Value In Non-small Cell Lung Cancer Research Articles

LncRNA-ATB Can Be a Biomarker for Diagnosis and Prognosis Evaluation of Non-Small Cell Lung Cancer

Objective: This study was set out to inquire into the expression and clinical significance of lncRNA activated by transforming growth factor β (LncRNA-ATB) and in cancer tissues of patients with non-small cell lung cancer (NSCLC). Methods: LncRNA-ATB in cancer tissues and adjacent tissues of 89 NSCLC patients was detected by quantitative real-time polymerase chain reaction (qRT-PCR), and its clinical diagnostic value in NSCLC was determined by receiver operating characteristic (ROC) curves. Based on the median expression of LncRNAATB in NSCLC tissues, 89 patients were allocated into high- and low-expression groups. The 3-year survival rate was calculated using Kaplan-Meier method and logrank test, and compared between lncRNA-ATB high and low expression groups by Log-rank test. Cox regression quantitatively analyzed the correlation between lncRNA-ATB and the clinical prognosis of NSCLC patients. Results: LncRNAATB presented notably higher expression in NSCLC tissues as compared to adjacent tissues (P < 0.05), and was linked to smoking history, TNM stage and greatest tumor diameter (GTD) (P < 0.05). The cut-off value, sensitivity and specificity, as well as the area under the curve (AUC) of NSCLC diagnosed by LncRNA-ATB were 2.103, 73.77%, 83.15% and 0.836, respectively. Patients with high lncRNA-ATB expression (40.00%) showed statistically lower than those with low lncRNA-ATB expression (63.36%). TNM stage, differentiation degree and lncRNA-ATB expression were independent variables affecting patient prognosis. Conclusions: LncRNA-ATB, with increased expression profiles in NSCLC tissues, is involved in NSCLC progression and associated with unfavourable prognosis of patients, which can be a plausible tumor marker for NSCLC patients.

Circulating Long Noncoding RNAs Act as Diagnostic Biomarkers in Non-Small Cell Lung Cancer.

Identification of novel effective early diagnostic biomarkers may provide alternative strategies to reduce the mortality for non-small cell lung cancer (NSCLC) patients. Circulating long non-coding RNAs (lncRNAs) have emerged as a new class of promising cancer biomarkers. Our study aimed to identify circulating lncRNAs for diagnosing NSCLC. A total 528 plasma samples were continuously collected and allocated to four progressive phases: discovery, training, verification, and expansion phases. The expression of candidate lung cancer related lncRNAs were detected using quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). We identified a 4-lncRNA panel (RMRP, NEAT1, TUG1, and MALAT1) that provided a high diagnostic value in NSCLC (AUC = 0.86 and 0.89 for training and verification phase, respectively). Subgroup analyses showed that the 4-lncRNA panel had a sensitivity of 78.95% [95% confidence interval (CI) = 62.22%–89.86%] in stage I-II patients and 75.00% (95% CI = 52.95%–89.40%) in patients with small tumor size (≤3cm). Notably, the sensitivity of 4-lncRNA panel was significantly higher than that of routine protein panels in adenocarcinoma (CEA, CA125, and CYFRA21-1, 86.30% vs. 73.96%). Adding 4-lncRNA to protein markers significantly improved the diagnostic capacity in both adenocarcinoma (AUC=0.85, 95% CI = 0.78–0.91) and squamous cell carcinoma (AUC=0.93, 95% CI = 0.86–0.97). In conclusion, we identified a plasma 4-lncRNA panel that has considerable clinical value in diagnosing NSCLC. The 4-lncRNA panel could improve the diagnostic values of routine tumor protein markers in diagnosing NSCLC. Circulating lncRNAs could be used as promising candidates for NSCLC diagnosis.

The Immune Cell Composition in Primary Lung Squamous Cell Carcinomas Influences Patterns of Recurrence and Survival

