Squamous Lung Cancer Research Articles

PAK5 promotes the cell stemness ability by phosphorylating SOX2 in lung squamous cell carcinomas

Growing evidences suggest that the overexpression of p21-activated kinase 5 (PAK5) plays an important role in various tumor progression. However, the role of PAK5 and its downstream target gene(s) in lung squamous cell carcinomas (LUSC) are waiting to be elucidated. TCGA data were utilized to evaluate the expression levels of PAK5 in LUSC. We then explored the role of PAK5 in maintaining the stem-like phenotype of lung squamous cancer cells through RT-PCR, flow cytometry, oncosphere-forming assay. In addition, co-immunoprecipitation, western blotting and immunofluorescence assays were used to determine SOX2 as a novel effector of PAK5. Xenograft models in nude mice were established to explore the roles of PAK5 in lung cancer growth. In this study, we have shown that PAK5 is overexpressed in LUSC tissues. The absence of PAK5 abolishes self-renewal ability of LUSC cells by decreasing the expression and phosphorylation of SOX2 in vitro and in vivo. In xenograft models, knockdown or pharmacological inhibition of PAK5 suppressed the tumor growth and metastasis of lung squamous cancer cells in vivo. Taken together, our findings suggest that the PAK5-mediated SOX2 phosphorylation promoted the cancer stem cell-like phenotype of LUSC cells. PAK5 inhibition may be a promising target in the treatment of SOX2 positive lung squamous cell cancer.

Multi-Institutional Prospective Validation of Prognostic mRNA Signatures in Early Stage Squamous Lung Cancer (Alliance).

Surgical resection is curative for some patients with early lung squamous cell carcinoma. Staging and clinical factors do not adequately predict recurrence risk. We sought to validate the discriminative performance of proposed prognostic gene expression signatures at a level of rigor sufficient to support clinical use. The two-stage validation used independent core laboratories, objective quality control standards, locked test parameters, and large multi-institutional specimen and data sets. The first stage validation confirmed a signature's ability to stratify patient survival. The second-stage validation determined which signature(s) optimally improved risk discrimination when added to baseline clinical predictors. Participants were prospectively enrolled in institutional (cohort I) or cooperative group (cohort II) biospecimen and data collection protocols. All cases underwent a central review of clinical, pathologic, and biospecimen parameters using objective criteria to determine final inclusion (cohort I: n= 249; cohort II: n= 234). Primary selection required that a signature significantly predict a 3-year survival after surgical resection in cohort I. Signatures meeting this criterion were further tested in cohort II, comparing risk prediction using baseline risk factors alone versus in combination with the signature. Male sex, advanced age, and higher stage were associated with shorter survival in cohort I and established a baseline clinical model. Of the three signatures validated in cohort I, one signature was validated in cohort II and statistically significantly enhanced the prognosis relative to the baseline model (C-index difference 0.122; p < 0.05). These results represent the first rigorous validation of a test appropriate to direct adjuvant treatment or clinical trials for patients with lung squamous cell carcinoma.

Differences in molecular epidemiology of lung cancer among ethnicities (Asian vs. Caucasian).

BackgroundDifferences in carcinogenesis and therapeutic efficacy according to ethnicity have been reported for lung cancer, and understanding differences in genetic mutation profiles among ethnicities is important for interpreting the results of clinical trials, preventing carcinogenesis, and individualizing treatment. However, no studies have focused on differences in mutation profiles among different ethnicities using large-scale genomic analysis data with detailed information on smoking history, the main cause of lung cancer.MethodsTo clarify the differences in genetic mutation profiles between Caucasian and Japanese subjects, we compared data from The Cancer Genome Atlas, which mainly included Caucasians, with results from the Japan Molecular Epidemiology for lung cancer study, which is an epidemiological study only involving Japanese subjects. We divided the participants into four groups according to smoking status and performed comparative analysis by tissue type (lung adenocarcinoma and squamous cell lung cancer).ResultsIn patients with lung adenocarcinoma, the frequency of EGFR mutations was lower in Caucasian subjects than in Japanese subjects (14.6% vs. 51.1%), whereas the frequencies of mutations in other genes, namely KRAS (32.9% vs. 9.3%), TP53 (45.2% vs. 20.7%), BRAF (9.6% vs. 1.3%), PIK3CA (5.9% vs. 2.6%), KEAP1 (17.8% vs. 0.5%), NF1 (10.9% vs. 0.5%), STK11 (17.8% vs. 0.7%), RBM10 (8.7% vs. 0.1%), and MET (7.8% vs. 0.1%), were higher in Caucasian subjects. Among patients with squamous cell carcinoma, TP53 (81.2% vs. 49.1%), PIK3CA (14.5% vs. 6.8%), KEAP1 (12.7% vs. 0.9%), and NFE2L2 mutations (15.8% vs. 13.6%) were more common in Caucasian subjects.ConclusionsEthnicity is an important and complex characteristic that must be recognized and considered, even in the era of precision medicine. We should collaborate to share data for different ethnicities and incorporate them into clinical practice and the design of global clinical studies. Carefully designed molecular epidemiological studies focusing on ethnic differences are warranted.

