Overall Survival In Non-small Cell Lung Cancer Patients Research Articles

GPR115 Contributes to Lung Adenocarcinoma Metastasis Associated With LAMC2 and Predicts a Poor Prognosis.

GPR115, a member of the adhesion G protein-coupled receptor family, is dysregulated in many cancers. However, the expression and function of GRP115 in non-small cell lung cancer (NSCLC) is not clear. Here, we examined the expression pattern, clinical significance, and function of GPR115 in NSCLC by analysis of clinical specimens and human cell lines and bioinformatics analysis. Immunohistochemical analysis of clinical samples showed that GPR115 was significantly upregulated in NSCLC tissues compares with normal lung epithelial tissue (P < 0.05). And GPR115 overexpression is an independent prognostic factor for 5-year overall survival of NSCLC patients [hazard ratio (HR)=1.625, P = 0.008]. Interestingly, higher expression of GPR115 was strongly correlation with differentiation level (P = 0.027), tumor size (P = 0.010), lymph node metastasis (P = 0.022), tumor-node-metastasis stage (P = 0.008), and poor prognosis of lung adenocarcinoma (LUAD, all P = 0.039), but not lung squamous cell carcinoma (LUSC, P > 0.05). Moreover, downregulation of GPR115 by RNA interference in human lung cancer lines inhibited cell proliferation, migration, and invasion. Preliminary bioinformatic analysis confirmed that GPR115 was closely associated with LAMC2 (Spearman correlation coefficient=0.67, P < 0.05), which was accumulated in ECM-receptor interaction and focal adhesion. Consistent with these findings, deceased of GPR115 was associated with E-cadherin, N-cadherin and Vimentin confirmed by western blot. In conclusion, these data suggest that GPR115 may play a role in the tumor growth and metastasis and may have utility as a diagnostic and prognostic marker for LUAD, but not LUSC.

Downregulation of Xeroderma Pigmentosum Complementation Group C Expression by 17-Allylamino-17-Demethoxygeldanamycin Enhances Bevacizumab-Induced Cytotoxicity in Human Lung Cancer Cells

Introduction: Xeroderma pigmentosum complementation group C (XPC) protein is an important DNA damage recognition factor involved in nucleotide excision repair and regulation of non-small-cell lung cancer (NSCLC) cell proliferation and viability. 17-Allylamino-17-demethoxygeldanamycin (17-AAG) blocks ATP binding to heat shock protein 90 (Hsp90), resulting in destabilization of Hsp90-client protein complexes. Vascular endothelial growth factor (VEGF) is a potent angiogenic growth factor expressed by many types of tumors. Bevacizumab (Avastin) is a humanized monoclonal antibody against human VEGF used as an antiangiogenesis agent in the therapy of many cancers, proving successful in increasing objective tumor response rate and prolonging overall survival in NSCLC patients. Methods: After the bevacizumab and/or 17-AAG treatment, the expressions of XPC mRNA were determined by quantitative real-time PCR analysis. Protein levels of XPC and phospho-AKT were determined by Western blot analysis. We used specific XPC small interfering RNA and PI3K inhibitor (LY294002) to examine the role of the AKT-XPC signal in regulating the chemosensitivity of bevacizumab and 17-AAG. Cell viability was assessed by the MTS assay and trypan blue exclusion assay. Results: In this study, bevacizumab decreased XPC expression in human lung squamous cell carcinoma H520 and H1703 cells via AKT inactivation. Enhancement of AKT activity by transfection with constitutively active AKT vectors increased XPC expression and cell survival after treatment with bevacizumab. In addition, 17-AAG synergistically enhanced bevacizumab-induced cytotoxicity and cell growth inhibition in H520 and H1703 cells, associated with downregulation of XPC expression and inactivation of AKT. Discussion/Conclusion: Together, these results may provide a rationale to combine bevacizumab with Hsp90 inhibitors in future to enhance therapeutic effects for lung cancer.

The microRNA-152/human leukocyte antigen-G axis affects proliferation and immune escape of non-small cell lung cancer cells.

ObjectiveTo investigate the role of human leukocyte antigen (HLA-G) on proliferation, invasion, and immune escape in non-small cell lung cancer (NSCLC).MethodsThe relationship between HLA-G and overall survival (OS) of NSCLC patients was analyzed using the KMPlot database. The expression of micro (mi)R-152 or HLA-G was modulated by transfecting synthetic oligonucleotides, and the impact of the miR-152/HLA-G axis on proliferation, invasion, colony formation in soft agar, and tolerance to natural killer (NK) cell cytolysis was measured.ResultsBioinformatics analysis showed that high HLA-G expression was correlated with poor OS in NSCLC patients. The tolerance of NSCLC cells to NK cytotoxicity was negatively correlated with HLA-G and positively correlated with miR-152 expression. Over-expressing miR-152 inhibited HLA-G expression in A549 cells and attenuated cell proliferation, migration, colony formation ability, and tolerance to NK cells. However, blocking HLA-G expression by small interfering RNA did not affect migration or colony formation, but only proliferation and tolerance to NK cells in vitro and in vivo. Blocking Ig-like transcript 2 on the surface of NK cells increased their killing effect in the presence of high HLA-G expression.ConclusionsmiR-152/HLA-G axis plays an oncogenic role in NSCLC by affecting cell proliferation and immune escape.

