Late-stage Non-small Cell Lung Research Articles

Next-Generation Sequencing with Liquid Biopsies from Treatment-Naïve Non-Small Cell Lung Carcinoma Patients.

Simple SummaryScreening for genomic alterations in treatment-naïve non-small cell lung carcinoma (NSCLC) is mainly done by tissue biopsy (TB), an invasive approach. However, it may not be possible to obtain a TB, the patient does not consent to it and/or the extracted nucleic acids are of poor quantity and/or quality for further genomic analyses, so a liquid biopsy (LB) is the only option to detect molecular target(s) for first-line treatment in these patients. However, a LB at diagnosis is still not often used in clinical centers since a TB is currently the gold standard approach for histological diagnosis, assessment of the PD-L1 status on tumor cells and evaluation of the molecular alterations. A number of different approaches are already available for the assessment of genetic abnormalities with LB, but next-generation sequencing (NGS) is the most promising. This review provides an overview of the main studies currently using LB NGS at diagnosis for NSCLC. We discuss its advantages and limitations in comparison with a TB and the perspectives for the future.Recently, the liquid biopsy (LB), a non-invasive and easy to repeat approach, has started to compete with the tissue biopsy (TB) for detection of targets for administration of therapeutic strategies for patients with advanced stages of lung cancer at tumor progression. A LB at diagnosis of late stage non-small cell lung carcinoma (NSCLC) is also being performed. It may be asked if a LB can be complementary (according to the clinical presentation or systematics) or even an alternative to a TB for treatment-naïve advanced NSCLC patients. Nucleic acid analysis with a TB by next-generation sequencing (NGS) is gradually replacing targeted sequencing methods for assessment of genomic alterations in lung cancer patients with tumor progression, but also at baseline. However, LB is still not often used in daily practice for NGS. This review addresses different aspects relating to the use of LB for NGS at diagnosis in advanced NSCLC, including its advantages and limitations.

P59.05 Multi-Omic Analysis Between Tumor Tissues from Early and Late Stage Non-Small Cell Lung Cancer Patients

P59.05 Multi-Omic Analysis Between Tumor Tissues from Early and Late Stage Non-Small Cell Lung Cancer Patients

1800P Concordance between treatment-naive tissue and circulating tumour DNA (ctDNA) in late stage non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC)

1800P Concordance between treatment-naive tissue and circulating tumour DNA (ctDNA) in late stage non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC)

Abstract 6011: Mesenchymal stem cells promote PD-L1 expression in lung cancer cells

Abstract Background Effects of mesenchymal stem cells (MSCs) on tumors have been extensively studied in recent years, but results still remain controversial. In this study we investigated the roles of MSCs on the expression of programmed death-ligand 1 (PD-L1) in lung cancer cells. Since overexpression of PD-L1 has been found to attenuate body's immune system against tumor cells, PD-1/PD-L1 blockages have become the most effective immunotherapy to the late stage non-small cell lung cancer (NSCLC). Therefore, understanding the effects of MSCs on PD-L1 expression is critical in managing immunotherapy for NSCLC patients. Method PD-L1 expressions were tested in two NSCLC cell lines, A549 and H1299 using qRT-PCR and Western Blot to qualitatively measure mRNA and protein levels, respectively. MSCs were isolated from umbilical cord blood and co-cultured with A549 and H1299 cells in complete medium using Transwell filters for 24-h, 48-h and 72-h. Then both A549 and H1299 cells were collected and PD-L1 expressions were tested. MSC-Conditioned Medium (MSC-CM) was generated by culturing 80% confluence of MSCs with serum-free medium. After 48 h, the medium was collected as MSC-CM. Both A549 and H1299 were then cultured in MSC-CM for 24-h, 48-h and 72-h, and PD-L1 expressions were measured from these two cell lines separately. Result Both MSCs and MSC-CM significantly increased the PD-L1 mRNA levels in the two NSCLC cells, while the MSC-CM had stronger stimulation to PD-L1 expressions (P < 0.0001) comparing to MSCs treatment (P<0.01). A549 cells reached the most significant effect at 48-h, while H1299 cells attended the same effect at 24-h treatment. The similar results were observed with PD-L1 protein levels. Both cell lines showed increased PD-L1 protein expressions after treating with MSCs and MSC-CM, but the augmented scales were smaller than the mRNA levels. The PD-L1 protein expressions in A549 cells increased significantly (p < 0.05) at 48-h when treating with MSCs, while increased much higher when treating with MSC-CM (P < 0.01). H1299 cells also showed the similar increase in PD-L1 protein expressions after 24-h treatment. Conclusion Both MSCs and MSC-CM significantly increase PD-L1 expressions in lung cancer cells, which ultimately attenuate the immune system against tumor cells. Citation Format: Hui Zhao, Hui Yang, Jinqiu Sun, Guoqiang Liang, Qianqian Zhao, Xiufang Zhang, Shengxian Liang, Rui Guo, Li Zhong. Mesenchymal stem cells promote PD-L1 expression in lung cancer cells [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 6011.

