Adenocarcinoma Lung Cancer Patients Research Articles

Investigating and Modeling the Differential DNA Methylation for Early Lung Adenocarcinoma Diagnosis

Background: Aberrant DNA methylations serve as rich sources of diagnostic biomarkers, but a further improvement in their accuracy and clinical utility is warranted. Methods: Large panel bisulfite sequencing was performed on paired normal and stage I/IV tumors from 226 lung adenocarcinoma cancer patients to characterize the differentially methylated regions (DMRs). Results: Random forest model achieved high prediction accuracy (sensitivity 96% and specificity 97.56%) to separate normal controls from both early and advanced cancer samples, which is superior to most previous prediction models tested in lung adenocarcinoma. Conclusion: Our results suggest that combining the random forest model with targeted bisulfite sequencing have great clinical potential to accurately predict and diagnose lung adenocarcinoma early during cancer screening.

Factors Affecting the Treatment Success of Short-Term Regimen for Drug Resistant Tuberculosis (DR TB) Patients at Dr. Saiful Anwar General Hospital Malang

Background: Epidermal growth factor (EGFR) is a transmembrane receptor that plays an important role in the development of cancer phenotypes. Some patients with lung cancer have genetic mutations. Several studies have found a close correlation of EGFR gene mutation with 15-20% of lung adenocarcinoma cases. This study aimed to determine the EGFR profile in adenocarcinoma lung cancer patients at Haji Adam Malik General Hospital.Methods: This was a prospective cohort study conducted at Haji Adam Malik General Hospital, Medan. This study used patients data for 3 years, starting from January 1, 2014 to D1ecember 31, 2016. The sample size in this study was 34 patients. The data were then analyzed using SPSS.Results: All patients were adenocarcinoma lung cancer patients with positive EGFR mutation. There were 9 subjects with EGFR mutation in exon 19; 11 subjects with exon 21 L858R mutation; and 3 subjects with exon 21 L861Q mutation. Meanwhile, there were 3 subjects with uncommon EGFR mutations, namely exon 18 mutation. Majority of subjects with exon 19 mutation were male, aged >60 years, smokers with mixed types of cigarettes and severe brinkmann index. In subjects with exon 21 L858R mutation, most of the patients were male, aged 50-60 years, smokers with mixed cigarette types and severe brinkmann index. The same characteristics were also observed in subjects with exon 18 mutation. However, for exon 21 L861Q mutation, the majority of subjects were female with varying ages, and were not smokers. Conclusion: Most of the study subjects profiles were male, aged over 60 years, smokers, with mixed types of cigarettes, and with severe Brinkman Index. The EGFR mutations most commonly occured in exon 21, followed by exon 19 (ins/del exon 19), exon 18, and a combination of 2 exons.

Abstract 5785: Tumors with higher heterogeneity were associated with poorer survival outcome in early stage lung adenocarcinoma cancer patients with low tumor mutational burden

Abstract Background: Response to immune checkpoint inhibitors is associated with tumor mutational burden (TMB). Pembrolizumab has recently been approved by the FDA for use in patients with high TMB. However low TMB patients are not considered good responders and could benefit from stratification. The predictive value of other biomarkers such as intratumor heterogeneity (ITH) has been studied across a range of cancer indications. The utility of ITH in Lung Adenocarcinoma (LUAD) has not been established. Here we evaluate the relationship between ITH and outcome in early stage low TMB patients and explore differences in immune cell enrichment and predicted driver genes. Stratifying early stage patients may yield an opportunity to improve Recurrence Free Survival (RFS) and lower the risk of progression to late stage disease. Methods: All molecular data was obtained from the Tumor Cancer Genome Atlas via the GDC Data Portal. Clinical data was from the PanCancer Atlas dataset through cBioPortal. We estimated ITH using PyClone, an algorithm that estimates the number of tumor clones. We generated our ITH values in triplicate across 3 separate days, yielding an average PyClone clone number. Clones with >2 mutations were counted. We used p-value minimization to choose 1.4 for the TMB cutoff and 2.6 for the PyClone cutoff. We generated 4 groups: HTHI, HTLI, LTHI, and LTLI, where H - high, L- low, T-TMB, and I-ITH. Immune cell enrichment analysis was obtained from the xCell webtool. Driver gene prediction was performed with OncoDriveClust via the R package maftools. Results: We found a statistically significant difference in RFS between early stage (Stage I and Stage II) LTHI (N = 18) and LTLI (N = 40) patients (16.69 months vs. 33.27 months, p-value = 0.0056). Interestingly the predicted driver genes did not significantly differ between these two groups. One driver (EGFR) and two drivers (EGFR and TP53) genes were predicted for LTHI and LTLI, respectively. Gene set enrichment analysis showed statistically significant differences in key immune subtypes. B-cells were depressed and Th1 cells were elevated in LTHI, both of which are associated with poorer survival outcomes. Conclusion: We found an association between high ITH and poor survival in low TMB early stage LUAD patients. Early stage patients can be stratified using heterogeneity metrics like PyClone. Given the presence of immune subtypes known to associate with poorer outcomes, early stage LTHI patients may benefit from additional adjuvant therapies. Citation Format: Stanislav Fridland, Young Kwang Chae. Tumors with higher heterogeneity were associated with poorer survival outcome in early stage lung adenocarcinoma cancer patients with low tumor mutational burden [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5785.

