Strategy For Non-small Cell Lung Cancer Research Articles

PD-L1 expression testing in non-small cell lung cancer.

In recent years, immunotherapy has revolutionized and changed the standard of care in patients with advanced non-small cell lung cancer (NSCLC). Immune checkpoint inhibitors, fundamentally those that act by blocking the programmed cell death receptor-1 (PD-1) and its ligand the programmed cell death ligand-1 (PD-L1) have emerged as novel treatment strategies in NSCLC, demonstrating undoubted superiority over chemotherapy in terms of efficacy. Several of these immune checkpoint modulators have recently gained regulatory approval for the treatment of advanced NSCLC, such as nivolumab, atezolizumab and pembrolizumab in first-line (only the latter) and second-line settings, and more recently, durvalumab as maintenance after chemoradiotherapy in locally advanced disease. There is consensus that PD-L1 expression on tumor cells predicts responsiveness to PD-1 inhibitors in several tumor types. Hence PD-L1 expression evaluated by immunohistochemistry (IHC) is currently used as a clinical decision-making tool to support the use of checkpoint inhibitors in NSCLC patients. However, the value of PD-L1 as the ‘definitive’ biomarker is controversial as its testing is puzzled by multiple unsolved issues such as the use of different staining platforms and antibodies, the type of cells in which PD-L1 is assessed (tumor versus immune cells), thresholds used for PD-L1-positivity, or the source and timing for sample collection. Therefore, newer biomarkers such as tumor mutation burden and neoantigens as well as biomarkers reflecting host environment (microbiome) or tumor inflamed microenvironment (gene expression signatures) are being explored as more reliable and accurate alternatives to IHC for guiding treatment selection with checkpoint inhibitors in NSCLC.

KRIBB11 accelerates Mcl-1 degradation through an HSF1-independent, Mule-dependent pathway in A549 non-small cell lung cancer cells

The Bcl-2 family protein, Mcl-1 is known to have anti-apoptotic functions, and depletion of Mcl-1 by cellular stresses favors the apoptotic process. Moreover, Mcl-1 levels are frequently increased in various cancer cells, including non-small cell lung cancer (NSCLC), and is implicated in resistance to conventional chemotherapy and in cancer metastasis. In this study, we demonstrated that KRIBB11 accelerates the proteasomal degradation of Mcl-1 in the NSCLC cell line, A549. While KRIBB11 is an inhibitor of HSF1, we found that KRIBB11 induced Mcl-1 degradation in an HSF1-independent manner. Furthermore, this process was triggered via increase ubiquitination by the E3 ligase, Mule, rather than via de-ubiquitination by USP9X. Additionally, we found that Mcl-1 levels were only transiently reduced by KRIBB11: Mcl-1 levels were gradually restored as KRIBB11 activity diminished. However, we found that this effect was blocked in BIS (Bcl-2 interacting cell death suppressor, also called BAG3)-depleted cells, and that BIS prevents Mcl-1 from undergoing HSP70-driven proteasomal degradation, through an interaction with HSP70. Taken together, our results suggest that targeting Mcl-1 with KRIBB11 treatment, while simultaneously downregulating BIS, could be a therapeutic strategy in NSCLC.

MEDI 4736 (durvalumab) in non-small cell lung cancer

ABSTRACTIntroduction: Immune checkpoint inhibitors (ICI) are now a therapeutic option for advanced non-small cell lung cancer (NSCLC) patients. ICI, such as the PD-1 inhibitors nivolumab and pembrolizumab and the PD-L1 inhibitor atezolizumab, have already been marketed for the treatment of pretreated patients with advanced NSCLC. Other notable PD-L1 inhibitors under development include avelumab and durvalumab.Areas covered: This article reviews literature on durvalumab development, from the preclinical data to the results of phase III clinical trials, whether published or presented at international scientific conferences. Ongoing clinical trials were also reviewed.Expert opinion: Early phase trials of durvalumab monotherapy (and in combination) have demonstrated activity in advanced NSCLC patients and it has demonstrated a good safety profile. The authors believe that durvalumab will likely play an important role in future treatment strategies for NSCLC. The PACIFIC trial assessing durvalumab after standard chemoradiotherapy for locally advanced NSCLC has already met its primary endpoint and the potential of durvalumab will be reinforced if phase III randomized studies of first-line (MYSTIC trial) and second or subsequent (ARCTIC trial) lines of therapy demonstrate superiority over the current standard of care.