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer with squamous cell carcinoma (SCCA) being a prominent histology. Emerging data suggests that immunological parameters have predictive and prognostic value in NSCLC. In this study we determine if the immune cell infiltrate (ICI) in primary lung SCCA tumors influences patterns of recurrence and survival. We retrospectively collected data from an IRB approved prospective tissue collection protocol, which profiled all tumors for gene expression with Affymetrix Hu-RSTA microarray chips (Affymetrix; Santa Clara, CA). We identified tumor samples from patients diagnosed with lung SCCA who underwent surgery between 1997 and 2010. The CIBERSORT deconvolution algorithm was employed to infer the composition of the immune cell infiltrate (22 distinct immune cell types) in tumors via gene expression. ICI presence was dichotomized at the median, where high refers to counts above the median. Time to event analyses were assessed with Kaplan-Meier methods and compared with log rank test. Cox regression was performed to evaluate the effect of predictors on defined outcomes. We identified 258 patients for this analysis. The median age was 70 years (range, 72-88) and the majority (67%) were male and had an ECOG performance status of 1 (65%). The median tobacco pack-years was 50 (range, 0-180), 33% were current and 64% were former smokers. Stages were IA (24%), IB (27%), IIA (8%), IIB (27%) and IIIA/B (14%). Nineteen (7 %) patients received NACT, 17% received adjuvant CT and 9% received adjuvant radiotherapy. The median follow-up time was 19 months with a 2-year OS of 76% and a 2-yr freedom from recurrence of 77%. ICI analysis identified that a high presence of activated CD4+ memory T cells predicts for recurrence in the lung vs other site (77% vs 23%, p = 0.01). Similarly, a high presence of activated NK cells predicts for locoregional recurrence (62% vs 38%, p = 0.03) and high resting NK cells predict for distant recurrence (66% vs 35%, p = 0.02). Predictors for OS were male gender (HR: 1.6, 95% CI 1.1-2.5, p = 0.03), and high presence of resting dendritic cells (HR: 1.5, 95% CI 1.0-2.2, p = 0.04) and eosinophils (HR: 1.6, 95% CI 1.0-2.4, p = 0.03) on multivariable analysis. Our results suggest that the immune cell composition of primary lung SCCA tumors may influence patterns of recurrence. This data may assist with novel means to optimize treatment selection and sequencing.

A novel tumor mutational burden estimation model as a predictive and prognostic biomarker in NSCLC patients

BackgroundTumor mutational burden (TMB) has both prognostic value in resected non-small cell lung cancer (NSCLC) patients and predictive value for immunotherapy response. However, TMB evaluation by whole-exome sequencing (WES) is expensive and time-consuming, hampering its application in clinical practice. In our study, we aimed to construct a mutational burden estimation model, with a small set of genes, that could precisely estimate WES-TMB and, at the same time, has prognostic and predictive value for NSCLC patients.MethodsTMB estimation model was trained based on genomic data from 1056 NSCLC samples from The Cancer Genome Atlas (TCGA). Validation was performed using three independent cohorts, including Rizvi cohort and our own Asian cohorts, including 89 early-stage and n late-stage Asian NSCLC patients, respectively. TCGA data were obtained on September 3, 2018. The two Asian cohort studies were performed from September 1, 2018, to March 5, 2019. Pearson’s correlation coefficient was used to assess the performance of estimated TMB with WES-TMB. The Kaplan-Meier survival analysis was applied to evaluate the association of estimated TMB with disease-free survival (DFS), overall survival (OS), and response to anti-programmed death-1 (PD-1) and anti-programmed death-ligand 1 (PD-L1) therapy.ResultsThe estimation model, consisted of only 23 genes, correlated well with WES-TMB both in the training set of TCGA cohort and validation set of Rizvi cohort and our own Asian cohort. Estimated TMB by the 23-gene panel was significantly associated with DFS and OS in patients with early-stage NSCLC and could serve as a predictive biomarker for anti-PD-1 and anti-PD-L1 treatment response.ConclusionsThe 23-gene panel, instead of WES or the currently used panel-based methods, could be used to assess the WES-TMB with a high relevance. This customized targeted sequencing panel could be easily applied into clinical practice to predict the immunotherapy response and prognosis of NSCLC.

Identification of Key circRNAs in Non-Small Cell Lung Cancer

BackgroundOur work aimed to identify the key differentially expressed circular RNAs (circRNAs) (DECs) in non-small cell lung cancer (NSCLC). Materials and MethodsAn integrated analysis based on public NSCLC datasets obtained from Gene Expression Omnibus was performed. DECs in NSCLC were subsequently identified. Bioinformatics analyses were utilized for describing the predictable functions of DECs including circRNA-miRNA network construction and pathway enrichment. The diagnostic value of candidate DECs among NSCLC and healthy individuals were preliminarily evaluated in GSE101586, GSEE101684, GSE112214 datasets. ResultsA total of 43 up-regulated and 78 down-regulated DECs in NSCLC were identified. The mTOR signaling pathway, ErbB signaling pathway and cell cycle were significantly enriched from the originated genes of DECs in NSCLC. In the circRNA-miRNA network, hsa-circ-0002702, hsa-circ-0049271, hsa-circ-0009150 and hsa-circ-0053958 had high connectivity with miRNAs, which respectively interacted with 122, 42, 41, and 39 miRNAs. hsa_circ_0003028, hsa_circ_0015278, hsa_circ_0043256, hsa_circ_0049657 and hsa_circ_0074930 could distinguish NSCLC patients from healthy individuals in GSE101586, GSEE101684, GSE112214 datasets. ConclusionsDECs including hsa_circ_0003028, hsa_circ_0015278, hsa_circ_0043256, hsa_circ_0049657 and hsa_circ_0074930 had diagnostic value in NSCLC. These DECs might play key roles in NSCLC pathogenesis through the mTOR signaling pathway, ErbB signaling pathway and cell cycle.

