Non-small Cell Lung Cancer Group Research Articles

Expression differences and analysis of Endostatin and VEGF in different efficacy NSCLC patients treated with rh-endostatin combined chemotherapy

Objective To investigate the clinical significance and difference in the expression of endostatin and vascular endothelial growth factor (VEGF) in non-small cell lung cancer (NSCLC) patients with different response to recombinant human endostatin (rh-endostatin) combined with chemotherapy. Methods Serum levels of endostatin and VEGF in peripheral blood of 30 patients with stage Ⅳ NSCLC (NSCLC group) and 30 healthy controls (control group) were determined by enzyme-linked immunosorbent assay. Two cycles of chemotherapy combined with rh-endostatin were provided to NSCLC patients to evaluate the efficacy of the regimen. Simultaneously, serum levels of endostatin and VEGF were measured before and after treatment. Results The level of serum endostatin was (37.96±9.01)ng/ml and (40.12±12.11)ng/ml in NSCLC patients and healthy controls, respectively, which was lower in the former than that of the latter, without statistical difference (P>0.05). Furthermore, the level of serum VEGF was (127.98 ±33.88)pg/ml and (36.33±15.43)pg/ml in NSCLC patients and healthy controls, respectively, which was higher in the former than that of the latter, with statistical difference (t=13.48, P 0.05). After two cycles of chemotherapy combined with rh-endostatin treatment, the level of serum endostatin in partial response (PR) or stable disease (SD) patients was (76.22±20.41)ng/ml, higher than that of progressive disease (PD) patients, which was (31.24±13.09)ng/ml (t=7.143, P<0.05). In addition, the level of serum VEGF in PR or SD patients was (93.28±21.33)pg/ml, which was lower than (155.81±48.38)pg/ml of the PD patients (t=3.503, P<0.05). Conclusions The levels of endostatin and VEGF are associated with the efficacy of anti-angiogenesis combined with chemotherapy in NSCLC patients. Key words: Carcinoma, non-small-cell lung/DT; Antineoplastic combined chemotherapy protocols/TU; Endostar/AD; Endostatins/ME; Vascular endothelial growth factors/ME

Abstract 617: Nivolumab efficacy and safety in veterans with and without HIV infection

Abstract The objective of this study was to compare the efficacy and safety of nivolumab in the HIV-infected and HIV-uninfected veteran populations. WHAT IS NEW: Most clinical trials highlighting checkpoint inhibitors excluded people living with HIV (PLWH). This is the first study to report equivalent nivolumab tumor response rates in PLWH, but higher rates of pneumonitis.METHODS: We searched the Corporate Data Warehouse (CDW) to identify all HIV-positive patients who used the Veterans Health Administration (VA) between 2000 and 2016. For this cohort of 46,916 PLWH, we obtained a 4:1 age-matched HIV-negative control cohort. We used pharmacy database to identify all nivolumab recipients through July 26, 2017. We reviewed patients' electronic medical records using Compensation and Pension Records Interchange (CAPRI) to obtain age, sex, geographic location, cancer type, number of nivolumab doses received, previous cancer therapy, adverse events, and response to therapy. We calculated summary descriptive measures for both groups and compared the proportion of patients experiencing adverse event or pneumonitis in groups by HIV status, using chi-square or Fisher's exact test. RESULTS: Sixteen PLWH and 68 HIV-negative controls received nivolumab during the study period. Only 51 controls had accessible data. Median age at nivolumab initiation was 65 years in both groups (range, 47-85 [IQR, 59-68] in PLWH; range, 42-79 [IQR 59-68] in controls); all patients were male. Overall, 63% received nivolumab for non-small cell lung cancer (NSCLC): 8/16 (50%) in PLWH, 34/51 (67%) in controls. Other indications in PLWH included renal cell carcinoma (RCC) (2/16 [13%]), Hodgkin lymphoma (HL) (2/16 [13%]), hepatocellular carcinoma (HCC) (2/16 [13%]). Other indications in controls were RCC (9/51 [18%]), melanoma (5/51, [10%]), and head and neck squamous cell carcinoma (3/51 [6%]).Regardless of HIV status, the median number of prior lines of therapy was 1, median number of nivolumab doses received was 6, and median progression-free survival (PFS) in NSCLC was 2.75 months. Half of each NSCLC group received radiation; fewer PLWH had COPD (50%, vs. 82% of controls). PLWH had a trend towards more adverse events (AEs) (6/15 [40%], vs. 13/51 [26%] of controls, p=0.28). A significantly higher proportion of PLWH experienced pneumonitis (4/15 [27%], vs. 2/51 [4%] of controls, p=0.007).Other AEs among PLWH were rash, hypothyroidism, and autoimmune diabetes mellitus. Other AEs among controls were fatigue, colitis, rash, hypothyroidism, arthritis, and hepatitis (1-3 patients each). CONCLUSION: Among veteran PLWH, rates of nivolumab administration, and PFS in NSCLC, were comparable to those of HIV-uninfected counterparts. Pneumonitis was significantly more frequent in PLWH. Further studies should investigate the mechanism of pneumonitis in this population, confirm the high frequency in larger cohorts of PLWH, and identify clinical risk factors for pneumonitis. Citation Format: Elaine Chang, Aaron P. Thrift, Donna L. White, Jennifer Kramer, Anita L. Sabichi, Christine Hartman, Kathryn E. Royse, Peter A. Richardson, Elizabeth Y. Chiao. Nivolumab efficacy and safety in veterans with and without HIV infection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 617.

