Lung Squamous Carcinoma Research Articles

Abstract 2212: Cellular and molecular changes associated with lung squamous premalignant lesion severity identified by single-cell RNA-seq

Abstract Background: Sampling the bronchial airway during bronchoscopies in screening populations at high risk for lung cancer has increased detection of squamous bronchial pre-malignant lesions (PML). The majority of these lesions will regress or remain stable without any clinical intervention. However, a subset of lesions will progress to invasive malignancy. In this work, we investigate the gene expression changes in epithelial cell states and immune cell proportion shifts associated with premalignant histologic stage using single-cell RNA sequencing to begin to identify molecular features associated with PML regression or progression. Methods: Cells (n=3,325) were isolated from 11 endobronchial biopsies from 9 patients, where biopsy histology range from normal, dysplasia, carcinoma in situ (CIS) and squamous cell carcinoma. Immune cells (CD45+) and epithelial cells (CD45− EPCAM+) were sorted into separate 96-well plates using fluorescence activated cell sorting (FACS) and sequenced using CEL-Seq2 single cell RNA seq protocol. Results: After filtering low quality cells, we profiled 2,998 cells (1,052 CD45+ and 1,946 CD45−) with an average of 1,190 genes per cell. Within the epithelial cells, we focused on basal cells as they are the airway progenitor cells. We observed distinct differences in expression of KRT5+ (basal) cells between different histologic grades, where KRT5+ cells present in a hyperplasic lesion expressed multiple secretory cell markers suggesting they were transitioning to a secretory cell phenotype and KRT5+ cells from a CIS lesion from a current smoker expressed higher levels of smoking inducible genes, such as GSTM1 and CYP1A1. Lastly, KRT5+ cells from a lung squamous carcinoma tumor expressed higher levels of many genes related to cell cycle progression. Among the immune populations, low-grade PMLs with non-dysplastic histology were enriched for CD8+ T cells, whereas higher-grade PMLs and an invasive tumor were enriched for myeloid cells. Conclusions: To date, we have begun to identify histology-associated cellular and molecular profiles in bronchial premalignancy and early-stage carcinoma. Future resampling of these patients and expansion of cases will allow us to discover biomarkers associated with lesion progression and molecular targets for lung cancer interception. Citation Format: Conor Shea, Lukas Kalinke, Kitty De Jong, Kate Gowers, Diane Ding, Sherry Zhang, Gang Liu, Jack Cunningham, Ipsita Dey-Guha, Mark Hennon, Sai Yendumuri, Christopher Stevenson, Steven Dubinett, Avrum Spira, Marc Lenburg, Samuel Janes, Mary Reid, Jennifer Beane, Sarah Mazzilli, Joshua D. Campbell. Cellular and molecular changes associated with lung squamous premalignant lesion severity identified by single-cell RNA-seq [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2212.

Anti-TIF1 gamma-positive IPAF patient developed stage IVB lung squamous carcinoma in 1 year: a case report

BackgroundPatients with connective tissue disease, such as dermatomyositis (DM), and positive anti-TIF1γ self-antibodies are commonly diagnosed with malignant tumors as a comorbidity. The relationship between anti-TIF1γ self-antibodies and existing malignant tumors has been confirmed by several reports. However, interstitial pneumonia with autoimmune features (IPAF) cases with a positive anti-TIF1γ self-antibody developing to solid malignant tumors are rarely reported now.Case presentationHerein, we presented an IPAF patient with anti-TIF1γ self-antibodies. No evidence of malignant tumors was found at the initial visit. However, the patient had developed stage IVB lung squamous cell carcinoma at the 1-year follow-up review.ConclusionsAltogether, this report described a rare case of IPAF patient with anti-TIF1γ self-antibodies developed to advanced lung squamous cell carcinoma in 1 year. The present case highlights more frequent imaging examinations to identify the occurrence of malignant tumors as early as possible in IPAF patients with positive anti-TIF1γ self-antibodies.

TMEM229A suppresses non‑small cell lung cancer progression via inactivating the ERK pathway.