The Effects of EGFR Exon 19 747–750 Deletion on the Risk of Developing Lung Cancer

Objective: Although smoking is the most significant factor in the etiology of lung cancer, other environmental pollutants and genetic predisposition also play major roles in its development. Histopathologically, lung cancers are divided into two major types, as small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). The latter accounts for almost 85% of all lung cancers with a very aggressive course and being associated with a high rate of mortality. Among the genetic mutations with prognostic value in NSCLC, the epidermal growth factor receptor (EGFR) mutation is most frequently found in 50 to 80% of cases. The EGFR is a transmembrane glycoprotein with tyrosine kinase activity which is associated with both normal cell growth and malignant transformations. Material and Methods: In the present study, we aimed to evaluate the effects of exon 19 747–750 deletion in the EGFR gene on the risk of developing lung cancer and to examine its potential relationship with the different histopathological types of lung cancer. The study sample comprised a total of 178 patients diagnosed with lung cancer at Mersin University, Medical Faculty, Oncology Clinics, and 192 age- and sex-matched healthy individuals as the control group. Deoxyribonucleic acid (DNA) isolation was performed using the standard salt-water precipitation method, while the mutation screening and genotyping analyses were carried out with a polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses. Results: The frequency of mutant EGFR exon-19 deletion in the control group was 15.1%, increasing to 36.9% in the lung cancer group, and increasing the risk of developing lung cancer by 2.64 times (p: 0.014). This increase did not significantly differ between the histopathological types of lung cancer (p: 0.76). Conclusion: Considering the distribution of lung cancer patients in different age groups, it is obvious that advanced age is a risk factor for the development of EGFR mutation and lung cancers (p<0.001).

Abstract 575: Adaptive CD4 Th1 response against telomerase in blood counteracts T-cell exhaustion in non-small cell lung cancer

Abstract Despite the critical roles played by IFN-γ+ CD4 Th1 response in tumor immunity, the translation of their potential in clinic remains challenging. Here, we evaluate the clinical significance of circulating anti-tumor CD4 Th1 response in non-small cell lung cancer (NSCLC) patients. 170 naïve-treatment patients were enrolled in this immune monitoring study. The antitumor adaptive Th1 response was assessed by IFN-γ ELISPOT assay in blood lymphocytes using a mixture of eight highly promiscuous and Th1-polarized HLA class II-restricted epitopes from telomerase (TERT). The presence of anti-TERT Th1 response was detected in 59/170 patients (35%). We found an opposite link between anti-TERT Th1 and hyper exhausted T cells co-expressing PD-1+ and TIM-3+ in NSCLC patients. In contrast to hyper exhausted CD8 T cells, the presence of anti-TERT Th1 response was associated with a low rate of hyper exhausted CD4 T cells. We showed that NSCLC stage dissemination is associated with a decrease of anti-TERT Th1 response but an increase of circulating hyper exhausted PD-1+/TIM-3+ CD4 T cells. Notably, anti-telomerase Th1 response and hyper exhausted CD4 T-cells displayed opposite prognosis value in NSCLC. While high level of anti-TERT Th1 cells play a protective role, hyper-exhausted PD-1+TIM-3+ CD4 T cells negatively affect patients’ survival. By using these two circulating immune factors, we stratified patients in distinct prognostic group. Patients with anti-TERT Th1high/CD4 PD-1 TIM-3low immune profile had better overall survival than anti-TERT Th1low/CD4 PD-1 TIM-3high group (median OS : not reached versus 4 months respectively p&amp;lt;0.0001). In conclusion, high circulating anti-TERT Th1 response plays a strong protective role in NSCLC by counteracting the deleterious effect associated with hyper exhausted T cells in blood. Our study provides a dynamic blood-based immunomonitoring tool allow patients stratification. Citation Format: Caroline Laheurte, Magalie Dosset, Dewi Vernerey, Elodie Lauret Marie Joseph, Laura Boullerot, Vincent Kaulec, Marion Jacquin, Laurie Cuche, Guillaume Eberst, Pascale Jacoulet, Virginie Westeel, Olivier Adotevi. Adaptive CD4 Th1 response against telomerase in blood counteracts T-cell exhaustion in non-small cell lung cancer [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 575.

Long non-coding RNAs function as novel predictors and targets of non-small cell lung cancer: a systematic review and meta-analysis.

