Non-smoker Non-small-cell Lung Cancer Research Articles

Immune checkpoint expression as prognostic biomarker candidates in non-small cell lung carcinoma patients.

Cancer immunotherapy has had an important role in oncologic therapeutics for patients with non-small cell lung cancer (NSCLC) using checkpoint inhibitors. We will explore the possible prognosis biomarker candidates such as: soluble OX40 (sOX40), OX40L (sOX40L), Glucocorticoid-induced tumor necrosis factor receptor family-related receptor (GITR), and their ligand (GITRL), 4-1BB or tumor necrosis factor receptor superfamily 9 (TNFRS9) and inducible T cell co-stimulator (ICOS) in peripheral blood of NSCLC patients. Fifty-eight patients were diagnosed with advanced NSCLC between January 2019 and March 2020. High sOX40 and low s4-1BB levels in smokers compared non-smoker NSCLC patients. Lower sOX40L levels were found in the male than female (p < 0.05). High sOX40 and sGITRL in stage III compared to the stage IV (p < 0.05). With follow-up at 21.4 months, 44.1% and 91.1% were alive in the sGITRhigh and sGITRlow groups, respectively (p = 0.02), and 73.3% and 27.7% were alive in the sGITRLhigh and sGITRLlow groups, respectively (p = 0.02). At 22 months, 38.7% and 92.3% were alive in the sOX40Lhigh and sOX40Llow groups, respectively (p = 0.01). sGITR, sGITRL, and sOX40L levels were potential prognostic biomarkers and could have an important role as new targets of immunotherapy in NSCLC patients. sGITR, sGITRL, sOX40L, and sOX40 levels were associated with smoking, sex, stage, and age in NSCLC.

Gene-environment interactions between clonal hematopoiesis of indeterminate potential and air pollution in non-small cell lung cancer among non-smokers.

10514 Background: Small particulate matter air pollution (≤ 2.5µm diameter; PM2.5) is a recognized driver of non-small cell lung cancer (NSCLC) among non-smokers. PM2.5 recruits pro-inflammatory macrophages to the lungs, which facilitate the clonal expansion of mutated epithelial cells and eventual transformation to NSCLC. Clonal hematopoiesis of indeterminate potential (CHIP) is a common age-related condition caused by the acquisition of somatic mutations in hematopoietic stem cells. CHIP and its accompanying pro-inflammatory phenotype have been linked with various age-related diseases, including incident NSCLC; however, the mechanisms underlying this relationship are unknown. In this study, we aimed to explore the interaction between CHIP and exposure to PM2.5 in the development of incident NSCLC. Methods: This study was conducted using data from participants in the UK Biobank (n = 451,095). CHIP status was determined from peripheral blood whole exome sequencing data, and defined as the presence of a somatic driver mutation in the blood at variant allele frequency (VAF) ≥2% (PMID: 36652671). Incident NSCLC was determined from UK cancer registry data, and PM2.5 exposure was determined based on ambient regional measures from 2010. Cox proportional hazard models were used to evaluate associations between CHIP, PM2.5, and NSCLC. Proteomics data was measured using OLINK proteomics in a subset of UK Biobank participants (n = 44,625). Results: CHIP was prevalent in 3.4% of UK Biobank participants (n = 15,633; never smokers: 3.0%, n = 6,916/227,133). There were 1983 incident cases of NSCLC (never smokers: n = 307). CHIP status was associated with incident NSCLC when adjusting for smoking status (HR = 1.73, 95% CI: 1.48–2.02) and analyzing exclusively non-smokers (HR = 2.01, 95% CI: 1.34-3.00). PM2.5 levels were not associated with NSCLC in non-smokers (HR = 1.00, 95% CI: 0.89–1.12 per µg/m3 increase in PM2.5); however, there was a significant interaction between PM2.5 and CHIP (HR = 1.46, 95% CI: 1.02–2.11, pint = 0.04), suggesting that it is the interplay between the two that drives incident NSCLC risk. We found no association between PM2.5 levels and increased CHIP prevalence or CHIP VAF. PM2.5 and CHIP were also found to interact to increase systemic inflammatory markers C-reactive peptide (pint = 0.01) and IL-6 (pint = 0.002). Conclusions: PM2.5 and CHIP act as a novel gene x environment interaction pair that play a key role in NSCLC etiology among non-smokers. Rather than acting in isolation to increase risk of NSCLC, PM2.5 intensifies the relationship between CHIP and NSCLC, presumably by exacerbating systemic inflammation and the hyper-inflammatory lung microenvironment. With PM2.5 levels in the UK among the lowest in the world, we posit that the CHIP x PM2.5 interaction contributes substantially to the global burden of NSCLC in non-smokers.