One Year Survival of Wild-Type Adenocarcinoma Lung Cancer Patients Receiving Chemotherapy at dr. Saiful Anwar Hospital, Malang

Background: Lung cancer is one of the most common malignancies that leads to mortality. In Indonesia, lung cancer ranks first in men and third in women. The most common histological type of lung cancer is adenocarcinoma. Adenocarcinoma lung cancer is divided into 2 types, namely EGFR mutations and no mutations (wild-type). Chemotherapy is the treatment of choice for advanced wild-type adenocarcinoma lung cancer. This study aimed to assess the one-year survival of wild-type adenocarcinoma lung cancer patients receiving chemotherapy. Method: This study used a cross-sectional study design.Data were taken from the medical records of cancer patients at Dr. Saiful Anwar Hospital Malang in 2018-2019. Data were processed and analyzed by chi-square test. Results: Of the 54 subjects, 24 patients received carboplatin/ pemetrexed (44.4%), 15 patients received carboplatin/paclitaxel (27.8%), 9 patients received carboplatin/gemcitabine (16.7%), 2 patients received pemetrexed (3.7%), and 4 patients received gemcitabine (7.4%). The chemotherapy drug regimen had no correlation with one-year survival (P=0.899). Conclusion: There wasno significant difference between one-year survival andchemotherapy drug regimens. This study required a larger sample to minimize bias.

Nucleolin Overexpression Predicts Patient Prognosis While Providing a Framework for Targeted Therapeutic Intervention in Lung Cancer

Simple SummaryDespite the clinical benefit of new anticancer therapies, such as immune checkpoint inhibitors, lung cancer remains the most frequent cause of cancer-related death worldwide, thus supporting the need to develop novel anticancer treatments. Endothelial cells of the tumor-associated vasculature are easily accessible to drugs administered intravenously, besides having greater genetic stability than neoplastic cells and thus lowering the risk of developing drug resistance. In this respect, the identification of alternative targets, and therapeutic strategies, within the tumor vasculature is of high relevance. Accordingly, this work aimed at characterizing nucleolin expression in patient-derived pulmonary carcinomas and further validating nucleolin as a novel target to mediate successful therapeutic interventions against human lung cancers. The highlighted prognostic value of nucleolin points towards the applicability of nucleolin-based targeting strategies against nucleolinhigh pulmonary carcinomas, present in every disease stage, in a clinical trial setting.Notwithstanding the advances in the treatment of lung cancer with immune checkpoint inhibitors, the high percentage of non-responders supports the development of novel anticancer treatments. Herein, the expression of the onco-target nucleolin in patient-derived pulmonary carcinomas was characterized, along with the assessment of its potential as a therapeutic target. The clinical prognostic value of nucleolin for human pulmonary carcinomas was evaluated through data mining from the Cancer Genome Atlas project and immunohistochemical detection in human samples. Cell surface expression of nucleolin was evaluated by flow cytometry and subcellular fraction Western blotting in lung cancer cell lines. Nucleolin mRNA overexpression correlated with poor overall survival of lung adenocarcinoma cancer patients and further predicted the disease progression of both lung adenocarcinoma and squamous carcinoma. Furthermore, a third of the cases presented extra-nuclear expression, contrasting with the nucleolar pattern in non-malignant tissues. A two- to twelve-fold improvement in cytotoxicity, subsequent to internalization into the lung cancer cell lines of doxorubicin-loaded liposomes functionalized by the nucleolin-binding F3 peptide, was correlated with the nucleolin cell surface levels and the corresponding extent of cell binding. Overall, the results suggested nucleolin overexpression as a poor prognosis predictor and thus a target for therapeutic intervention in lung cancer.

