Lung Squamous Carcinoma Research Articles

Abstract 4853: M6123 is an ADCC-enhanced and selective monovalent antagonist of FGFR1 showing antitumor activity in preclinical lung cancer models

Abstract Despite the considerable progress made with immunotherapies and targeted therapies, lung cancer remains a devastating disease with a high medical need for more effective and safer therapeutic options. Targeted therapies have delivered meaningful benefit to subsets of lung adenocarcinoma patients harboring specific oncogenic driver alterations but have not yet been extensively explored in other lung cancer subtypes. In the case of lung squamous carcinoma and small-cell lung cancer, amplification and overexpression of FGFR1 have been described as potential oncogenic drivers.Several small molecule tyrosine kinase inhibitors of the FGFR family are in clinical development but initial reports have suggested limited therapeutic benefit of these agents when applied as monotherapy in FGFR1-amplified lung cancer patients. To overcome toxicities associated with pan-FGFR inhibition, such as dysregulation of FGF23-mediated phosphate homeostasis, M6123 was developed as an FGFR1-selective biological antagonist. An immunization campaign in OmniRats® led to the identification of the fully human, potent and highly selective FGFR1 antibody A08 which was converted into the monovalent FGFR1 antagonist M6123 based on the proprietary SEED platform. Furthermore, M6123 was engineered for enhanced ADCC effector function. In contrast to bivalent FGFR1 antibodies, M6123 was well tolerated in mice, with no signs of body weight loss. Of note, M6123 did not interfere with hormonal FGF23 signaling in mice, a severe side effect of FGFR kinase inhibitors. Concentration-dependent inhibition of FGFR1 auto-phosphorylation was demonstrated in FGFR1-overexpressing lung cancer cells both in vitro and in vivo. Moreover, M6123 monotherapy had strong antitumor activity in specific FGFR1-expressing cell line- and patient-derived xenograft models, whilst combination with chemotherapy significantly enhanced efficacy. In summary, M6123 is a unique and clearly differentiated selective FGFR1 antagonist which warrants further testing in preclinical models to refine a predictive biomarker strategy for selection of potentially sensitive lung cancer patients. Citation Format: Christina Esdar, Christel Iffland, Astrid Zimmermann, Angela Lim, Michael Sanderson, Andree Blaukat. M6123 is an ADCC-enhanced and selective monovalent antagonist of FGFR1 showing antitumor activity in preclinical lung cancer models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4853.

Abstract 3234: PD-1 is intrinsically expressed by lung cancer cells with stemness features inhibited by PD-1 blockade

Abstract Purpose: Aim of this study is to explore the intrinsic expression, functional role and therapeutic modulation of PD-1 receptor in Non Small Cell Lung Cancer (NSCLC) cells. We previously reported (Sanlorenzo M et al. Clinical Cancer Res 2018) that PD-1+ melanoma cells may sustain disease relapse following treatment with BRAF/MEK inhibitors. We hypothesized that PD-1+ tumor cells may characterize a “stem-like” compartment also in NSCLC, sustaining chemo-resistance and disease relapse with potential therapeutic implications. Experimental procedures: The expression of PD-1 by NSCLC cells was explored by flow cytometry, western blot (WB) and RT-PCR. Its presence and role in lung cancer stem cells (CSC) was explored by sphere formation essays. CSC were further visualized by tumor-engineering with a lentiviral CSC-detector vector (LV-CSC) encoding eGFP under control of the OCT4 stem gene promoter. Selective PD-1 blockade and PD-1 stimulation with soluble ligand (PD-L1) were used to study the functional role and therapeutic modulation of intrinsic PD-1 in NSCLC. Results: PD-1 is intrinsically expressed by a small subset of NSCLC cells with stemness features. We found PD-1 consistently expressed on the membrane of a small tumor cell fraction (2% ± 0.3) within 6 NSCLC cell lines (H1975, EBC-1, H23, H820, HCC827), including a primary patient-derived NSCLC culture (SL1). Data were confirmed by RT-PCR and WB. Viable PD-1+ tumor cells were significantly enriched in NSCLC spheres generated in stem conditioned cultures, compared with the monolayer controls (10% [4-36] vs 2% [1-5] P<0,0001, n=6). The levels of PD-1 and stem gene OCT4 RNA comparably increased in NSCLC spheres (4.5 vs 4 fold, n=5). The tumor-intrinsic expression of PD-1 was confirmed by data mining in 67 adenocarcinoma (Affimetryx RNA value 4.348 [3.882-6.361]) and 28 squamous lung carcinoma (RNA value 4.338 [3.995-5.178]) cell lines (CCLE). The formation of NSCLC spheres was significantly inhibited (-30%±2, n=6 P=0.0004) by anti-PD-1 blocking antibody (100μg/μl), while enhanced (25% ± 2, n=3, P= 0.007) by soluble PD-L1 (50 μg/μl). Similar results were confirmed by selective PD-1 RNA-interference that revoked the pro-tumorigenic effect of soluble PD-L1. In vitro treatment with Cisplatin (IC50 dose) led to a relative enrichment of PD-1+ (2.5±0.3 fold, n=4) cells and OCT4+ CSC (3.5±0.3 fold, n=4). The sequential PD-1 inhibition significantly delayed NSCLC cell recovery (-45%±9.8, n=4) and sphere formation after cisplatin. Conclusions: PD-1 is intrinsically expressed by NSCLC cells with stemness features and mediates pro-tumorigenic activity. PD-1+ NSCLC cells are enriched after chemotherapy and may be inhibited by selective PD-1 blockade. We report a new, intrinsic, expression pattern of PD-1 in NSCLC, providing rationale to explore a lymphocyte-independent activity of anti-PD1 antibodies. Citation Format: Ramona Rotolo, Valeria Leuci, Chiara Donini, Martina Sanlorenzo, Igor Vujic, Giovanni Medico, Francesca Vita, Loretta Gammaitoni, Luisella Righi, Chiara Riganti, Elisa Vigna, Lorenzo D'Ambrosio, Giovanni Grignani, Giorgio V. Scagliotti, Silvia Novello, Massimo Aglietta, Dario Sangiolo. PD-1 is intrinsically expressed by lung cancer cells with stemness features inhibited by PD-1 blockade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3234.