Abstract 1335: Deubiquitinase OTUD6B isoforms are important regulators of growth and proliferation

Abstract Deubiquitinases (DUBs) are increasingly linked to the regulation of fundamental processes in normal and cancer cells, including DNA replication and repair, programmed cell death, and oncogenes and tumor suppressors signaling. Here evidence is presented that the deubiquitinase OTUD6B regulates protein synthesis in non-small cell lung cancer (NSCLC) cells, operating downstream from mTORC1. OTUD6B associates with the protein synthesis initiation complex and modifies components of the 48S preinitiation complex. The two main OTUD6B splicing isoforms seem to regulate protein synthesis in opposing fashions: the long OTUD6B-1 isoform is inhibitory, while the short OTUD6B-2 isoform stimulates protein synthesis. These properties affect NSCLC cell proliferation, since OTUD6B-1 represses DNA synthesis while OTUD6B-2 promotes it. Mutational analysis and downstream mediators suggest that the two OTUD6B isoforms modify different cellular targets. OTUD6B-2 influences the expression of cyclin D1 by promoting its translation while regulating (directly or indirectly) c-Myc protein stability. This phenomenon appears to have clinical relevance as NSCLC cells and human tumor specimens have a reduced OTUD6B-1/OTUD6B-2 mRNA ratio compared to normal samples. The global OTUD6B expression level does not change significantly between non-neoplastic and malignant tissues, suggesting that modifications of splicing factors during the process of transformation are responsible for this isoform switch. Because protein synthesis inhibition is a viable treatment strategy for NSCLC, these data indicate that OTUD6B isoform 2, being specifically linked to NSCLC growth, represents an attractive, novel therapeutic target and potential biomarker for early diagnosis of malignant NSCLC. Note: This abstract was not presented at the meeting. Citation Format: Maurizio Bocchetta, Clodia Osipo, Anna Sobol. Deubiquitinase OTUD6B isoforms are important regulators of growth and proliferation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1335. doi:10.1158/1538-7445.AM2017-1335

Abstract 151: Inhibition of AKT-1 for the treatment of human non-small cell lung cancer (NSCLC) in-vitro

Abstract AKT is a serine-threonine kinase implicated in tumorigenesis as a central regulator of cellular growth, proliferation, survival, metabolism, and migration. Activated AKT is overexpressed in 50-70% of NSCLC tumors and has exhibited an association with poor prognosis as well as chemotherapeutic resistance to platinum-based therapy. Accordingly, AKT inhibitors such as MK-2206 are currently undergoing clinical investigation for the treatment of human NSCLC however, these agents broadly target all three (1-3) AKT isoforms. Recent evidence suggests opposing roles of the AKT isoforms in tumorigenesis where loss of AKT-1 inhibits while the loss of AKT-2 enhances lung tumor development in transgenic mouse models. Based on these findings, we hypothesized that preferential inhibition of AKT-1 would warrant a more effective therapeutic strategy for NSCLC compared to the current clinical approach of broad AKT inhibition. WST-1 cell viability assays have revealed that a selective AKT-1 inhibitor A-674563 is a more potent regulator of survival in 6 NSCLC cell lines compared to a pan-AKT inhibitor MK-2206. Furthermore, off-target CDK2 inhibition likely contributes to the observed benefits of the AKT-1 inhibitor as the reduction in cell viability largely parallels the effects of a CDK2 inhibitor PHA-848125. In addition, the cell lines with higher endogenous CDK2 and p-CDK2 expression are more sensitive to the AKT-1 inhibitor relative to the pan-AKT inhibitor MK-2206. Basal protein and RNA levels in each of the cell lines have also shown that high AKT-3 expression may confer resistance to the pan-AKT inhibitor MK-2206. Thus, AKT-3 expression has the potential to serve as a predictive marker for patient response to AKT-1 versus broad AKT-inhibition. Cell cycle analysis demonstrated that the AKT-1 inhibitor decreases the proportion of cells in the Go/G1 phase and increases the proportion of cells in the S-phase, indicating a possible S-phase cell cycle arrest. These differences are also more significant in the cell lines with augmented sensitivity to the AKT-1 inhibitor. Therefore, altered cell cycle progression could be the major driver of the therapeutic benefits of the AKT-1 inhibitor. Overall, our findings suggest that AKT-1 inhibition is significantly more effective at reducing NSCLC cell viability in-vitro compared to pan-AKT inhibition. Furthermore, cell lines with higher CDK2 and AKT-3 expression have marginally increased sensitivity to the AKT-1 inhibitor A-674563 compared to the pan-AKT inhibitor MK-2206. Future research will focus on understanding the mechanism of action of the AKT-1 inhibitor through combined western blot, AKT antibody array, flow cytometry, and confocal microscopy data. Additionally, we will investigate the toxicity of these inhibitors on normal human small airway epithelial cells (HSAECs) to ensure preferential activity against malignant over somatic cells. Citation Format: Paige M. Chorner, Roger A. Moorehead. Inhibition of AKT-1 for the treatment of human non-small cell lung cancer (NSCLC) in-vitro [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 151. doi:10.1158/1538-7445.AM2017-151