Non-invasive Detection of Exosomal MicroRNAs via Tethered Cationic Lipoplex Nanoparticles (tCLN) Biochip for Lung Cancer Early Detection.

Circulating microRNAs carried by exosomes have emerged as promising diagnostic biomarkers for cancer because of their abundant amount and remarkable stability in body fluids. Exosomal microRNAs in blood are typically quantified using the RNA isolation-qRT-PCR workflow, which cannot distinguish circulating microRNAs secreted by cancer cells from those released by non-tumor cells, making it potentially less sensitive in detecting cancer-specific microRNA biomarkers. We have developed a sensitive and simple tethered cationic lipoplex nanoparticles (tCLN) biochip to detect exosomal microRNAs in human sera. The tCLN biochip allows the discrimination of tumor-derived exosomes from their non-tumor counterparts, and thus achieves higher detection sensitivity and specificity than qRT-PCR. We have demonstrated the clinical utility of the tCLN biochip in lung cancer diagnosis using sera from normal controls, therapy-naive early stage and late stage non-small cell lung cancer (NSCLC) patients. Total five microRNAs (miR-21, miR-25, miR-155, miR-210, and miR-486) were selected as the biomarkers. Each microRNA biomarker measured by tCLN assay showed higher sensitivity and specificity in lung cancer detection than that measured by qRT-PCR. When all five microRNAs were combined, the tCLN assay distinguished normal controls from all NSCLC patients with sensitivity of 0.969, specificity of 0.933 and AUC of 0.970, and provided much better diagnostic accuracy than qRT-PCR (sensitivity = 0.469, specificity = 1.000, AUC = 0.791). Remarkably, the tCLN assay achieved absolute sensitivity and specificity in discriminating early stage NSCLC patients from normal controls, demonstrating its great potential as a liquid biopsy assay for lung cancer early detection.

CNV Detection from Circulating Tumor DNA in Late Stage Non-Small Cell Lung Cancer Patients

While methods for detecting SNVs and indels in circulating tumor DNA (ctDNA) with hybridization capture-based next-generation sequencing (NGS) have been available, copy number variations (CNVs) detection is more challenging. Here, we present a method enabling CNV detection from a 150-gene panel using a very low amount of ctDNA. First, a read depth-based CNV estimation method without a paired blood sample was developed and cfDNA sequencing data from healthy people were used to build a panel of normal (PoN) model. Then, in silico and in vitro simulations were performed to define the limit of detection (LOD) for EGFR, ERBB2, and MET. Compared to the WES results of the 48 samples, the concordance rate for EGFR, ERBB2, and MET CNVs was 78%, 89.6%, and 92.4%, respectively. In another cohort profiled with the 150-gene panel from 5980 lung cancer ctDNA samples, we detected the three genes’ amplification with comparable population frequency with other cohorts. One lung adenocarcinoma patient with MET amplification detected by our method reached partial response to crizotinib. These findings show that our ctDNA CNV detection pipeline can detect CNVs with high specificity and concordance, which enables CNV calling in a non-invasive way for cancer patients when tissues are not available.