A novel prognostic signature based on smoking-associated genes for predicting prognosis and immune microenvironment in NSCLC smokers

BackgroundAs a highly heterogeneous tumor, non-small cell lung cancer (NSCLC) is famous for its high incidence and mortality worldwide. Smoking can cause genetic changes, which leading to the occurrence and progress of NSCLC. Nevertheless, the function of smoking-related genes in NSCLC needs more research.MethodsWe downloaded transcriptome data and clinicopathological parameters from Gene Expression Omnibus (GEO) databases, and screened smoking-related genes. Lasso regression were applied to establish the 7-gene signature. The associations between the 7-gene signature and immune microenvironment analysis, survival analysis, drug sensitivity analysis and enriched molecular pathways were studied. Ultimately, cell function experiments were conducted to research the function of FCGBP in NSCLC.ResultsThrough 7-gene signature, NSCLC samples were classified into high-risk group (HRG) and low-risk group (LRG). Significant difference in overall survival (OS) between HRG and LRG was found. Nomograms and ROC curves indicated that the 7-gene signature has a stable ability in predicting prognosis. Through the analysis of immune microenvironment, we found that LRG patients had better tumor immune activation. FCGBP showed the highest mutation frequency among the seven prognostic smoking related genes (LRRC31, HPGD, FCGBP, SPINK5, CYP24A1, S100P and FGG), and was notable down-regulated in NSCLC smokers compared with non-smoking NSCLC patients. The cell experiments confirmed that FCGBP knockdown promoting proliferation, migration, and invasion in NSCLC cells.ConclusionThis smoking-related prognostic signature represents a promising tool for assessing prognosis and tumor microenvironment in smokers with NSCLC. The role of FCGBP in NSCLC was found by cell experiments, which can be served as diagnostic biomarker and immunotherapy target for NSCLC.

Postoperative Tobacco Cessation Improves Quality of Life, Lung Function and Long-Term Survival in Non-Small-Cell Lung Cancer Patients.

About 90% of all non-small cell lung cancer (NSCLC) cases are associated with inhalative tabacco smoking. Half of patients continue smoking during lung cancer therapy. We examined the effects of postoperative smoking cessation on lung function, quality of life (QOL) and long-term survival. In total, 641 patients, who underwent lobectomy between 2012 and 2019, were identified from our single institutional data base. Postoperatively, patients that actively smoked at the time of operation were offered a structured 'smoking cessation' program. For this retrospective analysis, two patient groups (total n = 90) were selected by pair matching. Group A (n = 60) had no postoperative tobacco smoking. Group B (n = 30) involved postoperative continued smoking. Lung function (FEV1, DLCO) and QOL ('SF-36' questionnaire) were measured 12 months postoperatively. We compared long-term outcomes using Kaplan-Meier curves. The mean age in group A was 62.6 ± 12.5 years and that in group B was 64.3 ± 9.7 years (p = 0.82); 64% and 62%, respectively, were male (p = 0.46). Preoperative smoking habits were similar ('pack years': group A, 47 ± 31; group B, 49 ± 27; p = 0.87). All relevant baseline characteristics we collected were similar (p > 0.05). One year after lobectomy, FEV1 was reduced by 15% in both groups (p = 0.98). Smoking cessation was significantly associated with improved DLCO (group A: 11 ± 16%; group B: -5 ± 14%; p <0.001) and QOL (vitality (VT): +10 vs. -10, p = 0.017; physical role function (RP): +8 vs. -17, p = 0.012; general health perceptions (GH): +12 vs. -5, p = 0.024). Patients who stopped smoking postoperatively had a significantly superior overall survival (median survival: 89.8 ± 6.8 [95% CI: 76.6-103.1] months vs. 73.9 ± 3.6 [95% CI: 66.9-80.9] months, p = 0.034; 3-year OS rate: 96.2% vs. 81.0%, p = 0.02; 5-year OS rate: 80.0% vs. 64.0%, p = 0.016). The hazard ratio (HR) was 2.31 [95% CI: 1.04-5.13] for postoperative smoking versus tobacco cessation. Postoperative smoking cessation is associated with improved quality of life and lung function testing. Notably, a significant increase in long-term survival rates among non-smoking NSCLC patients was observed. These findings could serve as motivation for patients to successfully complete a non-smoking program.

