Carcinogenesis Of Lung Adenocarcinoma Research Articles

Integrating apaQTL and eQTL analysis identifies a potential causal variant associated with lung adenocarcinoma risk in the Chinese population

Alternative polyadenylation (APA) plays a crucial role in cancer biology. Here, we used data from the 3′aQTL-atlas, GTEx, and the China Nanjing Lung Cancer GWAS database to explore the association between apaQTL/eQTL-SNPs and the risk of lung adenocarcinoma (LUAD). The variant T allele of rs277646 in NIT2 is associated with an increased risk of LUAD (OR = 1.12, P = 0.015), lower PDUI values, and higher NIT2 expression. The 3′RACE experiment showed multiple poly (A) sites in NIT2, with the rs277646-T allele causing preferential use of the proximal poly (A) site, resulting in a shorter 3′UTR transcript. This leads to the loss of the hsa-miR-650 binding site, thereby affecting LUAD malignant phenotypes by regulating the expression level of NIT2. Our findings may provide new insights into understanding and exploring APA events in LUAD carcinogenesis.

MiR-1976/NCAPH/P65 axis inhibits the malignant phenotypes of lung adenocarcinoma.

Lung adenocarcinoma (LUAD) is a malignancy with an abysmal survival rate. High metastasis is the leading cause of the low survival rate of LUAD. NCAPH, an oncogene, is involved in the carcinogenesis of LUAD. However, the regulation of NCAPH in LUAD remains controversial. In this work, we identified an up-regulation of NCAPH in LUAD tissues. Patients who expressed more NCAPH had shorter overall survival (OS). Furthermore, NCAPH overexpression promoted LUAD cell migration while inhibiting apoptosis. MiR-1976 and miR-133b were predicted to target NCAPH expression by searching TargetScan and linkedomics databases. Following that, we confirmed that miR-1976 suppressed NCAPH by directly targeting a 7-bp region of NCAPH 3' untranslated regions (UTR). In addition, increased expression of miR-1976 decreased the proliferation & migration and promoted apoptosis of LUAD cells, and the re-introduction of NCAPH reversed these influences. Furthermore, the xenograft and metastasis mouse models also confirmed that miR-1976 inhibited tumor growth and metastasis in vivo by targeting NCAPH. Finally, we found that MiR-1976 targeting NCAPH blocked the activation of NF-κB. In conclusion, miR-1976 inhibits NCAPH activity in LUAD and acts as a tumor suppressor. The miR-1976/NCAPH/NF-κB axis may, in the future, represent crucial diagnostic and prognostic biomarkers and promising therapeutic options.

Abstract 3387: Spatial analysis revealed distinct molecular and immune landscape in genetic versus carcinogen-induced lung adenocarcinoma precancer mouse models

Abstract Introduction: Our understanding of early carcinogenesis of lung adenocarcinoma (LUAD) is still rudimentary. Single-cell technologies have unveiled the intricate interaction between tumor cells and tumor microenvironment (TME) with unprecedented precision. However, the spatial context of single-cell phenotypes within TME during LUAD early carcinogenesis remain poorly understood. Here, we sought to characterize human-relevant murine LUAD precancer models to elucidate the molecular and immune changes in the spatial context during early LUAD carcinogenesis to inform lung cancer interception strategies. Methods: We have developed genetically engineered mouse models (GEMMs) (129S4 KrasG12D) and carcinogen-induced LUAD precancer model (CITM) (129S4 Urethane). Lung tissues were collected at various time points after induction and analyzed using whole exome sequencing (WES), imaging mass cytometry (IMC), and spatial transcriptomics (10x Visium). Results: Pathological image analysis demonstrated a significantly higher tumor burden (p<0.01) in the 129S4 KrasG12D model compared to the 129S4 Urethane model across different time points. WES revealed a progressive increase in tumor mutation burden (TMB) from precancers to invasive LUADs in both models, with the CITM model having significantly higher TMB (p<0.05) at matched timepoints than GEMM model. IMC assessed 39 biomarkers in 284 regions of interest (ROIs) and identified 1.4 million individual cells with 17 major cell types. Macrophages were the most prevalent immune cell type in TME, representing 9.00% of total cells in GEMM and 8.29% in CITM. The percent immune cells of the total cells progressively decreased from early-stage hyperplasia to late-stage adenocarcinoma in both models. The CITM exhibited a significantly (p<0.05) higher immune proportion than the GEMM model at hyperplasia and adenoma stages, with no significant difference at the invasive adenocarcinoma tumor cores between these 2 models. On the other hand, CITM had a significantly higher overall immune proportion (p<0.05) and the CD8 T cell and B cell proportions than GEMM tumor adjacent lung tissues at the adenocarcinoma stage leading to a significantly higher immune cell gradient between tumor adjacent lung tissues and tumor cores in CITM models. Spatial transcriptomics (ST) data demonstrated an increased presence of immune response signaling at the tumor adjacent lung tissues compared to the tumor core in the latest stage. Conclusion: Spatial analysis revealed different molecular and immune landscapes in the genetic and carcinogen-induced LUAD precancer models, leading to distinct cellular spatial arrangements within the tumor microenvironment in each model. Overall, innate immunity, particularly macrophage, probably plays critical roles during early carcinogenesis of LUAD regardless of carcinogen exposure. Citation Format: Bo Zhu, Pingjun Chen, Jian-Rong Li, Muhammad Aminu, Chenyang Li, Hong Chen, Yanhua Tian, Frank R. Rojas, Parra Cuentas Edwin Roger, Ou Shi, Monique B. Nilsson, Alissa Poteete, Shawna M. Hubert, Khaja B. Khan, Wei Lu, Xingzhi Song, Jianhua Zhang, Don L. Gibbons, Luisa M. Solis Soto, Ignacio I. Wistuba, John V. Heymach, Chao Cheng, Jia Wu, Jianjun Zhang. Spatial analysis revealed distinct molecular and immune landscape in genetic versus carcinogen-induced lung adenocarcinoma precancer mouse models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3387.