Abstract 2363: Targeting squamous lung tumor heterogeneity with EZH2 inhibition to improve immunotherapy responses

Abstract Immunotherapies, including those targeting the PD1/PD-L1 immune checkpoint, have become popular treatment options, but they are effective for only 20% of squamous lung cancer patients. Work in our laboratory has suggested that inhibition of the histone methyltransferase EZH2 drives a basal-like transcriptional state which includes expression of the immunotherapy target PD-L1, and work from others has demonstrated that EZH2 inhibition could influence the tumor microenvironment. Therefore, we hypothesize that combining EZH2 inhibition with anti-PD1 therapy could boost response in squamous lung cancers by targeting both the tumor and the microenvironmental heterogeneity. Our laboratory uses genetically engineered mouse models of lung cancer and patient-derived cell lines and organoids to dissect the mechanistic effects of EZH2 inhibition on tumor cells and immune cells. First, we treated a panel of squamous and adeno-squamous human NSCLC cell lines with EZH2 inhibition and observed increased expression of PD-L1 and another basal cell marker, NGFR, by flow cytometry. Similar results were obtained by treating two patient-derived organoids of squamous lung cancer with EZH2 inhibitor, suggesting that EZH2 inhibition drives squamous tumor cells into a more basal-like state which may be more susceptible to anti-PD1 immunotherapy. In our mouse models of squamous lung cancer, we noted abundant tumor-associated neutrophils, which we believe may be inhibiting efficacy of immunotherapy by suppressing CD8 T cell function. In support of the idea that inhibiting EZH2 will ameliorate this phenotype, lung neutrophils isolated from EZH2 conditional knock-out mice had significantly lower levels of the T cell suppressive protein Arginase 1 than WT lung neutrophils or Lkb1/Pten TANs. Lastly, we treated mice harboring autochthonous squamous lung tumors with EZH2 inhibitor, and observed a striking increase in the proportion of tumor cells expressing PD-L1 but now defect in tumor growth. In contrast, two squamous lung tumor bearing mice treated with EZH2 inhibitor and anti-PD-1 immunotherapy had tumor regression that was durable up to 4 weeks post-treatment initiation. We plan to expand the cohorts of mice and test this promising combination therapy in a variety of lung cancer mouse models. Completion of these studies will solidify the efficacy of a promising therapeutic combination and uncover mechanisms by which tumor hierarchies and microenvironments are changed by EZH2 inhibitors in squamous lung cancers. Given that one arm of a Phase 1/2 clinical trial combining EZH2 inhibition with anti-PDL1 just began recruiting late stage non-small cell lung cancer patients, learning the phenotypes and mechanisms of responders and non-responders will be extremely timely for any Phase 2/3 trials that ensue. Funded by V Foundation Scholar Award, K22 CA201036 and KY LCRP to CFB Citation Format: Tanner J. DuCote, Christine Fillmore Brainson. Targeting squamous lung tumor heterogeneity with EZH2 inhibition to improve immunotherapy responses [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 2363.

Abstract 189: EGFR TKI resistance via role VEGFR2

Abstract The treatment paradigm for late stage non-small cell lung carcinoma (NSCLC) has shifted towards molecular targeted therapy like epidermal growth factor receptor (EGFR). Tyrosine kinase inhibitors (TKIs) against EGFR have proven to be effective; however, TKI resistance is commonly acquired by the T790M mutation in the EGFR kinase domain. Very few studies suggest the role of vascular endothelial growth factor receptor (VEGFR) in mediating erlotinib resistance. We suggest that overexpression of VEGFR-2 in tumors creates an autocrine loop that mediates TKI resistance. Thus, our purpose is to investigate the VEGF/VEGFR autocrine loop as a potential mechanism of TKI resistance via alternative signaling. H2170, H358, H1975 and H3255 NSCLC parental cells were obtained from ATCC. Parental cells were then treated with erlotinib to create resistant cells. DNA sequencing was used to exclude the presence of the T790M mutation. Quantitative real-time PCR (qPCR) was conducted to code for VEGF, VEGFR-2, and neuropilin-1 (NP-1) genes and immunoblotting (WB) was used to detect their protein expression. Fluorescence-activated cell sorting (FACS) analysis and immunofluorescence (IF) analysis was performed with antibodies against VEGFR-2 and NP-1. Secretion of VEGF from NSCLC cells was studied at 24/48 hours by ELISA. Cell proliferation assay (MTT) was conducted to assess the viability of resistant cells after treatment with VEGFR-2 inhibitor and VEGF neutralizing antibody in media with/without erlotinib. 48 and 31 tumor biopsies were immunostained (IHC) with anti-VEGFR-2 and anti-NP-1 antibodies, respectively. qPCR showed a 1.4 to 3.5-fold increase in expression of VEGF, VEGFR-2 and NP-1 in H2170 resistant cells and a 2.2 to 3.6-fold increase in expression in H358 resistant cells in comparison to their parental cells. WB results showed an increase in protein expression in resistant cells compared to parental cells. FACS analysis showed a 1.3 to 6-fold increase in expression of VEGFR and NP-1 in resistant cells which was confirmed by IF, showing higher expression of both VEGFR-2 and NP-1 in the resistant cells. ELISA studies showed that VEGF secretion increases over time. MTT assays showed that the VEGFR-2 inhibitor (ZM HCl 323-881) lowered cell death to 10-57% (p=0.05) with erlotinib in resistant cells compared to the erlotinib alone (72-75%). Kaplan-Meier analysis revealed shorter median survival with high vs low expression of total VEGFR-2 expression (14mo vs 21mo, p=0.05) in patients NSCLC tumors. Our results suggest that by upregulating the VEGF/VEGFR autocrine system as an alternative signaling pathway to promote proliferation, tumor cells may bypass the inhibitory effect of TKIs on the EGFR pathway, ultimately overcoming resistance. Furthermore, IHC demonstrated that high expression of VEGFR-2 is associated with poor prognosis in tumors, suggesting its potential as a prognostic biomarker for lung cancer. Citation Format: Chike Osude, Leo Lin, Neelu Puri. EGFR TKI resistance via role VEGFR2 [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 189.

