Overall Survival In Non-small Cell Lung Cancer Research Articles

Prognostic roles of the expression of sphingosine-1-phosphate metabolism enzymes in non-small cell lung cancer.

Sphingosine-1-phosphate (S1P), a bioactive lipid, is generally increased in human non-small cell lung cancer (NSCLC). Evidence has shown that the levels of enzymes in S1P metabolism were associated with clinical outcomes in patients with NSCLC. Nevertheless, the roles of mRNA expression of major enzymes (SPHK1, SPHK2 and SGPL1) in S1P metabolism for predicting outcomes in NSCLC patients have not been determined. "The Kaplan-Meier plotter" (the KM plotter) is an online database which contains gene expression and clinical data of 1,928 NSCLC patients. In this study, we analyzed the relationship between mRNA expression of major enzymes in S1P metabolism and overall survival (OS) in 1,926 NSCLC patients with the KM plotter. Further analyses stratified by smoking history, non-metastasis patents, clinical stages, negative surgical margin, chemotherapy and radiotherapy were also performed. High SPHK1 mRNA expression [hazard ratio (HR) 1.47, 95% confident interval (CI): 1.28-1.68, P=2.6e-08] was significantly correlated to worse OS, but high SPHK2 (0.66, 95% CI: 0.59-0.75, P=1.9e-10) or SGPL1 (HR 0.64, 95% CI: 0.55-0.75, P=8.7e-09) mRNA expression was in favor of better OS in NSCLC patients. The mRNA expression of SPHK1, SPHK2, and SGPL1 is potential predictor of outcomes in NSCLC patients.

MiR-650 promotes non-small cell lung cancer cell proliferation and invasion by targeting ING4 through Wnt-1/β-catenin pathway.

Non-small cell lung cancer (NSCLC) is the most frequent cancer worldwide with a poor 5-year survival. miR-650 acts as an oncogene and regulates tumor progress in various cancers. Molecular mechanisms of miR-650 in NSCLC cell proliferation and invasion was studied. The mRNA levels of miR-650 and special genes were calculated using RT-qPCR. MTT and transwell assays were applied to measure the proliferative and invasive ability. Kaplan-Meier method was used to assess the survival of NSCLC patients. miR-650 was upregulated in NSCLC and upregulation of miR-650 was associated with a poor overall survival of NSCLC, while the results of ING4 demonstrated the opposite results. miR-650 promoted proliferation and invasion through Wnt-1/β-catenin pathway by targeting inhibitor of growth 4 (ING4) in A549 cells. ING4 was a direct target gene of miR-650 and the expression of ING4 was mediated by exogenous altering the expression of miR-650. Remarkably, alterations of ING4 expression eliminated the functions of miR-650 on the proliferation and metastasis of NSCLC. miR-650 enhanced A549 cell proliferation and invasion through Wnt-1/β-catenin pathway by directly targeting the 3′-UTR of ING4 mRNA. The newly identified miR-650/ING4 axis provides a novel insight into the pathogenesis of NSCLC.

Integrin-α7 expression positively correlates with CD44 and CD133 expressions, and its high expression associates with advanced tumor features as well as poor survivals in non-small cell lung cancer patients.

BackgroundThis study aimed to explore the correlation of integrin-α7 (ITGA7) with common cancer stem cell marker (CD44 and CD133) expressions, clinicopathological characteristics and survival profiles in non-small cell lung cancer (NSCLC) patients.MethodsTwo hundred and seventy NSCLC patients who underwent resection were reviewed in this study and the expressions of ITGA7, CD44 and CD133 were detected using immunohistochemistry (IHC) assay in tumor tissue specimens. The clinical data (including age, gender, pathological grade, tumor size, lymph node metastasis (LYN), and TNM stage), survival data [including disease-free survival (DFS) and overall survival (OS)] were collected.ResultsThe numbers of NSCLC patients with ITGA7 high expression, CD44 high expression and CD133 high expression were 170 (63.0%), 241 (89.3%) and 37 (13.7%) respectively, and ITGA7 expression was positively associated with CD44 expression and CD133 expression. ITGA7 high expression was associated with higher pathological grade, larger tumor size, LYN and elevated TNM stage. However, there was no correlation of ITGA7 expression with age or gender. As for survival, patients with ITGA7 high expression presented reduced DFS and OS compared with those with ITGA7 low expression. In addition, multivariate Cox’s proportional hazards regression model analyses exhibited that ITGA7 high expression, higher pathological grade and LYN were independent risk factors for DFS and OS in NSCLC patients.ConclusionsITGA7 expression positively correlates with CD44 and CD133 expressions, and its high expression associates with advanced tumor features as well as poor survivals in NSCLC.

Tumor Transcriptome Reveals High Expression of IL-8 in Non-Small Cell Lung Cancer Patients with Low Pectoralis Muscle Area and Reduced Survival.