Algorithme thérapeutique des cancers bronchiques non à petites cellules étendus avec mutation de l’EGFR: Therapeutic algorithm of EGFR mutation driven advanced non-small cell carcinoma

EGFR mutations remain today the most frequently observed dependence mechanism in extensive non-small cell lung cancer (NSCLC), accounting for 12% of adenocarcinoma and 44% of NSCLC in non-smokers, in France. The “common” mutations of EGFR – deletions of exon 19 and L858R mutation, are observed in 89% of cases. However, the use of new molecular biology techniques confronts us frequently to the identification of “rare” EGFR mutations, but also to the observation of a molecular heterogeneity at diagnosis and during follow-up. This heterogeneity partly explains the variations in efficacy of EGFR tyrosine kinase inhibitors (EGFR-TKI) observed and on the other hand, leads to the emergence of resistant clones that direct the treatment during tumor progression. While 1st and 2nd line therapeutic strategies have been disrupted by the arrival of osimertinib, new evolutions are announced with the development of numerous trials associating different therapeutic classes such as chemotherapy, antiangiogenics but also targeted therapies i.e. small molecules, specific antibodies and conjugated antibodies.1877-1203/© 2023 SPLF. Published by Elsevier Masson SAS. All rights reserved.

Algorithme thérapeutique des cancers bronchiques non à petites cellules étendus avec mutation de l’EGR

EGFR mutations remain today the most frequently observed dependence mechanism in extensive non-small cell lung cancer (NSCLC), accounting for 12% of adenocarcinoma and 44% of NSCLC in non-smokers, in France. The «common» mutations of EGFR - deletions of exon 19 and L858R mutation, are observed in 89% of cases. However, the use of new molecular biology techniques confronts us frequently to the identification of «rare» EGFR mutations, but also to the observation of a molecular heterogeneity at diagnosis and during follow-up. This heterogeneity partly explains the variations in efficacy of EGFR tyrosine kinase inhibitors (EGFR-TKI) observed and on the other hand, leads to the emergence of resistant clones that direct the treatment during tumor progression. While 1st and 2nd line therapeutic strategies have been disrupted by the arrival of osimertinib, new evolutions are announced with the development of numerous trials associating different therapeutic classes such as chemotherapy, anti-angiogenics but also targeted therapies i.e. small molecules, specific antibodies and conjugated antibodies.1877-1203/© 2022 SPLF. Published by Elsevier Masson SAS. All rights reserved.

Characteristics and immune checkpoint inhibitor effects on non-smoking non-small cell lung cancer with KRAS mutation: A single center cohort (STROBE-compliant).

Kirsten rat sarcoma (KRAS) mutation (KRASm) is associated with poor prognosis in non-small cell lung cancer (NSCLC) patients. We have aimed to survey NSCLC patients harboring KRASm in Taiwan, where never-smoking lung adenocarcinoma predominates, and analyze the immune checkpoint inhibitor effect on NSCLC harboring KRASm.NSCLC patients with KRASm were enrolled and tested on programmed death-ligand 1 (PD-L1) expression using available tissue. We analyzed their clinical features, PD-L1 status, responses to ICIs, and overall survival (OS).We studied 93 patients with a median age 66.0 years, 23.7% of whom were women, and 22.6% were never-smokers. The results showed that G12C (36.6%) was the most common KRASm. In 47 patients with available tissue for PD-L1 testing, PD-L1 expression was positive in 66.0% of patients, while PD-L1 ≥50% was higher in ever-smokers (P = .038). Among 23 patients receiving ICI treatment, those with PD-L1 ≥50% experience a 45.5% response rate to ICI. There were benefits from ICI treatment on OS compared with no ICI treatment (median OS 35.6 vs 9.8 months, P = .002) for all of our patients, and for patients with PD-L1 ≥50% (median OS not-reached vs 8.4 months, P = .008). There were no differences in survival across different KRAS subtypes (P = .666).Never-smokers composed more than one-fifth of KRASm in NSCLC in Taiwan. A high PD-L1 expression was related to smoking history and responded well to ICI. ICI treatment improved the OS in NSCLC patients with KRASm, particularly those with PD-L1 ≥50%.