18P - Immunohistochemical expression of PD-L1 in early and late stage non-small cell lung cancer: Correlation with clinicopathological and molecular features

18P - Immunohistochemical expression of PD-L1 in early and late stage non-small cell lung cancer: Correlation with clinicopathological and molecular features

Therapeutic effects of Kanglaite injection combined with chemotherapy on advanced non-small cell lung cancer (NSCLC).

AbstractObjective: To study the therapeutic effect of Kanglaite injection combined with chemotherapy in the treatment of late stage non-small cell lung cancer (NSCLC), and also to observe the effect of the combination treatment on immune function. Methodology: 92 patients with advaned stage of non-small cell lung cancer who admitted to First Hospital of ZiBo city hospital from May 2017 to October 2018 were randomly divided into experimental group and control group, with 46 cases respectively. The control group was treated with chemotherapy only while the experimental group was treated with Kanglaite injection combined with chemotherapy which was the basic treatment for patients, and the total treatment effective rate, adverse reaction rate and immune index of the treatments on two groups were compared. Result: The total treatment effective rate of the experimental group was 80.43%, which was significantly higher than that of the control group, which was 63.04%. The incidence of adverse reactions in the experimental group was 36.96%, which was lower than that of the control group (78.26%). The immune indexes of the experimental group (CD3+, CD4+, IgG, IgA) were better than that of the control group, respectively. The differences between the two groups were statistically significant (P<0.05).Conclusion: During the chemotherapy of late stage or advanced non-small cell lung cancer, the addition use of Kanglaite injection has a significant effect on improving tumor control and reducing the side effects of chemotherapy, and helps to improve the immune function of patients, thus it is worth promoting.

Comparative efficacy and safety of licensed treatments for previously treated non-small cell lung cancer: A systematic review and network meta-analysis.

PurposeThis systematic review with network meta-analysis compared the efficacy and safety of currently licensed second-line treatments in patients with late stage non-small cell lung cancer (NSCLC).MethodsRandomised controlled trials (RCTs) of participants with advanced/metastatic NSCLC receiving second/third line treatments were screened. We searched electronic databases (MEDLINE; EMBASE; Web of Science) from January, 2000 to July, 2017.Two reviewers screened bibliographic records, extracted data, and assessed risk of bias of included studies. The outcomes were overall survival (OS), progression-free survival (PFS), and drug-related grade 3–5 adverse-events (AEs). We pooled study-specific hazard ratios (HR; for OS and PFS) and risk ratios (RR; for AEs) using conventional and network-meta-analyses, and ranked interventions by the surface under the cumulative ranking curve.FindingsWe included 11 RCTs (7,581 participants) comparing nine drugs. All drugs except for erlotinib significantly improved OS compared to docetaxel. Nivolumab was the highest ranking drug followed by atezolizumab and pembrolizumab. There was no significant difference in OS across these three drugs (HR = 0.98, 95% CI 0.79, 1.21 for nivolumab vs atezolizumab; HR = 0.98, 95% CI 0.77, 1.25 for nivolumab vs pembrolizumab). For PFS, ramucirumab + docetaxel and nivolumab were the drugs with the highest ranking. All interventions except ramucirumab + docetaxel had a reduced risk for severe drug-related AEs vs. docetaxel. Of the drugs with the highest ranking on AEs, nivolumab was significantly safer compared to atezolizumab (RR = 0.55, 95% CI 0.38, 0.79) or pembrolizumab (RR = 0.52, 95% CI 0.34, 0.81).ImplicationsNivolumab, pembrolizumab and atezolizumab exhibited superior benefit/risk balance compared to other licensed drugs used late stage NSCLC. Our results indicate that the use of immunotherapies in people diagnosed with non-specific late stage NSCLC should be promoted. The use of docetaxel may now be judged irrelevant as a comparator intervention for approval of new drugs for second line treatment of NSCLC.Study registration numberPROSPERO CRD42017065928.