Carboxypeptidase E mRNA: Overexpression predicts recurrence and death in lung adenocarcinoma cancer patients.

Effective biomarkers for prediction of recurrence of lung adenocarcinoma cancer (LADC) patients are needed to determine treatment strategies post-surgery to improve outcome. This study evaluates the efficacy of carboxypeptidase E (CPE) mRNA including its splice isoforms, CPE-ΔN, as a biomarker for predicting recurrence in adenocarcinoma patients. RNA was extracted from resected tumors from 86 patients with different stages of non-small cell LADC. cDNA was synthesized and qRT-PCR carried out to determine the copy numbers of CPE/CPE-ΔN mRNA. Patients were followed for 7years post-tumor resection to determine recurrence and death. ROC curve analysis showed the overall AUC for CPE/CPE-ΔN copy number was 0.563 in predicting recurrence and 0.562 in predicting death. Kaplan-Meier survival analysis showed statistical difference (p= 0.018), indicating that patients with high CPE/CPE-ΔN copy numbers had a shorter time of disease-free survival and also shorter time to death (p= 0.035). Subgroup analyses showed that association of disease-free survival time with CPE/CPE-ΔN copy number was stronger among stage I and II LADC patients (p= 0.047). CPE/CPE-ΔN mRNA is a potentially useful biomarker for predicting recurrence and death in LADC patients, especially in identifying patients at high risk of recurrence at early stages I and II.

Karakteristik Mutasi EGFR pada Pasien Kanker Paru Jenis Adenokarsinoma yang Diterapi dengan Tyrosien Kinase Inhibitor (TKI) di Rumah Sakit Paru Dr. H.A Rotinsulu Bandung

This descriptive study describes the proportion of Epidermal Growth Factor Receptor (EGFR) mutations in lung cancer Adenocarcinoma patients at Rumah Sakit Paru (RS Paru) Dr. H. A Rotinsulu Bandung in the range of 2018 to 2020. 
 EGFR mutations were examined using RGQ thera screen technology Real"‘Time Polymerase Chain Reaction from FFPE or cytology smear.
 There are 44 data point mutations after examining the EGFR mutation. 59% cases were 40"‘60 years old group dan 52 % were female.
 It was found that in this group 86% patient with single mutation and 16% had doube mutations. EGFR mutations occurred in Exon 21 L858R in 13 cases (30 %), Exon 21 L861Q in 1 case (2%), Exon 19del in 24 cases (55%), double mutation Exon 19 and Exon 20 T790M type in 2 cases (4%), double mutation Exon 19del and Exon 21 in 4 cases (9%).

KLRK1 as a prognostic biomarker for lung adenocarcinoma cancer

Lung cancer is one of the most common malignancy worldwide and causes estimated 1.6 million deaths each year. Cancer immunosurveillance has been found to play an important role in lung cancer and may be related with its prognosis. KLRK1, encoding NKG2D, is a homodimeric lectin-like receptor. However, there has not been one research of KLRK1 as a biomarker in lung cancer. Data including patients` clinical characteristics and RNAseq information of KLRK1 from TCGA were downloaded. A total of 1019 patients with lung cancer were included in this study, among which 407 patients were female and 611 patients were male. Evaluations of mRNA expression, diagnostic value by ROC (receiver operating characteristic) curves and prognostic value by survival curve, Cox model and subgroup analysis were performed. The level of KLRK1 expression in lung adenocarcinoma cancer tissues and normal lung tissues was detected by qRT-PCR. The CCK-8 assay investigated the proliferation rate and the wound healing assay assessed the migratory ability in vitro. The expression of KLRK1 in tumor was lower than that in normal tissue. KLRK1 expression was associated with gender, histologic grade, stage, T classification and vital status. Patients with high KLRK1 expression presented an improved overall survival (P = 0.0036) and relapse free survival (P = 0.0031). KLRK1 was found to have significant prognostic value in lung adenocarcinoma (P = 0.015), stage I/II (P = 0.03), older patients (P = 0.0052), and male (P = 0.0047) by subgroup overall survival analysis, and in lung adenocarcinoma (P = 0.0094), stage I/II (P = 0.0076), older patients (P = 0.0072), and male (P = 0.0033) by subgroup relapse free survival analysis. Lung adenocarcinoma cancer patients with high KLRK1 expression presented an improved overall survival (P = 0.015) and relapse free survival (P = 0.0094). In vitro studies indicated that KLRK1 inhibited tumor cell proliferation and migration. KLRK1 was an independent prognostic factor and high KLRK1 expression indicated a better overall and relapse free survival. KLRK1 may be a prognostic biomarker for lung adenocarcinoma cancer.