Abstract 6098: Spatial single-cell immune microenvironment analysis reveals the transition from innate to adaptive immune response during early lung adenocarcinoma carcinogenesis

Abstract Introduction: Lung cancer remains the leading cause of cancer death worldwide. Although prevention holds the promise to reduce lung cancer mortality, the results from randomized trials have been disappointing, likely due to our rudimentary knowledge of early lung cancer carcinogenesis. Cancer initiation and progression involve complex processes dependent on the dynamic interplay of individual cells within the tumor microenvironment (TME). Our understanding of the multicellular composition, interaction, spatial distribution, and functional dynamics during the early evolution from precancer to invasive LUAD is limited. Methods: We performed spatial single cell immune microenvironment analysis of 10 normal, 40 AAH, 22 AIS, 18 MIA, and 34 IAC by Imaging Mass Cytometry (IMC) and analyzed high-dimensional images of 1,618 regions-of-interest (ROIs) using a 34-plex marker. Using machine learning approaches, we classified 4,828,879 cells into 14 major cell types, which were subjected to further analysis for multicellular composition, interaction, spatial distribution, and functional dynamics in LUAD precursors of different stages. Results: From AAH to AIS, MIA, and IAC, we observed a gradual escalation in the density (number of cells/mm2) of all immune subsets. Proportionally, adaptive immune cells exhibited a progressive increase, whereas innate immune cells demonstrated a concurrent decrease with the neoplastic progression from normal lung to AAH, AIS, MIA, and ADC. Notably, macrophages emerged as the predominant immune cells within the immune microenvironment of these LUAD precursors. Upon closer examination of subtypes, protumor M2 macrophages showed a gradual increase from AAH to AIS, MIA, and IAC, whereas anti-tumor M1 macrophages increased from normal to AIH, AIS and subsequently decreased in AIS, MIA, and IAC stages. These alterations resulted in a progressive increase in M2/M1 ratios from AAH to IAC stages. Furthermore, M2 macrophages exhibited higher heterotypic interactions with epithelial cells than M1 across LUAD precursors of different stages, especially in the later stages. Co-expression analysis revealed 5 modules that distinguishes each histological stage. Tim-3 module was significantly enriched in AAH lesions, while NK cell-related features were enriched at AIS stage, B cell-related features enriched at MIA stage and Ki-67-related adaptive immune features enriched in IAC stage. Conclusion: The spatial single cell immune microenvironment analysis revealed a transition from innate to adaptive immunity during initiation and early progression of LUAD progression and innate immunity may play critical roles during early LUAD development. Therefore, reprograming innate immunity such as repolarization of M2/M1 may provide avenues to intercept LUAD precursors to prevent invasive LUAD. Citation Format: Bo Zhu, Pingjun Chen, Muhammad Aminu, Junya Fujimoto, Lingzhi Hong, Andre L. Moreira, Jian-Rong Li, Yanhua Tian, Luisa M. Solis Soto, Parra Cuentas Edwin Roger, Ou Shi, Hong Chen, Frank R. Rojas, Eduardo Vilar, Anirban Maitra, Ken Chen, Nicolas Navin, Cara L. Haymacker, Vamsidhar Velcheti, Daniel H. Sterman, P Andrew Futreal, Don L. Gibbons, Ignacio I. Wistuba, John V. Heymach, Chao Cheng, Harvey I. Pass, Jia Wu, Jianjun Zhang. Spatial single-cell immune microenvironment analysis reveals the transition from innate to adaptive immune response during early lung adenocarcinoma carcinogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6098.