Transmembrane protein 229A (TMEM229A) is a member of the TMEM family that plays an important role in tooth differentiation and development. However, the expression level and biological role of TMEM229A in cancer remains unknown. The present study aimed to investigate the expression level of TMEM229A in non-small cell lung cancer (NSCLC), as well as its effect and mechanism on NSCLC progression. Clinical specimens from patients with NSCLC were enrolled from the First People's Hospital of Huzhou (Huzhou, China). TMEM229A expression was detected using reverse transcription-quantitative PCR (RT-qPCR), western blotting and immunohistochemical analysis. The relationship between TMEM229A expression and the survival rate of patients with NSCLC was analyzed using Kaplan-Meier Plotter datasets. The effects of TMEM229A on cell proliferation, migration and invasion were detected using Cell Counting Kit-8, colony formation, soft agar, real-time cellular analysis and Transwell assays. The expression levels of epithelial-mesenchymal transition (EMT)-related proteins, as well as ERK and AKT phosphorylation were determined via RT-qPCR and western blot analysis. The results demonstrated that TMEM229A expression was significantly downregulated in human NSCLC tissues and in several cell lines compared with adjacent normal lung tissues and BEAS-2B cells, respectively. Survival analysis of lung adenocarcinoma and squamous cell lung carcinoma cases identified that low TMEM229A expression was associated with a poor prognosis. The in vitro assays indicated that overexpressing TMEM229A significantly inhibited cell proliferation, migration and invasion, while TMEM229A knockdown had the opposite effect. Mechanistically, TMEM229A overexpression effectively increased E-cadherin expression and reduced N-cadherin, snail family transcriptional repressor 1 and MMP2 expression, indicating that EMT was suppressed. In addition, overexpression of TMEM229A reduced the expression levels of phosphorylated (p)-ERK and p-AKT, and this effect was partially suppressed by the incorporation of specific ERK inhibitor PD98059. Collectively, the results of the present study demonstrated that the effects of TMEM229A on inhibiting cell proliferation, migration and invasion were partially mediated by inactivating the ERK signaling pathway, thereby providing a potential therapeutic target for the treatment of NSCLC.

A subset of lung cancer cases shows robust signs of homologous recombination deficiency associated genomic mutational signatures

PARP inhibitors are approved for the treatment of solid tumor types that frequently harbor alterations in the key homologous recombination (HR) genes, BRCA1/2. Other tumor types, such as lung cancer, may also be HR deficient, but the frequency of such cases is less well characterized. Specific DNA aberration profiles (mutational signatures) are induced by homologous recombination deficiency (HRD) and their presence can be used to assess the presence or absence of HR deficiency in a given tumor biopsy even in the absence of an observed alteration of an HR gene. We derived various HRD-associated mutational signatures from whole-genome and whole-exome sequencing data in the lung adenocarcinoma and lung squamous carcinoma cases from TCGA, and in a patient of ours with stage IVA lung cancer with exceptionally good response to platinum-based therapy, and in lung cancer cell lines. We found that a subset of the investigated cases, both with and without biallelic loss of BRCA1 or BRCA2, showed robust signs of HR deficiency. The extreme platinum responder case also showed a robust HRD-associated genomic mutational profile. HRD-associated mutational signatures were also associated with PARP inhibitor sensitivity in lung cancer cell lines. Consequently, lung cancer cases with HRD, as identified by diagnostic mutational signatures, may benefit from PARP inhibitor therapy.

Hypopituitarism as an Initial Presentation of Silent Squamous Cell Carcinoma of Lung