Evaluation of Serum Paired MicroRNA Ratios for Differential Diagnosis of Non-Small Cell Lung Cancer and Benign Pulmonary Diseases.

To clarify whether there are different expressions between lung cancer and benign pulmonary diseases, we studied seven microRNAs (miRNAs) in serum from patients with non-small cell lung cancer (NSCLC), benign pulmonary nodules and four pulmonary inflammation diseases. We detected the expression of miRNAs using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). We found that five miRNA ratios-miR-15b-5p/miR-146b-3p, miR-20a-5p/miR-146b-3p, miR-19a-3p/miR-146b-3p, miR-92a-3p/miR-146b-3p, and miR-16-5p/miR-146b-3p-show higher expression in the NSCLC group than the benign pulmonary nodule group, and 13 ratios of miRNAs were significantly upregulated in the NSCLC group compared with the pulmonary inflammation diseases group. Receiver operating characteristic (ROC) curve analysis was performed. For NSCLC and benign pulmonary nodules, the sensitivity and specificity were 0.70 and 0.90, respectively. For NSCLC and pulmonary inflammation diseases, the sensitivity and specificity were 0.81 and 0.71, respectively. The ratios of miRNAs can be used as potential non-invasive biomarkers for diagnosis of early-stage NSCLC and benign pulmonary diseases.

Association between EGFR mutation and expression of BRCA1 and RAP80 in non-small cell lung cancer.

Association between the epithelial growth factor receptor (EGFR) mutation and the expression of breast cancer 1 (BRCA1) and receptor-associated protein 80 (RAP80) in non-small cell lung cancer (NSCLC) was studied. From September 2014 to September 2016, 51 patients with NSCLC who were hospitalized in Department of Thoracic Surgery in The Affiliated Jiangyin Hospital of Southeast University Medical College and underwent biopsy or operation were selected. The pathological changes of lung tissue were detected by hematoxylin and eosin histopathological staining. The fluorescent expression of BRCA1 and RAP80 protein in the two groups was determined by immunofluorescence staining. Reverse transcriptase polymerase chain reaction method and western blot analysis were used to detect BRCA1 and RAP80 mRNA and protein expression. Then the EGFR gene mutation was detected and analyzed. The results show that non-small cell lung cancer has an association with smoking. Compared with the control, the lung tissue structure of the NSCLC group was damaged. The protein fluorescence expression of BRCA1 and RAP80 in non-small cell lung cancer group was significantly increased. The expression of BRCA1 and RAP80 mRNA and protein in NSCLC group was significantly increased. The difference in expression of BRCA1 and RAP80 in the control and the non-small cell lung cancer group was statistically significant (P<0.05). EGFR gene mutations detected 14 of the 51 patients with genetic mutations. Non-small cell lung cancer and smoking have certain relevance, and BRCA1 and RAP80 expression in the development and progression of NSCLC has a close relationship. EGFR mutation in non-small cell lung cancer significantly related to the mutation of EGFR and BRCA1 and RAP80 gene expression plays an important role in the diagnosis and treatment of NSCLC.

Genetic polymorphisms in the TERT gene and susceptibility to non-small cell lung cancer in a Chinese Han population.

BackgroundRecent studies have revealed that the TERT gene plays crucial roles in cancer initiation and development. Genome-wide analysis studies and case-control studies have demonstrated that polymorphisms in the TERT gene are associated with various cancers.Materials and methodsIn the current study, we analyzed the associations of eight single nucleotide polymorphisms (SNPs) in the TERT gene with non-small cell lung cancer (NSCLC) in a Chinese Han population. A total of 467 NSCLC patients and 526 healthy individuals were recruited for SNP genotyping using a TaqMan assay.ResultsOur results revealed that the allelic frequencies of rs2853677 and rs2853691 were significantly different between the NSCLC and control groups (P=0.004 and 0.001, respectively). Moreover, the T allele of rs2853677 and the A allele of rs2853691 might be the protective factors against NSCLC (OR=0.766; 95%CI: 0.639–0.918 and OR=0.714; 95%CI: 0.584–0.875, respectively). Additionally, stratified association analysis of the eight SNPs with the different pathological NSCLC stages (I+II and III+IV) and different pathological types (adenocarcinoma and squamous cell carcinoma) revealed that none of the SNPs were significantly different between patients with different pathological stages and pathological types.ConclusionOur results indicated that rs2853677 and rs2853691 in the TERT gene might be associated with NSCLC in this Chinese Han population.

