Purpose We performed a multi-institutional study to identify prognostic factors and determine outcomes for patients with ALK-rearranged non–small-cell lung cancer (NSCLC) and brain metastasis. Patients and Methods A total of 90 patients with brain metastases from ALK-rearranged NSCLC were identified from six institutions; 84 of 90 patients received radiotherapy to the brain (stereotactic radiosurgery [SRS] or whole-brain radiotherapy [WBRT]), and 86 of 90 received tyrosine kinase inhibitor (TKI) therapy. Estimates for overall (OS) and intracranial progression-free survival were determined and clinical prognostic factors were identified by Cox proportional hazards modeling. Results Median OS after development of brain metastases was 49.5 months (95% CI, 29.0 months to not reached), and median intracranial progression-free survival was 11.9 months (95% CI, 10.1 to 18.2 months). Forty-five percent of patients with follow-up had progressive brain metastases at death, and repeated interventions for brain metastases were common. Absence of extracranial metastases, Karnofsky performance score ≥ 90, and no history of TKIs before development of brain metastases were associated with improved survival (P = .003, < .001, and < .001, respectively), whereas a single brain metastasis or initial treatment with SRS versus WBRT were not (P = .633 and .666, respectively). Prognostic factors significant by multivariable analysis were used to describe four patient groups with 2-year OS estimates of 33%, 59%, 76%, and 100%, respectively (P < .001). Conclusion Patients with brain metastases from ALK-rearranged NSCLC treated with radiotherapy (SRS and/or WBRT) and TKIs have prolonged survival, suggesting that interventions to control intracranial disease are critical. The refinement of prognosis for this molecular subtype of NSCLC identifies a population of patients likely to benefit from first-line SRS, close CNS observation, and treatment of emergent CNS disease.

Lung cancer is the leading cause of cancer-related deaths worldwide. Despite advancements and improvements in surgical and medical treatments, the survival rate of lung cancer patients remains frustratingly poor. Local control for early-stage nonsmall cell lung cancer (NSCLC) has dramatically improved over the last decades for both operable and inoperable patients. However, the molecular mechanisms of NSCLC invasion leading to regional and distant disease spread remain poorly understood. Here, we identify microRNA-224 (miR-224) to be significantly up-regulated in NSCLC tissues, particularly in resected NSCLC metastasis. Increased miR-224 expression promotes cell migration, invasion, and proliferation by directly targeting the tumor suppressors TNFα-induced protein 1 (TNFAIP1) and SMAD4. In concordance with in vitro studies, mouse xenograft studies validated that miR-224 functions as a potent oncogenic miRNA in NSCLC in vivo. Moreover, we found promoter hypomethylation and activated ERK signaling to be involved in the regulation of miR-224 expression in NSCLC. Up-regulated miR-224, thus, facilitates tumor progression by shifting the equilibrium of the partially antagonist functions of SMAD4 and TNFAIP1 toward enhanced invasion and growth in NSCLC. Our findings indicate that targeting miR-224 could be effective in the treatment of certain lung cancer patients.ung cancer is the second most common cancer and the leading cause of cancer-related death worldwide. In 2013, there were an estimated 228,190 new cases of lung cancer and 159,480 deaths in the United States. Despite advancements and improvements in surgical and medical treatments, the 5-y survival rate of lung cancer patients remains frustratingly poor (1). Although local control for early-stage nonsmall cell lung cancer (NSCLC) has dramatically improved over the last decades for both operable and inoperable patients (2, 3), ∼20% of early-stage patients, however, are developing distant metastasis (4, 5), and

Purpose To determine the safety and efficacy of gefitinib, an EGFR tyrosine kinase inhibitor, in combination with radiation for newly diagnosed glioblastoma (GBM) patients. Methods and Materials Between 3/21/2002 and 5/03/2004 RTOG 0211 enrolled 31 and 147 GBM patients in the phase I and II arms respectively. Treatment consisted of daily oral gefinitnib started at the time of conventional cranial radiotherapy (RT) and continued post RT for 18 months or until progression. Tissue microarrays from 68 cases were analyzed for EGFR expression. Results The maximum tolerated dose (MTD) of gefitinib was determined to be 500 mg in patients on non enzyme-inducing anticonvulsant drugs (non-EIAEDs). All patients in the phase II component were treated at a gefitinib dose of 500mg; patients receiving EIADSs could be escalated to 750mg. The most common side-effects of gefitinib in combination with radiation were dermatologic and gastrointestinal. Median survival was 11.5 months for patients treated per protocol. There was no overall survival benefit for patients treated with gefitinib + RT when compared to a historical cohort of patients treated with RT alone, matched by RTOG RPA class distribution. Younger age was significantly associated with better outcome. Per protocol stratification, EGFR expression was not found to be of prognostic value for gefitinib + RT treated patients. Conclusions The addition of gefitinib to RT is well tolerated. Median survival of RTOG 0211 patients treated with radiation therapy with concurrent and adjuvant gefitinib was similar to a historical control cohort treated with radiation alone.