Survival associated with chronic obstructive pulmonary disease among elderly patients with non-small cell lung cancer.

e18107 Background: Lung cancer and chronic obstructive pulmonary disease (COPD) are among leading causes of morbidity and mortality worldwide. The association between pre-existing COPD and overall survival (OS) among patients with non-small cell lung cancer (NSCLC) remains unclear. We investigated the impact of pre-existing COPD and its subtypes: chronic bronchitis and emphysema on OS in elderly patients diagnosed with NSCLC at different stages. Methods: Using SEER-Medicare data, we identified patients diagnosed with NSCLC between January 1, 2006 and December 31, 2010, > 66 years of age, and continuously enrolled in Medicare Parts A and B in the 12 months prior to diagnosis. Pre-existing COPD in patients with NSCLC were identified using ICD-9 codes. Kaplan Meier method and log-rank tests were used to examine OS by COPD status and COPD subtype. Cox proportional hazards models were fit to assess the risk of death after cancer diagnosis while adjusting for baseline factors. Results: We identified 66,963 patients with NSCLC. Of these, 22,497 (33.60%) had documented COPD before NSCLC diagnosis. For each stage of NSCLC, median OS was shorter in the COPD compared to the non-COPD group (stage I: 692 vs 1130 days, P < 0.0001; stage II: 473 vs 627 days, P < 0.0001; stage III: 224 vs 229 days; P < 0.0001; stage IV: 106 vs 112 days, P < 0.0001). For COPD subtype, median OS for patients with pre-existing chronic bronchitis was shorter compared to emphysema across all stages of NSCLC (stage I: 672 vs 811 days, P < 0.0001; stage II 582 vs 445 days, P < 0.0001; stage III: 255 vs 229 days, P < 0.0001; stage IV: 105 vs 112 days, P < 0.0001). After multivariable adjustment, COPD patients exhibited an 11% shorter time to death compared to non-COPD patients (Hazard Ratio: 1.11, 95% Confidence Interval: 1.09—1.13). Conclusions: There were marked differences in early stage NSCLC, with a decrease in OS from stage I to stage IV in the COPD group. Patients with chronic bronchitis had shorter OS at every stage of NSCLC compared to emphysema. The results may help inform early detection strategies for NSCLC and treatment selection in early and advanced cancer.

Study of early stage non-small-cell lung cancer using Orbitrap-based global serum metabolomics

PurposeThe aim of the project was to apply ultra-high-performance liquid chromatography–quadrupole-Orbitrap-high-resolution mass spectrometry for serum metabolite profiling of non-small-cell lung cancer (NSCLC). This Orbitrap-based methodology has been applied for a study of NSCLC potential markers for the first time.MethodsAfter extraction using protein precipitation, sera were separated on the ACE Excel 2 C18-PFP (100 × 2.1 mm, 2.0 µm) column using gradient elution and analyzed within the range of 70–1000 m/z. Only patients with early stage disease (stages IA–IIB) were included in the study, providing opportunity to find biomarkers for early lung cancer detection. The resulting metabolite profiles were subjected to univariate and multivariate statistical tests.Results36 features were found significantly changed between NSCLC group and controls after FDR adjustment and 19 were identified using various metabolite databases (in-house library, HMDB, mzCloud). The study revealed a number of NSCLC biomarker candidates which belong to such compound classes as acylcarnitines, organic acids, and amino acids. Multivariate ROC curve built using 12 identified metabolites was characterized by AUC = 0.836 (0.722–0.946). There were no significant differences in the serum metabolite profiles between two most common histological types of lung cancer—adenocarcinoma and squamous cell carcinoma.ConclusionsThrough identification of novel potential tumor markers, Orbitrap-based global metabolic profiling is a useful strategy in cancer research. Our study can accelerate development of new diagnostic and therapeutic strategies in NSCLC. The metabolites involved in discrimination between NSCLC patients and the control subjects should be further explored using a targeted approach.

Targeted therapy in nonsmall cell lung cancer.