Background: Symptomatic metastasis to the pituitary gland is a rare, life-threatening condition. A better understanding of its clinical presentation could lead to early diagnosis and also improve quality of life. In this unusual case, we present a patient with hypopituitarism as the first manifestation of undiagnosed squamous cell carcinoma of the lung. Clinical Case: A 56-year-old Caucasian female with past medical history of pre-diabetes, hypertension, hyperlipidemia, and a chronic smoker for 40 years presented with recent onset of headache for two weeks and diplopia for three days. Initial CT head was negative but MRI of brain pituitary scan revealed a large heterogeneously enhancing sellar/suprasellar mass (1.5 x 2.8 x 2.4 cm) extending into the prepontine cistern with probable bilateral cavernous sinus invasion and contacting the optic chiasm without significant compression. Physical examination showed left abducens nerve (CN VI) palsy. Lab results showed elevated prolactin at 67.4 ng/ml (normal postmenopausal: 1.8 – 20.3 ng/ml), normal IGF-1 at 136 ng/ml (normal 48- 235 ng/ml), low TSH at 0.129 µIU/ml (normal 0.3 – 4.5 µIU/ml), low normal Free T4 at 0.50 ng/dl (normal 0.5 – 1.26 ng/dl), low ACTH at 5.1 pg/ml (normal 7.2 – 63.3 pg/ml), low am cortisol at 2.6 mcg/dl (normal 8.7 – 22.4 mcg/dl), low FSH at 2.6 mIU/ml (normal postmenopausal 23.0-116.3 mIU/ml) and low LH at 0.2 mIU/ml (normal postmenopausal 15.9 - 54.0 mIU/ml). She subsequently began treatment for hypopituitarism. Given her smoking history and unusual presentation of cranial nerve palsy, a chest x-ray was ordered and revealed a left upper lobe mass. CT chest confirmed the left upper lobe mass (4.0 x 3.3 x 2.9 cm), and biopsy reported poorly differentiated squamous cell carcinoma. Decompression of the pituitary macroadenoma was performed for symptomatic improvement but was complicated with the development of central diabetes insipidus. Pathology report disclosed metastatic pituitary carcinoma, compatible with primary lung carcinoma. Following surgical resection and radiation therapy to the pituitary gland, the patient remains stable and is currently tolerating all treatment. Conclusion: In a patient with occult malignancy, pituitary metastasis is an exceedingly rare and challenging diagnosis that carries a poor prognosis. The purpose of this abstract is to raise clinical suspicion for sellar metastasis in a patient presenting with hypopituitarism and cranial nerve palsy.

Use of GLP1RAs and occurrence of respiratory disorders: A meta-analysis of large randomized trials of GLP1RAs.

No large sample studies have been designed to evaluate the efficacy of glucagon-like peptide-1 receptor agonists (GLP1RAs) in the primary and secondary prevention of respiratory disorders. We aimed at evaluating the relationship between use of GLP1RAs and occurrence of 12 kinds of respiratory disorders. Large randomized placebo-controlled trials of GLP1RAs were included. We conducted meta-analysis using random effects model and measured heterogeneity using I2 . Treatment effect was presented as risk ratio (RR) and 95% confidence interval (CI). Seven trials including 55922 participants were included in meta-analysis. The occurrence rates of various respiratory disorders were low, with the minimum of 0.02% (pulmonary fibrosis) and the maximum of 2.31% (pneumonia). Although not reaching statistical significance, GLP1RAs versus placebo showed the reduced trends in the risks of nine kinds of respiratory disorders including pneumonia (RR 0.89, 95% CI 0.78-1.01), squamous cell carcinoma of lung (SCCL; RR 0.55, 95% CI 0.25-1.21), asthma (RR 0.82, 95% CI 0.51-1.32), and chronic obstructive pulmonary disease (COPD; RR 0.89, 95% CI 0.73-1.10), but the increased trend in interstitial lung disease (ILD; RR 1.89, 95% CI 0.87-4.08). GLP1RAs had neutral effects on two other respiratory disorders. Heterogeneity in any meta-analysis was absent or low. GLP1RAs show the reduced trends in the risks of nine kinds of respiratory disorders (eg, pneumonia, SCCL, asthma, and COPD), but the increased trend in the risk of ILD. However, these findings need to be validated by further studies due to the low incidence rates of all the respiratory disorders.

Potential Prospective Biomarkers for Non-small Cell Lung Cancer: Mini-Chromosome Maintenance Proteins.