An effective and accurate algorithm for measuring the tumor mutation burden of cancer patients plasma.

e14177 Background: Tumor Mutation Burden (TMB) is deemed to be one of the effective genomic biomarkers that are used to predict the response to PD-1/PD-L1 blockade immunotherapy. As an exciting biomarker for diagnose and an alternative assess method for tissue TMB (tTMB), blood TMB (bTMB) is measured to further distinguish the patients who may benefit from immunotherapy. However, there is no recognized rule to calculate bTMB so far. Methods: In this study, we developed an in-house algorithm called PTAB to calculate bTMB of plasma, all credible single nucleotide variants and insertion-deletions whose allele frequency is greater than 1% are selected in PTAB without synonymous mutations and drive mutations. We also constructed a 636 genes panel which region size is 1.95Mb to identify all the associated mutations, due to the high costs and long turnaround time of WES. Results: Meanwhile, we assessed a cohort of 50 lung cancer plasma samples and paired tissues to ensure the consistency of different algorithm used for tissue and plasma. The results show that Pearson correlation coefficient (R2) between bTMB and tTMB in 50 patients is 0.7128. In addition, we found that the consistency of lung squamous carcinomas obviously higher than that of lung adenocarcinoma (0.8932 vs. 0.5947). We also described the distribution of bTMB and tTMB value of Chinese patients, including 594 tissue samples and 561 plasma samples from 8 types of cancers, including lung cancer, colorectal cancer, breast cancer, et al. The overall comparative analysis of 1155 previous data suggested that the distribution modes of bTMB and tTMB were almost same in certain cancer type, though the level of bTMB was generally lower than that of tTMB. Conclusions: Preliminary data highlight that PTAB is an effective and accurate algorithm for measure the TMB of plasma. The applicability of bTMB testing on liquid biopsy needs further investigation and may be clarified within the ongoing clinical trials.

Immune-checkpoints inhibitors in metastatic non small cell lung cancer with rare histology.