Association between BIM Deletion Polymorphism and Efficacy of Osimertinib in Advanced EGFR T790M NSCLC Patients

BCL-2-like 11 (BIM) deletion polymorphism (BIM-del) has been found to be associated with resistance to first-generation epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs), such as gefitinib and erlotinib, and is a poor prognostic factor for non-small-cell lung cancer (NSCLC) patients with EGFR mutations. Nevertheless, the impact of the BIM-del in advanced EGFR T790M NSCLC patients treated with third-generation EGFR-TKI, osimertinib, remains undetermined. The aim of this study is to explore the relationship among BIM-del and therapeutic efficacy of osimertinib in NSCLC patients with EGFR T790M. We totally recruited 137 Chinese Han NSCLC patients with EGFR T790M receiving osimertinib treatment from December 2015 to December 2019. Peripheral blood samples from these patients were collected to detect BIM-del by polymerase chain reaction. Clinical outcomes among patients with and without BIM-del were analyzed by Cox proportional hazards models. BIM-del was detected in 17.5% of all patients (24/137). The majority of patients were no more than 60 years (66%, 91/137), female (57%, 78/137), never smokers (81%, 111/137), ECOG performance status (PS) 0-1 (91%, 124/137), and had central nervous system (CNS) metastases (55%, 75/137). There were no associations between the BIM-del and clinical characteristics including age, sex, histology, smoking status, stage, ECOG PS score, and CNS metastases. Patients with BIM-del had a poor objective response rate compared to those without (29% vs. 57%, P = 0.012). BIM-del was associated with shorter progress free survival (PFS) and overall survival (OS). (PFS, 8.3 vs. 10.4 months, P = 0.042; OS, 14.8 vs. 23.4 months, P = 0.005). Multivariate analysis indicated that BIM-del was the independent prognostic factor for both PFS and OS in NSCLC patients with EGFR T790M receiving osimertinib. BIM-del was associated with inferior efficacy and may serve as a negative predictive biomarker to osimertinib in EGFR T790M NSCLC patients. Genotype analysis of the BIM-del in EGFR mutant patients may be helpful for guiding appropriate treatment.

Long-Noncoding RNA CASC9 Promotes Progression of Non-Small Cell Lung Cancer by Promoting the Expression of CDC6 Through Binding to HuR.

ObjectiveThe long-noncoding RNAs (lncRNAs) have been identified as key players in diverse cellular processes in non-small cell lung cancer (NSCLC). However, the understanding of biological functions and detailed mechanisms of lncRNAs is still limited. Herein, the lncRNA cancer susceptibility candidate 9 (CASC9) on NSCLC progression is investigated.Materials and MethodsExpressions of CASC9, HuR and cell division cycle 6 (CDC6) in NSCLC tissues were detected with quantitative real-time polymerase chain reaction (qRT-PCR). The cell counting kit-8, transwell assays, and flow cytometry were used to examine cell proliferation, migration, and the cell cycle. Tumor growth in vivo was evaluated by xenograft tumor experiments and immunohistochemistry. RNA-binding protein immunoprecipitation (RIP) was used to identify the interaction between HuR and CDC6, and CASC9 and HuR.ResultsCASC9, CDC6 and HuR expression were found significantly upregulated in NSCLC tissues, which predicted poorer 5-year overall survival in NSCLC patients. Inhibition of CASC9 significantly reduced the malignancy of NSCLC cells, such as proliferation, migration and cell cycle. In vivo experiments further demonstrated that CASC9 knockdown reduced the tumor growth and the Ki-67 expression. Moreover, CASC9 knockdown inhibited the expression of CDC6 which was detected overexpressed in NSCLC tumor tissues. Then, up-regulation of CDC6 could partly reverse the negative effects of CASC9 on cell proliferation, migration and cell cycle. RIP assay and rescue experiment showed that CASC9 regulated CDC via binding to HuR.ConclusionOur results indicate that CASC9 conferred an aggressive phenotype in NSCLC and might be a pivotal target for this disease.

A fourteen-lncRNA risk score system for prognostic prediction of patients with non-small cell lung cancer.