Lung-MAP (SWOG S1400): Design, implementation, and lessons learned from a biomarker-driven master protocol (BDMP) for previously-treated squamous lung cancer (sqNSCLC).

9576 Background: S1400, a BDMP, was designed to address an unmet need in sqNSCLC, run within the National Clinical Trials Network of the National Cancer Institute using a public-private partnership (PPP). The goal of was to establish an infrastructure for biomarker-screening and rapid evaluation of targeted therapies in biomarker-defined groups leading to regulatory approval. Methods: S1400 included a screening part using the FoundationOne assay and a clinical trial part with biomarker-driven studies (BDS) and “non-match” studies (NMS) for patients not eligible for any BDS. Patients could be screened (SaP) at progression or pre-screened (PreS). Results: Between June 2014 and January 2019, 1864 patients enrolled (711 PreS, 1079 SaP), 1674 with biomarker results, and 653 registered to a study with 217 to BDS and 436 to NMS. Six BDS and 3 NMS were initiated in small subsets with all BDS and 2 NMS completed within 2-3 years (see Table). Completed BDS have not demonstrated activity with 0-2 responses. On S1400I, Nivolumab and ipilimumab did not improve survival. Response with durvalumab (S1400A) was 16%. Conclusions: Lung-MAP met its goal to quickly answer targeted and other novel therapy questions in rare sqNSCLC subpopulations, answering questions that likely would not have been otherwise feasible, thereby demonstrating value. Activated just prior to the success of PD-(L)1 therapies in sqNSCLC, the trial had to undergo major design changes. Lessons learned include the need to update based on new science and that the PPP collaboration was essential to success. Lung-MAP continues now with new BDS and NMS in all NSCLC as of January 2019. Clinical trial information: NCT02154490 . [Table: see text]

Podoplanin is a useful prognostic marker and indicates better differentiation in lung squamous cell cancer patients? A systematic review and meta-analysis

BackgroundThe CSC (cancer stem cell) markers often indicate poor prognosis and more cell invasion or migration of cancer patients. Podoplanin was assumed as a candidate CSC marker and predict poor prognosis among squamous cancers. Whereas, the prognostic value of podoplanin among lung squamous cancer (LUSC) patients remains controversial.MethodsA search of databases including PubMed, Embase and Web of Science was performed. Eligible articles studying the prognostic significance of podoplanin were selected. Odds ratio and HR (hazard ratio) were used to assess the relationships between podoplanin and clinical characteristics, as well as to quantify its prognostic role. The heterogeneity was estimated by I2 Statistic and P values from sensitivity analysis. Begg’s funnel plots were used to estimate possible publication bias.Results8 eligible studies containing 725 I-IV LUSC patients were included. Podoplanin expression showed no significant correlations with TNM stage, vascular invasion, lymphatic invasion, lymph node metastasis, pleural metastasis of tumor and gender of patients. However, podoplanin showed significant associations with better differentiation (pooled OR = 2.64, 95% CI 1.53–4.56, P = 0.0005, fixed effect) and better overall survival (HR = 2.14, 95% CI 1.45–3.15, P = 0.0001, fixed effect) and progression-free survival (HR = 1.73, 95% CI: 1.01–2.98, P = 0.05, fixed effect) of LUSC. Funnel plots illustrated no evidence of publication bias in our results.ConclusionsPodoplanin could be a useful prognostic marker and indicates better differentiation for LUSC patients, and the value of PDPN expression as a marker for cancer stem cells in LUSC should be critically evaluated in future studies.

Cancer genomics of lung cancer including malignant mesothelioma: A brief overview of current status and future prospects.

Cancer as a genetic disease is by now well recognized. Genomic analysis of cancer cells, therefore, has greatly enhanced our ability to identify genetic alterations associated with various cancer types, including both lympho-hematopoietic as well as solid tumors. Chronic myeloid leukemia (CML), based on the specific diagnostic genetic abnormality has served as a prototype disease to clearly demonstrate the significance of the genomic analysis of cancer in identifying targeted therapy. Such a success has provided extra ordinary opportunities to investigate the role of genetic abnormalities and the pathways amenable to targeted therapy, not only in blood cancers but solid tumors such as Lung, Brain, Colon, Renal, Breast cancers as well as other epithelial and mesenchymal tumors. The main focus of this presentation is to illustrate the role of genomic analysis in targeting lung cancer, based on abnormalities or the pathways deregulated in tumor cells from individual patients. Lung cancer is one of the most common epithelial cancers associated with chronic inflammation due to cigarette smoking and other environmental carcinogens, and includes four distinct histologic type; non-small cell lung cancer (NSCLC); small cell lung cancer (SCLC) and squamous cell lung cancer. According to current estimates, 1.3 million cases of lung cancer are expected to be diagnosed worldwide annually, resulting in one million deaths. Since the discovery that patient's tumors with specific mutations in the EGFR may be sensitive to targeted therapeutic approach and the subsequent realization that the such mutations in the gene are not as prevalent, several cancer centers including ours initiated intense efforts to find other mutations or genomic alterations, which may serve as targets of specific therapy. Such efforts have successfully resulted in a battery of genes such as KRAS, ALK, C-MET, HER-2/neu, ROS1, etc., which have helped oncologists to triage the patients for personalized therapies. A significant proportion of patients with lung cancer, however, do not show any of the above genetic abnormalities. Approximately 90% of lung cancers exhibit RB1 mutation/deletion and or KRAS mutations, therefore, the signaling pathways, which regulate multistep tumorigenesis in lung cancer, are important for the treatment of histologic subtypes of lung cancer, which includes NSCLC & SCLC. Equally important was the findings that similar signaling pathways are also shared by other solid tumor types. We have investigated the role of these pathways to target these cancers and develop new strategies to treat lung, brain and related cancers. In addition, our translational studies in other tumor types such as NF2 related malignancies, specifically, Malignant Mesothelioma (MM), in which NF2 related pathway amenable to targeted therapies was identified. Selected examples representing experimental approaches will be discussed to illustrate the critical role of translational research in developing novel therapeutics for the successful and durable responses in some of these cancer types.