Cachexia is a syndrome characterized by an ongoing loss of skeletal muscle mass associated with poor patient prognosis in non-small cell lung cancer (NSCLC). However, prognostic cachexia biomarkers in NSCLC are unknown. Here, we analyzed computed tomography (CT) images and tumor transcriptome data to identify potentially secreted cachexia biomarkers (PSCB) in NSCLC patients with low-muscularity. We integrated radiomics features (pectoralis muscle, sternum, and tenth thoracic (T10) vertebra) from CT of 89 NSCLC patients, which allowed us to identify an index for screening muscularity. Next, a tumor transcriptomic-based secretome analysis from these patients (discovery set) was evaluated to identify potential cachexia biomarkers in patients with low-muscularity. The prognostic value of these biomarkers for predicting recurrence and survival outcome was confirmed using expression data from eight lung cancer datasets (validation set). Finally, C2C12 myoblasts differentiated into myotubes were used to evaluate the ability of the selected biomarker, interleukin (IL)-8, in inducing muscle cell atrophy. We identified 75 over-expressed transcripts in patients with low-muscularity, which included IL-6, CSF3, and IL-8. Also, we identified NCAM1, CNTN1, SCG2, CADM1, IL-8, NPTX1, and APOD as PSCB in the tumor secretome. These PSCB were capable of distinguishing worse and better prognosis (recurrence and survival) in NSCLC patients. IL-8 was confirmed as a predictor of worse prognosis in all validation sets. In vitro assays revealed that IL-8 promoted C2C12 myotube atrophy. Tumors from low-muscularity patients presented a set of upregulated genes encoding for secreted proteins, including pro-inflammatory cytokines that predict worse overall survival in NSCLC. Among these upregulated genes, IL-8 expression in NSCLC tissues was associated with worse prognosis, and the recombinant IL-8 was capable of triggering atrophy in C2C12 myotubes.

Survival-based bioinformatics analysis to identify hub genes and key pathways in non-small cell lung cancer

BackgroundLung cancer is one of the leading causes of cancer mortality worldwide. Here, we performed an integrative bioinformatics analysis to screen hub genes and critical pathways in non-small cell lung cancer (NSCLC) based on the overall survival rate of differentially expressed genes (DEGs).MethodsFour datasets from the gene expression omnibus (GEO) were used to identify the DEGs. To obtain robust DEGs in NSCLC, only the DEGs that co-existed in the four datasets were selected for subsequent analysis. To identify the genes correlated with overall survival, the overall survival of these genes was then analyzed using the Kaplan-Meier plotter database. The genes significantly correlated with survival were used to perform gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis; next, these genes were used to construct a protein-protein interaction network. MCODE and CytoHubba were used to identify the clusters and hub genes. Finally, the hub genes were validated in the Cancer Genome Atlas (TCGA) and the Human Protein Atlas (HPA).ResultsWe found 522 up-regulated DEGs, and 989 down-regulated DEGs between the NSCLC and normal lung tissue, and 895 of them were correlated with a higher overall survival. GO analysis showed that the DEGs that were associated with a higher overall survival were enriched in cell division, cell cycle, DNA replication, angiogenesis, and cell migration. KEGG analysis was consistent with GO analysis and showed that p53 signaling pathway, pyrimidine metabolism, cGMP-PKG signaling pathway and renin secretion pathway were associated with overall survival in NSCLC. In the protein-protein analysis, we identified seven clusters and six hub genes which were BUB1B, CCNB1, CENPE, KIF18A, NDC10, and MAD2L1. Of these genes, CENPE and KIF18A had not been reported until now. Finally, the dysregulated expression of the six hub genes was validated by the data from the TCGA and HPA.ConclusionsWe identified the hub genes and potential mechanisms of NSCLC based on multiple-microarray analysis and overall survival; then, validated the hub genes in the TCGA and HPA database. These hub genes may serve as potential therapeutic targets.

Circular RNA hsa_circ_0023404 promotes proliferation, migration and invasion in non-small cell lung cancer by regulating miR-217/ZEB1 axis.

BackgroundCircular RNAs (circRNAs) have been considered as key regulators of cancer biology. However, the functional role of hsa_circ_0023404 in non-small cell lung cancer (NSCLC) and its regulatory mechanism are still almost unknown.MethodsThe expression of hsa_circ_0023404, miR-217 and zinc finger E-box-binding homeobox 1 (ZEB1) was evaluated by quantitative real-time polymerase chain reaction. The role of hsa_circ_0023404 in NSCLC progression was determined using cell count kit-8 assay, transwell migration and invasion assay. Luciferase reporter assay was performed to assess the interaction of hsa_circ_0023404, miR-217 and ZEB1 in NSCLC cells.ResultsThe expression of hsa_circ_0023404 was upregulated in NSCLC tissues, as well as in NSCLC cell lines. High hsa_circ_0023404 expression predicted short overall survival in NSCLC. Functionally, knockdown of hsa_circ_0023404 inhibited the proliferation, migration and invasion of NSCLC cells. In the further molecular mechanism study, hsa_circ_0023404 was shown to interact with miR-217/ZEB1 axis to contribute to the growth of NSCLC cells.Conclusionhsa_circ_0023404 promotes the proliferation, migration and invasion of NSCLC cells by regulating miR-217/ZEB1 axis, providing a fresh perspective on circRNAs in NSCLC development.

