The objective of this study was to summarize and determine the effectiveness of resistance training on exercise capacity in patients with chronic obstructive pulmonary disease (COPD). METHODS: We searched PubMed, EMBASE, Cochrane Library, and two Chinese databases (China National Knowledge Infrastructure and Wanfang Data) to identify articles written in English or Chinese and published from January 2000 to January 2019. Randomized controlled trials were included if they evaluated the effects of resistance training on exercise capacity in COPD patients. We assessed the quality of the trials using the Physiotherapy Evidence Database Scale. Data from these studies were pooled to calculate weighted mean difference (WMD) or standardized mean difference (SMD) with 95% confidence intervals (CI). RESULTS: Eleven studies with a total of 405 participants met the inclusion criteria. Compared with the non-exercise control group, resistance training significantly improved 6-min walking distance (WMD, 54.52; 95% CI 25.47-83.56; I2 = 43%; P = 0.14), transfer numbers for the 6-min pegboard and ring test (WMD, 25.17; 95% CI 10.17-40.16; I2 = 0%; P = 0.55), and tolerance time for the unsupported upper-limb exercise test (SMD, 0.41; 95% CI 0.03-0.79; I2 = 0%; P = 0.83). There were no significant differences in constant work rate endurance test results or in peak oxygen uptake between the two groups. CONCLUSIONS: Resistance training was an effective approach to improve functional exercise capacity, endurance exercise capacity, and peak exercise capacity in COPD patients.

Genome-wide copy number variation analysis identified ANO1 as a novel oncogene and prognostic biomarker in esophageal squamous cell cancer.

Copy number variations (CNVs) represent one of the most common genomic alterations. This study aimed to evaluate the roles of genes within highly aberrant genome regions in the prognosis of esophageal squamous cell cancer (ESCC). Exome sequencing data from 81 paired ESCC tissues were used to screen aberrant genomic regions. The associations between CNVs and gene expression were evaluated using gene expression data from the same individuals. Then, an RNA expression array profile from 119 ESCC samples was adopted for differential gene expression and prognostic analyses. Two independent ESCC cohorts with 315 subjects were further recruited to validate the prognostic value using immunohistochemistry tests. Finally, we explored the potential mechanism of our identified novel oncogene in ESCC. In total, 2003 genes with CNVs were observed, of which 76 genes showed recurrent CNVs in more than three samples. Among them, 32 genes were aberrantly expressed in ESCC tumor tissues and statistically correlated with CNVs. Strikingly, 4 (CTTN, SHANK2, INPPL1 and ANO1) of the 32 genes were significantly associated with the prognosis of ESCC patients. Patients with a positive expression of ANO1 had a poorer prognosis than ANO1 negative patients (overall survival rate: 42.91% versus 26.22% for ANO1-/+ samples, P < 0.001). Functionally, ANO1 promoted ESCC cell proliferation, migration and invasion by activating transforming growth factor-ß pathway. Knockdown of ANO1 significantly inhibited tumor progression in vitro and in vivo. In conclusion, ANO1 is a novel oncogene in ESCC and may serve as a prognostic biomarker for ESCC.

Integrating expression-related SNPs into genome-wide gene- and pathway-based analyses identified novel lung cancer susceptibility genes.

Traditional pathway analysis map single nucleotide polymorphisms (SNPs) to genes according to physical position, which lacks sufficient biological bases. Here, we incorporated genetics of gene expression into gene- and pathway-based analysis to identify genes and pathways associated with lung cancer risk. We identified expression-related SNPs (eSNPs) in lung tissues and integrated these eSNPs into three lung cancer genome-wide association studies (GWASs), including 12,843 lung cancer cases and 12,639 controls. We used SKAT-C for gene-based analysis, and conditional analysis to identify independent eSNPs of each gene. ARTP algorithm was used for pathway analysis. A total of 374,382 eSNPs in the GWAS datasets survived quality control, which were mapped to 5,084 genes and 2,752 pathways. In the gene-based analysis, nine genes showed significant associations with lung cancer risk. Among them, TP63 (3q28), RP11-650L12.2 (15q25.1) and CHRNA5 (15q25.1) were located in known lung cancer susceptibility loci. We also validated two newly identified susceptibility loci (RNASET2 and AL133458.1 in 6q27, and MPZL3 in 11q23.3). Besides, DVL3 (3q27.1), RP11-522I20.3 (9q21.32) and CCDC116 (22q11.21) were identified as novel lung cancer susceptibility genes. Pathway analysis showed that pathways involved in protein structure, the Notch signaling pathway and the nicotinic acetylcholine receptor-related pathways were associated with lung cancer risk. Combing eSNPs, gene- and pathway-based analysis identifies novel lung cancer susceptibility genes, which serves as a powerful approach to decipher biological mechanisms underlying GWAS findings.