Potential association of HSPD1 with dysregulations in ribosome biogenesis and immune cell infiltration in lung adenocarcinoma: An integrated bioinformatic approach.

Lung adenocarcinoma (LUAD) is a major histological subtype of lung cancer with a high mortality rate worldwide. Heat shock protein family D member 1 (HSPD1, also known as HSP60) is reported to be increased in tumor tissues of lung cancer patients compared with healthy control tissues. We aimed to investigate the roles of HSPD1 in prognosis, carcinogenesis, and immune infiltration in LUAD using an integrative bioinformatic analysis. HSPD1 expression in LUAD was investigated in several transcriptome-based and protein databases. Survival analysis was performed using the KM plotter and OSluca databases, while prognostic significance was independently confirmed through univariate and multivariate analyses. Integrative gene interaction network and enrichment analyses of HSPD1-correlated genes were performed to investigate the roles of HSPD1 in LUAD carcinogenesis. TIMER and TISIDB were used to analyze correlation between HSPD1 expression and immune cell infiltration. The mRNA and protein expressions of HSPD1 were higher in LUAD compared with normal tissues. High HSPD1 expression was associated with male gender and LUAD with advanced stages. High HSPD1 expression was an independent prognostic factor associated with poor survival in LUAD patients. HSPD1-correlated genes with prognostic impact were mainly involved in aberrant ribosome biogenesis, while LUAD patients with high HSPD1 expression had low tumor infiltrations of activated and immature B cells and CD4+ T cells. HSPD1 may play a role in the regulation of ribosome biogenesis and B cell-mediated immunity in LUAD. It could serve as a predictive biomarker for prognosis and immunotherapy response in LUAD.

COA6 promotes the oncogenesis and progression of breast cancer by oxidative phosphorylation pathway.

Mitochondrial oxidative phosphorylation (OXPHOS) has long been considered the primary energy source in breast cancer cells. Cytochrome c oxidase assembly factor 6 (COA6), which functions as a metal chaperone to transport copper to complex Ⅳ during the OXPHOS process, plays a crucial role in the carcinogenesis of lung adenocarcinoma. Nevertheless, its specific function in breast cancer is undefined. The present investigation aimed to clarify COA6's expression profile and regulatory functions in breast cancer, as well as to unveil its underlying mechanisms. Initially, our findings revealed a significant upregulation of COA6 in breast cancer, as evidenced by an analysis of the TCGA database and tissue microarrays. This upregulation correlated with tumor size and histological grade. Additionally, survival analysis revealed that elevated COA6 amounts were correlated with decreased overall survival (OS) in breast cancer. To delve deeper into the functions of COA6, both COA6-overexpressing and COA6-knockdown breast cancer cell models were established. These experiments demonstrated COA6 is pivotal in regulating cell proliferation, apoptosis, migration, and invasion, thereby promoting cancer progression in vitro. Notably, functional enrichment analysis indicated COA6 might be involved in breast cancer progression by modulating oxidative phosphorylation (OXPHOS). Collectively, this study reveals an overt tumorigenic role for COA6 in breast cancer and sheds light on its potential mechanisms, offering valuable therapeutic targets for breast cancer therapy.

GPR37-centered ceRNA network contributes to metastatic potential in lung adenocarcinoma: Evidence from high-throughput sequencing

The competing endogenous RNA (ceRNA)-based profiling has been extensively studied in carcinogenesis of lung adenocarcinoma (LUAD), while it has seldomly been applied to investigate the metastatic potential of LUAD. This study aims to examine the function and in-depth mechanism of GPR37-centered ceRNA network in LUAD. Cancer tissues and adjacent normal tissues from three LUAD patients were collected for high-throughput sequencing to screen for differentially expressed genes. A PPI network was constructed to screen the key gene GPR37, followed by analysis for the functions and pathways. Clinical data from LUAD patients were integrated with gene expression data in TCGA-LUAD dataset for survival analysis. Based on the miRNAs targeting_GPR37 and lncRNAs targeting_miRNAs, a lncRNA-miRNA-mRNA ceRNA network was established. GPR37 was up-regulated in LUAD tissue samples, and it may be a key gene involved in LUAD progression. GPR37 in LUAD was mainly enriched in the mitosis-related pathways. High GPR37 expression corresponded to poor prognosis in LUAD patients. Meanwhile, GPR37 could be used as an independent factor to predict the prognosis in LUAD patients. LncRNA DLEU1, up-regulated in LUAD tissue samples, may competitively bind to miR-4458 to up-regulate the expression of the miR-4458 downstream target GPR37. DLEU1 was associated with poor prognosis and tumor metastasis in LUAD patients. Altogether, our findings reveal a novel ceRNA network of DLEU1/miR-4458/GPR37 in LUAD growth and metastasis.