Abstract B42: Impact of EZH2 inhibition on the immune microenvironment of squamous lung cancers

Abstract Squamous cell carcinoma (SCC) is a subtype of non-small cell lung cancer (NSCLC) that often resists treatment, resulting in poor prognosis for the patient. Immunotherapies, including those targeting the PD1/PD-L1 immune checkpoint, have become popular treatment options, but they are effective for only 20% of squamous lung cancer patients. The likely cause of differential responses is tumor heterogeneity. This heterogeneity can present in the tumor cells, for example, if more or fewer tumor cells express PD-L1, and also in the microenvironment, for example, if the tumor is infiltrated by more immune-suppressive neutrophils or more immune-activating CD8+ T cells. In order to study SCC in immune-intact mice, our laboratory utilizes a mouse model whereby tumors are induced through biallelic deletion of Lkb1 (aka Stk11) and Pten in the mouse lung. These SCCs express PD-L1 at high levels on the surface of the tumor cells; however, treatment with anti-PD1 antibody caused only a delay in tumor growth, not a reduction. We believe the ineffectiveness of immunotherapy in this model is due to the presence of large populations of tumor-associated neutrophils (TANs). TANs have been shown to produce Arginase1, which has repressive effects on the T-cell populations by inhibiting proper signaling and activation. An inverse relationship between TAN and CD8+ T cell abundance has also been reported for squamous lung cancers. Finding a second therapy that can prime squamous lung cancers to respond to immunotherapy would be beneficial for a large number of patients. EZH2 is a histone methyl transferase that promotes differentiation of cells via lineage-specific gene repression. We hypothesize that using an anti-PD-1 immunotherapeutic in combination with an EZH2 inhibitor will increase the PD-L1 expression on the tumor cells, decrease the normal homing or T-cell suppression of neutrophil populations, and increase the efficacy of the immunotherapy. In order to test our hypothesis, we have treated lung cancer cells with EZH2 inhibitor both in vitro and in vivo. In vitro, treatment of lung cancer cell lines or patient-derived organoids of squamous lung cancers caused increased expression of PD-L1. In mice harboring Lkb1/Pten tumors, treatment with EZH2 inhibitor increased the proportion of tumor cells expressing PD-L1 but did not prevent the tumor from growing. In contrast, the mouse treated with EZH2 inhibitor and anti-PD-1 immunotherapy had tumor regression that was durable up to 8 weeks post-treatment initiation. We isolated TANs from untreated tumors and observed that they express very high levels of Arginase 1. Lung neutrophils isolated from EZH2 conditional knockout mice had significantly lower levels of Arginase 1 than WT lung neutrophils or Lkb1/Pten TANs. Together these data suggest that EZH2 inhibition will be an effective strategy to both increase the PD-L1+ tumor cell population, and to decrease TAN-mediate T cell suppression by decreasing TAN-derived Arginase 1 expression. Citation Format: Tanner J. DuCote, Xiulong Song, Christine F. Brainson. Impact of EZH2 inhibition on the immune microenvironment of squamous lung cancers [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr B42.

Characterization of Amplified Malate Dehydrogenase 1 (MDH1) in the Context of Squamous Cell Lung Cancer

