BACKGROUND AND OBJECTIVES

Cell lines serve as invaluable tools in studying lung cancer biology and developing new therapies to combat the disease. However, commercially available cell lines are typically of Caucasian origin and may be less representative of the local genetic background. To address this, our lab previously immortalized cells from pleural fluid of a Filipino non-small cell lung cancer (NSCLC) patient via CDK4 transduction. Copy number variations (CNVs) are a type of genetic variation which may affect physiology and disease by disrupting gene function or altering gene expression, and in cancer, these may be associated with patient outcomes. CNV profiling can be valuable for understanding the biology of our immortalized cells and identifying genes that could serve as potential targets for diagnostic, prognostic, and therapeutic interventions. This study aimed to characterize previously immortalized NSCLC-derived cells, GL01, in comparison with an established lung adenocarcinoma (LUAD) cell line, A549, through whole-genome microarray-based copy number profiling.

DNA was extracted from GL01 and A549 cells using a commercially-available silica-based DNA extraction kit. DNA extracts were quantified and normalized for microarray analysis. Whole-genome copy number profiling was done using the OncoScan CNV Plus Assay following the manufacturer’s protocols, and data was analyzed using the Chromosome Analysis Suite software. Functional analysis of genes identified to be involved in copy number aberrations was done using the PANTHER Classification System.

Copy number aberrations span 1,592,737,105 bp in GL01 and 1,715,708,552 bp in A549, with a high degree of concordance between the two. Large-scale and focal copy number aberrations previously identified to be recurrent in various LUAD cohorts were present in both GL01 and A549. Focal copy number aberrations associated with previously described lung cancer-related genes involve the PDE4D gene in GL01 and the SKIL and CDKN2A/CDKN2B genes in both GL01 and A549. PANTHER Pathway analysis of genes positively correlated with mRNA expression showed that the ubiquitin proteasome pathway was significantly overrepresented in both GL01 (FDR p = 0.000074) and A549 (FDR p = 0.000075), with 20 genes involved. Additionally, the KRAS:p.G12C/S:c.34G>T/A somatic mutation variant was detected in both GL01 and A549.

This study provides a method for identifying potentially clinically-relevant genes associated with a sample’s copy number aberrations and the pathways they represent, providing personalized mechanistic, prognostic, and therapeutic insights into the cancer biology of our cells.

INTRODUCTION

Most cancer-related deaths globally are caused by lung cancer. The diagnosis is typically made following the evaluation of respiratory symptoms such as chronic cough or incidental finding of pulmonary lesions such as nodules and mass. Cardiac metastasis occurs in 2-18% of lung cancers, but cardiac tamponade complicating malignant pericardial effusion is an extremely rare and life-threatening initial presentation of non-small cell lung carcinoma.

A 59-year-old-male with a smoking history of 72 pack-year presented at the emergency room with severe dyspnea and was assessed to be in cardiac tamponade. The patient arrested for 16 minutes but returned to spontaneous circulation after 650mL of serosanguinous fluid was removed by pericardiocentesis. The work-up for infectious and immunologic causes was negative. Chest CT scan with contrast did not reveal any pulmonary mass. However, the pericardial fluid cytology immunohistochemical stains pointed to a primary lung adenocarcinoma. PET scan was requested which confirmed hypermetabolic focus in the left lung base. included patients admitted at East Avenue Medical Center for DFU. The primary endpoint was major amputation of the lower extremities. Data were analyzed using Receiver Operating Characteristic (ROC) analysis and logistic regression.

This case showed an extremely rare situation where life-threatening cardiac tamponade was the initial presentation of non-small cell lung adenocarcinoma, which highlights the need for vigilance in atypical presentation. Comprehensive diagnostic approach, including PET scans and cytologic analysis, must be done when standard imaging is inconclusive.

INTRODUCTION

Lung cancer is the leading cause of cancer-related mortality worldwide with peak mortality rate occurring in patients aged 80 years and above. While NSCLC are often diagnosed at advanced stage when treatment options are few, access to treatment in elderly are even more limited due to treatment tolerability and potential toxicity. At present, Osimertinib is the first line treatment option for patients with metastatic NSCLC with EGFR mutations. Some adverse reactions are diarrhea, nausea, headaches, stomatitis, and rashes that lead to interruption or even stopping of the medication.

Here we present a case about an 89-year-old female with smoking history of 20 pack-years who initially presented at the emergency room with progressive shortness of breath. Chest radiograph showed right pleural effusion for which pigtail was inserted. Bronchoscopy revealed a completely obstructing mass at the right upper lobe. Her biopsy showed EGFR-mutated non-small cell lung adenocarcinoma. Patient underwent radiotherapy and was started on osimertinib 80mg daily. However, patient developed severe diarrhea for which her subsequent dosing was reduced to 40mg once daily. Repeat PET CT scan after 10 months showed significant reduction of the primary mass.