Algorithme thérapeutique des cancers bronchiques non à petites cellules étendus avec mutation de l’EGR: Therapeutic algorithm of EGFR mutation driven advanced non-small cell carcinoma

EGFR mutations remain today the most frequently observed dependence mechanism in extensive non-small cell lung cancer (NSCLC), accounting for 12% of adenocarcinoma and 44% of NSCLC in non-smokers, in France. The “common” mutations of EGFR − deletions of exon 19 and L858R mutation, are observed in 89% of cases. However, the use of new molecular biology techniques confronts us frequently to the identification of “rare” EGFR mutations, but also to the observation of a molecular heterogeneity at diagnosis and during follow-up. This heterogeneity partly explains the variations in efficacy of EGFR tyrosine kinase inhibitors (EGFR-TKI) observed and on the other hand, leads to the emergence of resistant clones that direct the treatment during tumor progression. While 1st and 2nd line therapeutic strategies have been disrupted by the arrival of osimertinib, new evolutions are announced with the development of numerous trials associating different therapeutic classes such as chemotherapy, antiangiogenics but also targeted therapies i.e. small molecules, specific antibodies and conjugated antibodies.© 2021 SPLF. Published by Elsevier Masson SAS. All rights reserved.

BAI1 nuclear expression reflects the survival of nonsmoking non-small cell lung cancer patients.

BackgroundSmoking‐ and nonsmoking‐associated lung cancers have different mechanisms of carcinogenesis. We divided non‐small cell lung cancer (NSCLC) patients into nonsmoking and smoking groups with the aim of trying to understand the utility of brain‐specific angiogenesis inhibitor 1 (BAI1) expression in the separate groups.MethodsClinicopathological data were obtained from 148 patients who had undergone surgery for NSCLC of the lung. Tissue microarray blocks were made of samples from NSCLC patients. Two pathologists graded the intensity of BAI1 expression as high or low expression in the cancer cells of patients in the smoking and nonsmoking groups.ResultsNSCLC nonsmokers with higher BAI1 nuclear expression had poor disease‐specific survival (DSS) (hazard ratio: 2.679; 95% confidence interval [CI]:1.022–7.022, p = 0.045). The Kaplan–Meier survival curve confirmed that higher BAI1 expression was significantly associated with poor DSS (p = 0.034) in the nonsmoking group.ConclusionsWe divided NSCLC patients into nonsmoking and smoking groups and found that nuclear BAI1 expression was related to patient survival in nonsmoking NSCLC patients. We suggest BAI1 expression as a predictive marker of nonsmoking‐associated NSCLC and recommend that it be evaluated as an AJCC staging criterion in the future.

Prognostic role of lipid phosphate phosphatases in non-smoker, lung adenocarcinoma patients

Sphingosine-1-phosphate (S1P), a bioactive sphingolipid, plays a crucial role in tumorigenesis. It mediates its function through S1P receptors. A few components of the S1P signaling pathway, such as sphingosine kinase 1 (SphK1) and S1P receptor 1 (S1PR1), have been shown to contribute to lung carcinogenesis. In the present study, using web-based computational tools, we assessed the prognostic roles of eight S1P metabolizing enzymes and five S1P receptors in non-small-cell lung cancer (NSCLC) patients. Except for SPHK1, low expression of S1P metabolizing enzymes was correlated with worse overall survival (OS) in NSCLC patients. Moreover, lower expression of lipid phosphate phosphatase-1 and - 3 (PLPP1 and PLPP3) was significantly associated with worse OS in lung adenocarcinoma (LUAD) and non-smoker NSCLC patients. Furthermore, the UALCAN database analysis showed that mRNA and protein expression of PLPP3 and S1PR1 are significantly down regulated in primary tumors due to hypermethylation of their respective promoters. Expression of PLPP3, S1PR1, and S1PR4 was positively correlated with tumor-infiltrating immune cells in NSCLC patients. These results indicate that S1P signaling genes play a critical prognostic role in LUAD patients. Therefore, this gene signature could be used to predict their prognosis more accurately.

Traitement des cancers bronchiques non à petites cellules de stades avancés mutés EGFR : quels inhibiteurs ? Quelles séquences thérapeutiques ?: EGFR mutation driven advanced non-small cell carcinoma: Which inhibitors? Which therapeutic sequences?

AbstractEGFR mutations remain today the most frequently observed dependence mechanism in extensive non-small cell lung cancer (NSCLC), accounting for 12% of adenocarcinoma and 44% of NSCLC in non-smokers, in France. The « common » mutations of EGFR – deletions of exon 19 and L858R mutation, are observed in 89% of cases. However, the use of new molecular biology techniques confronts us frequently to the identification of « rare » EGFR mutations, but also to the observation of a molecular heterogeneity at diagnosis and during follow-up. This heterogeneity partly explains the variations in efficacy of EGFR tyrosine kinase inhibitors (EGFR-TKI) observed and on the other hand, leads to the emergence of resistant clones that direct the treatment during tumor progression. Until recently, 1st and 2nd line therapeutic strategies were well established, but were disrupted by the arrival of the osimertinib in 2nd, then in 1st line.© 2020 SPLF. Published by Elsevier Masson SAS. All rights reserved.