Analysis of the Single-Cell Heterogeneity of Adenocarcinoma Cell Lines and the Investigation of Intratumor Heterogeneity Reveals the Expression of Transmembrane Protein 45A (TMEM45A) in Lung Adenocarcinoma Cancer Patients.

Simple SummaryNon-small cell lung cancer (NSCLC) is one of the main causes of cancer-related deaths worldwide. Intratumoral heterogeneity (ITH) is responsible for the majority of difficulties encountered in the treatment of lung-cancer patients. Therefore, the heterogeneity of NSCLC cell lines and primary lung adenocarcinoma was investigated by single-cell mass cytometry (CyTOF). Human NSCLC adenocarcinoma cells A549, H1975, and H1650 were studied at single-cell resolution for the expression pattern of 13 markers: GLUT1, MCT4, CA9, TMEM45A, CD66, CD274, CD24, CD326, pan-keratin, TRA-1-60, galectin-3, galectin-1, and EGFR. The intra- and inter-cell-line heterogeneity of A549, H1975, and H1650 cells were demonstrated through hypoxic modeling. Additionally, human primary lung adenocarcinoma, and non-involved healthy lung tissue were homogenized to prepare a single-cell suspension for CyTOF analysis. The single-cell heterogeneity was confirmed using unsupervised viSNE and FlowSOM analysis. Our results also show, for the first time, that TMEM45A is expressed in lung adenocarcinoma. Intratumoral heterogeneity (ITH) is responsible for the majority of difficulties encountered in the treatment of lung-cancer patients. Therefore, the heterogeneity of NSCLC cell lines and primary lung adenocarcinoma was investigated by single-cell mass cytometry (CyTOF). First, we studied the single-cell heterogeneity of frequent NSCLC adenocarcinoma models, such as A549, H1975, and H1650. The intra- and inter-cell-line single-cell heterogeneity is represented in the expression patterns of 13 markers—namely GLUT1, MCT4, CA9, TMEM45A, CD66, CD274 (PD-L1), CD24, CD326 (EpCAM), pan-keratin, TRA-1-60, galectin-3, galectin-1, and EGFR. The qRT-PCR and CyTOF analyses revealed that a hypoxic microenvironment and altered metabolism may influence cell-line heterogeneity. Additionally, human primary lung adenocarcinoma and non-involved healthy lung tissue biopsies were homogenized to prepare a single-cell suspension for CyTOF analysis. The CyTOF showed the ITH of human primary lung adenocarcinoma for 14 markers; particularly, the higher expressions of GLUT1, MCT4, CA9, TMEM45A, and CD66 were associated with the lung-tumor tissue. Our single-cell results are the first to demonstrate TMEM45A expression in human lung adenocarcinoma, which was verified by immunohistochemistry.

Cost-effectiveness analysis comparing companion diagnostic tests for EGFR, ALK, and ROS1 versus next-generation sequencing (NGS) in advanced adenocarcinoma lung cancer patients

BackgroundThe treatment of choice for advanced non–small cell lung cancer is selected according to the presence of specific alterations. Patients should undergo molecular testing for relevant modifications and the mutational status of EGFR and translocation of ALK and ROS1 are commonly tested to offer the best intervention. In addition, the tests costs should also be taken in consideration. Therefore, this work was performed in order to evaluate the cost-effectiveness of a unique exam using NGS (next generation sequencing) versus other routinely used tests which involve RT-PCR and FISH.MethodsThe target population was NSCLC, adenocarcinoma, and candidates to first-line therapy. Two strategies were undertaken, strategy 1 corresponded to sequential tests with EGFR RT-PCR, then FISH for ALK and ROS1. Strategy 2 differed from 1 in that ALK and ROS1 translocation testing were performed simultaneously by FISH. Strategy 3 considered single test next-generation sequencing, a platform that includes EGFR, ALK and ROS1 genes. A decision tree analysis was used to model genetic testing options. From the test results, a microsimulation model was nested to estimate survival outcomes and costs of therapeutic options.ResultsThe use of NGS added 24% extra true cases as well as extra costs attributed to the molecular testing. The ICER comparing NGS with sequential tests was US$ 3479.11/correct case detected. The NGS improved a slight gain in life years and QALYs.ConclusionOur results indicated that, although precise, the molecular diagnosis by NGS of patients with advanced stage NSCLC adenocarcinoma histology was not cost-effective in terms of quality-adjusted life years from the perspective of the Brazilian supplementary health system.