Impact of antibiotics and proton pump inhibitors on clinical outcomes of immune check point blockers in advanced non-small cell lung cancers and metastatic renal cell cancer.

e20520 Background: Studies have suggested that antibiotics can negatively influence efficacy of immune check-point blockers (ICB) but the results are inconsistent. The impact of concomitant proton pump inhibitors (PPI) is not known. We evaluated whether antibiotic and PPI use in patients with advanced non-small-cell lung cancer (NSCLC) and metastatic renal cell cancer (RCC) affects ICB efficacy. Methods: We identified advanced NSCLC and RCC patients treated with anti-PD1/PD-L1 at our institution between 5/2015 to 1/2019. Data regarding systemic antibiotics and PPI use < 1 month or during ICB treatment were collected. The primary outcome was progression free survival (PFS) per response evaluation criteria in solid tumors (RECIST 1.1). Secondary outcomes were overall survival (OS) and objective response rate (ORR). Logistic regression and cox proportional hazards model were used for statistical analysis. Results: 87/148 (58%) and 40/55 (72%) patients received antibiotics and 57/148 (39%) and 17/55 (31%) received PPI < 1 month or during ICI therapy in NSCLC and RCC respectively. In RCC, antibiotic use was associated with inferior PFS (2.9 v 5.0 months, HR = 2.3 95%CI 1.0-5.0; p = 0.04) but OS and ORR were not affected. In NSCLC, antibiotic exposure was associated with superior PFS (5.0 v 2.5 months, HR = 0.5, 95% CI 0.3-0.7; p < 0.01), OS (13.0 v 8.0 months, HR = 0.5, 95% CI 0.3-0.8; p < 0.01) and ORR (33% v 11%, OR = 4.6, p < 0.01). On univariate analysis, there was trend towards inferior OS in NSCLC (9.0 v 13.0 months, p = 0.05) with PPI use. PPI use was not associated with other primary or secondary outcomes in both the cohorts. The antibiotic effect remained significant for PFS in NSCLC and RCC in multivariate analysis. Conclusions: Antibiotic use significantly affected PFS in both NSCLC and RCC. PPI use did not affect outcomes. In contrast to previous studies, this is the first study to show that antibiotic use was associated with favorable outcomes in NSCLC. More studies are needed to explain this association. Future clinical trials with ICB should consider stratification of patients based on antibiotic exposure.

Genetic variants in RUNX3, AMD1 and MSRA in the methionine metabolic pathway and survival in nonsmall cell lung cancer patients.

Abnormal methionine dependence in cancer cells has led to methionine restriction as a potential therapeutic strategy. We hypothesized that genetic variants involved in methionine-metabolic genes are associated with survival in nonsmall cell lung cancer (NSCLC) patients. Therefore, we investigated associations of 16,378 common single-nucleotide polymorphisms (SNPs) in 97 methionine-metabolic pathway genes with overall survival (OS) in NSCLC patients using genotyping data from two published genome-wide association study (GWAS) datasets. In the single-locus analysis, 1,005 SNPs were significantly associated with NSCLC OS (p < 0.05 and false-positive report probability < 0.2) in the discovery dataset. Three SNPs (RUNX3 rs7553295 G > T, AMD1 rs1279590 G > A and MSRA rs73534533 C > A) were replicated in the validation dataset, and their meta-analysis showed an adjusted hazards ratio [HR] of 0.82 [95% confidence interval (CI) =0.75-0.89] and pmeta = 2.86 × 10-6 , 0.81 (0.73-0.91) and pmeta = 4.63 × 10-4 , and 0.77 (0.68-0.89) and pmeta = 2.07 × 10-4 , respectively). A genetic score of protective genotypes of these three SNPs revealed an increased OS in a dose-response manner (ptrend < 0.0001). Further expression quantitative trait loci (eQTL) analysis showed significant associations between these genotypes and mRNA expression levels. Moreover, differential expression analysis further supported a tumor-suppressive effect of MSRA, with lower mRNA levels in both lung squamous carcinoma and adenocarcinoma (p < 0.0001 and < 0.0001, respectively) than in adjacent normal tissues. Additionally, low mutation rates of these three genes indicated the critical roles of these functional SNPs in cancer progression. Taken together, these genetic variants of methionine-metabolic pathway genes may be promising predictors of survival in NSCLC patients.