Minichromosome maintenance proteins (MCMs) are considered to be essential factors coupling DNA replication to both cell cycle progression and checkpoint regulation. Previous studies have shown that dysregulation of MCMs are implicated in tumorigenesis of lung cancer. However, the distinct expression/mutation patterns and prognostic values of MCMs in lung cancer have yet to be systematically elucidated. In the present study, we analyzed the transcriptional levels, mutations, and prognostic value of MCM1-10 in non-small cell lung cancer (NSCLC) patients using multiple bioinformatics tools, including ONCOMINE, GEPIA, Kaplan–Meier Plotter, cBioPortal, and GESA. The analysis results from GEPIA dataset showed that MCM2/4/10 was significantly high expressed in both lung adenocarcinoma (LUAD) and squamous cell lung carcinomas (LUSCs). Meanwhile, the expression levels of MCM2/4/6/7/8 were associated with advanced tumor stages. Subsequent survival analysis using the Kaplan–Meier Plotter indicated that high expression levels of MCM1/2/3/4/5/6/7/8/10 were associated with worse overall survival (OS), while high expression level of MCM9 predicted better OS in these patients. Furthermore, we experimentally validated overexpression of MCM2 and MCM4 in NSCLC, thus the results from this study support a view that they may serve as potential prospective biomarkers to identify high-risk subgroups of NSCLC patients.

The tumor environment immune phenotype of LUSC by genome-wide analysis

PurposeTo compare the landscape of tumor microenvironment (TME) of lung squamous carcinoma (LUSC) in different immune pattern and explore potential factors on immune therapy and prognosis. Method and MaterialsWe have obtained the LUSC data from TCGA, GEO, and our department and classified them into 2 TME clusters by random forest model based on the infiltration pattern of 24 immune cell populations. We systemically compared the genomic significance, clinical characteristics, and immune infiltration pattern in 2 TME clusters. ResultsSamples were divided into 2 TME clusters based on the relative abundance of 24 immune cells, and a random forest classifier model was constructed. TME cluster B was a higher immune infiltration group with lower mutation load, richer co-infiltrate immune cells, upregulated immune-related cytokines, immune checkpoint molecules, and higher active immune cells. TME cluster was also an independent predictor in prognosis (B vs. A, p < 0.05) in patients from TCGA, GEO, and our department. ConclusionsOur study has described the microenvironment landscape of LUSC in different immune infiltration patterns and systemically analyzed genomic and clinical characteristics with distinct immunophenotypes, thus partly revealed the interaction between tumors and the immune microenvironment, which may guide a more precise and personalized immune therapeutic strategy for LUSC patients.

Clinical Significance of miR-183-3p and miR-182-5p in NSCLC and Their Correlation.

PurposeAccumulating evidence has indicated that dysregulated microRNAs (miRNAs) are involved in cancer progression. In this study, we evaluated the clinicopathologic significance of miR-183-3p and miR-182-5p, and the role of miR-183-3p in non-small-cell lung cancer (NSCLC) progression.Patients and MethodsSeventy-six NSCLC patients from Beijing Chest Hospital were included. The expression of miR-183-3p and miR-182-5p was evaluated by real-time quantitative polymerase chain reaction (RT-qPCR). Then, cell growth curve assays and colony formation assays were performed. Bioinformatics analysis of TCGA database was performed to explore the clinicopathological significance and prognostic value.ResultsmiR-183-3p and miR-182-5p were significantly increased in NSCLC tumor tissues (both P < 0.0001) and were positively correlated (r = 0.8519, P < 0.0001). miR-183-3p (P = 0.0444) and miR-182-5p (P = 0.0132) were correlated with tumor size. In addition, miR-183-3p (P = 0.0135) and miR-182-5p (P = 0.0009) were upregulated in normal lung tissues from smokers. In vitro, miR-183-3p was correlated with cell proliferation. In addition, bioinformatics analysis indicated that miR-183-3p was correlated with poor prognosis (P = 0.0466) and tumor size (P = 0.0017). In addition, miR-183-3p was higher in lung squamous carcinoma (LUSC) tissue (P < 0.0001) than in lung adenocarcinoma (LUAD) tissue, and miR-183-3p was higher in the tumor tissue of smokers (P = 0.0053) than in that of nonsmokers.ConclusionUpregulation of miR-183-3p and miR-182-5p may play an oncogenic role in NSCLC. miR-183-3p could be used as a potential prognostic biomarker and therapeutic target to manage lung cancer.