9106 Background: Immune-checkpoints inhibitors (ICIs) have clearly improved prognosis of metastatic lung squamous carcinoma and adenocarcinoma, while their benefit remains uncertain in patients (pts) with rare NSCLC histotypes (RH). The study aim was to evaluate ICIs efficacy in RH. Methods: We retrospectively collected data from consecutive metastatic NSCLC pts treated with ICIs at our Institution from 4/2013 to 12/2018. Objective response rate (ORR) and disease control rate (DCR) were assessed. Fisher’s exact test was used to compare ORR and DCR in RH versus not-RH (NRH). Univariate and multivariate survival analyses were estimated by Kaplan-Meier and Cox progression hazard models. Results: Of 268 pts, 31 (11.6%) had RH: 16 sarcomatoid, 7 pulmonary enteric adenocarcinoma, 4 large cell neuroendocrine carcinoma and 4 adenosquamous carcinoma. In RH group, median age was 67 years old (range 41-81), most were males (71%) and smokers (90.3%); ECOG PS was: 0 (16.1%), 1 (67.8%) and 2 (16.1%). PD-L1 < 1%, 1-49%, ≥50% and unknown expression were reported in 22.6%, 19.3%, 35.5% and 22.6% pts, respectively. Twelve pts received ICIs as first and 19 as second or further-line. ORR was 22.6% in RH, 20.3% in NRH (p = 0.81); DCR was 35.5% in RH, 53.1% in NRH (p = 0.08). After a median follow-up of 20 months (m) (95% CI 4.0 – 36.7 m), median progression-free survival (PFS) was 2.6 m (95% CI 1.9-3.3 m) in RH vs 2.6 m in NRH (95% CI 2.1 – 3.0 m); median overall survival (OS) was 4.6 m (95% CI 0.03-12.0 m) in RH vs 9.2 m (95% CI 7.4 – 10.9 m) in NRH. No statistically significant differences were seen between the two groups (p = 0.729 for PFS, p = 0.601 for OS). At multivariate analyses adjusted for age, sex, smoke, PS, PD-L1 status, line of therapy and histotype (RH vs NRH), only low PS and first line treatment showed better PFS and OS (p < 0.001 and p = 0.003, respectively) in overall population. Conclusions: Our analysis, limited by the small and heterogeneous RH sample size, reported no significant differences between RH and NRH in terms of ORR, DCR and survival. However, looking at OS and DCR data, RH seem to have worse outcome. Correlation between histotype and pts characteristics and survival analyses in a larger cohort of ICIs treated NSCLC pts is ongoing. Specific prospective trials are needed to evaluate ICIs role in RH.

Amplification of USP13 drives non-small cell lung cancer progression mediated by AKT/MAPK signaling

USP13 is emerging as a potential target in cancer therapy. However, the effect of USP13 on tumor progression is controversial. Here we focused on non-small cell lung cancer (NSCLC), a common cancer with high mortality, and studied the role of USP13 in tumor growth. By analysis of multi-level genetic database, we found USP13 is high expressed in heart among healthy primary tissues and is most amplified in lung cancer. Clinical samples of NSCLC showed tumor exhibited high USP13 level compared with adjacent normal tissues. We further utilized lung adenocarcinoma A549 and squamous carcinoma H226 cells as cell model and investigated USP13 effect by USP13 knockdown. As a results, downregulation of USP13 dramatically inhibited A549 and H226 cell proliferation by AKT/MAPK signaling and suppressed tumor growth in nude mice. Collectively, we identified USP13 as a tumor promoter in NSCLC and provide a promising target in cancer therapy.

Zator płucny i guz lewego przedsionka u pacjentki z nowotworem płuca — trudności diagnostyczno-terapeutyczne

A 64 year-old female with a history of hypertension and chronic obstructive pulmonary disease after radical treatment (radiotherapy and chemotherapy) of squamous cell carcinoma of the right lung in 2016, was admitted to the Depart- ment of Cardiology due to pulmonary embolism, confirmed in an angio-CT. Moreover, cancer recurrence on the right lung and a mass spreading through the right upper pulmonary vein to the left atrium (LA) that might be considered to be a thrombus, were demonstrated. The patient, for the week before admission, had suffered from exercise dyspnoea, pain in the lower limbs, and syncope. On admission, the patient presented in a serious general condition. In laboratory tests, elevated D-dimer and NT-proBNP (450 pg/ml) concentrations as well as mild normocytic anaemia were revealed. Echocardiography showed preserved systolic (ejection fraction = 55%) and impaired diastolic left ventricle function, without signs of valvular heart disease. In transoesophageal echocardiography (TEE) in the LA an additional echo of 18 × 12 mm, spreading through the right upper pulmonary vein that might be considered to be a thrombus was revealed. However, due to the location of neo- plastic lesions in the right lung, a cancerous tumour spreading throughout could not be excluded. An abdominal and pelvis CT did not reveal any metastasis. Low molecular weight heparin in a therapeutic dose and acetylsalicylic acid were incorporated into the therapy. A mul- tidisciplinary consultation (oncologist, cardiac surgeon, cardiologist, anaesthesiologist) was conducted and the mass in the LA was initially qualified for surgical resection. Prior to surgery, spirometry and the exclusion of thromboembolic material in the lower limb veins were required. Spirometry demonstrated moderate bronchial obstruction. In ultrasound of the lower limb, deep vein thrombosis was excluded, but a few hypoechal nodules in the shank muscles of the right lower limb were visualised. A suspicion of metastases was confirmed using angio-CT. In the TEE performed after five days of low molecular weight heparin therapy, the previously described mass in the LA reduced its echogenicity and dimensions (to 10 × 8 mm). The patient’s general condition improved slightly. The patient was re-consulted by the cardiac surgeon and, due to presence of distant metastases, was disqualified from surgery. Another oncologist consultation was performed, and further diagnostic and therapeutic aims were established. The therapeutic dose of heparin was maintained.