EGFR plasma mutation in prediction models for resistance with EGFR TKI and survival of non-small cell lung cancer

BackgroundThis study aims to clarify the prognostic role of epidermal growth factor receptor (EGFR) mutations in plasma of non-small cell lung cancer (NSCLC) for resistance to tyrosine kinase inhibitor (TKI), in correlation with clinical characteristics. A total of 94 Adenocarcinoma, clinical stage IV NSCLC patients with either E19del or L858R mutation were admitted to the prospective study from Jan-2016 to Jul-2018. EGFR mutations in plasma were detected by scorpions ARMS method. The Kaplan–Meier and Cox regression methods were used to estimate and test the difference of progression-free survival (PFS) and overall survival (OS) between groups. The prognostic power of each factor was appraised by the Bayesian Model Averaging (BMA) method.ResultsAmong 94 patients, 28 cases still are good responses according to the RECIST criteria and negative for EGFR mutations in plasma. Of 66 resistant patients, EGFR mutations were positive in plasma of 57 cases (86.4%) which was higher than the value of pre-treatment (48.5%). Of which, 17 patients (25.8%) have the occurrence of EGFR mutations in plasma earlier than progression 2.1 (0.6–7.9) months. The secondary T790M mutation was found in the plasma of 32 cases (48.5%). Median PFS and OS for the study subjects were 12.9 (11.0–14.2) and 29.5 (25.2–41.3) months, respectively. The post-treatment EGFR plasma test with brain and new metastasis were detected as independent prognostic factors for worse PFS (P = 0.008, 0.016 and 0.028, respectively). While EGFR plasma (P = 0.044) with bone metastasis at baseline (P = 0.012), new metastasis (P = 0.003), and high cfDNA concentration (P = 0.004) serve as the worse survival factors, surgery treatment helps to prolong OS in NSCLC treated with EGFR TKI (P = 0.012). BMA analysis identified EGFR plasma test as the strongest prognostic factor for both PFS and OS (possibility of 100% and 99.7%, respectively).ConclusionsEGFR plasma test is the powerfully prognostic factor for early resistance with EGFR TKI and worse survival in NSCLC regardless of clinical characteristics.

MiR-3607-3p suppresses non-small cell lung cancer (NSCLC) by targeting TGFBR1 and CCNE2.

Accumulating evidence indicates that miRNAs can be promising diagnostic and/or prognostic markers for various cancers. In this study, we identified a novel miRNA, miR-3607-3p, and its targets in non-small cell lung cancer (NSCLC). The expression of miR-3607-3p was measured and its correlation with patient prognosis was determined. Ectopic expression in NSCLC cells, xenografts, and metastasis models was used to evaluate the effects of miR-3607-3p on proliferation and migration of NSCLC. Luciferase assay and western blotting were performed to validate the potential targets of miR-3607-3p after preliminary screening by microarray analysis and computer-aided algorithms. We demonstrated that miR-3607-3p was downregulated in NSCLC tissues and that miR-3607-3p might act as an independent predictor for overall survival in NSCLC. Moreover, serum miR-3607-3p may be a novel and stable marker for NSCLC. We found that overexpression of miR-3607-3p inhibited cell proliferation, colony formation, migration and invasion, and hampered the cell cycle of NSCLC cell lines in vitro. Our results suggested that miR-3607-3p directly targets TGFBR1 and CCNE2. In accordance with in vitro studies, we confirmed that miR-3607-3p functions as a potent suppressor miRNA of NSCLC. We showed that miR-3607-3p agomir could reduce tumor growth and inhibit TGFBR1 and CCNE2 protein expression. Taken together, our findings indicate that miR-3607-3p can inhibit NSCLC cell growth and metastasis by targeting TGFBR1 and CCNE2 protein expression, and provide new evidence of miR-3607-3p as a potential non-invasive biomarker and therapeutic target for NSCLC.

Identification of Candidate Biomarkers Correlated With the Pathogenesis and Prognosis of Non-small Cell Lung Cancer via Integrated Bioinformatics Analysis.