Relevance of genetic polymorphisms in tobacco-related detoxifying enzymes in non-small cell lung carcinoma susceptibility

BackgroundTobacco smoking plays a role in the onset and development of non-small cell lung cancer (NSCLC). Biotransformation and detoxification of tobacco smoke carcinogens are brought about by xenobiotic-metabolizing enzymes (XME). Germline mutations in CYP1B1 (Ala119Ser) and GSTT1 (+/−) have been known to affect the functionality of these enzymes, thereby modifying their effect; amino acid substitution and complete gene deletion abolish enzyme activity. CYP1B1 (Ala119Ser) and GSTT1 (+/–) genotypic interactions and gene-smoking inteaction associated risk of NSCLC information is not documented in the literature. ObjectiveThe role of gene-gene and gene-smoking interaction with the clinicopathological parameters to estimate the risk of NSCLC among the North Indian population. MethodsCYP1B1 and GSTT1 genotypes were determined by the PCR-RFLP in 158 cases of NSCLC and 155 controls; demographical parameters and clinicopathological characteristics were recorded. ResultsGSTT1 (−/−) genotype demonstrated a four-fold risk of NSCLC and was significantly associated with squamous cell lung carcinoma (OR: 3.35). Synergistic interaction of CYP1B1 (Variant) and GSTT1 (−/−) enhanced the risk of NSCLC by 6.5 fold. CYP1B1 mutant genotypes in tobacco smokers showed a 3.5 fold risk, which was significantly (p = 0.02) higher than the non-smokers (OR: 1.67, p = 0.38). NSCLC risk with GSTT1 mutant genotype showed a similar trend of risk in both smokers (OR: 4.1) and non-smokers (OR: 3.8). ConclusionThe effect of CYP1B1 mutant appears to be a risk modifier in tobacco smokers, whereas deletions in GSTT1 (−/−) increases the risk of NSCLC in both smokers and non-smokers. A marked synergistic risk was evident when both mutations were present.

CYP3A4 genetic variants are associated with susceptibility of non-small cell lung cancer in a Shaanxi Han population

PurposeLung cancer (LC) is one of the fastest-growing malignant tumors in the world in terms of morbidity and mortality. CYP3A4 plays a crucial role in the occurrence of LC. Little is known about the contribution of CYP3A4 polymorphisms for non-small cell lung cancer (NSCLC) risk. This study aimed to explore the correlation of CYP3A4 genetic variants (rs3735451, rs4646440, rs35564277, and rs4646437) with NSCLC risk. MethodsFour single nucleotide polymorphisms (SNPs) were genotyped by Agena MassARRAY in this case-control study (507 NSCLC patients and 505 controls) among a Shaanxi Han population. Hardy-Weinberg equilibrium (HWE) of each SNP in controls was evaluated by exact test. The association of CYP3A4 polymorphisms with NSCLC risk was explored by calculating odds ratios (OR) and 95% confidence intervals (CI) using logistic regression analysis with adjustment for age and gender. ResultsOur research revealed that rs4646440 was significantly associated with an increased risk of NSCLC (OR 2.64, p = .005), while rs4646437 played a protective role in NSCLC risk (OR 0.48, p = 4.00 × 10−7). Stratified analyses indicated that rs4646440 significantly enhanced the susceptibility of NSCLC in BMI > 24 kg/m2, non-smokers and non-drinkers (OR 14.29, p = .012; OR 1.56, p = .023; OR 1.67, p = .031, respectively). Besides, we observed that rs3735451 exhibited an increased risk of NSCLC in BMI > 24 kg/m2 (OR 2.47, p = .030), whereas rs4646437 had a reduced risk of NSCLC in BMI ≤ 24 kg/m2 (OR 0.47, p = 5.17 × 10−5). We also found that rs35564277 was considered as a protective factor of NSCLC in non-smokers (OR 0.50, p = .032). ConclusionOur study indicated that CYP3A4 genetic variants were associated with NSCLC susceptibility in a Shaanxi Han population.