Genomic alterations and survival in young patients aged under 40 years with completely resected non-small cell lung cancer.

Young patients are rarely diagnosed with non-small cell lung cancer (NSCLC), and little is known about its predisposing genomic alterations and survival. This retrospective study was conducted to evaluate the genomic alterations, treatment and prognosis of young patients under 40 years old from a cohort of 640 lung adenocarcinoma and squamous carcinoma cases from Zhejiang Cancer Hospital. All patients were examined for EGFR, KRAS, NRAS, PIK3CA, BRAF and HER2 mutations and ALK, ROS1 and RET fusion genes. In total, 54 patients were aged under 40 years. The frequencies of genomic alterations in younger (≤40 years) and older (>40 years) patients were 68.5% and 54.8%, respectively (P=0.05). Younger patients harbored a higher frequency of fusion genes (22.2% vs. 4.1%, P<0.001) but not gene mutations (46.3% vs. 45.6%, P=0.92). There was a general trend toward shorter recurrence-free survival (RFS) (35.2 vs. 43.8 months, P=0.050), while no overall survival (OS) difference existed between younger and older patients (50.2 vs. 51.4 months, P=0.112). Younger patients with NSCLC had a higher frequency of gene fusions than older patients and had a trend of worse OS.

Genetic variants in RUNX3, AMD1 and MSRA in the methionine metabolic pathway and survival in nonsmall cell lung cancer patients.

Abnormal methionine dependence in cancer cells has led to methionine restriction as a potential therapeutic strategy. We hypothesized that genetic variants involved in methionine-metabolic genes are associated with survival in nonsmall cell lung cancer (NSCLC) patients. Therefore, we investigated associations of 16,378 common single-nucleotide polymorphisms (SNPs) in 97 methionine-metabolic pathway genes with overall survival (OS) in NSCLC patients using genotyping data from two published genome-wide association study (GWAS) datasets. In the single-locus analysis, 1,005 SNPs were significantly associated with NSCLC OS (p < 0.05 and false-positive report probability < 0.2) in the discovery dataset. Three SNPs (RUNX3 rs7553295 G > T, AMD1 rs1279590 G > A and MSRA rs73534533 C > A) were replicated in the validation dataset, and their meta-analysis showed an adjusted hazards ratio [HR] of 0.82 [95% confidence interval (CI) =0.75-0.89] and pmeta = 2.86 × 10-6 , 0.81 (0.73-0.91) and pmeta = 4.63 × 10-4 , and 0.77 (0.68-0.89) and pmeta = 2.07 × 10-4 , respectively). A genetic score of protective genotypes of these three SNPs revealed an increased OS in a dose-response manner (ptrend < 0.0001). Further expression quantitative trait loci (eQTL) analysis showed significant associations between these genotypes and mRNA expression levels. Moreover, differential expression analysis further supported a tumor-suppressive effect of MSRA, with lower mRNA levels in both lung squamous carcinoma and adenocarcinoma (p < 0.0001 and < 0.0001, respectively) than in adjacent normal tissues. Additionally, low mutation rates of these three genes indicated the critical roles of these functional SNPs in cancer progression. Taken together, these genetic variants of methionine-metabolic pathway genes may be promising predictors of survival in NSCLC patients.