Molecular characterization of lung squamous cell carcinoma tumors reveals therapeutically relevant alterations.

Introduction: Unlike lung adenocarcinoma patients, there is no FDA-approved targeted-therapy likely to benefit lung squamous cell carcinoma patients.Materials and Methods: We performed survival analyses of lung squamous cell carcinoma patients harboring therapeutically relevant alterations identified by whole exome sequencing and mass spectrometry-based validation across 430 lung squamous tumors.Results: We report a mean of 11.6 mutations/Mb with a characteristic smoking signature along with mutations in TP53 (65%), CDKN2A (20%), NFE2L2 (20%), FAT1 (15%), KMT2C (15%), LRP1B (15%), FGFR1 (14%), PTEN (10%) and PREX2 (5%) among lung squamous cell carcinoma patients of Indian descent. In addition, therapeutically relevant EGFR mutations occur in 5.8% patients, significantly higher than as reported among Caucasians. In overall, our data suggests 13.5% lung squamous patients harboring druggable mutations have lower median overall survival, and 19% patients with a mutation in at least one gene, known to be associated with cancer, result in significantly shorter median overall survival compared to those without mutations. Conclusions: We present the first comprehensive landscape of genetic alterations underlying Indian lung squamous cell carcinoma patients and identify EGFR, PIK3CA, KRAS and FGFR1 as potentially important therapeutic and prognostic target.

The Association of microRNA-34a With High Incidence and Metastasis of Lung Cancer in Gejiu and Xuanwei Yunnan.

The incidence and associated mortality of lung cancer in tin miners in Gejiu County and farmers in Xuanwei Country, Yunnan Province have been very high in the world. Current published literatures on the molecular mechanisms of lung cancer initiation and progression in Gejiu and Xuanwei County are still controversial. Studies confirmed that microRNA-34a (miR-34a) functioned as a vital tumor suppressor in tumorigenesis and progression. However, the role and precise mechanisms of miR-34a and its regulatory gene network in initiation and progression of lung cancer in Gejiu and Xuanwei County, Yunnan Province, have not been elucidated. In the current study, we first found that miR-34a was downregulated in Gejiu lung squamous carcinoma YTMLC-90, Xuanwei lung adenocarcinoma XWLC-05, and other non-small cell lung carcinoma (NSCLC) cell lines, and miR-34a overexpression inhibited cell proliferation, migration and invasion, as well as induced cell apoptosis in YTMLC-90 and XWLC-05 cells. Our findings revealed that miR-34a is critical and cannot be considered as the area-specific non-coding RNA in initiation and progression of lung cancer in Gejiu and Xuanwei County. Next we revealed that miR-34a overexpression suppressed lung cancer growth and metastasis partially via increasing PTEN but reducing CDK6 expression that might lead to subsequent inactivation of PI3K/AKT pathway. Furthermore, our findings demonstrated that YY1 functioned as a tumor suppressor gene in initiation and progression of lung cancer in Gejiu and Xuanwei County. In conclusion, our findings in the study confirmed that miR-34a overexpression could simultaneously suppress tumor growth and metastasis and play a vital role in tumorigenesis and progression of NSCLC via increasing PTEN and YY1 expression, but decreasing CDK6. Most interestingly, our findings also raised doubts about the current ideas about these area-specific diseases.

The effect and mechanism of miR-607/CANT1 axis in lung squamous carcinoma.