Activating mutations in the epidermal growth factor receptor gene and rearrangement of the anaplastic lymphoma kinase gene exemplify the molecular characterization of nonsmall cell lung cancer (NSCLC), particularly adenocarcinoma, and its therapeutic relevance. Several genetic alterations with prognostic and predictive role, including ROS, RET, MET, KRAS, have now been identified in adenocarcinoma and some such as DDR2 and fibroblast growth factor receptor 1 in squamous cell carcinoma. This has heralded the development of agents targeted against these aberrations. Better knowledge of tumor biology and development of targeted agents has ushered an era of personalized treatment strategies in NSCLC, leading to improvements not only in tumor control and duration of life but also in quality of life.

Function of follicular helper T cell is impaired and correlates with survival time in non-small cell lung cancer

Non-small cell lung cancer (NSCLC) represents one of the most common and aggressive cancers worldwide. The PD-1/PD-L1 interaction plays important roles in cancer immunology, and expression of PD-L1 has been discovered in NSCLC tumor cells. Since follicular helper T (Tfh) cells have characteristic high PD-1 expression, we therefore investigated the inflammatory status of Tfh in NSCLC. CD4+CXCR5+ T cell population was examined to define Tfh cells. Data showed that frequency of Tfh cells in peripheral blood was significantly lower in NSCLC patients than in healthy controls. In both primary and metastatic tumors, infiltration of Tfh cells was observed, suggesting that they participated in the antitumor immunity of NSCLC patients. Compared to other T cell subsets, the Tfh cells from the peripheral blood and the resected tumors of NSCLC patients presented elevated apoptosis and reduced proliferation capacity. The Tfh cells from NSCLC patients were also less effective at downregulating IgD and upregulating CD27 expression in naive B cells, and induced less IgM, IgG and IgA secretion, than those from healthy controls. We then found that the survival time from the date of surgery was positively correlated with the frequency of tumor-infiltrating Tfh cells in NSCLC subjects. Overall, the results from this study demonstrated that the Tfh cells were likely involved in the antitumor immunity and were associated with better clinical outcomes, but suffered strong immunosuppression in NSCLC. Enhancing the Tfh cell activity therefore represents a potential therapeutic strategy in NSCLC.

In pursuit of synergy: An investigation of the PI3K/mTOR/MEK co-targeted inhibition strategy in NSCLC.

Clinical PI3K inhibition has been somewhat disappointing, due to both inadequate patient stratification and compensatory cell signalling through bypass mechanisms. As such, investigation of PI3K-MEK co-targeted inhibition has been recommended. With high mortality rates and a clear need for new therapeutic intervention strategies, non-small cell lung cancer (NSCLC) is an important setting to investigate the effectiveness of this approach.Here, 174 NSCLC tumours were screened for 150 mutations by Fluidigm technology, with 15 patients being profiled for phosphoprotein expression. The effects of GDC-0941 (a pan PI3K inhibitor), GDC-0980 (a dual PI3K/mTOR inhibitor) and GDC-0973 (a MEK inhibitor) alone and in combination were assessed in 3 NSCLC cell lines.PIK3CA was mutated in 5.17% of NSCLC patients. GDC-0941 and GDC-0980 treatment induced anti-proliferative and pro-apoptotic responses across all NSCLC cell lines, while GDC-0973 treatment induced only anti-proliferative responses. GDC-0980 and GDC-0973 combined treatment led to significant increases in apoptosis and synergistic reductions in proliferation across the panel of cell lines.This study found that the PI3K/MEK co-targeted inhibition strategy is synergistic in all 3 molecular subtypes of NSCLC investigated. Consequently, we would advocate clinical trials for NSCLC patients combining GDC-0980 and GDC-0973, each of which are separately under clinical investigation currently.

P90 ribosomal S6 kinase: a potential therapeutic target in lung cancer.

A global survey of cancer has shown that lung cancer is the most common cause of the new cancer cases and cancer deaths in men worldwide. The mortality from lung cancer is more than the combined mortality from breast, prostate and colorectal cancers. The two major histological types of lung cancer are non-small cell lung cancer (NSCLC) accounting for about 85 % of cases and small cell lung cancer accounting for 15 % of cases. NSCLC, the more prevalent form of lung cancer, is often diagnosed at an advanced stage and has a very poor prognosis. Many factors have been shown to contribute to the development of lung cancer in humans including tobacco smoking, exposure to environmental carcinogens (asbestos, or radon) and genetic factors. Despite the advances in treatment, lung cancer remains one of the leading causes of cancer death worldwide. Interestingly, the overall 5 year survival from lung cancer has not changed appreciably in the past 25 years. For this reason, novel and more effective treatments and strategies for NSCLC are critically needed. p90 ribosomal S6 kinase (RSK), a serine threonine kinase that lies downstream of the Ras–MAPK (mitogen activated protein kinase) cascade, has been demonstrated to be involved in the regulation of cell proliferation in various malignancies through indirect (e.g., modulation of transcription factors) or direct effects on the cell-cycle machinery. Increased expression of RSK has been demonstrated in various cancers, including lung cancer. This review focuses on the role of RSK in lung cancer and its potential therapeutic application.