Background and Objective: Non-small cell lung cancer (NSCLC) accounts for 80–85% of all patients with lung cancer and 5-year relative overall survival (OS) rate is less than 20%, so that identifying novel diagnostic and prognostic biomarkers is urgently demanded. The present study attempted to identify potential key genes associated with the pathogenesis and prognosis of NSCLC.Methods: Four GEO datasets (GSE18842, GSE19804, GSE43458, and GSE62113) were obtained from the Gene Expression Omnibus (GEO) database. The differentially expressed genes (DEGs) between NSCLC samples and normal ones were analyzed using limma package, and RobustRankAggreg (RRA) package was used to conduct gene integration. Moreover, Search Tool for the Retrieval of Interacting Genes database (STRING), Cytoscape, and Molecular Complex Detection (MCODE) were utilized to establish protein–protein interaction (PPI) network of these DEGs. Furthermore, functional enrichment and pathway enrichment analyses for DEGs were performed by Funrich and OmicShare. While the expressions and prognostic values of top genes were carried out through Gene Expression Profiling Interactive Analysis (GEPIA) and Kaplan Meier-plotter (KM) online dataset.Results: A total of 249 DEGs (113 upregulated and 136 downregulated) were identified after gene integration. Moreover, the PPI network was established with 166 nodes and 1784 protein pairs. Topoisomerase II alpha (TOP2A), a top gene and hub node with higher node degrees in module 1, was significantly enriched in mitotic cell cycle pathway. In addition, Interleukin-6 (IL-6) was enriched in amb2 integrin signaling pathway. The mitotic cell cycle was the most significant pathway in module 1 with the highest P-value. Besides, five hub genes with high degree of connectivity were selected, including TOP2A, CCNB1, CCNA2, UBE2C, and KIF20A, and they were all correlated with worse OS in NSCLC. Conclusion: The results showed that TOP2A, CCNB1, CCNA2, UBE2C, KIF20A, and IL-6 may be potential key genes, while the mitotic cell cycle pathway may be a potential pathway contribute to progression in NSCLC. Further, it could be used as a new biomarker for diagnosis and to direct the synthesis medicine of NSCLC.

P3.15-12 Survival Impact of Patient Enrollment in Antineoplastic Drug Trials for Stage IV Non-Small Cell Lung Cancer (NSCLC)

Patient enrollment in antineoplastic drug trials (ADT) is essential for the development of effective therapies in stage IV NSCLC. An open question is whether NSCLC patients derive a survival benefit by enrolling in ADTs. We hypothesized that patient enrollment in ADT is associated with longer overall survival (OS) because of access to novel therapies and closer follow up, compared with no ADT enrollment. We reviewed electronic medical records of 193 patients diagnosed with biopsy-proven, stage IV NSCLC between 01/01/2007 and 12/31/2014, who received treatments at the Seattle Cancer Care Alliance. Other inclusion criteria were age > 18 and receipt of ≥ 1 antineoplastic agent within 180 days from diagnosis. We abstracted patient clinical characteristics, tumor histology, EGFR and ALK mutation status, receipt of chemotherapy, targeted therapies, and immunotherapy up to 5 consecutive lines, participation in ADTs, and OS from diagnosis to death. We fitted multivariate Cox regression models to estimate the risk-adjusted effect of ADT participation on survival, compared with no trial participation. Patients’ mean age was 63.4; 53.9% were female; 28.5% were never smokers; 75.1% had ECOG PS of 0-1; 95.3% had non-squamous histology; 27.5% had brain metastases; 20.7% and 4.1% had EGFR+ and ALK+ NSCLC. Fifty-three (27.5%) enrolled in ≥ 1 ADT, of whom 17 (32.0%) received trial drug(s) that later became FDA approved, and 44 (83.0%) and 13 (24.5%) enrolled in phases I/II or III trials. Adjusting for ECOG PS, smoking, EGFR mutation status, type of first line therapy, and number of treatment lines, participation in ≥ 1 ADT was associated with a hazard ratio for death of 0.62 (P = 0.02; Table 1). Participation in therapeutic trials is associated with longer OS in NSCLC. Besides supporting drug development, trial enrollment may improve patient outcomes. Efforts should focus on increasing patient enrollment in drug trials.

Genetic variant of IRAK2 in the toll-like receptor signaling pathway and survival of non-small cell lung cancer.

The toll-like receptor (TLR) signaling pathway plays an important role in the innate immune responses and antigen-specific acquired immunity. Aberrant activation of the TLR pathway has a significant impact on carcinogenesis or tumor progression. Therefore, we hypothesize that genetic variants in the TLR signaling pathway genes are associated with overall survival (OS) of patients with non-small cell lung cancer (NSCLC). To test this hypothesis, we first performed Cox proportional hazards regression analysis to evaluate associations between genetic variants of 165 TLR signaling pathway genes and NSCLC OS using the genome-wide association study (GWAS) dataset from the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial (PLCO). The results were further validated by the Harvard Lung Cancer Susceptibility GWAS dataset. Specifically, we identified IRAK2 rs779901 C > T as a predictor of NSCLC OS, with a variant-allele (T) attributed hazards ratio (HR) of 0.78 [95% confidence interval (CI) = 0.67-0.91, P = 0.001] in the PLCO dataset, 0.84 (0.72-0.98, 0.031) in the Harvard dataset, and 0.81 (0.73-0.90, 1.08x10-4 ) in the meta-analysis of these two GWAS datasets. In addition, the T allele was significantly associated with an increased mRNA expression level of IRAK2. Our findings suggest that IRAK2 rs779901 C > T may be a promising prognostic biomarker for NSCLC OS.