Lung squamous carcinoma (LUSC) is the second most frequent subtype of non-small cell lung cancer. Rarely gene alterations are identified in LUSC. Therefore, identifying LUSC-related genes to explain the relevant molecular mechanism is urgently needed. A potential biomarker, calcium-activated nucleotidase 1 (CANT1), was elevated in tissues of LUSC patients relative to normal cases based on the TCGA and/or GTEx database. CCK-8 and transwell tests were then implemented to measure the proliferative, invasive and migratory capacities, and showed that knockdown of CANT1 blocked LUSC cells proliferation. miR-607, predicted as an upstream factor for CANT1, was declined in LUSC using TargetScan analysis and luciferase activity test. Low miR-607 expression was related with unfavorable outcomes of LUSC patients. Moreover, miR-607 downregulation elevated cell viability, invasion and migration in LUSC cells, which was antagonized by si-CANT1. GEPIA website was accessed to estimate the relevance between CANT1 and epithelial-mesenchymal transition (EMT)-related positive factors. The protein levels of Fibronectin, Vimentin, Snail and β-catenin were altered due to the abnormal CANT1 and miR-607 expression. Together, these data unveiled that miR-607/CANT1 pair may exert a vital role in the progression of LUSC through mediating EMT process, which would furnish an available therapeutic therapy for LUSC.

TTN/TP53 mutation might act as the predictor for chemotherapy response in lung adenocarcinoma and lung squamous carcinoma patients.

BackgroundChemotherapy is the preferred treatment in many types of cancer including lung cancer. However, most of patients resist chemotherapy resulting in disease progressive and recurrence. Titin (TTN) mutation is proved as a beneficial role in lung squamous carcinoma (LUSC), but the predictive role on chemotherapy resistance of lung cancer is still limited and discussable.MethodsClinical information and related somatic mutation profiles were obtained from The Cancer Genome Atlas (TCGA) database and analyzed by R-Studio using R-package. Overall survival (OS) curve and the association between gene mutation and clinical features were determined by GraphPad 6.0 software.ResultsAvailable data including 563 lung adenocarcinoma (LUAD) and 505 LUSC subjects were included in this study. Among all patients, 205 out of 563 LUAD and 326 out of 505 LUSC patients displayed TTN gene mutation. When comparing the clinical features in TTN-mutated patients to patients without TTN mutation who received chemotherapy, the tumors were always located in the upper lung in LUAD patients with TTN mutation and most of TTN-mutated subjects were at low pathological stage, which was not observed in LUSC patients. However, patients with TTN-mutation, particularly missense mutation, had a higher chemosensitivity and longer OS period than that patients without TTN mutation in both LUAD and LUSC. Of note, LUAD and LUSC patients possessed favorable OS and better chemotherapy response benefiting from TTN/tumor protein 53 (TP53) double mutation compared to TTN and TP53 mutation alone, respectively. Additionally, TTN/TP53 double mutation-initiated high rate of chemotherapy response were largely concentrated within LUAD and LUSC patients whose anatomic neoplasm subdivision were located in the upper lung.ConclusionsCollectively, TTN/TP53 co-mutation is possibly served as an effective predictor for OS and chemotherapy response in lung cancer.

Human blood serum can donor-specifically antagonize effects of EGFR-targeted drugs on squamous carcinoma cell growth

Many patients fail to respond to EGFR-targeted therapeutics, and personalized diagnostics is needed to identify putative responders. We investigated 1630 colorectal and lung squamous carcinomas and 1357 normal lung and colon samples and observed huge variation in EGFR pathway activation in both cancerous and healthy tissues, irrespectively on EGFR gene mutation status. We investigated whether human blood serum can affect squamous carcinoma cell growth and EGFR drug response. We demonstrate that human serum antagonizes the effects of EGFR-targeted drugs erlotinib and cetuximab on A431 squamous carcinoma cells by increasing IC50 by about 2- and 20-fold, respectively. The effects on clonogenicity varied significantly across the individual serum samples in every experiment, with up to 100% differences. EGF concentration could explain many effects of blood serum samples, and EGFR ligands-depleted serum showed lesser effect on drug sensitivity.

P73.03 A Kinome CRISPR Screen in FGFR-Amplified Lung Cancer

Oncogenic alterations in fibroblast growth factor receptor (FGFR) are frequent, but therapeutic targeting of FGFR-driven cancers remains an unmet challenge in clinical oncology in that FGFR inhibitors as a single agent are inevitably limited by poor responsiveness and/or compensatory activation of various acquired resistance mechanisms. We performed kinome CRISPR/Cas9 screens to identify genetic determinants that limit efficacy of FGFR-targeted therapy in FGFR-mutant squamous lung carcinoma (SQLC). We identify several kinases including polo-like kinase I (PLK1) as key determinants of sensitivity to AZD4547, a selective FGFR inhibitor currently investigated in phase III clinical trials. We provided evidence showing that genetic and pharmacological inhibition of PLK1 synergistically enhances the effect of FGFR inhibitors in FGFR-amplified lung SQLC. CRISPR/Cas9 screening is a powerful tool to uncover genetic determinants underlying drug sensitivity in cancer.