Cancer cells often reroute or stimulate their metabolism to support proliferation, which requires a change in the activity or regulation of key metabolic residues. Malate dehydrogenase 1 (MDH1) catalyzes the NADH‐dependent conversion of oxaloacetate to malate, which can help drive glycolysis through the production of NAD+. MDH1 is amplified in many squamous cell lung carcinomas, the second leading cause of cancer related death, and this amplification is associated with a poor prognosis of 50% reduced survival. The precise role of MDH1 in driving squamous cell lung cancer and the mechanisms that regulate this protein are not yet well understood.Here, we develop a tool to examine the effect of MDH1 amplification in an in vitro model. We aim to examine MDH1 amplification in squamous cell lung carcinoma with an isogenic control. It has been previously shown that MDH1 methylation can suppress glutamine metabolism in pancreatic cancers, and that acetylation of MDH1 alters its activity. We will examine how MDH1 amplification affects squamous cell carcinomas by using biochemical kinetics and cellular methods. We have found that MDH1 amplification influences the production of metabolites in the TCA cycle. Through monoclonal single cell isolation, we will generate a squamous epithelial lung cancer cell line stably overexpressing MDH1. We hypothesize that MDH1 has cysteine residues that can be sensitive to oxidative stress which can alter the normal enzymatic activities of MDH1. Through site‐directed mutagenesis, we will mutate a cysteine residue to a serine residue at the 137 position to examine the possible effect of oxidative stress on MDH1. This residue has been shown to be sensitive to oxidation in the mouse form of MDH1. We will also heterologously express and purify MDH1 mutants and perform kinetic assays to examine the impact of amplified MDH1. By understanding the regulatory strategies of MDH1, we hope to identify new pathways that may be targeted therapeutically.Support or Funding InformationThis work was funded by a Research Scholar Grant, RSG‐19‐075‐01‐TBE, from the American Cancer Society (C.D.S.), National Institutes of Health R00 CA187594 (C.D.S.), U54CA132384 (SDSU) &amp; U54CA132379 (UC San Diego), and IMSD 5R25GM058906 (SDSU), as well as the California Metabolic Research Foundation (SDSU).

Association between Use of Aspirin and Risk of Lung Cancer: Results from Pooled Cohorts and Mendelian Randomization Analyses

Background: Massive studies have examined associations between use of aspirin and lung cancer risk. However, findings from previous studies are inconsistent and it remains unclear whether there is a causality between them. Thus, we conducted a meta-analysis and further carried out Mendelian randomization (MR) analyses to elucidate the associations between aspirin use with lung cancer risk. Methods: Eligible studies were identified by searching the PubMed and several databases up to November 2019. Cohort studies, nested case-control studies, and randomized controlled trials (RCTs) investigating the impact of aspirin exposure and lung cancer were included. Relative risk (RR) were evaluated for lung cancer risk. Subgroup analyses regarding gender, pathologic subtypes and smoking status were also executed. MR analyses were conducted using statistics obtained from Neale Lab and International Lung Cancer Consortium (ILCCO) to assess the possible causal relationship of aspirin on lung cancer risk. Findings: Sixteen eligible studies involving 1,522,687 patients were included. The combined RR of aspirin use for lung cancer risk was 0.95 (95% confidence interval (CI) 0.91-0.98). In terms of pathology, a significant protective effect of aspirin on lung cancer was observed in squamous cell lung cancer (RR=0.80; 95% CI 0.65-0.98). In subgroup meta-analyses, significant association between aspirin use and reduced lung cancer risk was only observed among men (RR=0.87; 95% CI 0.77-0.97). The MR risk analysis suggested a causal effect of aspirin on lung cancer risk, with evidence of a decreased risk for overall lung cancer (OR=0.042; 95% CI 0.003-0.564) and squamous cell lung cancer (OR=0.002; 95% CI 1.21×10-5-0.301). Interpretation: Our study provided evidence for a causal protective effect of aspirin on lung cancer risk among men, particularly on the squamous cell lung cancer risk. Funding Statement: National Key RD Key Project of Guangzhou Scientific Research Project; High-level university construction project of Guangzhou Medical University Declaration of Interests: The authors declare that there is no conflict of interest. Ethics Approval Statement: No patients participated in study design, patient recruitment and research conduct. Therefore, there is no need for ethical approval.

Thermal Analysis of Infrared Irradiation-Assisted Nanosecond-Pulsed Tumor Ablation

Nanosecond Pulsed Electric Fields (nsPEF) have the potential to treat a variety of cancer types including melanoma, pancreatic and lung squamous cancers. Recent studies show that nsPEF-based cancer therapy may be improved further with the assistance of moderate heating of the target. A feedback-looped heating system, utilizing a 980-nm fiber optic laser, was integrated into nsPEF electrodes for tumor ablation. The laser beam profile was determined to be Gaussian using a knife-edge technique. Thermal properties of the biological target were evaluated based on the treatment area, penetration depth and thermal distribution due to laser irradiation with or without nsPEF. Synergistic effects between nsPEF and the moderately elevated temperature at the target was observed, resulting in enhanced overall survival tumor regression up to 50% in the treatment of lung squamous cell cancer in mice.

Complete regression of pulmonary squamous carcinoma in IPF following gemcitabine plus cisplatin: a case report and literature review