Proportion of Mutation of Epidermal Growth Factor Receptor (EGFR) Genes from Tissue Biopsy and Liquid Biopsy ctDNA in Lung Adenocarcinoma

Backgrounds: In recent years, circulating tumor DNA (ctDNA) has emerged as a specific and sensitive blood-based biomarker to detect EGFR mutations. This study aims to determine the diagnostic accuracy of ctDNA in detecting EGFR gene mutations in adenocarcinoma lung cancer. 
 Methods: This study was a cross-sectional study with the subjects were adenocarcinoma lung cancer patients from histopathology or cytology examination and examined EGFR mutations from plasma tissue biopsy and ctDNA specimens from April 2018 to February 2019 in several hospitals in the Medan City.
 Results: There were 100 data have been collected, with male were 71 subjects and female were 29 subjects. Found 20 mutations, single mutations of tissue biopsy were 19 cases, del exon 19 were 12 cases, mutation in exon 21 (L858R) were 6 cases, mutation exon 21 (L861Q) was 1 case, del exon 19 and 21 (L861Q double mutations) was 1 case. From plasma ctDNA examination EGFR mutations were found 15 cases, del exon 19 were 12 cases and del exon 21 (L858R) were 3 cases. 
 Conclusions: The highest proportion of EGFR mutations by sex were women from tissue biopsy or ctDNA, the most often frequency of EGFR mutations from tissue biopsy and ctDNA in single mutations and exons19. (J Respir Indo. 2020; 40(3): 150-5)

Elemental and mutational analysis of lung tissue in lung adenocarcinoma patients.

This study aimed to observe the association between trace element concentrations in lung tissue from lung adenocarcinoma cancer (LADC) patients and mutations in the epidermal growth factor receptor (EGFR) and KRAS genes. LADC patients who had undergone lung resection were included in this study. Furthermore, twenty patients without lung cancer were included in this study as the control group. Samples were separately collected from both tumor and peritumor tissues. The mutational status was assessed for EGFR mutations, ALK rearrangements and KRAS mutations. Based on these analyses, patients were grouped into three groups: EGFR mutation, KRAS mutation and wild-type groups. The concentrations of various trace elements in the lung tissues were measured by a particle-induced X-ray emission (PIXE) system, and the results were analyzed for statistical significance. A total of 110 LADC patients were included in this study. The median age was 70 years, and 60% of the participants were female. Moreover, 18% and 20% of patients were EGFR- and KRAS-positive, respectively. Thirty-two trace elements were measured, and 18 trace elements were detectable. The concentrations of Fe, Co, Ni, Cu, Zn and Br were significantly higher in the KRAS mutation and wild-type groups than in the control group regardless of whether the samples were from tumor or peritumor tissues. For these 6 trace elements, the concentrations were significantly higher in smokers than in non-smokers. Considering the effect of smoking, differences in the trace element concentrations between each mutational group remained. Trace elements in the lung may play a role in development of LADC in both smokers and never-smokers. However, prospective studies with larger sample sizes are needed to support this hypothesis.

1740PD - Differential expression of B7-H4, VISTA, B7-H6, HHLA2, IDO-1, PD-L1 and CD8 in EGFR mutant and wild-type lung adenocarcinoma

