Plasma Lipidomics Profiling to Identify the Biomarkers of Diagnosis and Radiotherapy Response for Advanced Non-Small-Cell Lung Cancer Patients

Introduction: Lung cancer carries the highest cancer-related mortality rates worldwide. Despite all recent advances, the mortality from lung cancer is still rising. A better understanding of the risk factors may help us predict responses to chemotherapy for better management. Aim: To evaluate predictors of response to chemotherapy in advanced Non-Small Cell Lung Cancer (NSCLC) patients. Materials and Methods: This was a prospective cohort study conducted in the Department of Pulmonary Medicine at Government Medical College and Hospital, Chandigarh, India. A total of 60 confirmed cases of advanced (stage IIIB and IV) NSCLC patients were enrolled consecutively for a duration of two years. Baseline clinical parameters, routine blood tests, spirometry, exercise capacity using the 6 Minute Walk Test Distance (6MWTD), and Computed Tomography (CT)-based tumour size were recorded. Certain pre-defined patient, disease, and therapy-related factors (age, gender, dyspnoea, baseline blood tests, tumour size, histology, etc.) were evaluated for their possible role as predictors of treatment response in advanced NSCLC patients. A positive response was defined if the response to chemotherapy was Complete Response (CR) or Partial Response (PR), and a negative response if the response was Progressive Disease (PD) or Stable Disease (SD) as per revised RECIST (Response evaluation criteria in solid tumors) 1.1 criteria. Variables between the two groups were compared using the Mann-Whitney U test and Chi-square test. To find out the factors that may predict response to treatment, univariate and multivariate logistic regression analysis were used. Results: Out of a total of 60 confirmed cases of NSCLC patients, only 40 patients were able to complete the four cycles of chemotherapy. The mean age of the patients was 58.5±9.6 years. There were a total of 35 males (87.5%) and five females (12.5%) in the study. Out of 40 patients, 27 (67.5%) had squamous cell carcinoma and 13 (32.5%) had adenocarcinoma. On univariate analysis, Neutrophil-Lymphocyte Ratio (NLR) had a statistically significant association with tumour response (p<0.001). On multivariate analysis, advanced age (p=0.05) and high (>3.81) NLR (p=0.002) were found as independent predictors of poor response to chemotherapy. Conclusion: Pre-treatment high NLR and advanced age are significant factors for a poor response to chemotherapy treatment in advanced NSCLC patients.

Immune-Related Adverse Events and Outcomes in Patients with Advanced Non–Small Cell Lung Cancer: A Predictive Marker of Efficacy?

P2.01-037 Clinical Impact of Interstitial Lung Disease on Advanced Non-Small Cell Lung Cancer

Studies have shown that miRNA (miR) can be stably detected in serum, and aberrant expression of various miRNAs has shown diagnostic value in non-small cell lung cancer (NSCLC) patients. However, the role of miRNA in the context of prognosis has not been extensively investigated. Our previous study reported that miR-22, miR-125b, and miR-15b in serum had potential for use as tumor markers for auxiliary diagnosing of NSCLC. Therefore, the objective of this study was to detect the levels of miR-22, miR-125b, and miR-15b in serum from NSCLC patients and explore the potential prognostic significance of the three selected miRNAs. The relative expression of miR-22, miR-125b, and miR-15b in 74 patients with advanced NSCLC in pre- and post-chemotherapy were detected by real-time quantitative polymerase chain reaction. Serum level of miR-125b significantly decreased after chemotherapy (p < 0.05) and the levels of miR-15b significantly increased (p < 0.01), while there was no change in the level of serum miR-22 (Z = 0.716, p > 0.05). Compared with pre-chemotherapy, serum miR-125b expression in advanced NSCLC patients of responders (CR + PR) were significantly decreased post-chemotherapy (p < 0.05); serum miR-15b expression in advanced NSCLC patients of responders (CR + PR) were increased (p < 0.01). The chemotherapy sensitivity of advanced NSCLC patients with high expression of miR-125b was lower than that of NSCLC patients with low expression (p < 0.05). The chemotherapy sensitivity of advanced NSCLC patients with high expression of miR-15b was higher than that of NSCLC patients with low expression (p < 0.05). High levels of serum miR-125b and low levels of serum miR-15b were related to poor overall survival (p < 0.05). The serum levels of miR-125b and miR-15b in advanced NSCLC patients were changed pre- and post-chemotherapy and these changes were associated with chemotherapeutic response. Serum miR-125b and miR-15b have certain potential clinical value for chemotherapeutic response in advanced NSCLC. The serum levels of miR-125b and miR-15b in patients with advanced NSCLC before treatment may be used to estimate the overall survival.