The association of polymorphisms in miRNAs with nonsmall cell lung cancer in a Han Chinese population

BackgroundMicroRNAs (miRNAs) have been demonstrated to play important roles in cancer progression. Recently, studies have revealed that polymorphisms in miRNAs might be associated with cancer susceptibility.Materials and methodsIn the current study, we investigated the associations of single nucleotide polymorphisms (SNPs) in miRNAs (rs11134527 in pri-miR-218-2, rs74693964 in pri-miR-145, rs6062251 in pri-miR-133a-2, and rs4705343 in pri-miR-143) with nonsmall cell lung cancer (NSCLC) in a Han population from Yunnan Province, Southwest China using a binary logistic regression analysis. A total of 452 patients with NSCLC and 452 healthy individuals were recruited for polymorphism genotyping using the TaqMan assay.ResultsOur results showed that the allelic frequencies of rs11134527 and rs4705343 were significantly different between the NSCLC and control groups (P=0.025 and 0.029). Additionally, the genotypic frequencies of rs11134527 were significantly different between the NSCLC and control groups (P=0.045). The mode of inheritance analysis showed that genotypes A/G+G/G of rs11134527 were associated with a lower risk of NSCLC under the dominant model (OR=0.69; 95% CI: 0.51–0.94). In addition, genotypes 2C/C+C/T of rs4705343 were associated with an increased risk of NSCLC under the log-additive model (OR=1.25; 95% CI: 1.01–1.53). However, there was no significant difference in the other SNPs between the NSCLC and control groups (P>0.05). Moreover, the association analysis of these SNPs between adenocarcinoma and squamous cell carcinoma (SCC) showed that allele A of rs11134527 was associated with SCC (OR=0.65; 95% CI: 0.48–0.88).ConclusionOur results indicated that the A allele of rs11134527 might be a risk factor (OR=1.24; 95% CI: 1.03–1.50) and that the T allele of rs4705343 might be a protective factor (OR=0.80; 95% CI: 0.66–0.98) for NSCLC in a Han Chinese population.

The role of tumor-infiltrating lymphocytes (TILs) as a predictive biomarker of response to anti-PD1 therapy in patients with metastatic non-small cell lung cancer or metastatic melanoma.

Immunotherapy plays an important role in cancer treatment. Biomarkers that can predict response, including tumor-infiltrating lymphocytes (TILs), are in the spotlight of many studies. This cohort study was designed to evaluate the role of CD4+ and CD8+ TILs as predictive factors for response to anti PD-1 treatment in patients with metastatic non-small cell lung cancer (NSCLC) or metastatic melanoma. We evaluated the expression of CD4+ and CD8+ TILs in tissue samples of 56 patients with metastatic NSCLC or melanoma treated with anti-PD1 immunotherapy. The study included 30 patients with melanoma and 26 with NSCLC. An association was found between CD8+/CD4+ TILs ratio and response to anti-PD1 treatment in both cancers. Regarding melanoma patients, ratios of CD8+/CD4+ lower than 2 predicted lack of response to treatment (0%) (p=0.006), while CD8+/CD4+ ratios higher than 2.7 had an 81.3% response rate (p=0.0001). In addition, we found that the presence of more than 1900/mm2 of CD8+ lymphocytes in the melanoma tumor predicted a 90% response to therapy. In the metastatic NSCLC group, tumors with CD8+ lymphocyte count under 886/mm2 showed low response rates (16.7%, p=0.046). When the CD8+ lymphocyte count was in the range of 886-1899/mm2, the response rate was high (60%, p=0.017). In CD8+/CD4+ ratios lower than 2, the response rate was low (13.3%), and in ratios higher than 2, response rates ranged between 43 and 50% (p=0.035). The use of CD8+/CD4+ TILs ratios in tumor biopsies may predict response to anti-PD1 treatment in metastatic melanoma and NSCLC.

Prognostic value of interferon-gamma, interleukin-6, and tumor necrosis factor-alpha in the radiation response of patients diagnosed with locally advanced non-small-cell lung cancer and glioblastoma multiforme

Background/aim: This study aimed to investigate the effect of chemoradiotherapy (CRT) on interferon-gamma (IFN-γ), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α), which are critical markers of the clinical radiation response of patients with locally advanced non-small-cell lung cancer (NSCLC) and glioblastoma multiforme (GBM). Materials and methods: Thirty patients who were treated with CRT and 20 healthy controls were prospectively evaluated. Circulating levels of cytokines were measured by enzyme-linked immunosorbent assay procedure. Post-CRT and pre-CRT levels were compared. Results: Post-CRT, TNF-α and IFN-γ levels were significantly lower than pre-CRT levels in the NSCLC and GBM groups, respectively. The statistical analysis did not show any significant difference between the post- and pre-CRT IL-6 levels. However, the pre-CRT IL-6 levels in the GBM group and post-CRT IL-6 levels in the NSCLC group were significantly higher than those of the control group. Conclusion: CRT affected TNF-α levels in NSCLC and IFN-γ levels in GBM, with the levels of both decreasing significantly. The IL-6 levels of the post-CRT NSCLC group were higher than those of the post-CRT GBM group. Irradiation-induced IL-6 may be responsible for tumor regrowth. Therefore, treatment with IL-6 inhibitors could be a potential therapeutic strategy for sensitizing NSCLC to irradiation in the clinic.