ObjectivesNon-small cell lung cancer (NSCLC) is associated with high morbidity and mortality, leading the understanding the pathogenesis paramount. Recent studies suggest that long non-coding RNAs (lncRNAs) play an important role in NSCLC. We conducted a systematic review to examine the relationship between lncRNAs and NSCLC.Materials and MethodsWe calculated hazard ratios (HRs) and 95% confidence intervals (CIs) to estimate overall survival (OS), and odds ratios (ORs) and 95% CIs to assess clinicopathological parameters. Also, pooled sensitivity and specificity values were used to measure the diagnostic value of lncRNAs for NSCLC. Finally, we summarized the molecular mechanisms underlying the activity of lncRNAs in NSCLC.ResultsWe found that high expression of oncogenic lncRNAs was associated with a poor prognosis (OS: HR, 1.18; 95% CI, 1.14–1.22) and poor clinicopathological characteristics (tumor size: OR, 2.74 or 2.04; 95% CI, 1.66–4.52 or 1.09–3.79 based on the two classification criterias; lymph node metastasis: OR, 3.30; 95% Cl, 2.42–4.49), Also, high expression of tumor-suppressor lncRNAs was correlated with longer survival times (OS: HR, 0.54; 95% CI, 0.44–0.66) and improved clinical characteristics (tumor size: OR, 0.33 or 0.28; 95% CI, 0.14–0.75 or 0.18–0.45; lymph node metastasis: OR, 0.37; 95% Cl, 0.26–0.52). Furthermore, we found that lncRNAs could be used as serum biomarkers of NSCLC (sensitivity: 0.81; 95% CI, 0.72–0.87; specificity: 0.83; 95% CI, 0.73–0.90). Finally, lncRNAs regulated expression of key proteins, thereby mediating development of a malignant phenotype.ConclusionslncRNAs have significant clinical value in NSCLC and could function as novel predictors of disease and/or as therapeutic targets.

Prognostic value of total lesion glycolysis and metabolic active tumor volume in non-small cell lung cancer

IntroductionTo predict the outcome of patients with non-small cell lung cancer (NSCLC) the currently used prognostic system (TNM) is not accurate enough. The prognostic significance of the SUVmax measured by PET remains controversial. This study aims to evaluate the prognostic value in overall survival and progression free survival of SUVmax, the total lesion glycolysis (TLG) and the mean metabolic active volume (MATV) in NSCLC. MethodsWe retrospectively reviewed 105 patients (72 males, 33 females) with a new diagnosis of NSCLC (TNM stage I: 27.6%, II: 10.5%, III: 40.9% and IV: 21.0%) who underwent scanning with a PET/CT. For VOI definition a semi-automatic delineation tool was used. On PET images SUVmax, SUVmean and MATV of the primary tumor and the whole tumor burden were measured. TLG and MATV were measured by using a threshold of 50% of SUVmax. ResultsOS and PFS are found to be higher in patients with low-SUVTmax and low-TLGT values. OS and PFS were significantly higher for low-SUVWTBmax, low-MATVWTB and low-TLGWTB values of the whole-tumor burden.Multivariate analysis of the whole-tumor burden revealed that the most important prognostic factors for OS are high MATVWTB and TLGWTB values, increasing stage and male gender. TLGWTB and stage are also independent prognosticators in PFS. ConclusionOnly whole-body TLG is of prognostic value in NSCLC for both OS and PFS. Stratification of patients by TLGWTB might complement outcome prediction but the TNM stage remains the most important determinant of prognosis. MicroabstractIn order to predict the outcome of patients with non-small cell lung cancer (NSCLC) the currently used prognostic system (TNM) is not accurate enough. The prognostic significance of the standard uptake value (SUV) measured by PET remains controversial. This study aims to evaluate the prognostic value in overall survival (OS) and progression free survival (PFS) of the standard uptake value (SUV), the total lesion glycolysis (TLG) and the mean metabolic active volume (MATV) in NSCLC. The study reveals that TLG of the whole-tumor burden is an independent prognostic factor for OS and PFS in patients with NSCLC.

Activation of Janus kinase 1 confers poor prognosis in patients with non-small cell lung cancer.