A VEGF-A/SOX2/SRSF2 network controls VEGFR1 pre-mRNA alternative splicing in lung carcinoma cells

The splice variant sVEGFR1-i13 is a truncated version of the cell membrane-spanning VEGFR1 receptor that is devoid of its transmembrane and tyrosine kinase domains. We recently showed the contribution of sVEGFR1-i13 to the progression and the response of squamous lung carcinoma to anti-angiogenic therapies. In this study, we identify VEGF165, a splice variant of VEGF-A, as a regulator of sVEGFR1-i13 expression in these tumors, and further show that VEGF165 cooperates with the transcription factor SOX2 and the splicing factor SRSF2 to control sVEGFR1-i13 expression. We also demonstrate that anti-angiogenic therapies up-regulate sVEGFR1-i13 protein level in squamous lung carcinoma cells by a mechanism involving the VEGF165/SOX2/SRSF2 network. Collectively, our results identify for the first time a signaling network that controls VEGFR1 pre-mRNA alternative splicing in cancer cells.

Vascular Endothelial Growth Factor-D (VEGF-D) is Elevated in Bronchoalveolar Lavage Fluid of Patients with Lung Squamous Carcinoma.

Previous studies have found that vascular endothelial growth factor (VEGF) is associated with lung cancer, yet little is known about vascular endothelial growth factor-D (VEGF-D) in bronchoalveolar lavage fluid (BALF) of lung cancer patients. In this study, we aim to investigate the expression and evaluation of VEGF-D in BALF for lung cancer diagnosis. BALF samples were acquired from 81 patients: 40 with benign diseases and 41 with lung cancer. The expression of VEGF-D in BALF was measured using sandwich enzyme-linked immune sorbent assays (ELISA), and the evaluation of VEGF-D in BALF for lung cancer diagnosis was also investigated. In the BALF samples, the levels of VEGF-D in the lung cancer group were higher than in the benign disease group; however, there was no statistical significance between the two groups (p > 0.05). In the pathological classification of lung cancer, the levels of VEGF-D in the BALF differed significantly between the lung squamous carcinoma group and the benign disease group (p < 0.05). The diagnostic accuracies of VEGF-D in BALF for discrimination between patients with squamous cell carcinoma and benign disease were reasonable based on receiver operating characteristic (ROC curve) analysis, with a corresponding sensitivity of 64.7% and specificity of 60%, respectively. This study demonstrated that the detection of VEGF-D levels in BALF is a valuable diagnostic tool for lung squamous carcinoma.

A Rare Case of Synchronous Presentation of Acute Myeloid Leukemia and Lung Cancer

Background: There are very few publications on synchronous presentation of dual malignancies in the literature. Occurrence of second malignancy in a patient with a known malignant tumor is not uncommon, though synchronous primary malignancies are unusual. Case Presentation: A 77-year-old man was diagnosed elsewhere as squamous cell carcinoma of lung came to our oncology department for further management. He underwent bone marrow evaluation with the suspicious of metastasis in view of pancytopenia. On bone marrow evaluation, found to have prominence of blasts more than 20%, which were CD34 positive and CD117 positive and. Conclusion: Here we describe an unusual rare combination of synchronous presentation of acute myeloid leukemia and squamous cell carcinoma of lung.

Comparison of PANAMutyper and PNAClamp for Detecting KRAS Mutations from Patients With Malignant Pleural Effusion.

KRAS is one of the frequently mutated genes in human cancers and often relates with drug resistance and poor prognosis. PANAMutyper™ is a novel technology that integrates PNAClamp™ and PANA S-Melting™. In the present study, PANAMutyper™ and PNAClamp™ were compared for the detection of KRAS mutations using different samples of patients with malignant pleural effusion. A total of 103 patients (including 56 lung adenocarcinoma, 10 lung squamous carcinoma, 17 small cell lung cancer, 3 large cell lung cancer, 3 stomach cancer, 2 ovarian cancer, and others) with malignant pleural effusion were investigated using matched tumor tissue, cell block, and pleural effusion samples. The diagnostic performance of these two methods was compared. KRAS mutations were detected in 18 (17.5%) of 103 patients using tissue, cell block, and pleural effusion samples. All 18 patients with KRAS mutations were detected by PANAMutyper™ using any sample type, however, only 7 cases were detected by PNAClamp™. Among the subtypes of KRAS mutations, substitution in codon 12, 35G>T was the most frequent, followed by substitution in codon 12, 35G>A and codon 12, 34G>A. In pleural effusion specimens, PANAMutyper™ showed a better diagnostic performance compared to PNAClamp™. PANAMutyper™ had a diagnostic superiority for the detection of KRAS mutations in patients with malignant pleural effusion compared to PNAClamp™, although there was a concordance between PANAMutyper™ and PNAClamp™ results. Therefore, PANAMutyper™ can be used for a more sensitive and accurate detection of KRAS mutations.