A novel immunogenomic prognostic signature in lung squamous carcinoma.

Lung squamous carcinoma (LUSC) is a common subtype of lung cancer with limited available therapy and is thus associated with poor survival. Immune infiltrating cells and immune-related genes (IRGs) play a key role in the clinical outcomes of LUSC. In the present study, we aimed to develop a potential immunogenomic prognostic signature for patients with LUSC. The transcriptional profiles of 501 LUSC samples from The Cancer Genome Atlas (TCGA) and 2498 IRGs from the ImmPort database were used to develop the signature by Cox regression analysis. Ten differentially expressed and survival-associated IRGs were used to develop the risk signature, which could serve as an independent prognostic and predictive factor for patients with LUSC. Furthermore, this risk signature correlated with overall survival and clinical features, including age, in patients with LUSC. In addition, we identified 25 transcription factors that may regulate 15 survival-associated IRGs, using a regulatory network. Collectively, this immunogenomic signature could be a robust prognostic tool for patients with LUSC and holds great promise as individualized immunotherapy for LUSC.

Protein Glycopatterns in Bronchoalveolar Lavage Fluid as Novel Potential Biomarkers for Diagnosis of Lung Cancer.

Lung cancer is one of the most prevalent and life-threatening neoplasias worldwide due to the deficiency of ideal diagnostic biomarkers. Although aberrant glycosylation has been observed in human serum and tissue, little is known about the alterations in bronchoalveolar lavage fluid (BALF) that are extremely associated with lung cancer. In this study, our aim was to systematically investigate and assess the alterations of protein glycopatterns in BALF and possibility as biomarkers for diagnosis of lung cancer. Here, lectin microarrays and blotting analysis were utilized to detect the differential expression of BALF glycoproteins from patients with 80 adenocarcinomas (ADC), 77 squamous carcinomas (SCC), 51 small cell lung cancer (SCLC), and 73 benign pulmonary diseases (BPD). These 281 specimens were then randomly divided into a training cohort and validation cohort for constructing and verifying the diagnostic models based on the glycopattern abundances. Moreover, an independent test was performed with 120 newly collected BALF samples enrolled in the double-blind cohort to further assess the clinical application potential of the diagnostic models. According to the results, there were 15 (e.g., PHA-E, EEL, and BPL) and 14 lectins (e.g., PTL-II, LCA, and SJA) that individually showed significant variations in different types and stages of lung cancer compared to BPD. Notably, the diagnostic models achieved better discriminate power in the validation cohort and exhibited high accuracies of 0.917, 0.864, 0.712, 0.671, and 0.781 in the double-blind cohort for the diagnosis of lung cancer, early stage lung cancer, ADC, SCC, and SCLC, respectively. Taken together, the present study revealed that the abnormally altered protein glycopatterns in BALF are expected to be novel potential biomarkers for the identification and early diagnosis of lung cancer, which will contribute to explain the mechanism of the development of lung cancer from the perspective of glycobiology.

Identification of plasma SAA2 as a candidate biomarker for the detection and surveillance of non-small cell lung cancer.