The activation of Janus kinase 1 (JAK1) has been reported to occur in non-small cell lung cancer (NSCLC), activating the JAK/signal transducers and activators of transcription cascade. However, the association between JAK1 activation and the prognostic value in NSCLC remains unclear. The present study initially investigated the association between expression of the activated form of JAK1 (p-JAK1) and prognosis in patients with NSCLC. A cohort of 142 resected primary NSCLC tissue samples, including 74 adenocarcinoma (ADCC) and 68 squamous cell carcinoma samples, were analyzed. p-JAK1 expression status was determined by immunohistochemistry. Evaluation of epidermal growth factor receptor (EGFR) gene amplification by fluorescence in situ hybridization was subsequently performed in 74 ADCC samples. The prognostic significance of p-JAK1 expression and EGFR gene amplification were evaluated with univariate and multivariate survival analyses. Compared with normal lung tissue, p-JAK1 expression level was significantly increased in NSCLC (P<0.001). Positive p-JAK1 expression indicated a poor prognosis, particularly for patients in early stages (stage I/II, including tumor size <3 cm, Lymph node invasion N0/1; all P<0.05). p-JAK1 expression was an independent predictor of a poor prognosis (P=0.022). The overall survival time for patients with positive p-JAK1 expression and EGFR-amplified tumors was significantly shortened compared with patients with tumors negative for one or both features (both features present vs. neither feature present, P<0.001). The results provided clinical evidence that the activation of JAK1 was an independent prognostic factor, particularly in early stage NSCLC. The combination of EGFR gene amplification and p-JAK1 expression may be a novel target for the selection of individual therapy strategies and predicting the effects of therapy for NSCLC.

Association of long non-coding RNA H19 and microRNA-21 expression with the biological features and prognosis of non-small cell lung cancer.

In recent years, long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) have been shown to play important roles in tumor biological function. The aim of this study was to investigate the diagnostic and prognostic value of lncRNA H19 and miR-21 expression in non-small-cell lung cancer (NSCLC). H19 and miR-21 expression was measured in tumor tissues and corresponding non-tumor lung tissues from 200 patients by quantitative reverse transcription polymerase chain reaction. Moreover, the in vitro and in vivo effects of H19 knock out in A549 cells were investigated. Expression of both H19 and miR-21 was significantly higher in lung tissues from patients with NSCLC than in normal lung tissues. Increased expression of H19 and miR-21 was positively correlated with advanced tumor-node-metastasis stage and tumor size. miR-21 expression was highest in stage I and II NSCLC, whereas H19 expression was highest in stage III and IV NSCLC. Knockout of H19 significantly inhibited NSCLC cell proliferation both in vitro and in vivo. The results show that H19 may mainly contributes to the progression of NSCLC, and its expression levels can reflect the invasive and metastatic status to some extent. miR-21 expression more likely plays an important role in early stage NSCLC. Moreover, H19 and miR-21 interact in the regulation of NSCLC, and with greater expression of both H19 and miR-21, overall survival decreased. The combination of H19 and miR-21 may have diagnostic value in NSCLC and represent a target for new NSCLC treatments.

Over-expression of CircRNA_100876 in non-small cell lung cancer and its prognostic value

Circular RNAs (circRNAs) are a novel type of endogenous RNAs featuring stable structure and high tissue-specific expression. Recently, accumulating evidence has revealed that aberrant circRNAs expression plays important roles in carcinogenesis and tumor progression. However, the expression pattern and biological function of circRNAs in non small cell lung cancer (NSCLC) remain largely unknown. Therefore, the purpose of this study was to clarify the possible role of one of typical circRNAs, circRNA_100876 in NSCLC and to define its prognostic value in NSCLC. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of circRNA_100876 in tumor tissues and their adjacent nontumorous tissues in 101 patients with NSCLC. We found that the expression level of circRNA_100876 was significantly elevated in NSCLC tissues when compared with their adjacent nontumorous tissues (P=0.000). Moreover, there was a close correlation between the circRNA_100876 up-regulation expression and lymph node metastasis (P=0.001) and tumor staging (P=0.001) in NSCLC. In addition, Kaplan-Meier survival analysis demonstrated that the overall survival time of NSCLC patients with high circRNA_100876 expression was significantly shorter than those patients with low circRNA_100876 expression (P=0.000). In conclusion, our findings indicate that circRNA_100876 is closely related to the carcinogenesis of NSCLC and it might be served as a potential prognostic biomarker and therapeutic target for NSCLC.

The relationship between cardiac radiation dose and mediastinal lymph node involvement in stage III non-small cell lung cancer patients

PurposeThe results from Radiation Therapy Oncology Group (RTOG) 0617, a dose escalation trial that compared treatment with 60 Gy versus 74 Gy for patients with stage III non-small cell lung cancer (NSCLC), suggested that in these patients, the heart dose from radiation therapy correlates with survival. In particular, the study noted that patients with a high heart V5 and V30 had a poorer overall survival; however, the exact cause of this correlation is not known. We hypothesize that heart dose may be a surrogate for mediastinal nodal involvement, which has prognostic value in NSCLC. This study evaluates the relationship between heart dose and involvement of mediastinal lymph nodes in patients with stage III NSCLC treated with radiation therapy. Methods and materialsA total of 56 patients were identified and treated with definitive radiation therapy from 2007 to 2014. The heart was recontoured for every patient by a single physician, per the RTOG 1106 contouring atlas. We assessed lymph node station involvement using pretreatment data, and nodal coverage was confirmed on plan review. ResultsMean heart dose was found to be significantly higher in patients with multinodal station and level 7 involvement. On Spearman's rank correlation, level 7 was significantly associated with all heart parameters tested (P < .001). Patients who had 2 or more lymph node stations involved were found to have significantly higher heart doses for all parameters tested when compared with those who had only one station involved or no nodal involvement. ConclusionsOur findings suggest that heart dose may be a surrogate for other prognostic factors in stage III NSCLC rather than an independent predictor of outcome.