A risk score model based on lipid metabolism-related genes could predict response to immunotherapy and prognosis of lung adenocarcinoma: a multi-dataset study and cytological validation

BackgroundLipid metabolism is a key factor in tumorigenesis and drug resistance, and models related to lipid metabolism have shown potential to predict survival and curative effects of adjuvant therapy in various cancers. However, the relationship between lipid metabolism and prognosis and treatment response of lung adenocarcinoma (LUAD) are still unclear.MethodsWe enrolled seven bulk RNA-sequence datasets (GSE37745, GSE19188, GSE30219, GSE31547, GSE41271, GSE42127, and GSE72094) from the GEO database and one single-cell RNA-sequencing dataset (GSE117570) from the TISCH2 database. Non-negative matrix factorization (NMF) was utilized to construct the risk score model based on lipid score calculated by GSVA algorithm. Phs000452.v3, PMID: 26359337, PMID: 32472114, PRJEB23709 datasets were used to test the response to immunotherapy. Drug sensitivity analysis was assessed according to the GDSC database, and immunotherapy response was evaluated using the Wilcoxon test. Cellular function assays including clone formation, EDU assays and flow cytometry were implemented to explore the phenotype alteration caused by the knockdown of PTDSS1, which is one of key gene in risk score model.ResultsWe analyzed both bulk and single-cell RNA sequencing data to establish and validate a risk score model based on 18 lipid metabolism-related genes with significant impact on prognosis. After divided the patients into two groups according to risk score, we identified differences in lipid-related metabolic processes and a detailed portrait of the immune landscapes of high- and low-risk groups. Moreover, we investigated the potentials of our risk score in predicting response to immunotherapy and drug sensitivity. In addition, we silenced PTDSS1 in LUAD cell lines, and found that the proliferation of the cells was weakened, and the apoptosis of the cells was increased.ConclusionOur study highlights the crucial roles of lipid metabolism in LUAD and provides a reliable risk score model, which can aid in predicting prognosis and response to immunotherapy. Furthermore, we investigated the roles of PTDSS1 in LUAD carcinogenesis, which showed that PTDSS1 regulated proliferation and apoptosis of LUAD cells.

Targeting CD36 determines nicotine derivative NNK-induced lung adenocarcinoma carcinogenesis

Smoking carcinogen nicotine-derived nitrosamine ketone (NNK) is the most potent contributor to lung adenocarcinoma (LUAD) development, but the mechanism has not been fully elucidated. Here, we reported that fatty acid translocase CD36 was significantly overexpressed in both human LUAD tissues and NNK-induced A/J mice LUAD tumors. The overexpressed CD36 was positively correlated with Src kinase activation, smoking status, metastasis, and worse overall survival of patients with smoking history. Upon NNK binding with α7 nicotinic acetylcholine receptor (α7nAChR), sarcolemmal CD36 was increased and it interacted with surface α7nAChR and cytosol Src simultaneously, which in turn activated Src and downstream pro-carcinogenic kinase ERK1/2 and Akt, and finally caused LUAD cells to form subcutaneous and pulmonary metastatic tumors. This process could be blocked by CD36 knockdown and CD36 irreversible inhibitor SSO. Furthermore, the effect of NNK was inhibited obviously in CD36-/- A/J mice. Thus, targeting CD36 may provide a breakthrough therapy of LUAD.

The ethyl acetate extract of Wenxia Changfu Formula inhibits the carcinogenesis of lung adenocarcinoma by regulating PI3K-AKT signaling pathway