BackgroundLung cancer is one of the most common co-morbid conditions in patients with idiopathic pulmonary fibrosis (IPF) and negatively affects the prognosis of IPF; Current guidelines for the management of IPF do not give a clear statement on how to manage these patients, and traditional chemotherapy for lung cancer had a limited efficiency rate. Here, we present a rare case of primary lung squamous carcinoma in a patient with IPF whose tumor completely regressed following gemcitabine plus cisplatin therapy; the cancer was no longer detectable after 2 years upon follow-up.Case presentationSixty-seven year-old male patient with IPF was admitted to hospital due to acute onset hemoptysis. In addition to a definite usual interstitial pneumonia (UIP) pattern, a chest CT scan showed a non-enhancing nodular opacity in the right upper lobe and an enhancing nodule in the right lower lobe. Bronchoscopic biopsy of the nodule in the right lower lobe revealed squamous lung cancer. After 2 cycles of chemotherapy with gemcitabine and cisplatin, the tumor in the right lower lobe was no longer detectable after 2 years of follow-up; however, the nodule in the right upper lobe had increased significantly. Finally, Mycobacterium tuberculosis (MTB) was cultured from the bronchoalveolar (BAL) sample submitted at the last evaluation, and the patient was confirmed to have active pulmonary TB.ConclusionWe report the first documented case of complete pulmonary squamous carcinoma regression in IPF following gemcitabine plus cisplatin. Traditional chemotherapy is considered inadequate to cause the resulting regression of the tumor. The concomitant active pulmonary tuberculosis possibly underlies the mechanism.

Do systemic treatments delivered after Nivolumab result in better outcomes? A bicentric case-control study

BackgroundImmune checkpoint inhibitors (ICI) are now widely used at different stages of non-small cell lung cancers (NSCLC). Some clinical studies suggest that chemotherapy and immunotherapy have synergic activities, raising the question of the best therapeutic sequence. We studied the effect of chemotherapy in advanced NSCLC when administered after immunotherapy by nivolumab. MethodsWe performed a bicentric, retrospective, case-control study in two French hospitals. Patients with NSCLC treated with chemotherapy after nivolumab between January 2015 and January 2016 were included. Each case was matched on age and number of previous lines to one lung cancer patient who had not received nivolumab. Each CT-scanner has been reviewed and the objective response to chemotherapy was assessed for each patient according to the RECIST 1.1 criteria. ResultsThirty-one patients with advanced NSCL who had at least received one cycle of chemotherapy after progression under nivolumab in the inclusion period were matched to 31 controls. The median age for cases was 59 yo and the predominant tumoral histology was adenocarcinoma (77%). The progression free survival (PFS) was 2.95 months in the studied group vs 2.69 months (P=0.18) in the control group. At best response, disease control (DC=partial response and stable disease) was better in the case group than in the control group (58% vs 39%, P=0.127). Cases were about five times more likely to get objective response to best evaluation than controls (OR=5.043 [95% CI: 0.975–26.086]; P=0.054). The overall survival (OS) was 7.3 months in the case group and 3.3 months in the control group (P=0.074). Patients who have been treated with targeted therapy instead of chemotherapy and patients with squamous lung cancer had worst PFS and OS. ConclusionIn advanced NSCLC, the chemotherapy progression free survival does not seem higher when administered after nivolumab. However, when administered post-nivolumab, traditional chemotherapy has 5 times more chances to achieve objective response and seems to improve overall survival of cases. Pooled analysis with other similar studies might be interesting for a next step.

High-Throughput Drug Screening Identifies a Potent Wnt Inhibitor that Promotes Airway Basal Stem Cell Homeostasis

SUMMARYMechanisms underpinning airway epithelial homeostatic maintenance and ways to prevent its dysregulation remain elusive. Herein, we identify that β-catenin phosphorylated at Y489 (p-β-cateninY489) emerges during human squamous lung cancer progression. This led us to develop a model of airway basal stem cell (ABSC) hyperproliferation by driving Wnt/β-catenin signaling, resulting in a morphology that resembles premalignant lesions and loss of ciliated cell differentiation. To identify small molecules that could reverse this process, we performed a high-throughput drug screen for inhibitors of Wnt/β-catenin signaling. Our studies unveil Wnt inhibitor compound 1 (WIC1), which suppresses T-cell factor/lymphoid enhancer-binding factor (TCF/LEF) activity, reduces ABSC proliferation, induces ciliated cell differentiation, and decreases nuclear p-β-cateninY489. Collectively, our work elucidates a dysregulated Wnt/p-β-cateninY489 axis in lung premalignancy that can be modeled in vitro and identifies a Wnt/β-catenin inhibitor that promotes airway homeostasis. WIC1 may therefore serve as a tool compound in regenerative medicine studies with implications for restoring normal airway homeostasis after injury.

Identification of Potential Inhibitors from Traditional Chinese Medicine for Fibroblast Growth Factor Receptor 1 Based on Virtual Screening and Molecular Dynamics Analysis

AbstractFibroblast growth factor receptor 1 (FGFR1) is involved in the process of cell proliferation, differentiation and function, especially in bone skeletal development. It is a potential new therapeutic target for osteoporosis and other skeletal‐related diseases. In this study, we aim to identify FGFR1 inhibitors from Traditional Chinese Medicine (TCM) database by the structure‐based virtual screening and MM/GBSA scoring. First, 26450 compounds were filtered by Lipinski rules of five, followed by a hierarchy virtual screen workflow which includes glide docking, ranking, cluster analysis and visual inspection to afford 40 potential compound candidates. Finally, 6 candidates were selected based on MM/GBSA binding energy analysis of the with FGFR1 protein. Our modelling study would provide some clues to the discovery of novel FGFR1 inhibitors as potential therapeutic drugs for osteoporosis. In addition, given multipotency characteristic of FGFR1, these potential inhibitors might provide some reference to other related diseases, such as non‐small cell lung cancer and squamous cell lung cancer.