Identification of serum proteins and multivariate models for diagnosis and therapeutic monitoring of lung cancer.

Lung cancer is one of the most prevalent cancers and has very poor treatment outcome. Biomarkers useful for screening and assessing early therapeutic response may significantly improve the therapeutic outcome but are still lacking. In this study, serum samples from 218 non-small cell lung cancer (NSCLC) patients, 34 small cell lung cancer (SCLC) patients and 171 matched healthy controls from China were analyzed for 11 proteins using the Luminex multiplex assay. Eight of the 11 proteins (OPN, SAA, CRP, CYFRA21.1, CEA, NSE, AGP and HGF) are significantly elevated in NSCLC and SCLC (p = 10−5−10−59). At the individual protein level, OPN has the best diagnostic value for NSCLC (AUC = 0.92), two acute phase proteins (SAA and CRP) have AUC near 0.83, while CEA and CYFRA21.1 also possess good AUC (0.81 and 0.77, respectively). More importantly, several three-protein combinations that contain OPN and CEA plus one of four proteins (CRP, SAA, CYFRA21.1 or NSE) have excellent diagnostic potential for NSCLC (AUC = 0.96). Four proteins (CYFRA21.1, CRP, SAA and NSE) are severely reduced and three proteins (OPN, MIF and NSE) are moderately decreased after platinum-based chemotherapy. Therapeutic response index (TRI) computed with 3–5 proteins suggests that approximately 25% of the NSCLC patients respond well to the therapy and TRI is significantly correlated with pre-treatment protein levels. Our data suggest that therapeutic response in NSCLC patients can be effectively measured but personalized biomarkers may be needed to monitor different subsets of patients.

A Case Study in Advanced Lung Cancer Patients with Vimentin Over Expression.

Vimentin belongs to an intermediate filament (IF) family of proteins, mainly present in mesenchymal cells and has a crucial role in maintaining cellular integrity. Vimentin can induce epithelial to mesenchymal transition, and thus increase migration and invasion capacity of the cells. It has been shown to be a useful and reliable diagnostic and prognostic marker in several cancers including colon cancers, breast and hepatocellular cancers. Recent studies suggest that it may have a role in distant metastasis of non-small cell lung cancer (NSCLC) accounting for poor survival [1]. The aim of the study is to assess the impact of vimentin testing as a diagnostic and prognostic marker in NSCLC. This is a case study of 12 NSCLC patients who had vimentin testing as a part of their work up over the past five years at the University of Cincinnati. A total of 12 patients with advanced lung cancer were included in this case study as they were found to have strong vimentin expression. This was correlated with overall survival of this group of patients. Median survival of the patients was 4.66 months. This is 7.34 months less compared to the median survival of patients with stage IV NSCLC which is reported to be 12 months. More studies are warranted into the use of vimentin as an emerging useful marker for early diagnosis, aggressive transformation relapse, and prognostication of NSCLC. It may have therapeutic value in NSCLC as observed in other cancers.

Does Immunohistochemistry Represent a Robust Alternative Technique in Determining Drugable Predictive Gene Alterations in Non-Small Cell Lung Cancer?

Immunohistochemistry (IHC) is a widely-tested, low-cost and rapid ancillary technique available in all laboratories of pathology. This method is generally used for diagnostic purposes, but several studies have investigated the sensitivity and specificity of different immunohistochemical antibodies as a surrogate test in the determination of predictive biomarkers in non-small cell lung cancer (NSCLC), particularly for Epidermal Growth Factor Receptor (EGFR) gene mutations, Anaplastic Lymphoma Kinase (ALK) gene and ROS1 rearrangements. In this review, a critical examination of the works comparing the consistency of IHC expression and conventional molecular techniques to identify genetic alterations with predictive value in NSCLC is discussed. Summarizing, data on sensitivity and specificity of antibodies against ALK and ROS1 are very consistent and time has come to trust in IHC at least as a cost-effective screening tool to identify patients with rearranged tumors in clinical practice. On the other hand, mutant-specific antibodies against EGFR demonstrate a good specificity but a lowto- fair sensitivity, raising some cautions on their employment as robust predictive biomarkers. A brief comment on preliminary experiences with antibodies against BRAF, RET, HER2 and c-MET is also included.