BackgroundThe aim of this study was to describe the expression profiles of newly discovered B7 family (B7-H4, VISTA, B7-H6, HHLA2), IDO-1, PD-L1 and CD8 in resected lung adenocarcinoma tumor tissues and try to find potential immune target for lung cancer with EGFR mutation. MethodsA total of 372 adenocarcinoma lung cancer patients who underwent lung cancer resection were selected in the discovery cohort. The validation cohort contains another 231 adenocarcinoma lung cancer patients. Expression of B7-H4, VISTA, B7-H6, HHLA2, IDO-1, PD-L1 and CD8 was determined by immunohistochemical staining. ResultsIn the discovery cohort, 44.9% (167/372) of B7H4, 75.8% (282/372) of VISTA, 84.7% (315/372) of B7H6, 61.0% (227/372) of IDO-1 was positively stained in lung adenocarcinoma tissues. In the validation cohort, majority of cases was positive for B7H4 (41.6%, 96/231), VISTA (74.5%, 172/231), B7H6 (86.4%, 200/231) and IDO-1 (57.1%, 132/231), which was similar to the initial cohort. The positive expression rate of B7H4 in EGFR mutation group was significantly higher than that in wild type group in both cohort (P<0.05). However, expressions of IDO-1, PD-L1 and CD8 in wild type group were significantly higher than that in mutation group in discovery cohort, which was confirmed in the validation cohort. In the discovery cohort, the median IHC scores of B7H4 and HHLA2 for the EGFR mutation group were significantly higher than those in wild type group (P=0.045 for B7H4; P=0.003 for HHLA2). Whereas the median IHC scores of IDO-1 and PD-L1 in wild type group were significantly higher than those in mutation group (P=0.000 for IDO-1; P=0.000 for PD-L1). Results in the validation cohort confirmed observation above. In both discovery and validation set, the co-expressions of B7-H6, IDO-1, PD-L1 and CD8 were significantly higher in EGFR wild type group than those in EGFR mutation group. ConclusionsAberrant expressions of B7-H4 and HHLA2 were more obvious in EGFR mutation lung cancer. High co-expression of CD8 and IDO-1 in EGFR wild type patients reveals a potential good clinical efficacy of anti-IDO-1 therapy in patients with EGFR wild type. Legal entity responsible for the studyJifeng Feng. FundingThe National Natural Science Foundation of China. DisclosureAll authors have declared no conflicts of interest.

Proteomics analysis of human serum of patients with non-small-cell lung cancer reveals proteins as diagnostic biomarker candidates.

Non-small-cell lung carcinomas (NSCLC) is the most common type of lung cancer and it has a poor prognosis, because overall survival after 5 years is 20-25% for all stages. Thus, it is extremely important to increase the survival rate in the early stages NSCLC by focusing on novel screening tests of cancer identifying specific biomarkers expression associated with a more accurate tumor staging and patient prognosis. In this study, we focused our attention on quantitative proteomics of three heavily glycosylated serum proteins: AMBP,α2 macroglobulin, and SERPINA1. In particular, we analyzed serum samples from 20 NSCLC lung adenocarcinoma cancer patients in early and advanced stages, and 10 healthy donors to obtain a relative quantification through the MRM analysis of these proteins that have shown to be markers of cancer development and progression. AMBP, α2 macroglobulin, and SERPINA1 were chosen because all of them possess endopeptidase inhibitor activity and play key roles in cancer. We observe a variation in the expression of these proteins linked to the stage of the disease. Therefore, we believe that proteins like α2 macroglobulin, αmicroglobulin/bikunin, and SERPINA1 could be useful biomarkers for early detection of lung cancer and in monitoring its evolution.

Different next-generation sequencing pipelines based detection of tumor DNA in cerebrospinal fluid of lung adenocarcinoma cancer patients with leptomeningeal metastases

BackgroundThe nucleic acid mutation status in intracranial metastasis is markedly significant clinically. The goal of the current study was to explore whether the tumor-associated mutations can be detected by different next-generation sequencing (NGS) pipelines in paired cerebrospinal fluid (CSF) and plasma samples from lung adenocarcinoma (LAC) patients with leptomeningeal metastases (LM).MethodsPaired CSF cell free DNA (cfDNA), CSF cells, plasma and formalin-fixed and paraffin-embedded (FFPE) samples of primary tumors were collected from 29 LAC patients with LM to detect the mutations by different NGS pipelines.ResultsDNA libraries were generated successfully for 79 various samples in total for NGS sequencing, of which mutations were detected in 7 plasma samples (24.14%), 12 CSF cfDNA samples (66.67%), and 10 CSF cells (76.9%) samples. For the 26 patients with detected mutations, 8/26(30.77%) had mutations in plasma, which was significantly lower than that those from CSF cfDNA (12/15, 80.00%), CSF cells (10/11, 90.91%) and FFPE samples (13/17, 76.47%). When the input DNA of CSF cells was less than 20 ng, the cHOPE pipeline of NGS identified the most mutations for epidermal growth factor receptor (EGFR).ConclusionsNGS-based detection of mutations in cfDNA or cells from CSF provided more information than from plasma samples from LAC patients with LM. In addition, the cHOPE pipeline performed better than the other three NGS pipelines when input DNA from CSF cells was low.

Serum microRNA-139-5p is downregulated in lung cancer patients with lytic bone metastasis.