LP-300 as a potential first-in-class combination agent with tyrosine kinase inhibitors (TKIs) in non-smoker lung adenocarcinoma.

e21660 Background: Lung cancer in non-smokers (LCINS) is the seventh leading cause of death among solid tumors. LCINS is more frequent in women, and the histological incidence of adenocarcinoma is higher among non-smokers. The mutation pattern by smoking status in non-small cell lung cancer (NSCLC) is distinct with genomic alterations in EGFR, MET and ROS1 being more frequent in non-smokers. Non-smoking status is the strongest clinical predictor of benefit from the EGFR TKIs. Lantern Pharma is developing LP-300/Dimesna, a non-toxic, well-tolerated, water-soluble disulfide as a combination therapy agent potentially indicated in female never-smokers with adenocarcinoma. Retrospective subgroup analyses of prior phase 3 trial data showed substantial overall survival benefit for LP-300 in combination with standard chemotherapy in patients with advanced NSCLC, especially in the female non-smoker adenocarcinoma subgroup (13.4 months in control arm to 27 months in treatment arm). Purpose of this work conducted at BioNumerik Pharmaceuticals was to determine if LP-300 modulates activity of selected receptor tyrosine kinases relevant to non-smoker NSCLC. Methods: In vitro luminescence-based kinase assays were used to evaluate the effect of LP-300 and LP-300-derived mesna-disulfide heteroconjugates, Erlotinib, and Crizotinib, in combination and as single agents, on EGFR, MET, and ROS1 kinase activity. Results: LP-300 inhibited WT EGFR kinase and MET kinase. LP-300 had a time-dependent inhibitory effect on ROS1 kinase activity. LP-300 and LP-300-derived mesna-disulfide heteroconjugates stimulated Erlotinib-mediated inhibition of WT and T790M EGFR. T790M EGFR is normally insensitive to Erlotinib but when LP-300 was used with Erlotinib, inhibition was more prominent. LP-300 also stimulated Crizotinib-mediated inhibition of MET kinase and displayed time-dependent stimulation of Crizotinib-mediated inhibition of ROS1 kinase. Conclusions: LP-300 is a multi-targeted modulator of enzymatic activities of EGFR, MET and ROS1, and potentiates the activity of Erlotinib and Crizotinib. EGFR family members contain conserved extracellular structures that are stabilized by disulfide bonds. Compounds that disrupt extracellular disulfide bonds could inactivate EGFR and potentially downregulate EGFR-dependent proliferative signaling in cancer cells. We propose LP-300 as a Disulfide Bond Disrupting Agent (DDA) that may complement existing EGFR-targeted agents functioning through alternate mechanisms.

Traitement des cancers bronchiques non à petites cellules de stades avancés mutés EGFR: quelle(s) séquence(s) ?

EGFR mutations remain today the most frequently observed dependence mechanism in extensive non-small cell lung cancer (NSCLC), accounting for 12 % of adenocarcinoma and 44 % of NSCLC in non-smokers, in France. The « common » mutations of EGFR — deletions of exon 19 and L858R mutation, are observed in 89 % of cases. However, the use of new molecular biology techniques confronts us frequently to the identification of « rare » EGFR mutations, but also to the observation of a molecular heterogeneity at diagnosis and during follow-up. This heterogeneity partly explains the variations in efficacy of EGFR tyrosine kinase inhibitors (EGFR-TKI) observed and on the other hand, leads to the emergence of resistant clones that direct the treatment (T790M) during tumor progression. Until recently, 1st and 2nd line therapeutic strategies were well established, but were disrupted by the arrival of the osimertinib in 2nd, then in 1st line. In the absence of data concerning the impact on overall survival of osimertinib and on the secondary mechanisms of resistance at progression, it is important to update our knowledge about new molecules, but also strategies of already available treatments.

Abstract 582: Tumor immunoediting in a lung cancer mouse model harboring EGFR mutations