Abstract 2595: Role of VEGF/VEGFR autocrine in mediating EGFR TKI resistance in non-small cell lung cancer

Abstract The treatment paradigm for late stage non-small cell lung carcinoma (NSCLC) has shifted towards molecular targeted therapy like epidermal growth factor receptor (EGFR), one of the most studied targets. Tyrosine kinase inhibitors (TKIs) against EGFR have proven to be effective, however TKI resistance is common due to the acquisition of the T790M mutation in the EGFR kinase domain. There are currently very few studies suggesting the role of VEGFR in mediating erlotinib resistance. We suggest that over-expression of vascular endothelial growth factor receptors (VEGFRs) in tumors may create an autocrine loop that may mediate TKI resistance. The purpose of this study is to investigate the VEGF/VEGFR autocrine loop as a potential mechanism of TKI resistance via alternative signaling. H2170 NSCLC parental cells were obtained from ATCC. The resistant cells were obtained by treating parental cells with increasing concentrations of erlotinib. DNA sequencing was performed to exclude the presence of the T790M mutation. Quantitative real-time PCR was carried out for target genes coding for VEGF, VEGFR-2, and neuropilin-1 (NP-1). Fluorescence-activated cell sorting (FACS) analysis and immunofluorescence (IF) analysis were performed with antibodies against VEGFR-2 and NP-1. The viability of resistant cells was assessed after being treated with increasing concentrations of a VEGF inhibitor (VEGF neutralizing antibody) in medium with or without erlotinib using a cell proliferation assay (MTT). Thirty paraffin embedded tumor biopsies were immunostained (IHC) with an antibody against VEGFR-2. On the transcriptional level, real-time PCR revealed a 13.1, 3.4 and a 2.9 fold increase in gene expression for VEGF, VEGFR-2 and NP-1 respectively in the resistant cells compared to the parental cells. When looking at membrane bound receptors, FACS analysis showed a 2.1 and a 3.8 fold increase in the expression of VEGFR and NP-1 in resistant cells, respectively. IF studies correlated with the FACS results, showing significantly higher expression of both VEGFR-2 and NP-1 in the resistant cells. MTT assays demonstrated that the VEGF inhibitor was able to decrease cell death from 10 to 80% (0.1-10ug/ml) as evidenced by decreasing viability of resistant cells treated with increasing concentrations of VEGF inhibitor compared to the untreated control. Kaplan-Meier analysis revealed shorter median survival with high vs low expression of total VEGFR-2 expression (7mo vs 25mo, p=0.05) in NSCLC tumors in patients diagnosed with late stage NSCLC. Our results suggest that by up-regulating the VEGF/VEGFR autocrine system as an alternative signaling pathway to promote proliferation, tumor cells may bypass the inhibitory effect of TKI's on the EGFR pathway, leading to resistance. Furthermore, IHC demonstrated the presence of high expression of VEGFR-2 in tumors, suggesting its potential as a prognostic biomarker for lung cancer. Citation Format: Chike Osude, Leo Lin, Brad Foster, Neelu Puri. Role of VEGF/VEGFR autocrine in mediating EGFR TKI resistance 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 2595.