Construction of a transcription factor‑long non‑coding RNA‑microRNA network for the identification of key regulators in lung adenocarcinoma and lung squamous cell carcinoma.

The interactions of microRNAs (miRNAs), transcription factors (TFs) and their common target long non-coding RNAs (lncRNAs) can lead to the production of TF-miRNA-lncRNA (TML) network motifs. These motifs are functional regulators that perform a wide range of biological processes, such as carcinogenesis. However, TML network motifs have not been systematically identified, and their roles in lung adenocarcinoma (LUAD) and lung squamous carcinoma (LUSC) are largely unknown. In the present study, a computational integration approach was performed using multiple sources in order to construct a global TML network for LUAD and LUSC. The analysis revealed several dysregulated TML network motifs, which were common between the two lung cancer subtypes or specific to a single cancer subtype. In addition, functional analysis further indicated that the TML network motifs may potentially serve as putative biomarkers in LUAD and LUSC. The associations between drug treatments and dysregulated TML network motifs were also examined. Collectively, the present study elucidated the roles of TML network motifs in LUAD and LUSC, which may be beneficial for understanding the pathogenesis of lung cancer and its potential treatment.

Interleukin-7 promotes lung-resident CD14+ monocytes activity in patients with lung squamous carcinoma

Interleukin (IL)-7 enhances cytokines secretion by CD14+ monocytes, and induces recruitment of monocytes to endothelium. As an important regulator to different types of immune cells, the role of IL-7 in modulation of CD14+ monocytes is still not fully elucidated. Thus, the aim of current study was to investigate the immunoregulatory activity of IL-7 to peripheral and lung-resident CD14+ monocytes in lung squamous carcinoma patients. Thirty-seven lung squamous carcinoma patients and eighteen healthy individuals were enrolled. CD14+ monocytes and CD4+ T cells were purified from both peripheral bloods and bronchoalveolar lavage fluids (BALF). IL-7 expression in plasma and BALF was measured by ELISA, and CD127 expression in peripheral and lung-resident CD14+ monocytes was investigated by real-time PCR and flow cytometry, respectively. Cellular proliferation, cytokine production, and molecules in IL-7 signaling pathway was assessed in CD14+ monocytes in response to IL-7 stimulation. IL-7-induced CD14+ monocytes activity to CD4+ T cells was also assessed in direct and indirect contact co-culture system. There were no remarkable differences of plasma IL-7 concentration or CD127 level between healthy individuals and lung squamous carcinoma patients. However, IL-7 expression was significantly reduced in BALF from tumor site in squamous carcinoma patients, especially in stage III and IV. IL-7 stimulation not only promoted proliferation, cytokines secretion, and STAT-5 phosphorylation in lung-resident CD14+ monocytes, but also enhanced CD14+ monocytes-induced Th1 and T follicular helper cells activation, which presented as elevated interferon-γ and IL-21 secretion by CD4+ T cells. This process required direct cell-to-cell contact, and was dependent on IL-6 secretion. The current data revealed a potential immunopromotive property of IL-7 to lung-resident CD14+ monocytes in lung squamous carcinoma.

Cancer-associated fibroblasts suppress SOX2-induced dysplasia in a lung squamous cancer coculture.

Tumorigenesis depends on intricate interactions between genetically altered tumor cells and their surrounding microenvironment. While oncogenic drivers in lung squamous carcinoma (LUSC) have been described, the role of stroma in modulating tissue architecture, particularly cell polarity, remains unclear. Here, we report the establishment of a 3D coculture system of LUSC epithelial cells with cancer-associated fibroblasts (CAFs) and extracellular matrix that together capture key components of the tumor microenvironment (TME). Single LUSC epithelial cells develop into acinar-like structures with 0.02% efficiency, and addition of CAFs provides proper tumor-stromal interactions within an appropriate 3D architectural context. Using this model, we recapitulate key pathological changes during tumorigenesis, from hyperplasia to dysplasia and eventually invasion, in malignant LUSC spheroids that undergo phenotypic switching in response to cell intrinsic and extrinsic changes. Overexpression of SOX2 is sufficient to mediate the transition from hyperplasia to dysplasia in LUSC spheroids, while the presence of CAFs makes them invasive. Unexpectedly, CAFs suppress the activity of high SOX2 levels, restore hyperplasia, and enhance the formation of acinar-like structures. Taken together, these observations suggest that stromal factors can override cell intrinsic oncogenic changes in determining the disease phenotype, thus providing fundamental evidence for the existence of dynamic reciprocity between the nucleus and the TME of LUSC.