Background: Breast-cancer metastasis suppressor 1 (BRMS1) gene encodes for a predominantly nuclear protein that differentially regulates the expression of multiple genes, leading to suppression of metastasis without blocking orthotopic tumor growth. The aim of the present study was to evaluate for the first time the prognostic significance of BRMS1 promoter methylation in cell free DNA (cfDNA) circulating in plasma of non-small cell lung cancer (NSCLC) patients. Towards this goal, we examined the methylation status of BRMS1 promoter in NSCLC tissues, matched adjacent non-cancerous tissues and corresponding cfDNA as well as in an independent cohort of patients with advanced NSCLC and healthy individuals. Methods: BRMS1 promoter methylation was examined in 57 NSCLC tumors and adjacent non-cancerous tissues, in cfDNA isolated from 48 corresponding plasma samples, in cfDNA isolated from plasma of 74 patients with advanced NSCLC and 24 healthy individuals. Results: BRMS1 promoter was highly methylated both in operable NSCLC primary tissues (59.6%) and corresponding cfDNA (47.9%) but not in cfDNA from healthy individuals ( 0%), while it was also highly methylated in cfDNA from advanced NSCLC patients (63.5%). In operable NSCLC, Kaplan-Meier estimates were significantly different in favor of patients with non-methylated BRMS1 promoter in cfDNA, concerning both DFI (p=0.048) and OS (p=0.007). In advanced NSCLC, OS was significantly different in favour of patients with non-methylated BRMS1 promoter in their cfDNA (p=0.003). Multivariate analysis confirmed that BRMS1 promoter methylation has a statistical significant influence both on operable NSCLC patients' DFI time and OS and on advanced NSCLC patients' PFS and OS. Conclusions: BRMS1 promoter methylation in cfDNA isolated from plasma of NSCLC patients provides important prognostic information and merits to be further evaluated as a circulating tumor biomarker. Citation Format: Ioanna Balkouranidou, Maria Chimonidou, Georgia Milaki, Emily Tsaroucha, Stelios Kakolyris, Danny Welch, Vasilis Georgoulias, Evi Lianidou. Breast cancer metastasis suppressor-1 promoter methylation in cell free DNA provides prognostic information in non-small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3826. doi:10.1158/1538-7445.AM2014-3826

Objectives: Immune checkpoint inhibitors (ICIs) have changed the landscape of lung cancer therapy. However significant proportions of patients have primary or acquired resistance to ICIs. Molecular characterization is critical for patient selection and overcoming resistance to checkpoint inhibitors. The purpose of this study is to investigate the molecular characteristics associated with ICIs outcomes in advanced non-small cell lung cancer (NSCLC) patients. Materials and methods: All advanced stage NSCLC patients at City of Hope who received ICIs (pembrolizumab, nivolumab, atezolizumab, and durvalumab) were identified retrospectively. Overall survival (OS, from the start of the ICIs), Pathology and information on genomic alterations (GAs) including next-generation sequencing (NGS) data, tumor mutation burden (TMB), and Programmed death-ligand 1 (PD-L1) levels were collected. Chi-square and Fisher's exact test, Log-rank test were used for comparison of demographics, and survival curves respectively. Univariate and multivariate COX proportional hazards model was used for survival analysis. Results: 346 NSCLC patients were identified. Univariate and multivariate analysis found the association of OS with PD-L1 level ≥50% (Hazard ratio [HR], 0.19; 95% confidence interval [CI], 0.06-0.59; P&amp;lt;0.01), EGFR (HR 7.38; 95% CI, 1.15-47.42; P&amp;lt;0.05), and TET2 (HR 0.15; 95% CI, 0.03-0.90; P&amp;lt;0.05). The median OS was not reached [NR] for the 12 patients who had genomic alterations (GAs) in TET2 (12/108, 11%) versus (vs) 11.5 months in TET2 negative patients (98/108, 89%). Interestingly, GAs in TET2 and FANCA were mutually exclusive and patients who had GAs in FANCA gene (6%) had shorter OS (5.5 months vs 14.5 months, Log-rank test, P&amp;lt;0.05). Conclusions: We described the clinical and molecular features of NSCLC patients treated with ICIs. The association of GAs in TET2 with longer OS and its mutual exclusivity with FANCA GAs were insightful for developing novel therapeutic strategies to improve ICIs outcomes in NSCLC. Keywords: Lung cancer, molecular, next-generation sequencing (NGS), immune checkpoint inhibitors, TET2, FANCA, PD-L1, TMB, immunotherapy Citation Format: Dan Zhao, Isa Mambetsariev, Haiqing Li, Chen Chen, Jeremy Fricke, Patricia Fann, Prakash Kulkarni, Yan Xing, Peter Lee, Andrea Bild, Erminia Massarelli, Marianna Koczywas, Karen Reckamp, Ravi Salgia. Association of molecular characteristics with survival in advanced non-small cell lung cancer patients treated with checkpoint inhibitors [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3300.