Growing evidence has underscored long non-coding RNAs (lncRNAs) serving as potential biomarkers for cancer prognosis. However, systematic tracking of a lncRNA signature for prognosis prediction in non-small cell lung cancer (NSCLC) has not been accomplished yet. Here, comprehensive analysis with differential gene expression analysis, univariate and multivariate Cox regression analysis based on The Cancer Genome Atlas (TCGA) database was performed to identify the lncRNA signature for prediction of the overall survival of NSCLC patients. A risk-score model based on a 14-lncRNA signature was identified, which could classify patients into high-risk and low-risk groups and show poor and improved outcomes, respectively. The receiver operating characteristic (ROC) curve revealed that the risk-score model has good performance with high AUC value. Multivariate Cox’s regression model and stratified analysis indicated that the risk-score was independent of other clinicopathological prognostic factors. Furthermore, the risk-score model was competent for the prediction of metastasis-free survival in NSCLC patients. Moreover, the risk-score model was applicable for prediction of the overall survival in the other 30 caner types of TCGA. Our study highlighted the significant implications of lncRNAs as prognostic predictors in NSCLC. We hope the lncRNA signature could contribute to personalized therapy decisions in the future.

Circ_0000376, a Novel circRNA, Promotes the Progression of Non-Small Cell Lung Cancer Through Regulating the miR-1182/NOVA2 Network.

BackgroundHypoxia has been shown to induce the malignant progression of cancer, including non-small cell lung cancer (NSCLC). Circular RNA (circRNA) is considered to be an important regulator of cancer progression. However, the role of a newly discovered circRNA, circ_0000376, in the progression of NSCLC is unclear.MethodsThe relative expression levels of circ_0000376, miR-1182 and neuro-oncological ventral antigen 2 (NOVA2) were detected via quantitative real-time polymerase chain reaction (qRT-PCR). Glucose consumption and lactate production were determined using Glucose Assay Kit and Lactate Assay Kit, respectively. Moreover, the protein levels of glycolysis markers and NOVA2 were measured using Western blot (WB) analysis. Furthermore, 3-(4, 5-dimethyl-2 thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay was performed to assess cell viability, and transwell assay was employed to evaluate cell migration and invasion. The interaction between miR-1182 and circ_0000376 or NOVA2 was confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. In addition, animal experiments were conducted to assess the influence of circ_0000376 silencing on NSCLC tumor growth in vivo.ResultsCirc_0000376 was upregulated in NSCLC, and its high expression was related to the poor overall survival of NSCLC patients. Hypoxia could enhance circ_0000376 expression and promote the glycolysis, viability, migration, and invasion of NSCLC cells. However, silencing of circ_0000376 could inhibit the glycolysis, viability, migration, and invasion of hypoxia-induced NSCLC cells. Additionally, circ_0000376 could sponge miR-1182, and miR-1182 could target NOVA2. MiR-1182 silencing could reverse the inhibitory effect of circ_0000376 knockdown on NSCLC progression, and NOVA2 overexpression also could reverse the suppressive effect of miR-1182 overexpression on NSCLC progression. Meanwhile, miR-1182 inhibitor could invert the negative regulation effect of circ_0000376 silencing on NOVA2 expression. In addition, circ_0000376 knockdown inhibited the NSCLC tumor growth via regulating the miR-1182 and NOVA2 expression in vivo.ConclusionCirc_0000376 promoted NSCLC progression by regulating the miR-1182/NOVA2 axis, suggesting that circ_0000376 might be a potential biomarker for NSCLC treatment.

Mass spectrometry-based serum proteomic signature as a potential biomarker for survival in patients with non-small cell lung cancer receiving immunotherapy.

BackgroundVeriStrat test is a serum assay which uses a mass spectrometry (MS)-based proteomic signature derived from machine learning. It is currently used as a prognostic marker for patients with non-small cell lung cancer (NSCLC) receiving chemotherapy. However, little is known about its role for NSCLC patients receiving immune checkpoint inhibitors (ICIs).MethodsThis is a retrospective study that includes 47 patients with advanced stage NSCLC without an activating EGFR mutation, who underwent the VeriStrat test from 2016 to 2018. Spectra from blood samples were evaluated to assign patients into the VeriStrat ‘Good’ (VS-G) or VeriStrat ‘Poor’ (VS-P) risk group. The clinical outcomes of 32 patients who received programmed cell death 1 (PD-1) inhibitors nivolumab or pembrolizumab were analyzed by VeriStrat status.ResultsThe VS-G group demonstrated significantly higher progression-free survival (PFS) and overall survival (OS) compared to the VS-P group among overall NSCLC patients regardless of treatment (median PFS of 7.1 vs. 4.2 months, P=0.013, and median OS, not reached vs. 17.2 months, P=0.012). Among NSCLC patients treated with ICIs, VS-G classification was associated with significantly increased PFS in comparison to VS-P classification (median PFS of 6.2 vs. 3.0 months, P=0.012), while the differences in OS trended towards significance (median OS, not reached vs. 16.5 months P=0.076). Multivariate analysis showed that the VeriStrat status was significantly correlated with PFS and OS in NSCLC patients treated with ICIs (P=0.017, P=0.034, respectively).ConclusionsMS-based serum proteomic signature has potential as a biomarker for survival outcome in NSCLC patients receiving immunotherapy.