Predictive value of microRNA-10b expression in peripheral blood mononuclear cells in evaluating short- and long-term efficacy of chemotherapy for patients with advanced non-small-cell lung cancer.

The expression profile in peripheral blood mononuclear cells (PBMCs) from advanced non-small-cell lung cancer (NSCLC) patients and the role of microRNAs (miRs) in NSCLC remain unclear. Herein, the present study aims to investigate predictive value of miR-10b expression in PBMCs in evaluating short- and long-term efficacy of chemotherapy for NSCLC patients. A total of 194 advanced NSCLC patients were selected as the NSCLC group and 199 healthy individuals were recruited as a control group. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed to determine the miR-10b expression in PBMCs. The patients with advanced NSCLC were treated with chemotherapy, and the relationship between miR-10b expression in PBMCs and the clinicopathological characteristics, clinical response to chemotherapy and prognosis were analyzed. The values of miR-10b in diagnosis and prediction of clinical response to chemotherapy before chemotherapy were detected by receiver operating characteristic (ROC) curve analysis, the clinical response to chemotherapy by logistic regression analysis, and the risk factors of prognosis in NSCLC patients by Cox regression analysis. Compared with the control group, miR-10b in PBMCs was highly expressed in the NSCLC group. The area under the curve (AUC) of miR-10b expression in the diagnosis of NSCLC was 0.967, with sensitivity and specificity of 85.10% and 99.5%, respectively. MiR-10b expression was related to lymph node metastasis (LNM), distant metastasis, pathological types and differentiation. The AUC of miR-10b expression in the prognosis of advanced NSCLC was 0.793, with sensitivity and specificity of 75.20% and 76.62%, respectively. Logistic regression analysis showed that clinical response to chemotherapy was significantly influenced by miR-10b expression, distant metastasis and LNM. Cox regression analysis showed that the high miR-10b expression, smoking, LNM and distant metastasis were risk factors of prognosis for advanced NSCLC patients. The present study offers intriguing new perspectives based on evidence that miR-10b expression in PBMCs has predictive value for the tumor response to chemotherapy and prognosis for advanced NSCLC patients.

Expression level of long noncoding RNA H19 in plasma of patients with nonsmall cell lung cancer and its clinical significance.

This study was aimed to explore the expression level of long noncoding RNA H19 in the plasma of patients with nonsmall cell lung cancer (NSCLC) and its clinical significance. A total of 66 NSCLC patients (case group) and 31 patients with benign lung disease (control group) admitted from February 2015 to February 2017 were included in this study. Real-time polymerase chain reaction assay was applied to examine the relative expression level of long noncoding RNA H19 in the plasma of the two groups. The relationship between H19 expression and clinical, pathological features was explored. Receiver-operating characteristic (ROC) curve was applied to evaluate the clinical value of plasma H19 as a tumor marker in the auxiliary diagnosis of NSCLC. The relative expression levels of plasma H19 inpatients from NSCLC group and benign lung disease group were 5.62 ± 2.02 (ΔCt) and 7.74 ± 2.75 (ΔCt), respectively. The NSCLC group presented with significantly higher levels than that of the benign disease group (P < 0.05). According to the median of relative expression level of 5.54, the plasma H19 of NSCLC patients was classified into low expression group ≥5.54 (n = 34) and high expression group <5.52 (n = 32). The relationship between the patients' clinical, pathological features, and the expression level of H19 was analyzed. The expression of H19 was not significantly correlated with the gender, age, clinical staging, tumor diameter, and pathological type of the patients (Pall > 0.05). With the serum H19 as a diagnosis reference, the diagnostic sensitivity of NSCLC was 67.74%, and the specificity was 63.08%. The area under the ROC curve was 0.73, and the diagnostic cutoff value was 6.62. Plasma level of H19 in NSCLC patients was significantly increased, which could be applied as a serological marker for the auxiliary diagnosis of NSCLC.