Abstract Background: Anti-Programed cell death 1 (PD-1) antibodies such as nivolumab or pembrolizumab show promising treatment effects in smoking-related lung cancer. However, anti-PD-1 antibodies have very little effect on epidermal growth factor receptor (EGFR) mutant non-small cell lung cancer (NSCLC). EGFR mutations are the most frequent oncogenic drivers in non-smoking related NSCLC. Therefore, we aimed to explore the mechanism underlying tumor immunoediting in a lung cancer mouse model harboring EGFR mutations. Methods: We previously established two strains of transgenic lung cancer mouse (C57BL/6) harboring a mouse Egfr exon 19-deletion (mDEL) or a human EGFR L858R (hLR) mutation, driven by a surfactant protein (SP)-C promoter (Ohashi, Cancer Sci. 2008; Ohashi, Cancer Res. 2009). These mice developed EGFR-dependent multifocal lung adenocarcinomas from type II pneumocytes. We modified the two models by transplanting lung tumors subcutaneously into C57BL/6 (Ohashi, AACR. 2018). Evaluation of mice tumor lymphocyte profile was done through flow cytometry or immunohistochemistry (IHC). Effects of T cell depletion or anti-mouse PD-1 antibody (4H2) in vivo were also assessed. Comprehensive analysis of mRNA expression of immuno-checkpoint molecules in the tumors was performed using PanCancer Immune Profiling Panel (NanoString Technologies). Results: CD4+ T cells and/or CD8+ T cells were depleted in the subcutaneously transplanted tumor model followed by treatment with gefitinib (50 mg/kg/day, 5 days/week) for 2 weeks. As a result, after gefitinib discontinuation, rapid regrowth was seen in the tumors from groups with depletion of CD8+ T cells; depletion of CD4+ T cells had little effect on tumor regrowth, and tumor inhibition continued for 2 weeks after cessation of gefitinib. IHC showed relatively increased numbers of CD4+ T cells in the mice tumors whereas a decrease in the number of CD8+ T cells or Foxp3 positive cells was noted. PD-L1 expression was not observed in the tumors. Consistently, 4H2 showed little effect on tumor growth in vivo. The cancer immune panel showed increased expression of immune mediators such as Ccl6, Clec4n, and Lcn2. Conclusions: Our results suggest that CD8+ T cells can recognize mice tumors in vivo. New immuno-checkpoint molecules regulating CD8+ T cells from PanCancer Immune Profiling Panel were evaluated. Further, we plan to evaluate the efficacy of various immuno-checkpoint inhibitors in vivo. Citation Format: Kazuya Nishii, Kadoaki Ohashi, Kiichiro Ninomiya, Go Makimoto, Hiromi Watanabe, Hirohisa Kano, Naofumi Hara, Heiichiro Udono, Katsuyuki Kiura. Tumor immunoediting in a lung cancer mouse model harboring EGFR mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 582.

DNA methylation-based diagnostic and prognostic biomarkers of nonsmoking lung adenocarcinoma patients.

Currently, there are few studies on patients with nonsmoking lung adenocarcinoma, and the pathogenesis is still unclear. The role of DNA methylation in the pathogenesis of cancer is gradually being recognized. The purpose of this study was to determine the abnormal methylation genes and pathways involved in nonsmoking lung adenocarcinoma patients. Gene expression microarray data (GSE10072, GSE43458) and gene methylation microarray data (GSE62948) were downloaded from the Gene Expression Omnibus (GEO) database and differentially expressed genes were obtained through GEO2R. Next, we analyzed the function and enrichment of the selected genes using Database for Annotation, Visualization, and Integrated Discovery. The protein-protein interaction (PPI) networks were constructed using the Search Tool for the Retrieval of Interacting Genes database and visualized in Cytoscape. Finally, we performed module analysis of the PPI network using Molecular Complex Detection. And we obtained 10 hub genes by Cytoscape Centiscape. We analyzed the independent prognostic value of each hub gene in nonsmoking nonsmall cell lung cancer patients through Kaplan-Meier plotter. Seven hub genes (CXCL12, CDH1, CASP3, CREB1, COL1A1, ERBB2, and ENO2) were closely related to the overall survival time. This study provides an effective bioinformatics basis for further understanding the pathogenesis and prognosis of nonsmoking lung adenocarcinoma patients. Hub genes with prognostic value could be selected as effective biomarkers for timely diagnosis and prognostic of nonsmoking lung adenocarcinoma patients.

Metabolic gene NR4A1 as a potential therapeutic target for non-smoking female non-small cell lung cancer patients.