ZFHX3 mutation as a protective biomarker for immune checkpoint blockade in non-small cell lung cancer.

To date, immunotherapy has opened a new chapter in the treatment of lung cancer. Precise biomarkers can help to screen subpopulations of lung cancer to provide the best treatment. Multiple studies suggest that specific gene mutations may be predictive markers in guiding non-small cell lung cancer (NSCLC) immune checkpoint inhibitor (ICI) treatment. A published immunotherapy cohort with mutational and survival data for 350 NSCLC patients was used. First, the mutational data of the immunotherapy cohort were used to identify gene mutations related to the prognosis of ICI therapy. The immunotherapy cohort and TCGA-NSCLC cohort were further studied to elucidate the relationships between specific gene mutations and tumor immunogenicity, antitumor immune response capabilities, and immune cell and mutation counts in the DNA damage response (DDR) pathway. In the immunotherapy cohort (N = 350), ZFHX3 mutations were an independent predictive biomarker for NSCLC patients receiving ICI treatment. Significant differences were observed between ZFHX3-mutant (ZFHX3-MT) and ZFHX3-wild type (ZFHX3-WT) patients regarding the overall survival (OS) time (P < 0.001, HR = 0.26, 95% Cl 0.17-0.41). ZFHX3-MT is significantly associated with higher tumor mutation burden (TMB) and neoantigen load (NAL), and ZFHX3-MT positively correlates with known immunotherapy response biomarkers, including T-cell infiltration, immune-related gene expression, and mutation counts in the DDR pathway in NSCLC. ZFHX3-MT is closely related to longer OS in NSCLC patients treated with ICIs, suggesting that ZFHX3 mutations be used as a novel predictive marker in guiding NSCLC ICI treatment.

SMYD3 overexpression indicates poor prognosis and promotes cell proliferation, migration and invasion in non‑small cell lung cancer.

SET and MYND domain-containing protein 3 (SMYD3) is a lysine methyltransferase, and its aberrant expression has been implicated in several malignancies. However, its clinical and biological roles in non-small cell lung cancer (NSCLC) remain unclear. In the present study, it was revealed that SMYD3 was significantly upregulated in NSCLC tissues, as compared with paired adjacent normal tissues. A high SMYD3 expression was associated with aggressive clinicopathological characteristics, as well as poor disease-free survival and overall survival (OS) in NSCLC patients. Multivariate analysis revealed that SMYD3 overexpression was an independent predictor of poor OS in NSCLC patients. In addition, SMYD3 knockdown inhibited cell proliferation, triggered apoptosis, and blocked migration and invasion in NSCLC cells in vitro, whereas stable SMYD3 overexpression promoted NSCLC cell proliferation. Furthermore, the SMYD3-silenced NSCLC cells became more sensitive, whereas the SMYD3-overexpressed NSCLC cells became more resistant to the apoptosis induced by cisplatin. Mechanistic analysis revealed that SMYD3 knockdown led to the upregulation of Bim, Bak and Bax, and the downregulation of Bcl-2, Bcl-xl, MMP-2 and MMP-9 in NSCLC cells. In combination, the present findings indicated that SMYD3 has oncogenic potential in the context of NSCLC, providing evidence that may be exploited for both prognostic and therapeutic purposes in the future.

MiR-1299 Impedes the Progression of Non-Small-Cell Lung Cancer Through EGFR/PI3K/AKT Signaling Pathway.

BackgroundNon-small-cell lung cancer (NSCLC) is one of the most malignant tumors. In which, numerous miRNAs had been reported to participate in the pathogenesis. However, the expression and function of miR-1299 in NSCLC are not clear.MethodsTo explore the roles of miR-1299 in NSCLC, we detected the levels of miR-1299 in clinical samples of NSCLC and investigated the role of miR-1299 in the regulation of the NSCLC cells proliferation, metastasis, and EMT. Luciferase reporter assay was employed to verify the target of miR-1299. Additionally, the proliferation, metastasis, and EMT of A549 and H1299 cells were analyzed after the overexpression and knockdown of miR-1299.ResultsWe found that the miR-1299 expression negatively corresponded with the clinical stage and overall survival in NSCLC patients. Overexpression of miR-1299 inhibited the migration, invasion, and EMT of A549 and H1975 cells. Meanwhile, we proved that miR-1299 is the sponge of EGFR. Besides, our results suggested that miR-1299 inhibits the progression of NSCLC cells through the PI3K/Akt signal pathway.ConclusionWe demonstrated that miR-1299 inhibits the progression of NSCLC through the EGFR/PI3K/Akt signal pathway. Therapeutic intervention targeting the miR-1299 may provide a potential strategy for the treatment of NSCLC.