Abstract B17: Interleukin-17 and -22 expression in non-small cell lung cancer

Abstract Background: In non-small cell lung cancer (NSCLC) the TNM staging remains standard for prognostic assessment and therapy decisions. Nevertheless, stage-specific outcomes vary significantly, indicating a need for additional prognosticators. In this retrospective study we wanted to assess the role of the mediator interleukin-22 (IL-22), as IL-22 is known to be involved in various lung diseases and to be elevated in lung cancer patients. Moreover, we evaluated interleukin-17 (IL-17), which also depicts a proinflammatory cytokine, but seems to play a dual role in antitumor immunity. We asked the question of their link to prognosis, therapy response and recurrence. Methods: Tissue microarrays (TMAs) were generated from formalin-fixed, paraffin embedded tissue of 138 curatively resected patients with stage IA-IV NSCLC. TMAs included each three cores from the tumor center (CT) and invasive margin (IM), selected from areas with the most dense lymphocytic infiltrates. IL-22 and IL-17 expression of the tissue was analyzed by immunohistochemistry via cytoplasmic staining (double-staining: IL-22, CD3 resp. IL-17, CD3). Results: IL-22 expression status in adeno and SCC does not seem to have a prognostic impact on survival, regardless of the localization. However, a high IL-22 CT/IM ratio in adenocarcinoma is clearly linked to longer overall survival; this cannot be seen in SCC. For IL-17 a distinct tendency is visible for adeno and SCC in the invasive margin as well as in the tumor center—interestingly, this is more clear for SCC. In SCC a high CT/IM ratio for IL-17 seems to have a positive prognostic impact on overall survival. Patient numbers will be enlarged to validate these findings. Conclusion: The cytokines IL-22 and IL-17 seem—depending on histology and location—to have a prognostic impact on overall survival in NSCLC. In previous experiments we showed that multispectral assessment of CD8 and PD-L1 has a clear correlation with clinical outcome. Adding cytokines to this method might open novel avenues for predicting clinical outcome and therapeutic efficacy in NSCLC. Citation Format: Julia Stump, Yue Jiang, Clara Karches, Javier Suárez Gosálvez, Jens Neumann, Sebastian Kobold, Rudolf Hatz, Hauke Winter, Amanda Tufman, Rudolf M. Huber, Simone Reu. Interleukin-17 and -22 expression in non-small cell lung cancer [abstract]. In: Proceedings of the Fifth AACR-IASLC International Joint Conference: Lung Cancer Translational Science from the Bench to the Clinic; Jan 8-11, 2018; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(17_Suppl):Abstract nr B17.

Second malignancy in non-small cell lung cancer (NSCLC): prevalence and overall survival (OS) in routine clinical practice

Patients with non-small cell lung cancer (NSCLC) and a prior or synchronous second malignancy are generally excluded from clinical trials. Therefore, little is known on prevalence and prognosis of these patients. 1252 patients diagnosed with NSCLC in our center from 2006 to 2017 were studied. Overall survival (OS) of patients with a prior or synchronous malignancy was compared to controls including case-control analysis. 158 patients (12.6%) had a prior malignancy. The most common sites were prostate (17%), breast (16%), gastrointestinal tract (12%), head and neck (11%), bladder (10%), and lung (8%). Compared to controls, patients with prior malignancy were older (71.3 vs. 67.5years), but had otherwise better prognostic characteristics (stage I-III 63 vs. 53%). Survival was identical compared to controls [hazard ratio (HR) 1.017, CI 0.776-1.333]. A further 3.5% of patients had a synchronous malignancy including 34% prior lung cancer. Patients with a synchronous malignancy had an earlier stage (I-III 84%), and had longer median OS in unselected patients (38.6 vs. 16.2months, p = 0.021). However, the case-control analysis showed similar OS [hazard ratio (HR) 0.899, CI 0.497-1.621]. Prior or synchronous second malignancies are common at diagnosis of NSCLC. The sites reflect the high proportion of smokers in the population. The earlier stage of NSCLC with a second malignancy might be attributed to chance finding of NSCLC during follow-up. The second malignancy does not affect OS of NSCLC. Therefore, the exclusion of patients with second malignancies from NSCLC trials should be reconsidered.

Abstract 2657: Immune checkpoint status does not impact overall survival in NSCLC