BackgroundAlthough cigarette smoking is considered one of the key risk factors for lung cancer, 15% of male patients and 53% of female patients with lung cancer are non‐smokers. Metabolic changes are critical features of cancer. Therapeutic target identification from a metabolic perspective in non‐small cell lung cancer (NSCLC) tissue of female non‐smokers has long been ignored.ResultsBased on microarray data retrieved from Affymetrix expression arrays E‐GEOD‐19804, we found that the downregulated genes in non‐smoking female NSCLC patients tended to participate in protein/amino acid and lipid metabolism, while upregulated genes were more involved in protein/amino acid and carbohydrate metabolism. Combining nutrient metabolic co‐expression, protein–protein interaction network construction and overall survival assessment, we identified NR4A1 and TIE1 as potential therapeutic targets for NSCLC in female non‐smokers. To accelerate the drug development for non‐smoking female NSCLC patients, we identified nilotinib as a potential agonist targeting NR4A1 encoded protein by molecular docking and molecular dynamic stimulation. We also show that nilotinib inhibited proliferation and induced senescence of cells in non‐smoking female NSCLC patients in vitro.ConclusionsThese results not only uncover nutrient metabolic characteristics in non‐smoking female NSCLC patients, but also provide a new paradigm for identifying new targets and drugs for novel therapy for such patients.

Prise en charge thérapeutique des cancers bronchiques non à petites cellules de stades avancés mutés pour l’EGFR

EGFR mutations remain today the most frequently observed dependence mechanism in extensive non-small cell lung cancer (NSCLC), accounting for 12% of adenocarcinoma and 44% of NSCLC in non-smokers, in France. The “common” mutations of EGFR - deletions of exon 19 and L858R mutation, are observed in 89% of cases. However, the use of new molecular biology techniques confronts us frequently to the identification of “rare” EGFR mutations, but also to the observation of a molecular heterogeneity at diagnosis and during follow-up. This heterogeneity partly explains the variations in efficacy of EGFR tyrosine kinase inhibitors (EGFR-TKI) observed and on the other hand, it makes it possible to orient the treatment (T790M), during tumor progression. Until recently, 1st and 2nd line therapeutic strategies were well established, but were disrupted by the arrival of the osimertinib in 2nd, then in 1st line. In the absence of data concerning the impact on overall survival of osimertinib and on the secondary mechanisms of resistance at progression, it is important to update our knowledge about new molecules, but also strategies of already available treatments.

Abstract 560: Identification of a novel MUTYH mutation in non-small cell lung cancer

Abstract Introduction: Lung cancer is the leading cause of death from cancer in Hong Kong and worldwide. Nearly 80-85% of lung cancers are diagnosed with non-small cell lung cancer (NSCLC). Treatment strategies can be refined according to the distinct difference of NSCLC molecular profiling among different ethnic populations. Moreover, oncogenic driver mutations are closely linked to NSCLC in non-smokers and young people. Therefore, the urge of finding pathogenic mutations is inevitable. This study aims to discover novel mutations in NSCLC patients in Hong Kong by next-generation sequencing and characterize the clinical significance of these mutations. Methods: In order to identify mutations, DNA was purified from 6 pairs of lung tumor-normal tissues from 2 stage I adenocarcinoma (LUAD), 2 stage III LUAD and 2 stage I squamous cell carcinoma (LUSC) patients. Samples were sequenced by targeted sequencing with the TOMA OS-Seq 130 cancer gene panel. Sequence alignment and variant calling were performed by combining TOMA Stratus and in-house pipelines. Results: The mean aligned reads exceeded 14 million and the mean coverage of target region was over 1000X. MUTYH deletion was found in 66.7% (4/6) of the patients. Data from The Cancer Genome Atlas (TCGA) revealed that MUTYH deletion is significantly associated with poorer disease-free survival (DFS) (p=0.0063) and overall survival (p=0.022) in LUAD patients. TCGA data also showed that MUTYH copy number correlates with gene expression (r=0.473) and LUAD patients with MUTYH mRNA overexpression have better DFS (p=0.0205). However, such correlations were not observed in LUSC patients. MUTYH is a DNA glycosylase which involves in the base excision repair and it repairs oxidative DNA damage by preventing G:C to T:A transversion. Numerous studies indicated that biallelic mutations in MUTYH cause MUTYH-associated polyposis and increase the lifetime risk of colorectal cancer. MUTYH mutations could be associated with increased risk of endometrial and breast cancer. Conclusion: To elucidate the clinical significance of MUTYH in LUAD, an independent cohort will be sequenced to validate the primary result. Immunohistochemistry, RT-qPCR and functional assays will be performed to examine the protein expression, the gene expression and the functional role. All these findings suggested that MUTYH deletion may play a crucial role in LUAD and potentially lead to the development of biomarker and therapeutic target. Citation Format: Alvin Hong-Wai Fong, Eunice Yuen-Ting Lau, William Ka-Chun Cheung, William Chi-Shing Cho. Identification of a novel MUTYH mutation in non-small cell lung cancer [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 560.