Microwave Hyperthermia Combined with Gemcitabine Inhibits Proliferation and Induces Apoptosis of Human Lung Squamous Carcinoma Cells

背景与目的肺癌是全球发病率及死亡率最高的恶性肿瘤之一，寻找有效的抗肿瘤方法显得很重要，微波热疗作为一种新的治疗技术越来越受到重视。本研究旨在探讨微波热疗联合吉西他滨对人肺鳞癌细胞NCI-H1703和NCI-H2170体外增殖的影响及诱导凋亡的机制。方法CCK-8法分别检测微波热疗、吉西他滨对细胞增殖影响以及微波热疗与吉西他滨不同序贯方式对细胞增殖的影响；克隆形成实验观察阴性对照组、微波热疗组、吉西他滨组及热化联合组对细胞克隆形成的影响；流式细胞术检测不同处理组细胞的总凋亡率；Caspase-3、Caspase-8活性检测试验检测各组细胞Caspase-3、Caspase-8酶活性；CCK-8法检测AC-DEVD(Caspase-3抑制剂)对细胞增殖的影响；Western blot检测微波热疗和吉西他滨对肺鳞癌细胞凋亡相关蛋白的表达。结果微波热疗对肺鳞癌有增殖抑制作用；吉西他滨对两株细胞的IC50值分别为8.89 μmol/L和44.18 μmol/L；先化疗后微波热疗对两株肺鳞癌细胞有协同作用且可明显抑制细胞的克隆形成(P < 0.001)、促进细胞凋亡(P < 0.001)以及显著提高Caspase-3酶活性(P < 0.001)，但对Caspase-8酶活性影响不大(P > 0.05)；加入AC-DEVD(Caspase-3抑制剂)后，热化联合组细胞增殖率提高(P < 0.001)；Western blot检测显示微波热疗联合吉西他滨可上调p53、Caspase-3、Cleaved-Caspase-3、Cleaved-PARP以及Bax蛋白的表达。结论先吉西他滨后微波热疗可协同抑制人肺鳞癌细胞的体外增殖以及促进凋亡，其机制可能是通过活化p53、切割PARP蛋白诱导癌细胞发生Caspase-3依赖性凋亡，从而发挥其协同抑制癌细胞增殖的作用。

Label-Free Detection of Human Serum Using Surface-Enhanced Raman Spectroscopy Based on Highly Branched Gold Nanoparticle Substrates for Discrimination of Non-Small Cell Lung Cancer

Surface-enhanced Raman spectroscopy (SERS) is a good candidate for the development of fast and easy-to-use diagnostic tools, possibly used on serum in screening tests. In this study, a potential label-free serum test based on SERS spectroscopy was developed to analyze human serum for the diagnosis of the non-small cell lung cancer (NSCLC). We firstly synthesized novel highly branched gold nanoparticles (HGNPs) at high yield through a one-step reduction of HAuCl4 with dopamine hydrochloride at 60°C. Then, HGNP substrates with good reproducibility, uniformity, and high SERS effect were fabricated by the electrostatically assisted (3-aminopropyl) triethoxysilane-(APTES-) functionalized silicon wafer surface-sedimentary self-assembly method. Using as-prepared HGNP substrates as a high-performance sensing platform, SERS spectral data of serum obtained from healthy subjects, lung adenocarcinoma patients, lung squamous carcinoma patients, and large cell lung cancer patients were collected. The difference spectra among different types of NSCLC were compared, and analysis result revealed their intrinsic difference in types and contents of nucleic acids, proteins, carbohydrates, amino acids, and lipids. SERS spectra were analyzed by principal component analysis (PCA), which was able to distinguish different types of NSCLC. Considering its time efficiency, being label-free, and sensitivity, SERS based on HGNP substrates is very promising for mass screening NSCLC and plays an important role in the detection and prevention of other diseases.