Abstract LB-A05: Utilizing methylation and expression signatures to construct predictive models and personalized cancer treatments

Abstract Background: Data from high-throughput genomic arrays available through The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) have been helpful in elucidating the role of methylome and transcriptome level variations in the oncogenesis and progression of various cancers. Methylome remodeling has been observed across most cancers, and several studies have revealed a novel role for methylation in regulating alternative splicing. Alternative splicing contributes to the malignancy and aggressiveness of cancers, and intragenic methylation of alternatively spliced exons (ASEs) can serve as an effective biomarker for prognosis and personalized treatment. This study focuses on identifying and exploiting clinically relevant intragenic methylation signatures in cancer. Methods: In this study, methylation beta values from the Illumina HumanMethylation450 (HM450) platform, exon expression values from the Illumina HiSeq 2000 platform and patient survival data were downloaded from TCGA. Values were then sorted into methylated and unmethylated groups at each intragenic HM450 probe using an algorithm for dichotomizing microarray data. Probes were selected if their methylation status significantly affected both exon expression and patient mortality and sorted by the difference in patient mortality between the two groups. Biomarkers with beta values falling within the lower mortality group were considered “positive biomarkers.” A series of univariate analyses were used to determine groupings of patients with similar outcomes based on the total number of positive biomarkers in a sample. This methodology was applied to multiple TCGA cohorts including breast cancer, non-small cell lung cancer (NSCLC) and metastatic melanoma. Prognostic models were constructed using multivariate regressions for each TCGA cohort and independently validated with data from GEO. Using clinical data from TCGA and Broad GDAC Firehose, each group of patients was also analyzed based on treatment including surgery, radiation and chemotherapy. Results: In each type of cancer, an unweighted index was created from the top biomarkers. In breast cancer nine probes were included in the index. Patients with seven to nine positive biomarkers had similar overall survivals (HR=1.000), as did patients with four to six positive biomarkers (HR=2.336, 95% CI=1.507-3.621) and zero to three positive biomarkers (HR=8.223, 95% CI=4.103-16.482). Benefits of radiation therapy were seen in the group with four to six positive biomarkers (p&amp;lt;.01). On average, normal breast tissue had more positive biomarkers than tumor tissue (p&amp;lt;.0001). In NSCLC, shorter survival rates were associated with patients in the top biomarker group treated with radiation therapy (p&amp;lt;.05). Patients in the second NSCLC group responded best to cisplatin (p&amp;lt;.05) while patients in the third group responded best to carboplatin (p&amp;lt;.05). The full results of the study will be made available online through an interactive web tool for physicians and researchers. Conclusions: This study has identified biomarkers of genes and pathways involved in alternative splicing in various cancers. These biomarkers are powerful predictors of prognosis and therapeutic response in the tested cohorts and may ultimately be useful for predicting personalized patient prognosis and guiding effective therapy. Citation Format: Caleb Reagor, Rebecca Conway. Utilizing methylation and expression signatures to construct predictive models and personalized cancer treatments [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr LB-A05.

Genetic variation in CDH13 gene was associated with non-small cell lung cancer (NSCLC): A population-based case-control study.

Cadherin 13 (CDH13, T-cadherin, H-cadherin) has been identified as an anti-oncogene in various cancers. Recent studies have reported that downregulation of H-cadherin in cancers is associated with CDH13 promoter hypermethylation, which could be affected by the single nucleotide polymorphisms (SNPs) near CpG sites in the CDH13 promoter. In the current study, we investigated and analyzed the association of seven SNPs (rs11646213, rs12596316, rs3865188, rs12444338, rs4783244, rs12051272 and rs7195409) with non-small cell lung cancer (NSCLC) using logistic regression analysis. SNPs rs11646213, rs12596316, rs3865188 and rs12444338 are located in the promoter region, rs4783244 and rs12051272 are located in intron 1, and rs7195409 is located in intron 7. A total of 454 patients with NSCLC were placed into a NSCLC group and 444 healthy controls were placed into a control group, all participants were recruited to genotype the SNPs using Taqman assay. Our results showed that the allelic frequencies of rs11646213 were significantly different between NSCLC and control groups (P = 0.006). In addition, the association analysis of these SNPs stratified into NSCLC pathologic stages I+II and III+IV showed that the allelic frequencies rs7195409 had a significant difference between NSCLC pathologic stages I+II and III+IV (P = 0.006). Our results indicated that the rs11646213 and rs7195409 in CDH13 could be associated with NSCLC or its pathologic stages in the Chinese Han population.