Bone remodeling can be interrupted by tumor cells which leads to an inappropriate balance of osteoblasts and osteoclasts. As the progenitors of osteoblasts, mesenchymal stem cells (MSCs) have been reported to exhibit an abnormal osteogenic differentiation potential in some cancer‑related bone lesions. However, the evidence is very limited in terms of the biological alterations of MSCs in the bone metastasis of non‑small cell lung cancers (NSCLC). We investigated the expression and function of miR‑139‑5p in MSC osteogenic differentiation invitro in normal and NSCLC-exposed condition. Then, we compared the serum miR‑139‑5p in stageIV lung adenocarcinoma cancer patients with and without lytic bone metastasis. We found that MSCs exhibited a significant increase in miR‑139‑5p expression after exposure to osteogenic differentiation induction medium. However, Notch1, which was confirmed as a target of miR‑139‑5p by luciferase and western blot assays, showed a marked downregulated expression together with its pathway downstream factors during MSC osteogenesis. miR‑139‑5p positively regulated MSC osteogenic differentiation but this effect was abrogated significantly by Notch1 knockdown. After exposure to conditions of lung cancer cells A549 and L9981, MSCs exhibited significant downregulation of miR‑139‑5p expression and early osteogenic marker ALP activity. Furthermore, we demonstrated that the expression of serum miR‑139‑5p from lung adenocarcinoma patients with lytic bone metastasis was significantly lower compared to that in patients with metastases in other organs. The potential roles of miR‑139‑5p as a biomarker and treatment target in monitoring and controlling bone metastasis in lung cancer patients are worthy of being further explored.

Association between clinical characteristics and the diagnostic accuracy of circulating single-molecule amplification and resequencing technology on detection epidermal growth factor receptor mutation status in plasma of lung adenocarcinoma.

Lung cancer is the leading cause of cancer-related mortality in the world. Circulating single-molecule amplification and resequencing technology (cSMART) can successfully detect epidermal growth factor receptor (EGFR) mutation in non-small cell lung cancer (NSCLC). However, few studies have investigated the association between clinical characteristics and the diagnostic accuracy of cSMART technique in lung adenocarcinoma. We enrolled 95 patients, which included paraffin embedded tumor tissues and matched plasma samples. Retrospectively analyzed the correlation between clinical characteristics and sensitivity, specificity, accuracy, positive predictive value (PPV) and negative predictive value (NPV) of cSMART. Of the 95 lung adenocarcinoma cancer patients, 49 (51.5%) and 40 (42.1%) harbored EGFR mutations respectively in tissue and plasma. In younger than 60years group, sensitivity, specificity and consistency for cSMART were 81.0%, 100%, and 90.9% (P<.001). In metastasis group, sensitivity, specificity, and consistency for cSMART were 92.9%, 77.8%, and 87.0% (P=.001). By univariate analysis, younger than 60years (OR=5.938; 95% confidence interval: 1.835-19.210; P=.001); metastasis group (OR=4.482; 95% confidence interval: 1.432-14.024; P=.007) were significantly correlated with a higher accuracy. By multivariate analysis, younger than 60years (P=.003) and metastasis (P=.004) were confirmed as independent factors for diagnostic accuracy of EGFR mutation in plasma through cSMART. cSMART is feasible for detection EGFR mutation in plasma when tissue is unavailable. Age and metastasis might be considered as independent factors in diagnostic accuracy of cSMART in lung adenocarcinoma.

Detect T790M in cell free tumor DNA of Chinese advanced non-small cell lung cancer adenocarcinoma patients by different platforms and evaluate clinical outcomes of T790M positive patients with osimertinib monotherapy.

TPS9104 Background: EGFR T790M mutation occurs in approximately 50-60% of non-small cell lung cancer adenocarcinoma (NSCLC) patients with acquired EGFR-TKI resistance, based on tumor re-biopsies using an invasive clinical procedure. Recently, Cell free tumor DNA (ctDNA) has emerged as a specific and sensitive blood-based biomarker and studies have demonstrated ctDNA as a feasible and minimally invasive alternative to tissue biopsy. Data on different technology platforms used for EGFR T790M detection in blood in China is limited. We aim to compare the methods currently available in hospital practise, including cobas EGFR Mutation Test (Roche Molecular Systems), super-ARMS, digital PCR and NGS, to compare each platform and clinically validate each as companion diagnostic to osimertinib. Methods: This is an open-label, multi-center study in 250 locally advanced or metastatic NSCLC patients with documented EGFR sensitizing mutation and progression on previous EGFR-TKI. T790M mutation in plasma ctDNA will be tested by four methods: cobas, super-ARMS, digital PCR and NGS in order to evaluate the concordance, sensitivity and specificity of T790M testing in plasma between the cobas test and the other platforms. T790M positive patients by any of the four platforms will receive osimertinib treatment (administered orally as one 80 mg tablet once a day in ASTRIS study, NCT02474355) and the clinical outcomes (PFS, ORR, OS) will be followed. Patients will continue to receive osimertinib until disease progression (PD), as assessed by investigators. Digital PCR and NGS will be used to monitor the molecular evolution of T790M and C797S in plasma from NSCLC patients during osimertinib treatment. NGS will also be used to explore acquired resistance mechanisms before osimertinib treatment and after PD. 23 of planned 250 patients have been enrolled in the study as of January 2017. Clinical trial information: NCT02997501.