Exosomal lipids for classifying early and late stage non-small cell lung cancer

Lung cancer is the leading cause of cancer deaths in the United States. Patients with early stage lung cancer have the best prognosis with surgical removal of the tumor, but the disease is often asymptomatic until advanced disease develops, and there are no effective blood-based screening methods for early detection of lung cancer in at-risk populations. We have explored the lipid profiles of blood plasma exosomes using ultra high-resolution Fourier transform mass spectrometry (UHR-FTMS) for early detection of the prevalent non-small cell lung cancers (NSCLC). Exosomes are nanovehicles released by various cells and tumor tissues to elicit important biofunctions such as immune modulation and tumor development. Plasma exosomal lipid profiles were acquired from 39 normal and 91 NSCLC subjects (44 early stage and 47 late stage). We have applied two multivariate statistical methods, Random Forest (RF) and Least Absolute Shrinkage and Selection Operator (LASSO) to classify the data. For the RF method, the Gini importance of the assigned lipids was calculated to select 16 lipids with top importance. Using the LASSO method, 7 features were selected based on a grouped LASSO penalty. The Area Under the Receiver Operating Characteristic curve for early and late stage cancer versus normal subjects using the selected lipid features was 0.85 and 0.88 for RF and 0.79 and 0.77 for LASSO, respectively. These results show the value of RF and LASSO for metabolomics data-based biomarker development, which provide robust an independent classifiers with sparse data sets. Application of LASSO and Random Forests identifies lipid features that successfully distinguish early stage lung cancer patient from healthy individuals.

Efficacy and safety of crizotinib in patients with anaplastic lymphoma kinase-positive stage IV non-small cell lung cancer.

e20516 Background: Crizotinib is an oral small-molecule tyrosine kinase inhibitor targeting ALK, MET, and ROS1 tyrosine kinases. It was first approved by the Food and Drug Administration (FDA) in 2011 for treatment of patients in late-stage (locally advanced or metastatic) non-small cell lung cancer (NSCLC) with abnormal anaplastic lymphoma kinase (ALK) gene as detected by an FDA-approved test, as it confers improved progression-free survival comparison with chemotherapy. Methods: This is a retrospective chart review study of patients with stage IV ALK-positive NSCLC who attended the medical oncology department of Kuwait Cancer Control Centre (Kuwait) and King Fahad Specialist Hospital (Saudi Arabia) between January 2013 and May 2016. The efficacy and safety of crizotinib (250 mg orally, twice daily, with or without food.) were evaluated based on overall survival (OS), progression-free survival (PFS), Objective response rate (ORR), time to objective response, duration of response, and dose reduction or cessation because of crizotinib toxicity. Results: Twenty-three patients were involved in this study with a median age of 55.5 years and male-to-female ratio of 2.4:1 The median OS from initiation of crizotinib has not been reached; 1-year overall survival was 71.2%. Crizotinib treatment demonstrated median PFS of 9.6 months (95% CI, 3.0-24.1 months). The ORR was 70.9%. Complete response was achieved in 4.2% of patients, and partial response was achieved in 66.7% of patients. The most frequently reported adverse effects of crizotinib (Grade 1 / 2) were muscle ache, fatigue, nausea, vomiting, diarrhea, constipation, edema, elevated transaminases, bradycardia, and vision disorders. Grade 3 / 4 diarrhea was reported in 12.5% of patients. The proportion of patients who required dose reduction because of Crizotinib-induced bradycardia was 8.3% of patients. Conclusions: Crizotinib appears to be a very favorable option for treating patients with stage IV ALK-positive NSCLC. Crizotinib showed a very tolerable toxicity profile.

Influence of PD-1 inhibitor pembrolizumab on T lymphocyte subsets and NK cells in patients with late stage non-small cell lung cancer