Dapagliflozin Inhibits Cell Adhesion to Collagen I and IV and Increases Ectodomain Proteolytic Cleavage of DDR1 by Increasing ADAM10 Activity.

Dapagliflozin, empagliflozin, tofogliflozin, selective inhibitors of sodium-glucose cotransporter 2 (SGLT2), is used clinically to reduce circulation glucose levels in patients with type 2 diabetes mellitus by blocking the reabsorption of glucose by the kidneys. Dapagliflozin is metabolized and inactivated by UGT1A9. Empagliflozin is metabolized and inactivated by UGT1A9 and by other related isoforms UGT2B7, UGT1A3, and UGT1A8. Tofogliflozin is metabolized and inactivated by five different enzymes CYP2C18, CYP3A4, CYP3A5, CYP4A11, and CYP4F3. Dapagliflozin treatment of HCT116 cells, which express SGLT2 but not UGT1A9, results in the loss of cell adhesion, whereas HepG2 cells, which express both SGLT2 and UGT1A9, are resistant to the adhesion-related effects of dapagliflozin. PANC-1 and H1792 cells, which do not express either SGLT2 or UGT1A9, are also resistant to adhesion related effects of dapagliflozin. On the other hand, either empagliflozin or tofogliflozin treatment of HCT116, HepG2, PANC-1, and H1792 cells are resistant to the adhesion-related effects as observed in dapagliflozin treated HCT116 cells. Knockdown of UGT1A9 by shRNA in HepG2 cells increased dapagliflozin sensitivity, whereas the overexpression of UGT1A9 in HCT116 cells protected against dapagliflozin-dependent loos of cell adhesion. Dapagliflozin treatment had no effect on cellular interactions with fibronectin, vitronectin, or laminin, but it induced a loss of interaction with collagen I and IV. In parallel, dapagliflozin treatment reduced protein levels of the full-length discoidin domain receptor I (DDR1), concomitant with appearance of DDR1 cleavage products and ectodomain shedding of DDR1. In line with these observations, unmetabolized dapagliflozin increased ADAM10 activity. Dapagliflozin treatment also significantly reduced Y792 tyrosine phosphorylation of DDR1 leading to decrement of DDR1 function and detachment of cancer cells. Concomitant with these lines of results, we experienced that CEA in patients with colon cancer, which express SGLT2 but not UGT1A9, and type 2 diabetes mellitus treated by dapagliflozin in addition to chemotherapy was decreased (case 1). CEA in patients with colon cancer, which express SGLT2 but not UGT1A9, and type 2 diabetes mellitus was treated by dapagliflozin alone after radiation therapy was decreased but started to rise after cessation of dapagliflozin (case 2). CA19-9 in two of patients with pancreatic cancer and type 2 diabetes mellitus was resistant to the combination therapy of dapagliflozin and chemotherapy (case 3 and 4 respectively). PIVKAII in patients with liver cancer and type 2 diabetes mellitus, and CYFRA in patients with squamous lung cancer and type 2 diabetes mellitus was also resistant the combination therapy of dapagliflozin and chemotherapy (case 5 and 6 respectively). Taken together, these data suggest a potential role for dapagliflozin anticancer therapy against colon cancer cells that express SGLT2, but not UGT1A9.

Genetic determinants of the molecular portraits of epithelial cancers

The ability to characterize and predict tumor phenotypes is crucial to precision medicine. In this study, we present an integrative computational approach using a genome-wide association analysis and an Elastic Net prediction method to analyze the relationship between DNA copy number alterations and an archive of gene expression signatures. Across breast cancers, we are able to quantitatively predict many gene signatures levels within individual tumors with high accuracy based upon DNA copy number features alone, including proliferation status and Estrogen-signaling pathway activity. We can also predict many other key phenotypes, including intrinsic molecular subtypes, estrogen receptor status, and TP53 mutation. This approach is also applied to TCGA Pan-Cancer, which identify repeatedly predictable signatures across tumor types including immune features in lung squamous and basal-like breast cancers. These Elastic Net DNA predictors could also be called from DNA-based gene panels, thus facilitating their use as biomarkers to guide therapeutic decision making.