MicroRNA‑361‑5p suppresses cancer progression by targeting signal transducer and activator of transcription 6 in non‑small cell lung cancer.

The incidence of non-small cell lung cancer (NSCLC) has significantly increased in China, while the prognosis of affected patients is poor. The pathogenesis of NSCLC is thought to be regulated by microRNAs (miRs). The present study used a miR array in order to determine the expression of miR-361-5p, which was significantly lower in NSCLC tissues compared with that in adjacent tissues, indicating a crucial role of miR-361-5p during the progression of NSCLC. Furthermore, the effects of transfection-induced upregulation of miR-361-5p on the NSCLC cell line H23 were assessed. Overexpression of miR-361-5p inhibited the proliferation and colony formation ability of H23 cells. In addition, apoptosis of H23 cells was induced by upregulation of miR-361-5p. Furthermore, signal transducer and activator of transcription 6 (STAT6) was confirmed as a direct target of miR-361-5p by a dual-luciferase reporter assay. Moreover, inhibition of STAT6 by small interfering RNA or miR-361-5p also decreased the expression of B-cell lymphoma extra large (Bcl-xL). In vivo, miR-361-5p significantly reduced tumor growth in a nude mouse xenograft model, and suppressed STAT6 and Bcl-xL expression. In conclusion, the present study indicated that miR-361-5p may represent a novel molecular tool for therapeutic and diagnostic strategies in NSCLC.

Abstract 5355: Oncolytic viral therapy with immune modulation is an effective novel treatment strategy for non-small cell lung cancer

Abstract Background: Lung cancer is the leading cause of cancer mortality worldwide. Non-small cell lung cancer (NSCLC) represents approximately 85% of these diagnoses and despite incremental improvements with cytotoxic chemotherapy and targeted therapeutics, the five-year overall survival is only 17%. Recently improved understanding of immune checkpoint pathways and immunosuppresive cellular populations such as myeloid-derived suppressor cells (MDSCs) have led to a resurgence in immunotherapeutic strategies. Reovirus (RV) is an oncolytic virus that only targets cancer cells with certain aberrant signaling pathways such as Ras while avoiding normal cells. Furthermore, its role in stimulating an anti-tumor immune response has been recently described. Additionally, Sunitinib (S), a multitargeted tyrosine kinase inhibitor (anti-VEGFR, C-Kit and PDGFRa) also affects tumor microenvironment Tregs and MDSC numerically, which could consequentially augment RV immunotherapeutic potential. Hypothesis: RV and S combination therapy is an effective treatment modality for NSCLC via direct oncolysis, tumor immunosuppression reversal, and host anti-tumor immune response stimulation. Methods: To determine the relative susceptibility of selected NSCLC cell lines to RV, S and combination therapy, a WST-1 cell viability assay was conducted. Four human NSCLC cell lines A549, H460, H1299, and H1975 and the mouse Lewis lung carcinoma (LLC) cell line LL2 were tested, and the combination index (CI) was calculated using Calcusyn software to determine in vitro synergy. The LLC syngeneic C57BL/6 immunocompetent mouse model bearing LL2 cells in the right hind-flank was treated with PBS, S, RV, UV-inactivated RV (DV), or combination therapies (N = 6 mice/group). Mice were sacrificed on day 27. Spleens, blood, and tumors were harvested for analysis. Splenocytes were subject to flow cytometry CD11b and Gr-1 detection for MDSC quantification. Splenocyte CD8+ enrichment was performed, co-cultured with LL2, RV, DV, S or combinations and subject to IFN-γ quantification via ELISA to determine immune recognition. Results: Treatment resulted in efficacy and synergy of RV/S in A549, H460, and H1299 in vitro as indicated by CI values < 1. LL2 showed additive to synergistic responses at varying treatment concentrations. When LLC tumors in a C57BL/6 immunocompetent murine model were treated with the combination of RV/S, a significant tumor reduction occurred compared to monotherapies and untreated controls (non-overlapping 95% confidence intervals). Furthermore, an increase in anti-tumor immunity stimulating IFN-γ was detected in mice samples upon RV and combination exposure at 0.13μg/mL and 0.09μg/mL compared to PBS treated mice at 0μg/mL. Both S and RV/S treated mice had a significant splenocyte MDSC reduction. These results indicate that Reovirus and Sunitinib combination therapy holds promise as a novel treatment strategy for NSCLC. Citation Format: Jianrui Liu, Jason Spurrel, Zhong Qiao Shi, WenQian Chen, Don G. Morris. Oncolytic viral therapy with immune modulation is an effective novel treatment strategy for 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 5355. doi:10.1158/1538-7445.AM2015-5355