Comparative Network Analysis of Patients with Non-Small Cell Lung Cancer and Smokers for Representing Potential Therapeutic Targets

Cigarette smoking is the leading cause of lung cancer worldwide. In this study, we evaluated the serum autoantibody (AAb) repertoires of non-small cell lung cancer (NSCLC) patients and smokers (SM), leading to the identification of overactivated pathways and hubs involved in the pathogenesis of NSCLC. Surface- and solution-phase biopanning were performed on immunoglobulin G purified from the sera of NSCLC and SM groups. In total, 20 NSCLC- and 12 SM-specific peptides were detected, which were used to generate NSCLC and SM protein datasets. NSCLC- and SM-related proteins were visualized using STRING and Gephi, and their modules were analyzed using Enrichr. By integrating the overrepresented pathways such as pathways in cancer, epithelial growth factor receptor, c-Met, interleukin-4 (IL-4) and IL-6 signaling pathways, along with a set of proteins (e.g. phospholipase D (PLD), IL-4 receptor, IL-17 receptor, laminins, collagens, and mucins) into the PLD pathway and inflammatory cytokines network as the most critical events in both groups, two super networks were made to elucidate new aspects of NSCLC pathogenesis and to determine the influence of cigarette smoking on tumour formation. Taken together, assessment of the AAb repertoires using a systems biology approach can delineate the hidden events involved in various disorders.

Establishment of cut-off value of serum pro-gastrin-releasing peptide for diagnosis of small cell lung cancer and evaluation on the clinical diagnosis efficiency

Objective: To determine critical reference value (cut-off value) of serum pro-gastrin-releasing peptide (ProGRP) and neuron specific enolase(NSE) in the diagnosis of small cell lung cancer(SCLC). To evaluate the clinical significance of serum levels of ProGRP and NSE in diagnosis and differential diagnosis in SCLC. Methods: Three hundred and fifty-two SCLC patients, 163 non small cell lung cancer(NSCLC)patients , 193 benign pulmonary disease patients and 140 healthy people visiting in National Cancer Hospital were analyzed retrospectively from January 2014 to July 2017.The levels of serum ProGRP and NSE of people were determined using electrochemiluminescent immunoassay respectively . Reference value ranges of the makers were determined by using the method of ROC curves. Results: In NSCLC group, benign lung disease group, healthy control group and mixed group (NSCLC+ lung benign diseases+ healthy control group) as a reference, the cut-off values were 58.3, 62.3, 57.8, 61.3 ng/L. In the diagnosis and differential diagnosis of SCLC and NSCLC, benign lung diseases, healthy controls and mixed group, AUC of ProGRP was 0.940 (0.919-0.961), 0.941 (0.921-0.960), 0.959 (0.944-0.975), 0.946 (0.928-0.963) respectively. The sensitivities of ProGRP were 86.4%, 84.9%, 86.4% and 84.7% respectively. The specificities of ProGRP were 95.7%, 96.9%, 99.3%, 98% respectively. In all groups the Youden's index of ProGRP and NSE were 0.821 vs 0.612, 0.818 vs 0.674, 0.857 vs 0.810, 0.827 vs 0.674. In healthy controls, no statistically significant difference was found between ProGRP and NSE (P>0.05) in the diagnosis of AUC. However, in the remaining 3 groups, the ProGRP diagnosis of AUC was significantly greater than that of NSE (P<0.01). Compared with single marker detection, the sensitivity of combined detection of ProGRP and NSE in diagnosis of SCLC increased to 95.5%, 94%, 96.6% and 94% in each group. There was no significant difference between ProGRP and ProGRP+ NSE in the diagnosis of AUC when compared with the NSCLC group and the mixed group (P>0.05). However, when combined with a healthy control group and a benign lung disease group, the ProGRP+ NSE combination was the highest for AUC diagnosis, compared with ProGRP and NSE (P<0.01). In the SCLC ED group serum ProGRP and NSE levels[776.33(3 103.4)ng/L, 52.14(60.59)μg/L]were higher than those in the SCLC LD group[295.59(799.65)ng/L, 23.36(22.97)μg/L], respectively (all P<0.001). The serum ProGRP levels of N0, N1, N2 and N3 in TNM staging were 113.0(343.65), 167.04(724.56), 427.42(1 388.62), 735.99(1 709.95)ng/L respectively (all P<0.001). Serum ProGRP and NSE levels were not statistically different between the sex groups and the age groups (all P>0.05). Conclusion: To establish the cut-off value of serum ProGRP is helpful for the diagnosis and differential diagnosis of SCLC.

Human epididymis protein 4 may not be a reliable screening biomarker for detecting lung carcinoma patients.

Human epididymis protein 4 (HE4) acts as a protease inhibitor. It has been detected in the serum of patients with lung cancer patients and its utility for lung cancer screening was found in different studies. Nevertheless, little is known regarding the expression of HE4 in lung carcinoma subtypes. The aim of the present study was to investigate whether HE4 expression is a reliable marker for detecting any lung carcinoma subtypes, including small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC) and adenocarcinoma (AC). In total, 141 lung carcinoma patients were enrolled in the study. Biopsy samples were obtained from bronchoscopic biopsy. The tumors were classified as SCLC (group 1, 54 cases) or NSCLC (group 2, 87 cases) based on histology and immunohistochemistry. The latter was sub-grouped as adenocarcinoma (group 2a, AC) and squamous cell carcinoma (group 2b, SCC). The immunohistochemical expression of HE4 was compared between the groups. The study revealed that the majority of the SCLC and SCC cases were devoid of HE4 (90.1 and 89.65%, respectively). Approximately 10% of cases had HE4 immune expression and the staining was focal and moderate throughout the tumor tissue. On the other hand, 78.8% of AC expressed HE4 and the staining was diffuse and strong. The overall HE4 expression in the lung cancer patients was 33.7%. In conclusion, the results of the present study have shown that HE4 is expressed in adenocarcinoma of the lung but it is not frequent in SCC and SCLC. The value of HE4 as a screening biomarker for lung cancer is limited but its presence exclusively in adenocarcinoma may provide new insight for targeted therapy.