Abstract Checkpoint blockade has altered standard of care for non-small cell lung cancer (NSCLC) expressing PD-L1, with trials demonstrating favorable prognosis in patients whose tumors were PD-L1 positive and were subsequently paired with anti-PD-1 therapy. However, the published literature remains controversial on whether PD-L1 expression confers prognostic significance independent of immunotherapy in NSCLC. Recent meta-analyses suggest that PD-L1 expression is associated with worse outcomes in NSCLC although other such studies associate PD-L1 expression with favorable prognosis. Moreover, few studies in NSCLC have simultaneously assessed PD-1 expression on tumor-infiltrating leukocytes (TILs), an important contributor to tumor checkpoint status. To assess the prognostic significance of PD-L1/PD-1 status in NSCLC we assembled 280 NSCLC FFPE tissues from lung cancer cases with detailed clinical and questionnaire data available who were recruited from the Metropolitan Detroit region. Tissues were compiled into duplicate tissue-microarrays and were assayed for PD-L1 (223C), PD-1 (NBP1-88104), and whole-transcriptome mRNA expression. PD-L1 staining was assessed by an experienced pathologist and was reported as the percent of tumor cells with membrane positive 223C staining to mirror clinical reporting of PD-L1 status. PD-1 staining was also assessed by the pathologist and was reported as the percent of TILs with membrane positive NBP1-88104 staining. RNA was extracted from FFPE samples and assayed on the Affymetrix Whole-Transcriptome 2.1 array. Array data was batch normalized and Log2 transformed prior to analysis. Immune marker effects on overall survival (OS) were measured with a Cox proportional hazards model, adjusting for tumor stage and histology. 36% of NSCLC tumor samples were PD-L1 positive, defined as ≥ 1% of tumor cells demonstrating positive 223C membrane staining. PD-L1 status was not associated with tumor stage or histology but was strongly correlated with PD-L1 (CD274) mRNA expression (OR=2.37, p=1.1x10-7), adjusting for stage and histology. PD-L1 positive NSCLC were found to have similar OS when compared to tumors with no detectable PD-L1 tumor staining (HR=0.94, p=0.78). Likewise, tumors containing PD-1 positive TILs did not differ in OS when compared to tumors with no detectable PD-1 positive TILs (HR=0.68, p=0.14). Furthermore, there were no detectable differences in OS when combined PD-L1/PD-1 status was assessed (PD-L1+/PD-1+ HR=0.72 p=0.36, PD-L1+/PD-1- HR=1.39 p=0.49, PD-L1-/PD-1+ HR=0.79 p=0.49, PD-L1-/PD-1- Referent). Our findings suggest that neither PD-L1 tumor or PD-1 TIL status are significant prognostic markers in NSCLC independent of immunotherapy. Citation Format: Donovan Watza, Valerie Murphy, Chrissy Lusk, Angie S. Wenzlaff, Lonardo Fulvio, Christine Neslund-Dudas, Gerold Bepler, Ann G. Schwartz. Immune checkpoint status does not impact overall survival in NSCLC [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 2657.

Prognostic and clinicopathological significance of Beclin-1 in non-small-cell lung cancer: a meta-analysis.

BackgroundAutophagy plays a key role in the development of non-small-cell lung cancer (NSCLC). Beclin-1 is essential for the initiation and regulation of autophagy. Accumulated studies have investigated the prognostic role of Beclin-1 in NSCLC, but conclusions remain controversial. Therefore, we conducted this meta-analysis to assess the potential significance of Beclin-1 in NSCLC.Materials and methodsPubMed and Embase databases were searched for eligible studies published before December 31, 2017. Odds ratio (OR) was pooled to evaluate the clinicopathological significance of Beclin-1 in NSCLC. Hazard ratio (HR) was adopted to assess the association of Beclin-1 with overall survival (OS).ResultsEight studies involving 1,159 patients were included in this meta-analysis. The pooled results showed that high Beclin-1 expression was significantly correlated with earlier tumor grade (OR=0.54, 95% CI: 0.36–0.81, P=0.003), less nodal involvement (OR=0.56, 95% CI: 0.37–0.86, P=0.007), earlier TNM stage (OR=0.62, 95% CI: 0.43–0.89, P=0.010), smaller tumor size (OR=0.54, 95% CI: 0.36–0.81, P=0.003), better differentiation (OR=0.48, 95% CI: 0.36–0.64, P<0.001), and less recurrence (OR=0.24, 95% CI: 0.14–0.41, P<0.001). Moreover, high level of Beclin-1 was significantly associated with better OS in NSCLC (HR=0.41, 95% CI: 0.26–0.64, P<0.001).ConclusionOur meta-analysis suggests that high Beclin-1 expression predicts a better clinicopathological status and a better prognosis in NSCLC. Beclin-1 might act as a promising prognostic biomarker for NSCLC.

Negative association of antibiotics on clinical activity of immune checkpoint inhibitors in patients with advanced renal cell and non-small-cell lung cancer