e21669 Background: In the new era of immunotherapy, the regimen based on bevacizumab is still one of the standard options for treatment of advanced non-small cell lung cancer (NSCLC) patients without driver mutations. However, the prognostic factors for bevacizumab are still missing. We aimed to determine the integrative value of computed tomography (CT), epigenetic modifications, clinicopathological and systemic inflammatory factors for predicting the survival of advanced NSCLC patients with bevacizumab. Methods: Clinicopathological parameters, dynamic systemic inflammatory factors, radiomics features, and DNA methylation profiling in advanced non-squamous NSCLC patients receiving first- or second- line bevacizumab plus chemotherapy were included in this study. The prognostic radiomics signature were constructed by least absolute shrinkage and selection operator (LASSO) Cox analysis. A multi-omics prediction nomogram for progression-free survival (PFS) based on radiomics, clinicopathological and systemic inflammatory features was established and independently validated. Furthermore, radiomics signature-related DNA methylation were submitted to functional enrichment analysis. Results: Total of 272 patients were included in analysis, 224 in training cohort and 48 in validation cohort. Five radiomics features, including Information Measure Corr1, Inverse Variance, Local Std Max, Gauss Area, Spherical Disproportion, were finally selected to construct radiomics signature with the AUC of 0.71. Smoking history, anatomical feature, liver metastasis, LDH4, NLR2 and radiomics signature were found to be independent prognostic factors for PFS. A multi-omics nomogram was developed based on these features in training cohort with the C-index of 0.76, and external validated with the C-index of 0.75. The tissue slices of 20 patients receiving bevacizumab were used for DNA methylation profiling. Functional enrichment analysis indicated widespread and statistically significant associations between radiomics features and DNA methylation changes which involved in several pathways related to angiogenesis and immune system, such as Notch signaling pathway, small GTPase Rho signal transduction, TGFβ signal transduction. Conclusions: This multi-omics nomogram integrating radiomics, clinicopathological, systemics inflammatory features improved the prediction of PFS in advanced NSCLC patients receiving bevacizumab. And the radiomics signature were found to be related to angiogenesis and immune status.

Background:Breast-cancer metastasis suppressor 1 (BRMS1) gene encodes for a predominantly nuclear protein that differentially regulates the expression of multiple genes, leading to suppression of metastasis without blocking orthotropic tumour growth. The aim of the present study was to evaluate for the first time the prognostic significance of BRMS1 promoter methylation in cell-free DNA (cfDNA) circulating in plasma of non-small cell lung cancer (NSCLC) patients. Towards this goal, we examined the methylation status of BRMS1 promoter in NSCLC tissues, matched adjacent non-cancerous tissues and corresponding cfDNA as well as in an independent cohort of patients with advanced NSCLC and healthy individuals.Methods:Methylation of BRMS1 promoter was examined in 57 NSCLC tumours and adjacent non-cancerous tissues, in cfDNA isolated from 48 corresponding plasma samples, in cfDNA isolated from plasma of 74 patients with advanced NSCLC and 24 healthy individuals.Results:The BRMS1 promoter was highly methylated both in operable NSCLC primary tissues (59.6%) and in corresponding cfDNA (47.9%) but not in cfDNA from healthy individuals (0%), while it was also highly methylated in cfDNA from advanced NSCLC patients (63.5%). In operable NSCLC, Kaplan–Meier estimates were significantly different in favour of patients with non-methylated BRMS1 promoter in cfDNA, concerning both disease-free interval (DFI) (P=0.048) and overall survival (OS) (P=0.007). In advanced NSCLC, OS was significantly different in favour of patients with non-methylated BRMS1 promoter in their cfDNA (P=0.003). Multivariate analysis confirmed that BRMS1 promoter methylation has a statistical significant influence both on operable NSCLC patients' DFI time and OS and on advanced NSCLC patients' PFS and OS.Conclusions:Methylation of BRMS1 promoter in cfDNA isolated from plasma of NSCLC patients provides important prognostic information and merits to be further evaluated as a circulating tumour biomarker.