Lung adenocarcinoma is the most common type of lung cancer. With a rise in new cases worldwide each year, early diagnosis and treatment are very important. Network pharmacology provides the effective way to evaluate poly-pharmacological effects and anticancer molecular mechanisms of drugs. The aim of the present study was to explore the anti-tumor mechanism of ethyl acetate extract of Wenxia Changfu Formula (WFEA) in lung adenocarcinoma by using analytical chemistry, network pharmacology and molecular biology. A total of 193 compounds were identified from WFEA, mainly including esters, phenols, ketones and alkaloids. Totally, 374 targets were regarded as potential targets of WFEA against lung adenocarcinoma. Interestingly, PI3K-AKT was found to be one of the significantly enriched signaling pathways of targets of WFEA against lung adenocarcinoma. AKT1, MMP3, CASP3 and BCL2 had strong binding effect with compound molecules of WFEA. Some combinations with the best docking binding were identified, including quercetin/oleanolic_acid/emodin/aloe_emodin/catechin-AKT1 and quercetin-MMP3. In lung adenocarcinoma cells, the WFEA inhibited the proliferation, migration and invasion, and promoted the apoptosis. Moreover, the WFEA inhibited the mRNA expression of MMP3 and Bcl-2 and promoted the mRNA expression of Caspase3. In addition, WFEA inhibited the protein phosphorylation of AKT and PI3K. The WFEA had a significant inhibitory effect on lung adenocarcinoma cells, which could inhibit cell proliferation, invasion and metastasis, and induce cell apoptosis. The mechanism of action of WFEA may be involved in the regulation of the PI3K-AKT signaling pathway in the lung adenocarcinoma.

LncRNA-AC009948.5 promotes invasion and metastasis of lung adenocarcinoma by binding to miR-186-5p.

BackgroundLong non-coding RNAs (LncRNAs) has been confirmed to play a crucial role in the development and progression of various cancer types. Here we evaluated the expression profiles of LncRNAs in Lung adenocarcinoma (LUAD) tissues and identified a novel LncRNA, termed LncRNA-AC009948.5. However, the role and potential molecular mechanisms of this novel LncRNA in LUAD carcinogenesis is unknown.MethodsRegarding the public databases and based on integrating bioinformatics analyses, we determined whether LncRNA-AC009948.5 exerts its oncogenic functions via sponging miR-186-5p in LUAD. Furthermore, we determined whether NCAPG2 was a downstream target of miR-186-5p. Moreover, the expression level and biological function of LncRNA-AC009948.5 in LUAD were determined by qRT-PCR, cell apoptosis, Edu, transwell, wound healing and western blot assays. Besides, xenograft mice were established for validation. We explored the expression of LncRNA-AC009948.5 and its roles in the prognosis of LUAD.ResultsLncRNA expression microarray data indicate that LncRNA-AC009948.5 is upregulated in LUAD samples. The present study confirmed the upregulation of LncRNA-AC009948.5 in LUAD tissues and cells. Encreased expression of LncRNA-AC009948.5 was correlated with tumor size, lymph nodes, distant metastasis and histological grade, and poor prognosis.LncRNA-AC009948.5 knockdown significantly inhibited cell proliferation, migration, and invasion in vitro, as well as tumorigenesis and metastasis in vivo. Conversely, LncRNA-AC009948.5 upregulated had opposite effects. Mechanistically, we elucidated that LncRNA-AC009948.5 could directly bind to miR-186-5p and subsequently suppress expression of the target gene of NCAPG2.ConclusionsLncRNA-AC009948.5 promotes lung adenocarcinoma cells metastasis via the miR-186-5p/NCAPG2 axis and activation of the EMT process. Which may serve as potential targets for the treatment of LUAD in the future.

MiR-382-3p downregulation contributes to the carcinogenesis of lung adenocarcinoma by promoting AKT SUMOylation and phosphorylation.

Lung adenocarcinoma (LA), the primary histological type of non-small cell lung cancer, is still incurable; its diagnosis and treatment remain a major clinical challenge. A previous study by our group examined the microRNA (miRNA/miR) expression profile in the extracellular vesicles from patients with LA and healthy controls and indicated that miR-382-3p levels were reduced in patients with LA. However, the precise roles of miR-382-3p in LA have so far remained elusive. In the present study, the miR-382-3p levels in tumor and adjacent non-tumor control samples from 78 patients with LA were examined and it was identified that miR-382-3p expression was reduced in LA tumor samples compared with that in adjacent non-tumor control tissues (P=0.022). Furthermore, miR-382-3p overexpression inhibited LA growth in a xenograft mouse model. Prediction results indicated that miR-382-3p may regulate protein ubiquitination and SUMOylation. Small ubiquitin-like modifier (SUMO)1 activating enzyme subunit 1 (SAE1), one of the key components of the SUMO-activating complex, was identified as a direct target of miR-382-3p via dual-luciferase and immunoblotting assays. In patients with LA, miR-382-3p expression was negatively correlated with SAE1 protein levels (r=-0.39, P<0.05) and higher SAE1 expression contributed to poor prognosis (P<0.01). Using immunoprecipitation, it was identified that miR-382-3p reduction-induced SAE1 overexpression upregulated AKT SUMOylation, which further promoted AKT phosphorylation and activated the AKT signaling pathway. miR-382-3p inhibition promoted proliferation and inhibited apoptosis in LA cell lines, which was restored by SAE1 knockdown. In conclusion, the present study revealed that downregulation of miR-382-3p contributed to the carcinogenesis of LA via upregulation of SAE1 and promotion of AKT SUMOylation, providing a candidate target for LA treatment.