Co-Expression Analysis Reveals Mechanisms Underlying the Varied Roles of NOTCH1 in NSCLC

IntroductionNotch receptor family dysregulation can be tumor promoting or suppressing depending on cellular context. Our studies shed light on the mechanistic differences that are responsible for NOTCH1’s opposing roles in lung adenocarcinoma and lung squamous cell carcinoma. MethodsWe integrated transcriptional patient-derived datasets with gene co-expression analyses to elucidate mechanisms behind NOTCH1 function in subsets of NSCLC. Differential co-expression was examined using hierarchical clustering and principal component analysis. Enrichment analysis was used to examine pathways associated with the underlying transcriptional networks. These pathways were validated in vitro and in vivo. Endogenously epitope-tagged NOTCH1 was used to identify novel interacting proteins. ResultsNOTCH1 co-expressed genes in lung adenocarcinoma and squamous carcinoma were distinct and associated with either angiogenesis and immune system pathways or cell cycle control and mitosis pathways, respectively. Tissue culture and xenograft studies of lung adenocarcinoma and lung squamous models with NOTCH1 knockdown showed growth differences and opposing effects on these pathways. Differential NOTCH1 interacting proteins were identified as potential mediators of these differences. ConclusionsRecognition of the opposing role of NOTCH1 in lung cancer, downstream pathways, and interacting proteins in each context may help direct the development of rational NOTCH1 pathway-dependent targeted therapies for specific tumor subsets of NSCLC.

SUMO1 promotes the proliferation and invasion of non-small cell lung cancer cells by regulating NF-κB.

BackgroundOur previous study showed that SUMO1 expression is closely related to progression in non‐small cell lung cancer (NSCLC); however, the function of SUMO1 in NSCLC has not yet been well elucidated.Methods SUMO1 was enhanced or silenced in two NSCLC cell lines by using either forced SUMO1 expression or short hairpin RNA against SUMO1 lentiviral vectors, respectively. The biological functions of SUMO1 in NSCLC were investigated through colony‐formation, cell proliferation, and invasion assays, and cell cycle analysis. NF‐κB expression was detected in the overexpressed and silenced SUMO1 cell lines. Immunohistochemistry was used to detect an association between SUMO1 and NF‐κB in the cancer and adjacent tissues of 168 patients with lung cancer.ResultsOverexpressed SUMO1 promoted the proliferation rate, colony formation ability, invasion, and NF‐κB expression in an A549 cell line. Conversely, SUMO1 depletion inhibited the cell growth rate, colony formation ability, invasion, and NF‐κB expression in a Calu‐1 cell line. SUMO1 expression was significantly correlated with NF‐κB expression in lung adenocarcinoma and squamous carcinoma patients (r > 0.5, P < 0.001).ConclusionOur results provide evidence that SUMO1 promotes the proliferation and invasion of NSCLC cells by regulating NF‐κB.

Expression pattern and level of ING5 protein in normal and cancer tissues.

Inhibitor of growth family 5 (ING5) functions as a type-II tumor suppressor gene and exerts an important role in DNA repair, apoptotic induction, proliferative inhibition, chromatin remodeling and the invasion process. In the present study, immunohistochemistry was performed to characterize the expression profile of ING5 protein on a tissue microarray containing mouse and human normal tissues, and human cancer tissues, including hepatocellular (n=62), renal clear cell (n=62), pancreatic (n=62), esophageal squamous cell (n=45), cervical squamous cell (n=31), breast (n=144), gastric (n=196), colorectal (n=96), endometrial (n=96) and lung carcinoma (n=192). In the mouse tissues, ING5 expression was detected in the cytoplasm of neurons, the nephric tubule and glomerulus, alveolar epithelium, gastrointestinal glands, squamous epithelium of the skin and skeletal muscles. By contrast, ING5 was localized to the cell nucleus in breast tissues. In human tissues, ING5 protein was primarily localized in the cytoplasm. However, ING5 was detected in the cytoplasm and nucleus in various types of normal tissues, including the tongue, stomach, intestine, lung and breast. In total, ING5 expression was detected in 400/986 cancer tissues (40.6%). In the majority of cases, ING5 expression was observed to be restricted to the cytoplasm. However, ING5 was also detected in the nucleus in a number of cancer tissues, including gastric, colorectal and lung carcinoma. Notably, ING5 was more frequently expressed in breast (79.9%), colorectal (56.3%) and endometrial carcinoma (50.0%). The incidence of ING5 expression in hepatocellular carcinoma (14.5%) and pancreatic carcinoma (22.6%) was low. These findings indicate that ING5 may be involved in cell regeneration and be associated with colorectal carcinogenesis.