High circ-SEC31A expression predicts unfavorable prognoses in non-small cell lung cancer by regulating the miR-520a-5p/GOT-2 axis.

Dysregulation of circular RNAs (circRNAs) has recently been shown to play important regulatory roles in cancer development and progression, including non-small cell lung cancer (NSCLC). However, the roles of most circRNAs in NSCLC are still unknown. In this study, we found that hsa_circ_0001421 (circ-SEC31A) was upregulated in NSCLC tissues and cell lines. Increased circ-SEC31A expression in NSCLC was significantly correlated with malignant characteristics and served as an independent risk factor for the post-surgical overall survival of NSCLC patients. Reduced circ-SEC31A expression in NSCLC decreased tumor cell proliferation, migration, invasion, and malate-aspartate metabolism. Mechanistically, we demonstrated that silencing circ-SEC31A downregulated GOT-2 expression by relieving the sponging effect of miR-520a-5p, which resulted in significantly reduced malate-aspartate metabolism in NSCLC cells. Taken together, these results revealed the important role of circ-SEC31A in the proliferation, migration, invasion, and metabolic regulation of NSCLC cells, providing a new perspective on circRNAs in NSCLC progression.

The long non‑coding RNA CASC7 inhibits growth and invasion of non‑small cell lung cancer cells through phosphatase and tensin homolog upregulation via sequestration of miR‑92a.

Accumulating evidence has demonstrated the crucial roles of long non-coding RNAs (lncRNAs) in various human cancers, including non-small cell lung cancer (NSCLC). However, to the best of our knowledge, the role of the lncRNA cancer susceptibility candidate 7 (CASC7) in NSCLC has not been clearly determined. The aim of the present study was to investigate the involvement of CASC7 in NSCLC. Marked downregulation of CASC7 was observed in NSCLC tissues and cell lines, and this downregulation of CASC7 was closely associated with distant metastasis, lymph node involvement and poor overall survival in NSCLC patients. Furthermore, overexpression of CASC7 significantly suppressed the proliferation, invasion and migration of the NSCLC cells A549 and H358, and promoted cell apoptosis in vitro. In addition, CASC7 was shown to act as a competing endogenous RNA by sponging miR-92a, which was proven to be an oncogenic miRNA in our previous study. The expression of miR-92a was upregulated in NSCLC tissues and cell lines, and was found to be inversely associated with CASC7 expression in NSCLC tissues. It was also demonstrated that CASC7 upregulated the expression of the tumor suppressor gene phosphatase and tensin homolog (a well-known target of miR-92a) by sequestration of miR-92a. Moreover, the tumor-suppressive effects of CASC7 were partly reversed by miR-92a overexpression in NSCLC cells. Collectively, the results of the present study indicated that CASC7 may act as a tumor-suppressive lncRNA that inhibits NSCLC progression by sponging miR-92a. These findings may improve our understanding of the potential mechanisms through which gain of CASC7 expression represses NSCLC progression.

Upregulated lncRNA CASC9 Contributes to Progression of Non-Small Cell Lung Cancer Through Inhibition of miR-335-3p and Activation S100A14 Expression.

IntroductionNon-small cell lung cancer (NSCLC) is a deadly cancer type worldwide and the main sub-type of lung cancer. Cancer susceptibility candidate-9 (CASC9) was reported to be a key player in cancer progression. However, its function and underlying mechanism in NSCLC remain unclear.Materials and MethodsExpression level of CASC9 in NSCLC tissues and cells was measured with RT-qPCR. Biological roles of CASC9 in NSCLC were analyzed with a series of in vitro experiments. Potential mechanisms of CASC9 in NSCLC were analyzed by predicting and validating the possible targets of CASC9 in NSCLC.ResultsIn this study, we found CASC9 expression was upregulated in NSCLC tissues and cell lines. High CASC9 expression was identified as a predictor for poorer overall survival of NSCLC patients. Furthermore, functional assays showed CASC9 knockdown suppressed NSCLC cell proliferation, migration, and invasion, while CASC9 overexpression caused opposite effects. We also found microRNA-335-3p (miR-335-3p) could act as a target of CASC9 in NSCLC and the inhibition effect of CASC9 knockdown on NSCLC progression required the activity of miR-335-3p. In addition, we identified S100 calcium-binding protein A14 (S100A14) acts as a target of miR-335-3p.DiscussionTaken together, our study suggested CASC9 could promote NSCLC progression via miR-335-3p/S100A14 axis. The CASC9/miR-335-3p/S100A14 regulatory triplets identified in this work might provide new therapeutic strategies for NSCLC treatment.