Methylation- and homologous recombination deficiency-related mutant genes predict the prognosis of lung adenocarcinoma.

BackgroundLung adenocarcinoma (LUAD) is a lung cancer subtype with poor prognosis. We investigated the prognostic value of methylation‐ and homologous recombination deficiency (HRD)‐associated gene signatures in LUAD.MethodsData on RNA sequencing, somatic mutations, and methylation were obtained from TCGA database. HRD scores were used to stratify patients with LUAD into high and low HRD groups and identify differentially mutated and expressed genes (DMEGs). Pearson correlation analysis between DMEGs and methylation yielded methylation‐associated DMEGs. Cox regression analysis was used to construct a prognostic model, and the distribution of clinical features in the high‐ and low‐risk groups was compared.ResultsPatients with different HRD scores showed different DNA mutation patterns. There were 272 differentially mutated genes and 6294 differentially expressed genes. Fifty‐seven DMEGs were obtained; the top 10 upregulated genes were COL11A1, EXO1, ASPM, COL12A1, COL2A1, COL3A1, COL5A2, DIAPH3, CAD, and SLC25A13, while the top 10 downregulated genes were C7, ERN2, DLC1, SCN7A, SMARCA2, CARD11, LAMA2, ITIH5, FRY, and EPHB6. Forty‐two DMEGs were negatively correlated with 259 methylation sites. Gene ontology and pathway enrichment analysis of the DMEGs revealed enrichment of loci involved in extracellular matrix‐related remodeling and signaling. Six out of the 42 methylation‐associated DMEGs were significantly associated with LUAD prognosis and included in the prognostic model. The model effectively stratified high‐ and low‐risk patients, with the high‐risk group having more patients with advanced stage disease.ConclusionWe developed a novel prognostic model for LUAD based on methylation and HRD. Methylation‐associated DMEGs may function as biomarkers and therapeutic targets for LUAD. Further studies are needed to elucidate their roles in LUAD carcinogenesis.

Methyltransferase SETD2 inhibits tumor growth and metastasis via STAT1-IL-8 signaling-mediated epithelial-mesenchymal transition in lung adenocarcinoma.

Lung adenocarcinoma (LUAD) is a major subtype of non–small‐cell lung cancer, which is the leading cause of cancer death worldwide. The histone H3K36 methyltransferase SETD2 has been reported to be frequently mutated or deleted in types of human cancer. However, the functions of SETD2 in tumor growth and metastasis in LUAD has not been well illustrated. Here, we found that SETD2 was significantly downregulated in human lung cancer and greatly impaired proliferation, migration, and invasion in vitro and in vivo. Furthermore, we found that SETD2 overexpression significantly attenuated the epithelial–mesenchymal transition (EMT) of LUAD cells. RNA‐seq analysis identified differentially expressed transcripts that showed an elevated level of interleukin 8 (IL‐8) in STED2‐knockdown LUAD cells, which was further verified using qPCR, western blot, and promoter luciferase report assay. Mechanically, SETD2‐mediated H3K36me3 prevented assembly of Stat1 on the IL‐8 promoter and contributed to the inhibition of tumorigenesis in LUAD. Our findings highlight the suppressive role of SETD2/H3K36me3 in cell proliferation, migration, invasion, and EMT during LUAD carcinogenesis, via regulation of the STAT1–IL‐8 signaling pathway. Therefore, our studies on the molecular mechanism of SETD2 will advance our understanding of epigenetic dysregulation at LUAD progression.