BackgroundThe composition of gut microbiota affects antitumor immune responses, preclinical and clinical outcome following immune checkpoint inhibitors (ICI) in cancer. Antibiotics (ATB) alter gut microbiota diversity and composition leading to dysbiosis, which may affect effectiveness of ICI. Patients and methodsWe examined patients with advanced renal cell carcinoma (RCC) and non-small-cell lung cancer (NSCLC) treated with anti-programmed cell death ligand-1 mAb monotherapy or combination at two academic institutions. Those receiving ATB within 30 days of beginning ICI were compared with those who did not. Objective response, progression-free survival (PFS) determined by RECIST1.1 and overall survival (OS) were assessed. ResultsSixteen of 121 (13%) RCC patients and 48 of 239 (20%) NSCLC patients received ATB. The most common ATB were β-lactam or quinolones for pneumonia or urinary tract infections. In RCC patients, ATB compared with no ATB was associated with increased risk of primary progressive disease (PD) (75% versus 22%, P < 0.01), shorter PFS [median 1.9 versus 7.4 months, hazard ratio (HR) 3.1, 95% confidence interval (CI) 1.4–6.9, P < 0.01], and shorter OS (median 17.3 versus 30.6 months, HR 3.5, 95% CI 1.1–10.8, P = 0.03). In NSCLC patients, ATB was associated with similar rates of primary PD (52% versus 43%, P = 0.26) but decreased PFS (median 1.9 versus 3.8 months, HR 1.5, 95% CI 1.0–2.2, P = 0.03) and OS (median 7.9 versus 24.6 months, HR 4.4, 95% CI 2.6–7.7, P < 0.01). In multivariate analyses, the impact of ATB remained significant for PFS in RCC and for OS in NSCLC. ConclusionATB were associated with reduced clinical benefit from ICI in RCC and NSCLC. Modulatation of ATB-related dysbiosis and gut microbiota composition may be a strategy to improve clinical outcomes with ICI.

Prognostic Impact of Missed Chemotherapy Doses During Chemoradiation Therapy for Non-Small Cell Lung Cancer.

The aim of this study is to investigate the impact of missed chemotherapy administrations (MCA) on the prognosis of non-small cell lung cancer (NSCLC) patients treated with definitive chemoradiation therapy (CRT). In total, 97 patients with NSCLC treated with definitive CRT were assessed for MCA due to toxicities. Logistic regression was used to determine factors associated with MCA. Kaplan-Meier curves, log-rank tests, and Cox Proportional Hazards models were conducted. MCA occurred in 39% (n=38) of the patients. Median overall survival was 9.6 months for patients with MCA compared with 24.3 months for those receiving all doses (P=0.004). MCA due to decline in performance status was associated with the worst survival (4.6 mo) followed by allergic reaction (10.0 mo), hematologic toxicity (11 mo), and esophagitis (17.2 mo, P=0.027). In multivariate models, MCA was associated with higher mortality (hazard ratio, 1.97; P=0.01) and worse progression-free survival (hazard ratio, 1.96; P=0. 009). MCA correlated with worse prognosis and increased mortality. Methods to reduce toxicity may improve administration of all chemotherapy doses and increase overall survival in NSCLC treated with CRT.

115P Heart and pulmonary artery radiation doses in non-small cell lung cancer

Background: Heart and pulmonary artery (PA) radiation doses following radical (chemo) radiotherapy to non-small cell lung cancer (NSCLC) correlate with overall survival (OS). We aimed to quantify radiation doses delivered to heart and PA during radical (chemo) radiotherapy in NSCLC, and to analyse dose volume histogram (DVH) based parameters that were associated with poor OS. Methods: NSCLC patients who underwent radical (chemo) radiotherapy using 3-D conformal radiotherapy between 2015 and 2017 were identified from our institutional radiotherapy electronic database. Heart and PA were contoured according to the Radiotherapy Oncology Group Trial 1106 contouring atlas. Maximum, minimum and mean radiation doses delivered to heart and PA were quantified. DVH based parameters, volume of heart received 5 Gy (V5), and volume of PA received 40, 45, 50, 55 and 60 Gy (V40, V45, V50, V55 and V60 respectively) were obtained from radiotherapy plans and compared with values that showed correlation with poor OS in NSCLC. Results: Out of a total of nineteen patients, 63% had stage III tumours. Twelve patients were treated with 60 Gy/30 (over 6 weeks) and seven were treated with 55 Gy/20 (over 4 weeks) radiation dose-fractionation schedules. For heart, average radiation doses of mean, minimum and maximum radiation doses were 7.49, 0.33 and 44.18 Gy and corresponding PA radiation doses were 27.14, 5.37 and 52.67 Gy. Evidence suggests mean radiation dose of heart >7.8 Gy and heart V5 of >72.3% correlate with poor OS. In our study, we observed mean heart dose and V5 were less than these values in 74% and 89% patients respectively. PA DVH based parameters, V40 ≥ 80%, V45 > 70%, V50 ≥ 45%, V55 ≥ 32% and V60 > 37 correlates with poor OS. In our cohort of patients, V40, V45, V50, V55 and V60 were less than these values in 95%, 89%, 79%, 74% and 89% patients. Conclusions: In this retrospective study, we found heart and PA DVH based radiation dose parameters in NSCLC radical (chemo) radiotherapy were less than those of published values that were associated with poor OS. It is possible to achieve this in 3-D conformal radiotherapy planning for most patients. Including these DVH based parameters during radiotherapy planning would ensure radiation doses to these critical organs are optimised. Legal entity responsible for the study: N/A Funding: Has not received any funding Disclosure: All authors have declared no conflicts of interest.