A phase Ib study of utomilumab (PF-05082566) in combination with mogamulizumab in patients with advanced solid tumors

BackgroundExpressed on activated T and natural killer cells, 4-1BB/CD137 is a costimulatory receptor that signals a series of events resulting in cytokine secretion and enhanced effector function. Targeting 4-1BB/CD137 with agonist antibodies has been associated with tumor reduction and antitumor immunity. C-C chemokine receptor 4 (CCR4) is highly expressed in various solid tumor indications and associated with poor prognosis. This phase Ib, open-label study in patients with advanced solid tumors assessed the safety, efficacy, pharmacokinetics, and pharmacodynamics of utomilumab (PF-05082566), a human monoclonal antibody (mAb) agonist of the T-cell costimulatory receptor 4-1BB/CD137, in combination with mogamulizumab, a humanized mAb targeting CCR4 reported to deplete subsets of regulatory T cells (Tregs).MethodsUtomilumab 1.2–5 mg/kg or 100 mg flat dose every 4 weeks plus mogamulizumab 1 mg/kg (weekly in Cycle 1 followed by biweekly in Cycles ≥2) was administered intravenously to 24 adults with solid tumors. Blood was collected pre- and post-dose for assessment of drug pharmacokinetics, immunogenicity, and pharmacodynamic markers. Baseline tumor biopsies from a subset of patients were also analyzed for the presence of programmed cell death-ligand 1 (PD-L1), CD8, FoxP3, and 4-1BB/CD137. Radiologic tumor assessments were conducted at baseline and on treatment every 8 weeks.ResultsNo dose-limiting toxicities occurred and the maximum tolerated dose was determined to be at least 2.4 mg/kg per the time-to-event continual reassessment method. No serious adverse events related to either treatment were observed; anemia was the only grade 3 non-serious adverse event related to both treatments. Utomilumab systemic exposure appeared to increase with dose. One patient with PD-L1–refractory squamous lung cancer achieved a best overall response of partial response and 9 patients had a best overall response of stable disease. No patients achieved complete response. Objective response rate was 4.2% (95% confidence interval: 0.1–21.1%) per RECIST 1.1. Depletion of Tregs in peripheral blood was accompanied by evidence of T-cell expansion as assessed by T-cell receptor sequence analysis.ConclusionsThe combination of utomilumab/mogamulizumab was safe and tolerable, and may be suitable for evaluation in settings where CCR4-expressing Tregs are suppressing anticancer immunity.Trial registrationClinicalTrials.gov identifier: NCT02444793.

Abstract A002: Discovery of bis(hydroxymethyl)benzopyrrolo[2,1-a]phthalazine derivatives as orally bioavailable antitumor agents with anti-angiogenic and DNA cross-linking activities

Abstract Angiogenesis plays a critical role in sustaining the microenvironment, growth, and metastatic potential of a variety of tumors, such as lung cancer, colorectal cancer, and renal cancer. However, the rapid development of resistance to anti-angiogenesis agents has limited the clinical benefits. To improve the therapeutic efficacy, the designed dual functional inhibitors, composed of benzophthalazine (anti-angiogenesis moiety) and bis(hydroxymethyl)pyrrole (DNA interstrand crosslinking moiety), were synthesized. We first demonstrated that 1,2-bis(hydroxymethyl)benzopyrrolo[2,1-a]phthalazines displayed dual functions, inducing DNA crosslinking and inhibiting angiogenesis. By aid of DIVAA, we confirmed the in vivo anti-angiogenetic activity. They were cytotoxic to a panel of cancer cell lines, including leukemia, colorectal, renal, and lung cancer cells. Among them, small cell lung cancer (SCLC) cells were the most susceptible cells with the IC50 values ranged from subnanomolar to micromolar. Significantly, we observed tumor suppression by treatment of compound 8 in a given formulation at the dose of 20 mg/kg (10 times in 2 weeks) in SCLC H526, squamous lung cancer H520 and renal cancer 786-O xenograft models. In addition, orally admission of 60 mg/kg compound 8 daily for 14 days was also more potent than cisplatin in suppression of growth of SCLC H526 xenografts. Importantly, compound 8 at the dose given did not cause body weight lost. In summary, our results indicate that bis(hydroxymethyl)benzopyrrolo[2,1-a]phthalazine hybrids are potent than anticancer agents against a wide spectrum of cancers. They deserve our further development. Citation Format: Tai-Lin Chen, Vicky Jain, Anilkumar S. Patel, Yi-Wen Lin, Tsann-Long Su, Te-Chang Lee. Discovery of bis(hydroxymethyl)benzopyrrolo[2,1-a]phthalazine derivatives as orally bioavailable antitumor agents with anti-angiogenic and DNA cross-linking activities [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A002. doi:10.1158/1535-7163.TARG-19-A002

A rationale for surgical debulking to improve anti-PD1 therapy outcome in non small cell lung cancer

Anti-PD1 immunotherapy has emerged as a gold-standard treatment for first- or second-line treatment of stage IV NSCLC, with response rates ranging from 10 to 60%. Strategies to improve the disease control rate are needed. Several reports suggested that debulking surgery enhances anti-tumor immunity. We aimed at examining tumor burden as a predictive factor of anti-PD1 tretment efficacy and to evaluate the role of cytoreductive surgery in anti-PD1 treated NSCLC. Immunocompetent DBA/2 mice engrafted with various amount of allogeneic lung squamous cancer KLN-205 cells were treated with anti-PD1 monoclonal antibody. Mice engrafted with two tumors also underwent a debulking surgery or a sham procedure. Tumor volume was monitored to assess treatment efficacy. Tumor infiltrating lymphocytes were assessed by flow cytometry. In a retrospective study of 48 stage IV NSCLC patients treated with Nivolumab who underwent a 18-FDG PETscan before treatment onset, the prognostic role of metabolic tumor volume was analysed. Anti-PD1 treatment effect was greater in mice bearing smaller tumors. Treatment with higher doses of anti-PD1 antibody did not improve the outcome, independently of the size of the tumor. In mice bearing 2 tumors, excision of 1 tumor improved the anti-PD1 treatment effect on the remaining tumor. In 48 NSCLC patients receiving anti-PD1 treatment, high metabolic tumor volume was associated with poor overall survival and the absence of clinical benefit. Treg infiltration, but not effector T cells, was positively correlated to tumor volume. Taken together, our results suggest that tumor volume is a predictive factor of anti-PD1 efficacy in NSCLC. Additionally, an experimental murine model suggests that tumor debulking may improve control of residual tumor.