Abstract 3501: EGFR tyrosine kinase inhibitors combined with cytotoxic drugs for treatment of NSCLC with EGFR gene mutations: Efficacy and mechanisms

Abstract Background: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have improved overall survival in non-small-cell lung cancer (NSCLC) patients whose tumors have EGFR activating mutations. After treatment with EGFR-TKIs, NSCLC tumors with EGFR activating mutations become resistant to EGFR-TKIs, and development of novel treatment strategies is needed to improve survival in such patients.Next-generation EGFR-TKIs have been developed, and a clinical trial showed clinical activity of 3rd generation EGFR-TKIs in patients with acquired resistance, especially secondary mutation T790M to EGFR-TKIs. Moreover, clinical trials demonstrated survival benefit of combination with cytotoxic agents and EGFR-TKIs in patients with EGFR-mutated NSCLC. In contrast, the IMPRESS study showed that continuation of gefitinib in addition to cisplatin and pemetrexed didn't show survival benefit in patients with acquired resistance to gefitinib.Several basic studies reported that efficacy of targeted therapy combined with chemotherapy is schedule dependent. We, therefore, conducted an in vitro study to develop a novel therapeutic strategy for NSCLC with EGFR mutations. Methods: In vitro effects of docetaxel and an EGFR-TKI, gefitinib (AstraZeneca) or AZD9291 (Selleckchem), were examined in five lung cancer cell lines (PC9, PC9GR, HCC827, HCC827GR, and H1975) using a 3D culture model (PrimeSurface 96, SUMITOMO BAKELITE CO.). The NSCLC cells were treated with the combinationeither concurrently or sequentially (EGFR-TKIs after docetaxel or docetaxel after EGFR-TKIs). Combination Index (CI) values were calculated at various levels of cytotoxicity (the Chou-Talalay method). We evaluated changes in MAPK pathway activation and in expressions of Bcl2 family proteins and cleaved PARP (Cell Signaling) by western blot analysis. Results: Docetaxel followed by EGFR-TKIs weremore effective at CI, compared with EGFR-TKIs followed by docetaxel in two of the five cell lines with EGFR activating mutations, PC9GR (EGFR19del+T790M) and HCC827GR (EGFR19del+METamp) cells. The HCC827GR cells were likely to be more sensitive than PC9GR to the combination with docetaxel and gefitinib. The combo led to inactivation of the MAPK pathway and alteration of the Bcl2 family proteins. d) Conclusion: In conclusion, our results suggest that the sequential combination of docetaxel followed by EGFR-TKIs was effective against NSCLC with acquired resistance to EGFR-TKIs. Citation Format: Ryohei Yoshida, Synsuke Okumura, Takaaki Sasaki, Yoshinobu Ohsaki. EGFR tyrosine kinase inhibitors combined with cytotoxic drugs for treatment of NSCLC with EGFR gene mutations: Efficacy and mechanisms. [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 3501. doi:10.1158/1538-7445.AM2015-3501

Abstract B08: Kicking the addictions: Therapeutic strategies for NSCLC

Abstract The discovery of cancer cells' dependency on the self-cannibalization program autophagy has spawned the investigation of its efficacy as a therapeutic target. However, the use of lysomotropic agents to inhibit autophagy as single-agent therapeutics has proven less than optimal. Recent investigations into the design of sequential treatment strategies targeting pathways preferentially used by cancer cells have shown promise. This novel spin on personalized medicine seeks to exploit the kinetics and mechanisms by which cancer cells become dependent on proliferation and survival pathways. Oncogenic KRAS-driven non-small cell lung cancer (NSCLC) is known to depend on mTORC1-driven growth and autophagy for survival; such autophagy addiction is beneficial to cancer cells, as it eliminates the need for exogenous nutrient supplies. We aim to elucidate the mechanism and kinetics of autophagy addiction in NSCLC, in order to design sequential treatment strategies against this disease. Citation Format: Abigail R. Solitro, Jeffrey P. MacKeigan. Kicking the addictions: Therapeutic strategies for NSCLC. [abstract]. In: Proceedings of the AACR Special Conference: Targeting the PI3K-mTOR Network in Cancer; Sep 14-17, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(7 Suppl):Abstract nr B08.