This study aimed to measure the expression of SAA2 in plasma and to assess its diagnostic efficacy as a biomarker for non-small cell lung cancer (NSCLC). The gene expression of SAA2 in NSCLC was analyzed based on a database. Then, SAA2 expression was detected by immunohistochemistry in lung tissue and by enzyme-linked immunosorbent assay in 90 patients with NSCLC and 61 normal controls. Finally, the diagnostic performance was assessed in terms of accuracy, sensitivity, and specificity. At the gene and protein levels, the SAA2 expression was significantly higher in the NSCLC group than in the control group (p<0.01). It was higher in lung squamous carcinoma than in lung adenocarcinoma and in males than in females, and this trend was also observed in the lung squamous carcinoma group. Of note, the expression of SAA2 increased with increasing disease stage. Receiver operating characteristic (ROC) curve analysis revealed that the sensitivity of SAA2 was 83.61%, the specificity was 91.11%, and the area under the curve (AUC) was 0.9252. Its accuracy was 68.89%, which was higher than that of other conventional diagnostic biomarkers, and the combined application can effectively improve the diagnostic efficiency. Based on the results, SAA2 expression was positively correlated with the disease stage of NSCLC. Notably, SAA2 is more concerning in male patients with lung squamous carcinoma, and it can help in the screening and diagnosis of NSCLC. SAA2 may represent a novel diagnostic biomarker in NSCLC.

Comprehensive Characterization of Tumor Microenvironment Differences in Cancer between Male and Female Patients

Background: Sex has been recognized as a key variable that affects cancer initiation, progression, response to therapy, and prognosis outcome. The sex differences of the tumor microenvironment (TME) characterization associated with cancer immunotherapy remains poorly understood. A comprehensive analysis to investigate sex-biased TME related features across a broad range of cancer types is required. Methods: We collected 19 cancer types from The Cancer Genome Atlas (TCGA), and remained 1,771 male and 1,137 female samples with immune infiltration. The sex-based differences of TME related features, including tumor mutational burden (TMB), immune scores, stromal scores, tumor purity, immune cells, immune checkpoint genes, and functional pathways were evaluated in pan-cancer and individual cancer types. Findings: We discovered that male patients possess higher tumor mutational burden (TMB) and tumor purity, but lower immune and stromal scores than female patients in pan-cancer. We then revealed that lung adenocarcinoma, lung squamous carcinoma, kidney papillary carcinoma, and head and neck squamous carcinoma possess significant sex bias in immune cells, immune checkpoint genes, and functional pathways. We further identified male- and female-specific prognostic biomarkers based on immune cells in male and female patients of lung adenocarcinoma and validated our results in two independent datasets. Interpretation: This study provides a comprehensive perspective of sex differences for TME related features in diverse cancers and suggests sex may affect the tumor microenvironment, and which should be considered in cancer immunotherapy. Funding: This work was supported by National Natural Science Foundation of China and Natural Science Foundation of Heilongjiang. Declaration of Interest: None to declare.

Ubiquilin 1 suppresses the cancer stem cell–like traits of non-small cell lung cancer cells by regulating reactive oxygen species homeostasis

ABSTRACT Cancer stem cell (CSC) has been confirmed to trigger tumor occurrence and progression and CSC can develop strategies to maintain a lower reactive oxygen species (ROS) level compared to cancer cells. However, the mechanisms contributing to ROS homeostasis in CSC are still lacking key elements. In the current study, we found that reductive redox states and ROS levels were suppressed in non-adherent spheres formed by non-small cell lung cancer (NSCLC) cells, which were confirmed to hold CSC-like traits. However, mitochondria DNA content and cellular oxygen consumption rate analyses revealed fewer numbers of mitochondria in NSCLC spheres. Further exploration attributed this result to decreased mitochondrial biogenesis, likely resulted from the accelerated degradation of peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α). Mechanistic studies indicated that Ubiquilin 1 (UBQLN1) increased PGC1α protein stability via reducing the ubiquitination of PGC1α protein. Moreover, UBQLN1 was lowly expressed in NSCLC spheres compared to that in parental NSCLC cells and UBQLN1 overexpression suppressed the CSC-like traits of NSCLC cells, which was characterized as the decrease of ALDH1 activity, sphere-formation ability, and CSC marker expression. Finally, clinical investigations further demonstrated that UBQLN1 level was positively correlated with patient’s survival of lung adenocarcinoma, but not squamous cell carcinoma of lung. Taken together, our results revealed a novel mechanism involving ROS homeostasis and mitochondrial biogenesis in non-small cell lung CSCs, which may provide novel potential targets and methods for NSCLC patients.