RASSF1A and SIRT6 in non-small cell lung cancer: Relationship with clinical outcome.

This study investigated the expression of RASSF1A and SIRT6 in non-small cell lung cancer (NSCLC) and its relationship with clinical prognosis. The expression in 122 cases of NSCLC tissues (NSCLC group) and 122 cases of normal lung tissues (NOR group) during the same period were detected by immunohistochemical Super Pic Ture™ Polymer two-step method, and the relationship between its expression and the clinicopathological features and prognosis of patients was analyzed. The positive expression rates of RASSF1A and SIRT6 in NSCLC group were lower than those in the normal group (55.74 vs. 84.43% and 52.46 vs. 82.49%, P<0.01). The differences in expression intensity of RASSF1A in NSCLC tissues between different tumor pathological types, tumor differentiation degrees and lymph node metastases were statistically significant, and the differences in expression intensity of SIRT6 between different TNM stages, tumor differentiation degrees and lymph node metastases were statistically significant. There was a positive correlation between the expression of RASSF1A and SIRT6 in NSCLC group (r=0.532, P<0.01). The 3-year survival rate of patients with high-expression of RASSF1A was higher than in those with low-expression of RASSF1A (81.33 vs. 39.45%, log-rank χ2=19,102, P<0.01); the 3-year survival rate of patients with high-expression of SIRT6 was higher than in those with low-expression of SIRT6 (83.51 vs. 42.43%, log-rank χ2=17,180, P<0.01). The low expression of RASSF1A and SIRT6 and lymph node metastasis were the risk factors affecting the prognosis of NSCLC patients. There is a better correlation between the expression of RASSF1A and SIRT6 in NSCLC tissues, and the detection of their expression is of great significance in the judgement of clinicopathological features and prognosis of NSCLC patients.

Inter-heterogeneity and intra-heterogeneity of αvβ3 in non-small cell lung cancer and small cell lung cancer patients as revealed by 68Ga-RGD2 PET imaging

Integrin αvβ3 is the therapeutic target of the anti-angiogenic drug cilengitide. The objective of this study was to compare αvβ3 levels in non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) patients, by using the positron emission tomography (PET) tracer 68Ga-labeled dimerized-RGD (68Ga-RGD2). Thirty-one patients with pathologically confirmed lung cancer were enrolled (21 were NSCLC and 10 were SCLC). PET/CT images were acquired using 68Ga-RGD2.18F-FDG PET/CT images were also acquired on the consecutive day as reference. The standard uptake values (SUV) and the tumor/nontarget (T/NT) values were quantitatively compared. Expression of the angiogenesis marker αvβ3 in NSCLC and SCLC lesions was analyzed by immunohistochemistry. The 18F-FDG SUVmax and the SUVmean were not significantly different between NSCLC and SCLC patients. The 68Ga-RGD2 uptake of SCLC patients was at background levels in both SUV and T/NT measurements and was significantly lower than that of NSCLC patients. The range value of 68Ga-RGD2 SUVmean was 4.5 in the NSCLC group and 2.2 in the SCLC group, while the variation coefficient was 36.2% and 39.3% in NSCLC and SCLC primary lesions, respectively. Heterogeneity between primary lesions and putative distant metastases was also observed in some NSCLC cases. Immunostaining showed that αvβ3 integrin was expressed in the cells and neovasculature of NSCLC lesions, while SCLC samples had negative expression. The uptake of 68Ga-RGD2 in SCLC patients is significantly lower than that in NSCLC patients, indicating a lower αvβ3 target level for cilengitide in SCLC. Apparent intra-tumor heterogeneities of αvβ3 also exist in both NSCLC and SCLC. Such inter- and intra-heterogeneity of αvβ3 may potentially improve current applications of αvβ3-targeted therapy and diagnostic imaging in lung cancer.