MS05.04 Innate Immune Mediators in Lung Cancer

Non-small-cell lung cancer (NSCLC) accounts for about 85% of total lung cancer cases. The major types of NSCLC—squamous cell carcinoma and adenocarcinoma—harbor distinct histopathologies, biomarker expression, genomic alterations, and response to therapy [1,2]. Recent studies have shown that there are also differences in their tumor immune microenvironments [3–6]. Specifically, adenocarcinomas have increased infiltration of tumor-associated macrophages, while squamous lung tumors exhibit an enrichment in tumor-associated neutrophils (TANs) in both mouse and human lung tumors. The two major subtypes of NSCLC are also associated with distinct lineage-specific master regulators: SOX2 is commonly amplified and up-regulated in the vast majority of squamous tumors anddrives the squamous fate, whereas NKX2-1 is highly expressed in adenocarcinoma and governs adenocarcinoma fate [2]. We developed novel genetically engineered mouse models (GEMMs) of squamous lung cancer on the basis of overexpression of the transcription factor Sox2 and loss of the tumor suppressor Lkb1 (SL mice) (Mukhopadhyay et al, Cell Rep, 2014 [7]). SL tumors recapitulated gene expression and immune infiltrate features of human squamous NSCLC, including an enrichment of TANs and a decrease in expression of NKX2-1. Deletion of Nkx2-1in SL mice (SNL) revealed that NKX2-1 suppresses SOX2-driven squamous tumorigenesis by repressing adeno-to-squamous transdifferentiation. We further employed multiple GEMMs to elucidate the role of SOX2 and NKX2-1 in tumor cell fate and TAN recruitment. In Kras-driven adenocarcinomas,mis-expression of Sox2 or loss of Nkx2-1 led to TANrecruitment. SOX2 recruits, whereas NKX2-1 suppresses, TANs at least partly through inverse regulation of the chemokineCxcl5. Tumor-derived CXCL5 is sufficient to recruit TANs. Single cell RNA sequencing (scRNA-seq) revealed that TANs exhibit tumor-promoting features, including production of reactive oxygen species (ROS), and distinct gene expression profiles compared to blood neutrophils (Mollaoglu et al, Immunity, 2018 [8]). Depletion of TANs through LY6G blocking antibodies or CXCR2 inhibitors in SNL mice reduced squamous tumors, suggesting that TANs foster squamous cell fate. Furthermore, TAN depletion coupled with scRNA-seq suggests that TANs regulate distinct aspects of tumor cell fate. Together, these data suggest that lineage-defining transcription factors determine the tumor immune microenvironment, which in turn can impact the nature of the tumor. References 1 Langer, C.J. et al. (2016) Incremental Innovation and Progress in Advanced Squamous Cell Lung Cancer: Current Status and Future Impact of Treatment. J. Thorac. Oncol.11, 2066–2081. 2 Campbell, J.D.et al. (2016) Distinct patterns of somatic genome alterations in lung adenocarcinomas and squamous cell carcinomas. Nat. Genet.48, 607–616. 3 Kargl, J. et al. (2017) Neutrophils dominate the immune cell composition in non-small cell lung cancer. Nat. Commun.8, 14381 4 Nagaraj, A.S. et al. (2017) Cell of Origin Links Histotype Spectrum to Immune Microenvironment Diversity in Non-small-Cell Lung Cancer Driven by Mutant Kras and Loss of Lkb1. Cell Reports 18, 673–684. 5 Xu, C. et al. (2014) Loss of Lkb1 and Pten Leads to Lung Squamous Cell Carcinoma with Elevated PD-L1 Expression. Cancer Cell 25, 590–604. 6 Ferone G., et al. SOX2 Is the Determining Oncogenic Switch in Promoting Lung Squamous Cell Carcinoma from Different Cells of Origin. Cancer Cell. 2016;30(4):519-532. https://doi.org/10.1016/J.CCELL.2016.09.001. 7 Mukhopadhyay, A. et al. (2014) Sox2 Cooperates with Lkb1 Loss in a Mouse Model of Squamous Cell Lung Cancer. Cell Reports 8, 40–49. 8 Mollaoglu, G. et al. (2018) The Lineage-Defining Transcription Factors SOX2 and NKX2-1 Determine Lung Cancer Cell Fate and Shape the Tumor Immune Microenvironment. Immunity 49, 764-779.e9. squamous, neutrophil, SOX2