Paraneoplastic Leukocytosis and Thrombocytosis as Prognostic Biomarkers in Non-small Cell Lung Cancer

Background and ObjectivesSearch for inexpensive laboratory markers have identified associations between blood counts and lung cancer outcomes. In this study, we evaluated the prognostic value of paraneoplastic leukocytosis (p-Leukocytosis) and paraneoplastic thrombocytosis (p-Thrombocytosis) in patients with non-small cell lung cancer (NSCLC). We also studied their relation to the expression of commonly detected molecular markers. MethodsWe conducted a retrospective chart review on 571 consecutive NSCLC patients over a 10 year period. Blood counts were recorded at the time of cancer diagnosis. Kaplan-Meier survival curves were used to compare overall survival (OS) between patients with and without p-Leukocytosis (or) p-Thrombocytosis (p-Leuko/Thrombocytosis). Cox regression was used to determine if leukocytosis/thrombocytosis was a predictor of OS in NSCLC.ResultsPatients with p-Leukocytosis and p-Thrombocytosis had a significantly poorer survival compared patients with normal blood counts (P < 0.001). In a multivariate survival analysis, both continued to correlate even when adjusted for histology, gender, stage and chemotherapy (P < 0.01, 0.03 respectively). Stage Ⅰ and Ⅱ NSCLC with p-Leuko/Thrombocytosis did not perform poorly compared to stage Ⅰ/Ⅱ NSCLC patients without paraneoplasia. Patients with the combined leukothrombocytosis syndrome did not have worse outcomes compared to those with either paraneoplastic syndrome alone.Conclusionsp-Leuko/Thrombocytosis is an accessible laboratory parameter of prognostic value in NSCLC. Evidence of p-Leuko/Thrombocytosis portends poor survival. The role of various cytokines in tumor pathobiology provides a rationale for identifying cytokine factors responsible for the paraneoplasia and administering anti-cytokine therapies alongside traditional chemotherapy in an attempt to improve survival outcomes in these subset of patients.

CPA4 is a Novel Diagnostic and Prognostic Marker for Human Non-Small-Cell Lung Cancer.

Background. Carboxypeptidase A4 (CPA4) belongs to a member of the metallocarboxypeptidase family, and its expression in lung cancer samples and clinical significance are still not investigated until now. In this study, we aimed to evaluate the level of CPA4 in non-small-cell lung cancer (NSCLC) samples and correlate its level with clinical outcome.Methods. CPA4 gene expression in lung cancer tissues were analyzed by using the Oncomine database (www.oncomine.org). The expression of CPA4, Survivin and VEGF in lung cancer and adjacent normal tissues were evaluated by IHC using the corresponding primary antibodies on two different commercial tissue arrays (Shanghai Biochip Co., Ltd., Shanghai, China). Their levels in serum were determined by using commercial human enzyme-linked immunosorbent assay kits. We also examined their relations to clinicopathologic parameters, and explored the diagnostic and prognostic value in NSCLC.Results. We identified an elevation of CPA4 in mRNA level and gene amplification in lung cancer tissues in comparison to normal lung tissues. High CPA4 expression was observed in 120/165 (72.7%) NSCLC samples, and significantly correlated with Tumor size, Depth of invasion, Lymph Node Metastasis, Stage, VEGF level and Survivin level. High CPA4 expression is associated with poor prognosis of NSCLC patients. Multivariable Cox regression analysis demonstrated that CPA4 expression was an independent prognostic factor. Furthermore, serum CPA4 level was also significantly higher in NSCLC patients than in healthy controls. Logistic regression analysis revealed that serum CPA4 and CYFRA21-1 level were the significant parameters for detecting NSCLC. Receiver operating characteristic curves (ROC) in NSCLC patients versus normal people yielded the optimal cut-off value was 2.70 ng/ml for CPA4 and 19 ng/ml for CYFRA21-1, respectively. The area under ROC curve (AUC) was 0.830 for the combination of the two tumor markers.Conclusion. Our results demonstrated that overexpression of CPA4 in NSCLC is associated with an unfavorable prognosis, and serum CPA4 level combining with serum CYFRA21-1 level could be used to aid early detection of NSCLC.