The Role of a Cachexia Grading System in Patients with Non-Small Cell Lung Cancer Treated with Immunotherapy: Implications for Survival

Objective The association between cancer-induced weight-loss (CIWL) and poor clinical outcomes in patients treated with immunotherapy is scarcely understood. We evaluated the use of a cachexia-grading system in IO-treated non-small cell lung cancer (NSCLC) patients in order to predict clinical outcomes. Materials 300 patients with NSCLC, who received immunotherapy during any line of therapy, were included. All patients were graded according to a previously validated cachexia scale, which takes into consideration body mass index (BMI) and weight loss, stratifying patients into five risk categories (0 [pre-cachexia] − 4 [refractory cachexia]). Primary endpoint was overall survival (OS). Results Ninety-one (30.3%) patients were classified in the low risk category, 176 (58.6%) were classified in the intermediate risk category and 33 (11%) were in the high risk category. Patients classified as low-risk had a significantly longer OS compared with those with intermediate or high risk (22.4 mo, [95%CI: 16.6–NR] vs. 17.1 [95%CI: 13.5–22.4] vs. 8.0 [3.9–18.4]; p < 0.001). In the multivariate analysis, after adjusting for age, hemoglobin and ORR, hazard of death increased as per the cachexia risk scale (Hazard ratio: 1.62 [1.22–2.16]; p = 0.001). Conclusion Cachexia is independently associated with worse OS in NSCLC patients who receive immunotherapy, highlighting the role for nutritional assessment.

Abstract D088: Analysis of tumor mutational burden, cancer immunotherapy use and outcomes based on genomic ancestry in non-small cell lung cancer patients treated in community practice in the United States

Abstract Introduction Racial disparities in cancer care and outcome are multifactorial and well documented, being attributed to social determinants, access to and quality of care. In addition, variation in biologic drivers by self-reported race/ethnicity or genetic ancestry may contribute to inequities in outcomes. Differences in tumor mutational burden (TMB), an important predictor of response to cancer immunotherapy (CIT), have been reported in non-small cell lung cancer (NSCLC) patients of different genomic ancestries. The impact of differing TMB-levels on outcomes and underlying cause is unknown, though smoking is associated with increased TMB. This study seeks to describe the receipt of CIT-containing regimens and the distribution and relationship of TMB with overall survival in NSCLC patients across ancestral groups. Methods 3962 patients with advanced NSCLC diagnosed between 1/1/11 and 4/30/19 were selected from the de-identified Flatiron Health-Foundation Medicine (FMI) linked clinicogenomics database. TMB was categorized as high or low (16mutations (mut)/megabase (Mb) cut off). Genomic ancestry was classified as previously described by FMI. CIT use and TMB status was described by ancestry and the association of TMB status with overall survival (OS) from diagnosis was estimated using Cox proportional hazards models. Analyses were further stratified by genomic ancestry (European vs. Non-European) and receipt of CIT at any point after advanced diagnosis date. Results The study population was 72% European ancestry (EUR), 8% African (AFR), 5% American (AMR), and 3% Asian (ASN). Genomic ancestry was largely correlated with self-reported race. The use of CIT-containing regimens was similar across ancestries, from 36% - 45% for those of ASN and AFR ancestry, respectively. TMB levels varied across ancestry groups; patients of AFR ancestry had the highest median TMB level (9.6 mut/Mb, [Interquartile Range: 4.4 – 15.7]), whereas ASN had lowest (3.5 mut/Mb, [2.6, 13.0]). Median TMB for EUR patients was 7.8 mut/Mb [3.5 – 13.9]. Similar trends were observed among patients with a history of smoking, though overall TMB levels were higher. TMB-high patients accounted for 24% of those of AFR, 20% for EUR, and 27% and 5% for AMR and ASN ancestry respectively. TMB-high was associated with better survival in patients who had ever received CIT (0.63, [95% CI: 0.52, 0.77]). These results were consistent when stratified by EUR (0.62 [0.50, 0.77]) and non-EUR ancestry (0.59 [0.34, 1.02]). Conclusions We observed different levels of TMB across ancestral groups. Differences in prevalence of smoking may be a contributing factor. TMB-high was equally predictive of improved OS among patients receiving CIT for those of EUR and non-EUR ancestry. There were no observed differences in receipt of CIT based on genomic ancestry in this population. These results suggest that addressing reported inequitable access to TMB testing among African American NSCLC patients may improve racial/ethnic disparities in NSCLC outcomes. Citation Format: Nicole Richie, Patricia Luhn, Uzor Ogbu, Gerren Wilson, William Capra, Mark Lee, Otis Brawley. Analysis of tumor mutational burden, cancer immunotherapy use and outcomes based on genomic ancestry in non-small cell lung cancer patients treated in community practice in the United States [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr D088.