Study of early stage non-small-cell lung cancer using Orbitrap-based global serum metabolomics

PurposeThe aim of the project was to apply ultra-high-performance liquid chromatography–quadrupole-Orbitrap-high-resolution mass spectrometry for serum metabolite profiling of non-small-cell lung cancer (NSCLC). This Orbitrap-based methodology has been applied for a study of NSCLC potential markers for the first time.MethodsAfter extraction using protein precipitation, sera were separated on the ACE Excel 2 C18-PFP (100 × 2.1 mm, 2.0 µm) column using gradient elution and analyzed within the range of 70–1000 m/z. Only patients with early stage disease (stages IA–IIB) were included in the study, providing opportunity to find biomarkers for early lung cancer detection. The resulting metabolite profiles were subjected to univariate and multivariate statistical tests.Results36 features were found significantly changed between NSCLC group and controls after FDR adjustment and 19 were identified using various metabolite databases (in-house library, HMDB, mzCloud). The study revealed a number of NSCLC biomarker candidates which belong to such compound classes as acylcarnitines, organic acids, and amino acids. Multivariate ROC curve built using 12 identified metabolites was characterized by AUC = 0.836 (0.722–0.946). There were no significant differences in the serum metabolite profiles between two most common histological types of lung cancer—adenocarcinoma and squamous cell carcinoma.ConclusionsThrough identification of novel potential tumor markers, Orbitrap-based global metabolic profiling is a useful strategy in cancer research. Our study can accelerate development of new diagnostic and therapeutic strategies in NSCLC. The metabolites involved in discrimination between NSCLC patients and the control subjects should be further explored using a targeted approach.

Prognostic Value of Osteoprotegerin and sRANKL in Bronchoalveolar Lavage Fluid of Patients with Advanced Non-small Cell Lung Cancer.

Osteoprotegerin (OPG) is a soluble decoy receptor for receptor activator of nuclear factor-kappa B ligand (sRANKL). OPG promotes endothelial cell survival and neoangiogenesis. Dysregulation of the OPG/RANKL system has been detected in several tumors. In the present study, we evaluated the clinical usefulness of OPG and sRANKL assessment in bronchoalveolar lavage fluid (BALF) of patients with advanced non-small cell lung cancer (NSCLC). We measured the concentration of OPG and sRANKL in BALF of 44 NSCLC patients and 15 healthy volunteers taken as control subjects. The OPG content was higher in the NSCLC group than that in controls [0.48 (0.12-1.45) vs. 0.23 (0.14-0.75) pmol/l; p=0.0001]. There were no significant differences in sRANKL content between the NSCLC and control groups [1.22 (0.74-23.00) vs. 1.12 (0.79-4.39) pmol/l; p=0.67]. However, we found that the greater the level of sRANKL in NSCLC patients, the shorter the overall survival. We found a correlation between the content of sRANKL and the percentage of lymphocytes in BALF of NSCLC patients (r=0.52; p=0.041). We conclude that NSCLC patients have a higher content of OPG in BALF than healthy people. A high level of sRANKL in BALF of NSCLC patients may predict worse survival.

P1.06-025 Analysis of Risk Factors for Development of Skeletal-Related Events in Women with Bone Metastases from NSCLC and Breast Cancer

Bone metastases (BM) are common (up to 50% of cases) in patients with advanced non-small cell lung cancer (NSCLC) and other malignancies, including prostate cancer and breast cancer (BC). In patients with BMs, the onset of skeletal-related events (SREs), such as pathological fracture, malignant hypercalcemia, or spinal cord compression requiring surgery or radiation therapy, seriously affects the quality of life of patients and overall survival. The purpose of this study was to analyze the risk factors (RFs) for development of SREs in women with advanced NSCLC and BC, with the aim of highlighting the differences (if any) between the two groups of patients. The medical records of 16 women with BMs from NSCLC (Group A) and 15 women with BMs from luminal-type BC (Group B) were reviewed. The following RFs have been considered: age >65 years, ECOG performance status (PS) <2, the presence of extra-skeletal metastases (ESM) or hypercalcemia (>2.65 mmol/L), and number of BMs >1. Odds ratio (OR) estimates and the relative 95% confidence interval (CI) were calculated. A p-value level <0.05 was considered statistically significant. During follow-up, 5 (33.3%) Group A and 111 (68.7%) Group B patients developed SREs (OR=4.40, p=0.04), respectively. The results are reported in the Table. No significant difference (p=NS) was found between groups in relation to ECOG-PS, ESM or hypercalcemia, and number of BMs. Only the age >65 years (OR=0.22, p=0.04) represented a weak significant risk factor.Tabled 1ParameterNSCLCBCOR95% CIp-valueNo. of patients1516---Skeletal-related events33.3%68.7%4.400.97-19.850.04Age >65 years73.3%37.5%0.220.05-1.010.04ECOG-Performance status >240.0%18.8%0.340.07-1.760.25Extra-skeletal metastases26.7%43.7%2.140.47-9.700.32Malignant hypercalcemia26.7%12.5%0.390.06-2.550.39Multiple bone metastases53.5%37.5%0.520.12-2.200.38 Open table in a new tab Women with BMs from NSCLC has a reduced risk for development of SREs compared to those with BC, but elderly (>65 years) patients require a closer surveillance, and a precocious bisphosphonate treatment could be suggested.