Basic fibroblast growth factor shows prognostic impact on survival in operable non-small cell lung cancer patients.

BackgroundThe important role of angiogenesis displaying in tumor development and metastasis has been generally realized. Vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and endostatin (ES) are critical members of angiogenesis modulating the balance between pro-angiogenenic and anti-angiogenenic factors. The aim of this study was to evaluate the circulating level of these factors in serum and explore their prognostic significance in 96 operable non-small cell lung cancer (NSCLC) patients.MethodsPre-operational serum VEGF, bFGF, and ES were determined by commercially available enzyme-link immunosorbent assay for 96 NSCLC patients and compared to a cohort of healthy controls (n = 51). Values were correlated with clinicopathological features and overall survival (OS).ResultsThe pretreatment serum levels of VEGF, bFGF and ES in NSCLC were significantly higher than in the healthy control (P < 0.001, P = 0.009 and P = 0.016, respectively). Univariate survival analysis showed that a high bFGF level correlated with shorter OS and remained an independent factor in multivariate analysis (hazard ratio [HR] = 1.918, 95% confidence interval [CI], 1.061–3.464). In the squamous subtype, a high bFGF indicated a particularly poor prognosis (HR = 2.609, 95% CI, 1.188–5.729).ConclusionsbFGF is an independent predictor of poor survival in patients with NSCLC. For patients with high serum bFGF, aggressive antitumor treatments should be given after surgery. Approaches targeting the bFGF signaling pathway should be considered as potentially promising therapeutic strategies in NSCLC, especially for the squamous subtype.

Abstract 1704: Combination of Pirfenidone and Cisplatin as a potential therapeutic strategy in NSCLC

Abstract Lung cancer is the leading cause of cancer death in the United States. Due to limited efficacy of current conventional chemotherapy, the majority of patients face poor prognosis and eventually succumb the disease. Therefore, the study of combination therapy using traditional anticancer drugs with novel therapeutic approaches is needed. Pirfenidone is a small molecule that is being used in Phase II and III clinical trials for idiopathic pulmonary fibrosis (IPF). Although it has already been approved for IPF in Europe, there is no known mechanism of action known for the drug. Recently we discovered that Pirfenidone has a detrimental effect on cancer associated fibroblasts (CAFs) as well as in non-small cell lung cancer (NSCLC) cells. Our hypothesis is that Pirfenidone, in combination with standard chemotherapy, will act synergistically and proffer a more potent therapeutic strategy for NSCLC. Our studies show that in a co-culture model there is a decrease in viability on CAFs and NSCLC cells when the combination of Pirfenidone and Cisplatin is used. Furthermore, Pirfenidone and Cisplatin were able to have an anti-tumor effect in a xenograft model of NSCLC and CAFs. In addition, insights in the mechanism by which Pirfenidone and Cisplatin induce synergy in NSCLC cells are being elucidated. Our studies show that the combination of drugs leads to an increase in phosphorylated ERK as well as a decrease in phosphorylated Akt. Thus, involving the PI3K and MAPK/ERK pathways in Pirfenidone/Cisplatin induced cell death. Our lab is the first to recognize the therapeutic potential of combining Pirfenidone and Cisplatin, as well as elucidating the mechanism of action for both drugs in NSCLC. Citation Format: Melanie Mediavilla-Varela, David Noyes, Kingsley Boateng, Scott Antonia. Combination of Pirfenidone and Cisplatin as a potential therapeutic strategy in NSCLC. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1704. doi:10.1158/1538-7445.AM2014-1704

Abstract 1716: Targeting the insulin-like growth factor receptor/Insulin receptor and Src signaling network for the treatment of non-small cell lung cancer