Cyclooxygenase inhibitor use during checkpoint immunotherapy and effect on time to progression and overall survival for metastatic melanoma and non-small cell lung cancer patients.

e22034 Background: Immune checkpoint inhibitors (ICIs) improve survival outcomes in metastatic melanoma and non-small cell lung cancer (NSCLC). Preclinical evidence suggests that overexpression of cyclooxygenase-2 (COX2) in tumors facilitates immune evasion through prostaglandin E2 production and that COX inhibition synergizes with ICIs to promote antitumor T-cell activation. This study investigates whether concurrent COX inhibitor (COXi) use during ICI treatment is associated with improved time-to-progression (TTP), objective response rate (ORR) and overall survival (OS) in metastatic melanoma and NSCLC patients. Methods: We retrospectively reviewed 90 metastatic melanoma and 37 metastatic NSCLC patients, treated only with ICI between 2011 and 2019. Differences in TTP and OS by COXi use were compared using Kaplan-Meier and Cox regression. Interaction between COXi use and neutrophil-lymphocyte ratio (NLR) was examined. Independent radiology review per RECIST v1.1 was performed. Results: For melanoma patients, median TTP was significantly prolonged in COXi users (245 v 100.5 days, p = .0002). On multivariate analysis (MVA), COXi use associated with increased TTP (HR 0.36, 95% CI: 0.2 – 0.66, p = 0.001) when adjusted for age, pretreatment NLR, and gender. For NSCLC patients, COXi use also associated with increased TTP on MVA (HR 0.45; 95% CI: 0.21 – 0.97; p = 0.042) adjusted for age. ORR at 6-months was significantly higher in patients who received concurrent COXi compared to those that did not in both melanoma (58.6% v 19.2%, p = 0.0005) and NSCLC (73.7% v 33.3%, p = 0.036) cohorts. COXi use was associated with improved OS in NSCLC patients on MVA (HR 0.3; 95% CI: 0.12 – 0.78; p = 0.013), but not in melanoma patients. In the melanoma cohort, high pretreatment NLR of &gt; 5 associated with decreased TTP on MVA (HR 3.21, 95% CI: 1.64 – 6.3; p = 0.0007). However, for melanoma patients with high NLR &gt; 5, COXi use significantly associated with increased TTP (HR 0.08, 95% CI: 0.03 – 0.25); but in melanoma patients with low NLR ≤5, there was no effect on TTP (HR 0.65, 95% CI: 0.32-1.32). Similar outcomes were found in an adjusted melanoma cohort after RECIST review. Conclusions: Our study suggests that use of COXi concurrent with ICI is significantly associated with longer TTP and improved ORR at 6-months in metastatic melanoma and NSCLC patients. NSCLC patients using COXi had prolonged OS. Furthermore, COXi use appears to reverse the negative prognostic effect of a high NLR by prolonging TTP in melanoma patients.

PTPRD/PTPRT mutations as potential positive predictor for response of immune checkpoint inhibitors in non-small cell lung cancer (NSCLC).

e15112 Background: Immune checkpoint inhibitors (ICIs) have demonstrated positive results in non-small cell lung cancer (NSCLC) patients, with durable responses and prolonged overall survival (OS). Nevertheless, the response rate to immunotherapy is still limited. It is necessary to identify clinically useful biomarkers that can distinguish patients who can respond to ICIs. PTPRD/PTPRT are the phosphatases of JAK-STAT signaling, which may be associated with response to ICIs. Here we aimed to demonstrate the association between PTPRD/PTPRT and ICIs. Methods: Genomic and survival data of NSCLC patients administrated with anti–PD-1/PD-L1 or anti–CTLA-4 antibodies (Rizvi2015; Hellmann2018; Rizvi2018 Samstein2019) were retrieved from publicly accessible data. Genomic, survival and mRNA data of 1007 patients with NSCLC was obtained from The Cancer Genome Atlas (TCGA). Association between PTPRD/PTPRT mutation and progression-free survival (PFS) and overall survival (OS) were analyzed. Gene set enrichment analysis (GSEA) was used to determine potentially relevant gene expression signatures between specific subgroups. Results: PTPRD/PTPRT mutations were significantly associated with better PFS in Rizvi2015 cohort (HR = 0.16; 95% CI, 0.02-1.17; P = 0.03), Hellmann2018 cohort (HR, 0.49; 95% CI, 0.26-0.94; P = 0.03) and Rizvi2018 cohort (HR = 0.64; 95% CI, 0.44-0.92; P = 0.01). PTPRD/PTPRT mutation was also significantly associated with better OS in Samstein2019 cohort (HR, 0.66; 95% CI, 0.45-0.97; P = 0.03). In TCGA, no association between PTPRD/PTPRT mutations and OS was observed (P = 0.91), suggesting that PTPRD/PTPRT mutations were not prognostic factor. PTPRD/PTPRT mutations were associated with increased TMB (P &lt; 0.0001). The mRNA expression of STAT1 and CD4 was higher in patients with PTPRD/PTPRT mutant type than PTPRD/PTPRT wild type. Gene Set Enrichment Analysis revealed prominent enrichment of signatures related to inflammatory response, interferon gamma response and antigen processing and presentation in patients with PTPRD/PTPRT mutation. Conclusions: Our results suggest that PTPRD/PTPRT mutation is associated with better PFS and OS in NSCLC patients receiving ICIs by increasing immune-related gene signatures. The role of PTPRD/PTPRT in immunotherapy is needed to be further studied.