Objective To observe the changes of T lymphocyte subsets and natural killer (NK) cells in patients with late stage non-small cell lung cancer (NSCLC) before and after treatment with PD-1 inhibitor and its clinical effect. Methods Totally 23 patients with NSCLC in Guangzhou Modern Hospital from January 2015 to January 2016 were collected. All patients were given 6 cycles of PD-1 inhibitor treatment after chemotherapy or targeted drug treatment failure. Peripheral venous blood was collected before and after treatment to detect the percentage of CD3+ , CD4+ , CD8+ and NK cells in peripheral blood lymphocytes. The curative effects were evaluated by chest CT after treatment of 2, 4, 6 cycles. Results Compared with before treatment, the proportions of CD3+ (69.56%±7.81% vs. 63.91%±6.43%, t=2.679, P=0.005), CD4+ (39.01%±4.98% vs. 36.09%±4.77%, t=2.031, P=0.024) and CD4+ /CD8+ (1.82±0.48 vs. 1.49±0.32, t=2.743, P=0.004) were increased after treatment, with significant differences. While compared with before treatment, the proportions of CD8+ (24.08%±5.13% vs. 26.04%±6.44%, t=1.142, P=0.130) and NK cells (22.68%±9.56% vs. 21.45%±10.01%, t=0.426, P=0.337) had little changes, with no significant differences. There were 3 patients with complete remission, 10 patients with partial remission, 8 patients with stable disease and 2 patients with progressive disease when completing 6 cycles of PD-1 inhibitor treatment. Ten patients showed untoward effects such as mild sleepiness, thirst, tussis, pruritus and rash, and they were well tolerable. Conclusion PD-1 inhibitor can improve the patient′s cellular immune function, and can achieve a more satisfactory short-term efficacy and acceptable adverse reactions, which maybe bring new hopes for patients with NSCLC. Key words: Immunosuppressive agents; Carcinoma, non-small-cell lung; T lymphocyte subsets; Treatment outcome; PD-1 inhibitor

QUALITY OF LIFE OF LATE STAGE NON-SMALL CELL LUNG CANCER PATIENTS

Background: Lung cancer is the most common malignancies and remains the leading cause of cancer-related deaths in Viet Nam. Majority of cases present initially at late stage. Palliative chemotherapy is the standard treatment for these situations to prolong survival and improve quality of life for the patient. Purpose: To appreciate quality of life in patients in late stage non-small cell lung cancer pre-post, during palliative chemotherapy and to determine the factors affecting on post-chemotherapy quality of life. Patients and Methods: A prospective, descriptive study, eligible patients included 65 late stage non-small cell lung cancer patients from Hue University Hospital from 1/2014 to 6/2016. The EORTC QLQ-C30 and Lung cancer Questionnaire EORTC QLQ-LC13 were used to assess quality of life. T-test was used to compare quality of life score at two assessed times. T-test, ANOVA, Mann Whitney, Kruskal Wallis were used to determine the correlation between 2 factors. Pearson and Spearman Coefficient were used to measure the strength of relationship between the factors. Results: The most effected age group was 54.4 ± 11.3. The global health scale before treatment was 47.3± 23.6, the functional scales as well as disease-related symptom scales improved clearly after the 2nd cycle (64.8 ± 16.0) and were relatively stable at the 4th cycle of chemotherapy (62.2 ± 19.3). Meanwhile, the toxicities including hair loss and peripheral neuropathy rose gradually after chemotherapy cycles. Age, occupation, nationality, religion, weight loss, PS, pathology, stage of disease, chemotherapy regimen, progressive disease, disease-related symptoms and treatment-related toxicities associated post-chemotherapy quality of life. Conclusions: This study showed that there were an improvement of quality of life after chemotherapy. Epidemiologic, clinical, treatment factors had effects on post-chemotherapy quality of life. Key words: lung cancer, quality of life, palliative chemotherapy

Abstract 1350: Genetic variations in apoptosis pathway are associated with survival in late stage non-small cell lung cancer.

Abstract Dysregulation of apoptosis pathway plays an essential role in carcinogenesis, tumor progression, and resistance to chemotherapy. We hypothesized that genetic variations in genes involved in the apoptosis pathway may affect the survival in late stage non-small cell lung cancer (NSCLC) patients. To test this hypothesis, we genotyped 583 tagging SNPs in 598 late stage NSCLC patients and analyzed their associations with survival individually and jointly. Seven SNPs were statistically significantly associated with overall survival after correction for multiple comparisons (q<0.1). The most significant SNP was rs17592236 on FOXO1 gene under the dominant model (OR: 2.54, 95%CI: 1.71-3.78, p=4.15x10−6). We also observed a significant cumulative effect. Compared to the reference group (patients with no more than two risk alleles), those having five or more risk alleles were much more likely to die within seven months (HR: 3.58, 95% CI: 2.55-5.04, p<0.001). Notably, the median survival time (MST) was significantly reduced by up to 15 months in high risk patients when compared to the reference group (MST: 22.2 months, p<0.001). Survival tree analysis revealed potential higher order gene-gene interactions among these SNPs. Our results provided evidence that genetic variations in the apoptosis pathway modulate NSCLC survival and may serve as biomarkers to differentiate NSCLC patients with different prognosis. Citation Format: Xiang Shu, Charles Lu, Joe Y. Chang, Jack A. Roth, Xifeng Wu. Genetic variations in apoptosis pathway are associated with survival in late stage non-small cell lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1350. doi:10.1158/1538-7445.AM2013-1350