7535 Background: The epithelial to mesenchymal transition (EMT) is a fundamental biological process during which epithelial cells change to a mesenchymal phenotype, it has a profound impact on cancer progression and treatment. The purpose of this study was to investigate the role of EMT to predict the efficacy of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) in advanced non-small cell lung cancer (NSCLC) patients. Methods: We evaluated the correlation between EMT and sensitivity to EGFR-TKIs in advanced NSCLC patients. Immunohistochemistry was used to examine the expression of EMT markers, E-cadherin, fibronectin, N-cadherin and vimentin and ARMs was used to detect the EGFR mutation in tumor specimens obtained before the treatment. The EMT phenotype status was determined according to the expression of E-cadherin, fibronectin, N-cadherin and vimentin. Results: 101 patients enrolled into this study and 97 had enough tissue to perform the EMT examination. The median age was 59 years old, male/female:56/45, never smoker/smoker: 71/30, adenocarcinoma/non-adeno: 73/28, PS 0-1/2-3: 83/18, Stage IIIB/IV: 2/99, 1st /2nd-4th line: 13/88. The response rate and progression free survival(PFS) in the whole population were 45.5% and 7.6 months respectively. EMT test revealed that 46(47.4%) samples were epithelial phenotype, while 51(52.6%) were mesenchymal phenotype. 38 of 79 patients harbour activating EGFR mutation. Among the patients with EGFR wild type or unknown status, patients with an epithelial phenotype had a higher response rate(40% Vs 17.6%, P=0.056) and significantly longer PFS(7.6 m Vs 3.8 m, P=0.045) than these with a mesenchymal phenotype. However, the response rate(85.7% Vs 64.7%, P=0.249) and PFS(15.2 m Vs 12.2 m, P=0.420) were similar in the patients with activated EGFR mutation. Conclusions: Patients with an epithelial phenotype got more benefit from the treatment of EGFR-TKIs in the advanced EGFR wild type NSCLC patients, which indicated that the EMT might be a potential marker to guide the individual therapy of EGFR-TKIs in this subpopulation.

Adoptive cellular immunotherapy (ITx) targeting various types of carcinoma including the lung is widely used, yet the clinical effectiveness has not been evaluated in large cases of lung carcinoma. So we carried out a case-control study to investigate the effect of ITx for the treatment of advanced non-small cell lung cancer (NSCLC) patients in Japanese population. Between 2001 and 2006, 163 patients were identified as having advanced NSCLC with ECOG performance status (PS) 0/1 who underwent ITx with or without standard chemotherapy (CTx). The controls were 232 advanced NSCLC patients with PS 0/1 who had standard CTx or BSC alone. Overall survival (OS) and 1- and 2-year survival were obtained with Kaplan-Meyer survival curves and compared using log-rank and generalized Wilcoxon analysis. Multivariate analysis with Cox proportional hazard models was also performed to analyze survival. The mean age of total population was 64.8 + 10.6 years. There were 251 males and 144 females and 295 adenocarcinoma (Ad) and 74 squamous cell carcinoma (Sq) patients. The mean period and frequency of ITx was 10.2 months and 8.7 times, respectively. Median OS for CTx and ITx + CTx was 15.7 and 20.8 months, respectively. Multivariate analysis with Cox proportional hazard model showed that sex (male vs female) (HR=0.33, p&amp;lt;0.001) and histology (Ad vs Sq) (HR=0.14, p=0.045) were independent predictors of OS. It is revealed that ITx when administered as monotherapy was effective compared to BSC in male with Sq (HR=0.47, p=0.038) and female with Ad (HR=0.53, p=0.016). Additional effect of ITx on CTx was obtained in Ad histology, especially in female (p=0.039). The present findings suggest that ITx may contribute to better survival for advanced or metastatic NSCLC patients under certain circumstances. These results should be confirmed in large prospective clinical trials. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5523. doi:10.1158/1538-7445.AM2011-5523