Membrane progesterone receptor \u03b1 (mPR\u03b1) enhances hypoxia-induced vascular endothelial growth factor secretion and angiogenesis in lung adenocarcinoma through STAT3 signaling

Lung cancer remains a huge challenge to public health because of its high incidence and mortality, and lung adenocarcinoma (LUAD) is the main subtype of lung cancer. Hypoxia-induced vascular endothelial growth factor (VEGF) release and angiogenesis have been regarded as critical events in LUAD carcinogenesis. In the present study, membrane progesterone receptor α (mPRα) is deregulated within LUAD tissue samples; increased mPRα contributes to a higher microvessel density (MVD) in LUAD tissues. mPRα knockdown in A549 and PC-9 cells significantly inhibited STAT3 phosphorylation, as well as HIF1α and VEGF protein levels, decreasing cancer cell migration and invasion. The in vivo xenograft model further confirmed that mPRα enhanced the aggressiveness of LUAD cells. Furthermore, mPRα knockdown significantly inhibited hypoxia-induced upregulation in HIF1α and VEGF levels, as well as LUAD cell migration and invasion. Under the hypoxic condition, conditioned medium (CM) derived from mPRα knockdown A549 cells, namely si-mPRα-CM, significantly inhibited HUVEC migration and tube formation and decreased VEGF level in the culture medium. In contrast, CM derived from mPRα-overexpressing A549 cells, namely mPRα-CM, further enhanced HUVEC migration and tube formation and increased VEGF level under hypoxia, which was partially reversed by STAT3 inhibitor Stattic. In conclusion, in LUAD cells, highly expressed mPRα enhances the activation of cAMP/JAK/STAT3 signaling and increases HIF1α-induced VEGF secretion into the tumor microenvironment, promoting HUVEC migration and tube formation under hypoxia.

Transcriptomic analysis of tumor tissues and organoids reveals the crucial genes regulating the proliferation of lung adenocarcinoma

BackgroundAccumulative evidence shows that an organoid is a more practical and reliable tool in cancer biology research. This study aimed to identify and validate crucial genes involved in non-small cell lung cancer carcinogenesis and development using the transcriptomic analysis of tumor tissues and organoids.MethodsGene set enrichment analysis (GSEA) of tumor tissues, tumor organoids, and normal tissues was performed to reveal the similar and different mechanisms involved in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) carcinogenesis and progression. Differentially expressed gene analysis, prognostic analysis, and gene co-expression network analysis were further used to identify hub genes involved in LUAD and LUSC carcinogenesis and development. Finally, LUAD cell lines and organoids were used to validate these findings.ResultsGSEA analysis was performed to reveal the similar mechanisms involved in LUAD and LUSC carcinogenesis and development, such as P53 signaling pathway, base mismatch repair, DNA replication, cAMP signaling pathway and PPAR pathway. However, comparing with LUSC organoids, LUAD organoids showed downregulation of immune-related pathways, inflammation-related pathways, MAPK signaling pathways, and Rap1 signaling pathways, although these pathways were downregulated in LUAD and LUSC tissues by comparing with normal lung tissues. Further gene co-expression network analysis and prognostic analysis indicated CDK1, CCNB2, and CDC25A as the hub tumor-promoting genes in LUAD but not in LUSC, which were further validated in other datasets. Using LUAD cell lines and organoid models, CDK1 and CCNB2 knockdown were found to suppress LUAD proliferation. However, CDC25A knockdown did not inhibit LUAD cell line proliferation but could effectively suppress LUAD organoid growth, indicating that an organoid could be used as an effective tool to study cancer biology in LUAD.ConclusionsThe results revealed CDK1, CCNB2, and CDC25A as the hub genes involved in LUAD carcinogenesis and development, which could be used as the potential biomarkers and targets for LUAD.

Integrative analyses identified ion channel genes GJB2 and SCNN1B as prognostic biomarkers and therapeutic targets for lung adenocarcinoma

ObjectivesAbnormal expressions of ion channel genes are associated with the occurrence and progression of tumors. At present, their roles in the carcinogenesis of lung adenocarcinoma (LUAD) are not clear. Materials and methodsDifferentially expressed (DE) genes in the tumorigenesis were identified from 328 ion channel genes in 102 LUAD and paired adjacent normal samples. Similar analyses were performed between 177 metastatic and 286 non-metastatic LUAD samples to identify DE ion channel genes in the progression of LUAD. Independent prognostic factors selected from DE ion channel genes were used to construct a prognostic model. Correlation analysis and drugs-drug targets interaction network were used to screen the potential drugs for LUAD patients stratified by GJB2 or SCNN1B. ResultsSix ion channel genes (GJB2, CACNA1D, KCNQ1, SCNN1B, SCNN1G and TRPV6) were continuous differentially expressed in the tumorigenesis and progression of LUAD. The survival analysis in four datasets with 522 LUAD samples showed that GJB2 and SCNN1B were independent prognostic biomarkers. Patients with overexpression of GJB2 or underexpression of SCNN1B had shorter overall survival. Moreover, multi-omics analysis showed that hypomethylation of GJB2 and hypermethylation of SCNN1B in the promoter region may contribute to their aberrant expressions. KEGG enrichment analysis showed that the overexpressed genes in the group with high GJB2 or low SCNN1B were enriched in cancer-related pathways, while the underexpressed genes were enriched in metabolism-related pathways. The prognostic model with GJB2 and SCNN1B can stratify all LUAD patients into two groups with significantly different survival. Correlation analysis and drugs-drug targets interaction network suggested that GJB2 and SCNN1B expression might have indicative therapeutic values for LUAD patients. Finally, pan-cancer analysis in other eight cancer types showed that GJB2 and SCNN1B might be also potential prognostic factors for KIRC. ConclusionsGJB2 and SCNN1B were identified as prognostic biomarkers and therapeutic targets for LUAD.