Final Results of a Phase II Study of Bevacizumab, Cisplatin and Pemetrexed as First-Line Therapy for Patients with Advanced Non-Squamous Non-Small Cell Lung Cancer

Background: Efficacy and safety data for cisplatin and pemetrexed plus bevacizumabinnon squamousnon non-small cell lung cancer (NSCLC) are still limited. Nevertheless, either bevacizumab plus platinum doublet or pemetrexed plus platinum is approved options for first line therapy. Predictive factors for bevacizumab are needed. KRAS is one of the most common oncogenic drivers in lung cancer. Its prognostic and predictive value in NSCLC is under investigation. Patients and methods: This trial evaluates the addition of bevacizumab 7.5 mg/kg to cisplatin 75 mg/m2 plus pemetrexed 500 mg/m2 as first line treatment in stage IV non-squamous NSCLC patients. Maintenance bevacizumab was received as monotherapy until progressive disease, unacceptable toxicityor consent with drawal. The primary objective was progression free survival (PFS). Secondary objectives included overall survival (OS), safety, global objective responses and the determination of KRAS mutation at baseline. Results: From March 2009 to March 2012, 31 patients were enrolled. Mean age was 59.19 (standard deviation (SD) 8.53). From all the patients included in this trial, 67.70% were men. KRAS was wild type in 19 patients (58.06%); in 7 (22.58%) was mutated and was unknown in 6 patients (19.35%). Median PFS for KRAS mutated patients was 4 months, whereas for the KRAS wild type it was 7.9 months (P = 0.0031). Median OS was 4 months for the KRAS population, and 16.1 months for the KRAS wild type (P = 0.0032). Twenty four patients (77.42%) experienced at least a grade 3 - 4 adverse event. The most common grade 3 - 4 toxicity was asthenia. Conclusions: Both PFS and OS were statistically longer for the KRAS wild type patients compared with the KRAS mutated population (P = 0.0031). Median OS was shorter than the reported in previous trials with bevacizumab. Nevertheless, focussing on the OS for KRAS wild type patients, this achieves a result or 16.1 months. Therefore, this would be a consistent data supporting to qualify this parameter as a predictive factor before starting treatment for NSCLC.

Abstract 4331: Menacalc as an independent prognostic factor and predictor of metastasis in Non-Small Cell Lung Cancer

Abstract The Mena protein has been shown to be critical for breast cancer metastasis, and, in pre-clinical models two of its isoforms MenaINV and Mena11a are expressed in invasive and non-invasive tumor cell subpopulations, respectively. We have previously developed a quantitative immunofluorescence approach to assess the fraction of Mena protein lacking Mena11a called Menacalc as a method to infer presence of invasive tumor cells. Menacalc is determined by subtracting the population-based Z-score (quantifies the relationship to the mean) for Mena11a quantitative expression score from the population-based Z-score for total Mena quantitative expression score. The resulting Menacalc scores range from -4 to +4 with high scores reflecting tumors with relatively high total Mena (all isoforms) but low Mena11a. We showed that high MENAcalc scores associated significantly with poor disease-specific survival in breast cancer patients, including node-negative and positive cases. A recent study reported that a semi-quantitative approached to Mena assessment shows prognostic value in early Non-Small Cell Lung Cancer (NSCLC) (Bria et. al 2014). Here, we asked whether the quantitative Menacalc assay has prognostic value in NSCLC and if it can predict metastatic disease. We determined Menacalc on a cohort of 201 predominantly early-stage NSCLC cases using multiplexed quantitative immunofluorescence (MQIF) and then assessed Menacalc with respect to clinical and pathological features, as well as disease-specific survival. There was no significant difference in Menacalc by stage, sex, or age. However, Menacalc was significantly higher (P = 0.001) in squamous cell carcinoma (N = 32) as compared to other histological subtypes. High levels of Menacalc were associated (10% significance level) with decrease 5-year disease specific survival in all patients [HR = 1.78 (95%CI: 0.92-3.43); P = 0.08], and were significantly associated with survival when either corrected for histological subtype [HR = 2.10 (95%CI: 1.04-4.26); P = 0.04] or in the squamous-only population [HR = 6.60 (95%CI: 1.22-53.75); P = 0.04]. There was no difference in survival as a function of stage nor in adenocarcinomas. Furthermore, consistent with Mena isoform status being associated with the metastatic phenotype, Menacalc was 2-fold higher (P = 0.09) in metastatic patients, but the sample size with available clinical data was limited (N = 9). Taken together, these data demonstrate that Menacalc has potential prognostic value especially in squamous cell carcinoma patients. These data also preliminarily suggest that Menacalc can predict the metastatic phenotype. Further studies examining Mena isoform expression in squamous and later-stage metastatic NSCLC patients is thus warranted and currently ongoing. Citation Format: Mark Gustavson, Wendy Davis, Oscar Bronsther, Frank Gertler. Menacalc as an independent prognostic factor and predictor of metastasis in Non-Small Cell Lung Cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4331. doi:10.1158/1538-7445.AM2015-4331