Histone deacetylase inhibitor BG45-mediated HO-1 expression induces apoptosis of multiple myeloma cells by the JAK2/STAT3 pathway.

Multiple myeloma (MM) is a hematological malignancy that is characterized by the clonal expansion of plasma cells in the bone marrow. Histone deacetylases (HDACs) represent a new type of molecular targeted therapy for different types of cancers and promising targets for myeloma therapy. We showed that HDAC3 mRNA and protein levels of CD138 mononuclear cells from MM patients were higher than those in healthy donors. Therefore, we investigated the effects of a novel class I HDAC inhibitor BG45 on MM cells in vitro. BG45 downmodulated heme oxygenase 1 (HO-1) when class I HDACs decreased in MM cells. HO-1 is a target for the treatment of MM. Moreover, BG45 induced hyperacetylation of histone H3 and inhibited the growth, especially the apoptosis of MM cell lines. Treatment with BG45 induced apoptosis by downregulating bcl-2 and Bcl-xl, upregulating Bax and other antiapoptotic proteins and activating poly(ADP-ribose)polymerase, and decreasing protein levels of p-JAK2 and p-STAT3. These effects were partly blocked by HO-1. Correspondingly, BG45 led to an accumulation in the G0/G1 phase, accompanied by decreased levels of CDK4 and phospho-retinoblastoma protein, an increased level of p21, and a moderately reduced level of CDK2. Clinical use of single agents was limited because of toxic side effects and drug resistance. However, combining BG45 with lenalidomide exerted synergistic effects. In conclusion, we verified the potent antimyeloma activity of this novel HDAC inhibitor and that the combination of BG45 and lenalidomide is a new method for MM treatment. Thus, BG45 may be applicable to the treatment of MM and other hematological malignancies.

Induction of heme oxygenase-1 by Na+-H+ exchanger 1 protein plays a crucial role in imatinib-resistant chronic myeloid leukemia cells.

Resistance toward imatinib (IM) and other BCR/ABL tyrosine kinase inhibitors remains troublesome in the treatment of advanced stage chronic myeloid leukemia (CML). The aim of this study was to estimate the reversal effects of down-regulation of Na(+)/H(+) exchanger 1 (NHE1) on the chemoresistance of BCR-ABL-positive leukemia patients' cells and cell lines. After treatment with the specific NHE1 inhibitor cariporide to decrease intracellular pH (pHi), the heme oxygenase-1 (HO-1) levels of the K562R cell line and cells from IM-insensitive CML patients decreased. HO-1, as a Bcr/Abl-dependent survival molecule in CML cells, is important for the resistance to tyrosine kinase inhibitors in patients with newly diagnosed CML or IM-resistant CML. Silencing PKC-ß and Nrf-2 or treatment with inhibitors of p38 pathways obviously blocked NHE1-induced HO-1 expression. Furthermore, treatment with HO-1 or p38 inhibitor plus IM increased the apoptosis of the K562R cell line and IM-insensitive CML patients' cells. Inhibiting HO-1 enhanced the activation of caspase-3 and poly(ADP-ribose) polymerase-1. Hence, the results support the anti-apoptotic role of HO-1 induced by NHE1 in the K562R cell line and IM-insensitive CML patients and provide a mechanism by which inducing HO-1 expression via the PKC-ß/p38-MAPK pathway may promote tumor resistance to oxidative stress.