EFHD2 contributes to non-small cell lung cancer cisplatin resistance by the activation of NOX4-ROS-ABCC1 axis

Recurrence and metastasis remain the major cause of cancer mortality. Even for early-stage lung cancer, adjuvant chemotherapy yields merely slight increase to patient survival. EF-hand domain-containing protein D2 (EFHD2) has recently been implicated in recurrence of patients with stage I lung adenocarcinoma. In this study, we investigated the correlation between EFHD2 and chemoresistance in non-small cell lung cancer (NSCLC). High expression of EFHD2 was significantly associated with poor overall survival of NSCLC patients with chemotherapy in in silica analysis. Ectopic EFHD2 overexpression increased cisplatin resistance, whereas EFHD2 knockdown improved chemoresponse. Mechanistically, EFHD2 induced the production of NADPH oxidase 4 (NOX4) and in turn the increase of intracellular reactive oxygen species (ROS), consequently activating membrane expression of the ATP-binding cassette subfamily C member 1 (ABCC1) for drug efflux. Non-steroidal anti-inflammatory drug (NSAID) ibuprofen suppressed EFHD2 expression by leading to the proteasomal and lysosomal degradation of EFHD2 through a cyclooxygenase (COX)-independent mechanism. Combining ibuprofen with cisplatin enhanced antitumor responsiveness in a murine xenograft model in comparison with the individual treatment. In conclusion, we demonstrate that EFHD2 promotes chemoresistance through the NOX4-ROS-ABCC1 axis and therefore developing EFHD2-targeting strategies may offer a new avenue to improve adjuvant chemotherapy of lung cancer.

Circ_100565 promotes proliferation, migration and invasion in non-small cell lung cancer through upregulating HMGA2 via sponging miR-506-3p

BackgroundCircular RNAs (circRNAs) play a vital role in the development of various cancers. Circ_100565 was found to be a highly expressed circRNA in non-small cell lung cancer (NSCLC) tissues screened by microarray profiles of circRNAs. However, the role of circ_100565 in NSCLC still remains unknown.MethodsMicroarray analysis was used to screen for differentially expressed circRNAs in NSCLC tissues. The expression levels of circ_100565, microRNA-506-3p (miR-506-3p) and high mobility group AT-hook 2 (HMGA2) were measured by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was detected by cell counting kit-8 (CCK-8) and colony formation assays. Transwell assay was used to determine the migration and invasion of cells. Besides, Western blot (WB) analysis was performed to assess the levels of proliferation and metastasis-related proteins and HMGA2 protein. Moreover, animal experiments were used to confirm the effect of circ_100565 on NSCLC tumor growth in vivo. In addition, the interaction between miR-506-3p and circ_100565 or HMGA2 was confirmed by dual-luciferase reporter, RNA immunoprecipitation (RIP) assay or biotin-labeled pull-down assay.ResultsCirc_100565 was upregulated in NSCLC, and its high expression was positively associated with the poor overall survival of NSCLC patients. Silencing of circ_100565 suppressed the proliferation, migration and invasion of NSCLC cells in vitro and reduced the tumor growth of NSCLC in vivo. Circ_100565 could sponge miR-506-3p, and miR-506-3p could target HMGA2. Moreover, miR-506-3p inhibitor or HMGA2 overexpression could reverse the inhibition effect of circ_100565 knockdown on NSCLC progression.ConclusionCirc_100565 increased HMGA2 expression to promote proliferation, migration and invasion in NSCLC via absorbing miR-506-3p. Our findings provided a new biomarker for NSCLC therapy.