Immune evolution from preneoplasia to invasive lung adenocarcinomas and underlying molecular features

The mechanism by which anti-cancer immunity shapes early carcinogenesis of lung adenocarcinoma (ADC) is unknown. In this study, we characterize the immune contexture of invasive lung ADC and its precursors by transcriptomic immune profiling, T cell receptor (TCR) sequencing and multiplex immunofluorescence (mIF). Our results demonstrate that anti-tumor immunity evolved as a continuum from lung preneoplasia, to preinvasive ADC, minimally-invasive ADC and frankly invasive lung ADC with a gradually less effective and more intensively regulated immune response including down-regulation of immune-activation pathways, up-regulation of immunosuppressive pathways, lower infiltration of cytotoxic T cells (CTLs) and anti-tumor helper T cells (Th), higher infiltration of regulatory T cells (Tregs), decreased T cell clonality, and lower frequencies of top T cell clones in later-stages. Driver mutations, chromosomal copy number aberrations (CNAs) and aberrant DNA methylation may collectively impinge host immune responses and facilitate immune evasion, promoting the outgrowth of fit subclones in preneoplasia into dominant clones in invasive ADC.

Prevalence of the Brazilian TP53 Founder c.1010G>A (p.Arg337His) in Lung Adenocarcinoma: Is Genotyping Warranted in All Brazilian Patients?

In Southern and Southeastern Brazil, there is a germline pathogenic variant with incomplete penetrance located in the oligomerization domain of TP53, c.1010G>A (p.Arg337His). Due to a founder effect, the variant is present in 0.3% of the general population of the region. Recently, this variant was identified in 4.4 and 8.9% of two apparently unselected, single center case series of Brazilian lung adenocarcinoma (LUAD) patients from the Southeastern and Central regions of the country, respectively. In the present study, our aim was to examine TP53 c.1010G>A allele and genotype frequencies in LUAD samples obtained from patients diagnosed in Southern Brazil. A total of 586 LUAD samples (tumor DNA) recruited from multiple centers in the region were tested, and the mutant allele was identified using TaqMan® assays in seven cases (7/586, 1.2%) which were submitted to next generation sequencing analyses for confirmation. Somatic EGFR mutations were more frequent in TP53 c.1010G>A carriers than in non-carriers (57.1 vs. 17.6%, respectively). Further studies are needed to confirm if TP53 c.1010G>A is a driver in LUAD carcinogenesis and to verify if there is a combined effect of EGFR and germline TP53 c.1010G>A. Although variant frequency was higher than observed in the general population, it is less than previously reported in LUAD patients from other Brazilian regions. Additional data, producing regional allele frequency information in larger series of patients and including cost-effectiveness analyses, are necessary to determine if TP53 c.1010G>A screening in all Brazilian LUAD patients is justified.

Evolution of DNA methylome from precancerous lesions to invasive lung adenocarcinomas

The evolution of DNA methylome and methylation intra-tumor heterogeneity (ITH) during early carcinogenesis of lung adenocarcinoma has not been systematically studied. We perform reduced representation bisulfite sequencing of invasive lung adenocarcinoma and its precursors, atypical adenomatous hyperplasia, adenocarcinoma in situ and minimally invasive adenocarcinoma. We observe gradual increase of methylation aberrations and significantly higher level of methylation ITH in later-stage lesions. The phylogenetic patterns inferred from methylation aberrations resemble those based on somatic mutations suggesting parallel methylation and genetic evolution. De-convolution reveal higher ratio of T regulatory cells (Tregs) versus CD8 + T cells in later-stage diseases, implying progressive immunosuppression with neoplastic progression. Furthermore, increased global hypomethylation is associated with higher mutation burden, copy number variation burden and AI burden as well as higher Treg/CD8 ratio, highlighting the potential impact of methylation on chromosomal instability, mutagenesis and tumor immune microenvironment during early carcinogenesis of lung adenocarcinomas.