Purpose We performed genome-wide microRNA-sequencing (miRNA-seq) in primary cancer tissue from lung adenocarcinoma patients to identify markers for the presence of lymph node metastasis. Experimental Design Markers for lymph node metastasis identified by sequencing were validated in a separate cohort using QPCR. After additional validation in the TCGA dataset, functional characterization studies were performed in vitro. Results MiR-31 was upregulated in lung adenocarcinoma tissues from patients with lymph node metastases compared to those without lymph node metastases. We confirmed miR-31 to be up-regulated in lymph node positive patients in a separate patient cohort (p=0.009, t-test), and to be expressed higher in adenocarcinoma tissue than in matched normal adjacent lung tissues (p<0.0001, paired t-test). MiR-31 was then validated as a marker for lymph node metastasis in an external validation cohort of 233 lung adenocarcinoma cases of the TCGA (p=0.031, t-test). In vitro functional assays showed that miR-31 increases cell migration, invasion, and proliferation in an ERK1/2 signaling dependent manner. Of note, miR-31 was a significant predictor of survival in a multivariate cox regression model even when controlling for cancer staging. Exploratory in silico analysis showed that low expression of miR-31 is associated with excellent survival for T2N0 patients. Conclusions We applied microRNA-seq to study microRNomes in lung adenocarcinoma tissue samples for the first time and identified potentially a microRNA predicting the presence of lymph node metastasis and survival outcomes in lung adenocarcinoma patients.

MicroRNAs regulate several aspects of tumorigenesis and cancer progression. Most cancer tissues are archived formalin-fixed and paraffin-embedded (FFPE). While microRNAs are a more stable form of RNA thought to withstand FFPE-processing and degradation there is only limited evidence for the latter assumption. We examined whether microRNA profiling can be successfully conducted on FFPE cancer tissues using SOLiD ligation based sequencing. Tissue storage times (2–9 years) appeared to not affect the number of detected microRNAs in FFPE samples compared to matched frozen samples (paired t-test p>0.7). Correlations of microRNA expression values were very high across microRNAs in a given sample (Pearson’s r = 0.71–0.95). Higher variance of expression values among samples was associated with higher correlation coefficients between FFPE and frozen tissues. One of the FFPE samples in this study was degraded for unknown reasons with a peak read length of 17 nucleotides compared to 21 in all other samples. The number of detected microRNAs in this sample was within the range of microRNAs detected in all other samples. Ligation-based microRNA deep sequencing on FFPE cancer tissues is feasible and RNA degradation to the degree observed in our study appears to not affect the number of microRNAs that can be quantified.

We conducted a pilot study to assess the feasibility and the potential implications of detecting promoter (p)-mutant cell-free tumor-derived DNA (tDNA) in the cerebrospinal fluid (CSF) and plasma of glioblastoma patients. Matched CSF and plasma samples were collected in 60 patients with glial tumors. The CSF collection was obtained during surgery, before any surgical manipulation of the tumor. The extracted tDNA and corresponding tumor DNA samples were analyzed for p and isocitrate dehydrogenase ( hotspot mutations. In addition, the variant allele frequency (VAF) of p mutation in the CSF-tDNA was correlated with tumor features and patients' outcome. Thirty-eight patients had p-mutant/ wild-type glioblastomas. The matched p mutation in the CSF-tDNA was successfully detected with 100% specificity (95% CI, 87.6-100%) and 92.1% sensitivity (95% CI, 78.6-98.3%) ( = 35/38). In contrast, the sensitivity in the plasma-tDNA was far lower [ = 3/38, 7.9% (95% CI, 1.6-21.4%)]. We concordantly observed a longer overall survival of patients with low VAF in the CSF-tDNA when compared with patients with high VAF, irrespective of using the lower quartile VAF [11.45%; 14.0 mo. (95% confidence interval, CI, 10.3-17.6) vs. 8.6 mo. (95% CI, 4.1-13.2), = 0.035], the lower third VAF [13%; 15.4 mo. (95% CI, 11.6-19.2) vs. 8.3 mo. (95% CI, 2.3-14.4), = 0.008], or the median VAF [20.3%; 14.0 mo. (95% CI, 9.2-18.7) vs. 8.6 mo. (95% CI, 7.5-9.8), = 0.062] to dichotomize the patients. This pilot study highlights the value of CSF-tDNA for an accurate and reliable detection of p mutations. Furthermore, our findings suggest that highp mutation VAF levels in the CSF-tDNA may represent a suitable predictor of poor survival in glioblastoma patients. Further studies are needed to complement the findings of our exploratory analysis. .

No abstract

Purpose We employed a metabolomics-based approach with the goal to better understand the molecular signatures of glioblastoma (GBM) cells and tissues, with an aim towards identifying potential targetable biomarkers for developing more effective and novel therapies. Experimental Design We used liquid chromatography coupled with mass spectrometry (LC-MS/Q-TOF and LC-MS/QQQ) for the discovery and validation of metabolites from primary and established GBM cells, GBM tissues, and normal human astrocytes. Results We identified tryptophan, methionine, kynurenine, and 5-methylthioadenosine as differentially regulated metabolites (DRMs) in GBM cells compared to normal human astrocytes (NHAs). Unlike NHAs, GBM cells depend on dietary methionine for proliferation, colony formation, survival, and to maintain a deregulated methylome (SAM:SAH ratio). In methylthioadenosine phosphorylase (MTAP) deficient GBM cells, expression of MTAP transgene did not alter methionine dependency, but compromised tumor growth in vivo. We discovered that a lack of the kynurenine metabolizing enzymes kynurenine monooxygenase and/or kynureninase promotes the accumulation of kynurenine, which triggers immune evasion in GBM cells. Insilico analysis of the identified DRMs mapped the activation of key oncogenic kinases that promotes tumorigenesis in GBM. We validated this result by demonstrating that the exogenous addition of DRMs to GBM cells in vitro, results in oncogene activation as well as the simultaneous downregulation of Ser/Thr phosphatase PP2A. Conclusion We have connected a four-metabolite signature, implicated in the methionine and kynurenine pathways, to the promotion and maintenance of GBM. Together, our data suggest that these metabolites and their respective metabolic pathways serve as potential therapeutic targets for GBM.