Poster - Thur Eve - 53: Analysis of the distribution of dose delivery during respiratory-gated step-and-shoot IMRT for lung cancer radiotherapy.

Respiratory motion is a large source of dosimetric error when treating lung cancer with Intensity Modulated Radiation Therapy (IMRT). The asynchronicity of the tumour motion and the multileaf collimator (MLC) used to modulate the radiation beam intensity, leads to the interplay effect. One method to account for this effect is respiratory gating. Treatment planning optimization for gated IMRT is performed on a subset average 4D-CT which includes the phases surrounding end exhalation. However, this assumes that the beam delivery will be evenly distributed amongst those phases. This study investigates the distribution of beam delivery during gated step-and-shoot IMRT (SS-IMRT) for both early and late stage non-small cell lung cancer (NSCLC). Four Stage I NSCLC patients, prescribed a dose of 54 Gy in 3 fractions, and five Stage III NSCLC patients, prescribed a dose of 60 Gy in 30 fractions, were retrospectively planned with high and low modulation beams-IMRT, and delivered using the QUASAR™ Programmable Respiratory Motion Platform with 15 mm and 20 mm peak-to-peak sinusoidal motion and real patient breathing motion. The percent monitor units delivered at each phase were compared. For Stage I patients, the monitor units delivered were evenly distributed over the gating window due to a high number of monitor units delivered per control point. For Stage III patients, as the complexity of SS-IMRT increases, there were more monitor units delivered in the initial gating phase. This dose discrepancy could potentially lead to geographic miss of the tumour and should be taken into account during treatment planning.

Fluid biopsy for circulating tumor cell identification in patients with early-and late-stage non-small cell lung cancer: a glimpse into lung cancer biology

Circulating tumor cell (CTC) counts are an established prognostic marker in metastatic prostate, breast and colorectal cancer, and recent data suggest a similar role in late stage non-small cell lung cancer (NSCLC). However, due to sensitivity constraints in current enrichment-based CTC detection technologies, there are few published data about CTC prevalence rates and morphologic heterogeneity in early-stage NSCLC, or the correlation of CTCs with disease progression and their usability for clinical staging. We investigated CTC counts, morphology and aggregation in early stage, locally advanced and metastatic NSCLC patients by using a fluid-phase biopsy approach that identifies CTCs without relying on surface-receptor-based enrichment and presents them in sufficiently high definition (HD) to satisfy diagnostic pathology image quality requirements. HD-CTCs were analyzed in blood samples from 78 chemotherapy-naïve NSCLC patients. 73% of the total population had a positive HD-CTC count (>0 CTC in 1 mL of blood) with a median of 4.4 HD-CTCs mL−1 (range 0–515.6) and a mean of 44.7 (±95.2) HD-CTCs mL−1. No significant difference in the medians of HD-CTC counts was detected between stage IV (n = 31, range 0–178.2), stage III (n = 34, range 0–515.6) and stages I/II (n = 13, range 0–442.3). Furthermore, HD-CTCs exhibited a uniformity in terms of molecular and physical characteristics such as fluorescent cytokeratin intensity, nuclear size, frequency of apoptosis and aggregate formation across the spectrum of staging. Our results demonstrate that despite stringent morphologic inclusion criteria for the definition of HD-CTCs, the HD-CTC assay shows high sensitivity in the detection and characterization of both early- and late-stage lung cancer CTCs. Extensive studies are warranted to investigate the prognostic value of CTC profiling in early-stage lung cancer. This finding has implications for the design of extensive studies examining screening, therapy and surveillance in lung cancer patients.

Treatment of Late Stage Non-small Cell Lung Cancer: We Have More Work to Do

Treatment of Late Stage Non-small Cell Lung Cancer: We Have More Work to Do