JCES01.09 A Comparison of ddPCR and ARMS for Detecting EGFR T790M Status from Advanced NSCLC Patients with Acquired EGFR-TKI Resistance

e21159 Background: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) are the standard therapy for EGFR-mutated non-small cell lung cancer (NSCLC) patients. Unfortunately, patients eventually develop resistance to EGFR-TKI and disease progression. Re-exposure after a drug-free interval may be an alternative to overcome tumor resistance. We performed a systematic review and meta-analysis to assess EGFR-TKI retreatment’s efficacy in advanced NSCLC. Methods: We systematically searched PubMed, Embase, and Cochrane databases for clinical trials and observational cohort studies evaluating EGFR-TKI retreatment in advanced NSCLC patients. We aimed to assess the objective response rate (ORR), disease control rate (DCR), and survival outcomes. Subgroup analyses were performed according to the type of EGFRmutation and the TKI drug used in retreatment. We further stratified studies to assess the efficacy of rechallenging with the same drug used initially or a different one. Heterogeneity was assessed using the Cochran Q test, and I2 statistics and random effects models were fitted. Results: We included 16 studies (7 prospective clinical trials, and 9 retrospective cohorts) with 806 patients. Most of them had adenocarcinoma (70.8%), were females (58.1%), and were non-smokers (74.5%). The most frequently TKI given as the initial treatment was gefitinib (58.1%), whereas in the rechallenge it was erlotinib (36.6%) followed by gefitinib (31.6%). In a pooled analysis of patients who were retreated with TKI, the median PFS was 4.1 months (95%CI 3.0 - 4.4), and OS was 12.6 months (95%CI 10.2 - 12.6). ORR was 16% (95%CI 10 - 22%) and DCR was documented in 63% (95%CI 0.54 - 0.72%). Patients harboring a sensitive EGFR mutation had a significantly higher DCR, compared to patients with EGFRT790M mutation, and those with unknown mutational status, (DCR: 70%, 62%, and 48%, respectively, p = 0.02). Patients rechallenged with gefitinib, erlotinib, or afatinib had a similar DCR of 60%, while patients re-exposed to osimertinib had a greater DCR of 70% (95%CI 59 - 80%), (p &lt; 0.01). Regarding ORR, no significant difference was observed amongst the groups defined by the type of EGFR mutation (p = 0.74) or type of TKI used (p = 0.05). Rechallenge using the same versus a different TKI resulted in similar ORR and DCR. In a subgroup analysis of 102 patients who had disease control with the first TKI, 62% (95%CI 45 – 79%) achieved disease control with TKI rechallenge. Conclusions: Our meta-analysis suggests that a subgroup of advanced EGFR-mutated NSCLC patients who failed TKI treatment benefit from rechallenge with an EGFR-TKI after a TKI-free interval. Re-exposure with either the same or a different TKI was shown to be equally effective. Patients treated with osimertinib and those with EGFR-sensitive mutations have better responses to the treatment.

Symptom burden and quality of life in advanced non-small cell lung cancer patients in France and Germany

Association of molecular characteristics with survival in advanced non-small cell lung cancer patients treated with checkpoint inhibitors

Objective To study the effects of Huangqi Injection on hematological system in advanced nonsmall-cell lung cancer (NSCLC)patients after chemotherapy. Methods A total of 100 NSCLC patients were randomly recruited into a control group (40 patients) and a treatment group (60 patients). The control group was treated with MVP project (chemotherapy with mitomycin, vindesine, and platinol) and orally taken with ondansetron, batilol, and leucogen; while the treatment group was treated with Huangqi Injection on the basis of the control group. Therapeutic effect was compared between the two groups after the treatment. Results There was a significant decrease of white blood cell count and platelet count in the treatment group. The occurrence of aneamia in the treatment group was significantly lower than the control group (P＜0.05) . Conclusion Huangqi Injection has a obvious effects in reducing toxic and side effects of chemotherapy in NSCLC patients. Key words: Huangqi injection; Advanced nonsmall cell lung cancer; Hematological system