Cost-effectiveness analysis comparing companion diagnostic tests for EGFR, ALK and ROS-1 versus next-generation sequence (NGS) in advanced adenocarcinoma lung cancer patients.

9068 Background: Success of a target therapy is directly correlated with the accuracy of its companion diagnostic test. Without a corresponding biomarker, target therapy may yield shorter survival, waste time, increase burdens and costs. As important as conducting cost-effectiveness studies for therapies, it is also valuable to compare different molecular tests. In lung cancer, the mutational status of EGFR and translocation of ALK and ROS-1 are commonly tested to offer the best intervention. Our objective is to evaluate the cost-effectiveness of a unique exam using NGS versus other routinely used tests such as the ones which involve RT-PCR and FISH. Methods: Target population is NSCLC, adenocarcinoma, and candidates to first-line therapy. Strategy 1: test EGFR mutation if EGFR test is negative, individual follow to FISH for ALK; if FISH is negative, follow to FISH for ROS-1. Strategy 2: differs from 1 since FISH for ALK and ROS are requested together. Strategy 3: all individuals are submitted to NGS (multicomplex platform which include EGFR, ALK, ROS-1 and other genes). Tests could also be classified as unknown due to the quality of tumor sample. Prevalence of biomarker, test accuracy and proportion of unknown results were used to calculate each decision tree branch. Sensitivity and specificity was obtained from literature review using Sanger as reference standard for RT-PCR tests and NGS. The study was analyzed from a healthcare-payer perspective based on Brazilian private sector. Costs were based on data from diagnostic companies, ANS and AMB-CBHPM 2016. Results: The use of NGS added 24% extra cases correct identified as well as extra costs (US$ 800.76; PPP 2015) attributed to the molecular testing. The ICER comparing NGS with sequential tests was US$ 3,381.82/correct case detected. Comparing strategy 2:1, the ICER was US$937.86/correct case detected. Conclusions: this study is part of an effort to integrate companion diagnostics discussions on precision medicine and covered drugs in the Brazilian health system. These findings can also subsidy cost-effectiveness studies that incorporates subsequent treatments and survival.

Is Leukocyte Telomere Length Related with Lung Cancer Risk?: A Meta-Analysis.

Background:Epidemiological studies have probed the correlation between telomere length and the risk of lung cancer, but their findings are inconsistent in this regard. The present meta-analysis study has been carried out to demonstrate the association between relative telomere length in peripheral blood leukocytes and the risk of lung cancer using an established Q-PCR technique.Methods:A systematic search was carried out using PubMed, EMBASE, and ISI before 2015. A total of 2925 cases of lung cancer and 2931 controls from 9 studies were employed to probe the relationship between lung cancer and telomere length. ORs were used at 95% CI. Random-effects models were used to investigate this relationship based on the heterogeneity test. Heterogeneity among studies was analyzed employing subgroup analysis based on type studies and the year of publication.Results:Random-effects meta-analysis revealed that patients with lung cancer were expected to have shorter telomere length than the control (1.13, 95% CI: 0.82-1.81, P=0.46). The summary of the pooled ORs of telomere length in adenocarcinoma lung cancer patients was 1 (95%CI=0.68-1.47, I2=93%) compared to patients with squamous cell lung cancer, which was 1.78 (95% CI=1.25-2.53, I2=3.9%). The meta-regression revealed that the effect of telomere length shortening, decreased and increased with the year of publication and the age of risks to lung cancer, was clearly related to short telomeres lengths.Conclusion:Lung cancer risks clearly related with short telomeres lengths. In patients with breathing problems, lung cancer risk can be predicted by telomere length adjustment with age, sex, and smoking.