Abstract Insulin-like growth factor I receptor (IGF-1R)-mediated signaling plays an important role in the proliferation, survival, and metastasis of cancer cells. The IGF-1R-targeting anticancer agents including monoclonal antibodies and small molecule tyrosine kinase inhibitors (TKIs) have been developed, but their antitumor effects have been marginal and limited in clinical trials. Therefore, the mechanism underlying resistance to the IGF-1R-targeting therapies and the rational combination strategies to overcome potential drug resistance need to be investigated. In this study, we demonstrated the association of Src with the resistance to the IGF-1R TKI in non-small cell lung cancer (NSCLC). We found the co-activation of IGF-1R/IR and Src in various human NSCLC cell lines, The mRNA expression and phosphorylation of IGF-1R and Src were also significantly correlated with each other in NSCLC databases from a public dataset and a tissue microarray (n=353). Next, we found Src can be activated through multiple pathways including EGFR and integrin β3 signaling and function as an alternative kinase for phosphorylation of IGF-1R, especially at Tyr1135/36, but not Tyr1131. Src is activated in lung cancer cells possessing both primary and acquired resistance to an IGF-1R TKI. Consistent with the results, inhibition of Src significantly attenuated activation of IGF-1R/IR and co-targeting of IGF-1R and Src effectively suppressed cell proliferation, colony formation, and tumor growth in vitro and in vivo. Taken together, these results suggest that Src causes resistance to IGF-1R TKIs by functioning as a key downstream modulator of multiple signaling pathways for IGF-1R phosphorylation and thus co-targeting IGF-1R and Src could be an effective therapeutic strategy for NSCLC. Citation Format: Hye-Young Min, Hye Jeong Yun, Hyo-Jong Lee, Jaebeom Cho, Hyun-Ji Jang, Kyung Min Kim, Woo-Young Kim, Seung-Hyun Oh, Diane Liu, J. Jack Lee, Waun Ki Hong, Ignacio I. Wistuba, Ho-Young Lee. Targeting the insulin-like growth factor receptor/Insulin receptor and Src signaling network for the treatment of non-small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1716. doi:10.1158/1538-7445.AM2014-1716

Tumor biomarker evaluation of 6,785 patients for combination treatment strategies in NSCLC.

e19012 Background: Non-small cell lung cancer (NSCLC) exhibits activation of multiple tumor pathways. Presence of multiple aberrations may account for drug resistance as well as strategies for combination therapies. We examined concurrent aberrations of biomarkers in NSCLC to present an overview of potential patient cohorts who may benefit from such combinations. Methods: 6,785 NSCLC cases were evaluated. Specific testing was performed and included a multiplatform approach: sequencing (Sanger, NGS), protein expression (IHC) and gene amplification (CISH/FISH). Results: The EGFR mutation (MT) rate in this population was 12.7% (135/1059), of which 57% (55/96) overexpressed EGFR (IHC) and 61% (169/277) had EGFR gene amplification (FISH). This demonstrates the dependence on the EGFR pathway and potential importance of dual inhibition with cetuximab and EGFR TKIs. 66% (63/96) and 7% (6/90) of EGFR MT patients were MET high (IHC) and amplified (CISH), respectively, suggesting potential benefit from dual targetin...

PIK3CA Mutations Frequently Coexist with EGFR/KRAS Mutations in Non-Small Cell Lung Cancer and Suggest Poor Prognosis in EGFR/KRAS Wildtype Subgroup

Purpose PIK3CA gene encoding a catalytic subunit of the phosphatidylinositol-3-kinase (PI3K) is mutated and/or amplified in various neoplasia, including lung cancer. Here we investigated PIK3CA gene alterations, the expression of core components of PI3K pathway, and evaluated their clinical importance in non-small cell lung cancer (NSCLC).Materials and methodsOncogenic mutations/rearrangements in PIK3CA, EGFR, KRAS, HER2, BRAF, AKT1 and ALK genes were detected in tumors from 1117 patients with NSCLC. PIK3CA gene copy number was examined by fluorescent in situ hybridization and the expression of PI3K p110 subunit alpha (PI3K p110α), p-Akt, mTOR, PTEN was determined by immunohistochemistry in PIK3CA mutant cases and 108 patients without PIK3CA mutation.Results PIK3CA mutation was found in 3.9% of squamous cell carcinoma and 2.7% of adenocarcinoma. Among 34 PIK3CA mutant cases, 17 tumors harbored concurrent EGFR mutations and 4 had KRAS mutations. PIK3CA mutation was significantly associated with high expression of PI3K p110α (p<0.0001), p-Akt (p = 0.024) and mTOR (p = 0.001), but not correlated with PIK3CA amplification (p = 0.463). Patients with single PIK3CA mutation had shorter overall survival than those with PIK3CA-EGFR/KRAS co-mutation or wildtype PIK3CA (p = 0.004). A significantly worse survival was also found in patients with PIK3CA mutations than those without PIK3CA mutations in the EGFR/KRAS wildtype subgroup (p = 0.043)Conclusions PIK3CA mutations frequently coexist with EGFR/KRAS mutations. The poor prognosis of patients with single PIK3CA mutation in NSCLC and the prognostic value of PIK3CA mutation in EGFR/KRAS wildtype subgroup suggest the distinct mutation status of PIK3CA gene should be determined for individual therapeutic strategies in NSCLC.