In patients with metastatic EGFR-mutated lung adenocarcinoma, Osimertinib proves to be an effective option and is associated with improved overall survival even on a low-dose. This dose reduction strategy may be an option especially for elderly patients with tolerability issues. Nonetheless, treatment choices should prioritize patients' functional status and comorbidities over age, underscoring the importance of personalized approaches despite chemotherapy's inherent risks.

BACKGROUND: In China, lung cancer remains the cancer with the highest incidence and mortality rate. Among early-stage lung adenocarcinomas (LUAD), the micropapillary (MPP) component is prevalent and typically exhibits high aggressiveness, significantly correlating with early metastasis, lymphatic infiltration, and reduced five-year survival rates. Therefore, the study is to explore the similarities and differences between MPP and non-micropapillary (non-MPP) components in malignant pulmonary nodules characterized by GGOs in early-stage LUAD, identify unique mutational features of the MPP component and analyze the relationship between the ZNF469 gene, a member of the zinc-finger protein family, and the prognosis of early-stage LUAD, as well as its correlation with immune infiltration. METHODS: A total of 31 malignant pulmonary nodules of LUAD were collected and dissected into paired MPP and non-MPP components using microdissection. Whole-exome sequencing (WES) was performed on the components of early-stage malignant pulmonary nodules. Mutational signatures analysis was conducted using R packages such as maftools, Nonnegative Matrix Factorization (NMF), and Sigminer to unveil the genomic mutational characteristics unique to MPP components in invasive LUAD compared to other tumor tissues. Furthermore, we explored the expression of the ZNF469 gene in LUAD using The Cancer Genome Atlas (TCGA) database to investigate its potential association with the prognosis. We also investigated gene interaction networks and signaling pathways related to ZNF469 in LUAD using the GeneMANIA database and conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Lastly, we analyzed the correlation between ZNF469 gene expression and levels of immune cell infiltration in LUAD using the TIMER and TISIDB databases. RESULTS: MPP components exhibited a higher number of genomic variations, particularly the 13th COSMIC (Catalogue of Somatic Mutations in Cancer) mutational signature characterized by the activity of the cytidine deaminase APOBEC family, which was unique to MPP components compared to non-MPP components in tumor tissues. This suggests the potential involvement of APOBEC in the progression of MPP components in early-stage LUAD. Additionally, MPP samples with high similarity to APOBEC signature displayed a higher tumor mutational burden (TMB), indicating that these patients may be more likely to benefit from immunotherapy. The expression of ZNF469 was significantly upregulated in LUAD compared to normal tissue, and was associated with poor prognosis in LUAD patients (P<0.05). Gene interaction network analysis and GO/KEGG enrichment analysis revealed that COL6A1, COL1A1, COL1A2, TGFB2, MMP2, COL8A2 and C2CD4C interacted with ZNF469 and were mainly involved in encoding collagen proteins and participating in the constitution of extracellular matrix. ZNF469 expression was positively correlated with immune cell infiltration in LUAD (P<0.05). CONCLUSIONS The study has unveiled distinctive mutational signatures in the MPP components of early-stage invasive LUAD in the Asian population. Furthermore, we have identified that the elevated expression of mutated ZNF469 impacts the prognosis and immune infiltration in LUAD, suggesting its potential as a diagnostic and prognostic biomarker in LUAD.
Humans ; Lung Neoplasms/genetics* ; Adenocarcinoma of Lung/genetics* ; China ; Prognosis ; Transcription Factors

Ground-glass nodule (GGN) lung cancer often progresses slowly in clinical and there are few clinical studies on long-term follow-up of patients with operable GGN lung cancer treated with stereotactic body radiation therapy (SBRT). We present a successful case of GGN lung cancer treated with SBRT, but a new GGN was found in the lung adjacent to the SBRT target during follow-up. The nodule progressed rapidly and was confirmed as lung adenocarcinoma by surgical resection. No significant risk factors and related driving genes were found in molecular pathological findings and genetic tests. It deserves further study whether new GGN is related to the SBRT. This case suggests that the follow-up after SBRT should be vigilant against the occurrence of new rapidly progressive lung cancer in the target area and adjacent lung tissue.
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Humans ; Lung Neoplasms/pathology* ; Radiosurgery ; Retrospective Studies ; Adenocarcinoma of Lung/surgery* ; Lung/pathology*

Background and Objectives: Cell lines serve as invaluable tools in studying lung cancer biology and developing new therapies to combat the disease. However, commercially available cell lines are typically of Caucasian origin and may be less representative of the local genetic background. To address this, our lab previously immortalized cells from pleural fluid of a Filipino non-small cell lung cancer (NSCLC) patient via CDK4 transduction. Copy number variations (CNVs) are a type of genetic variation which may affect physiology and disease by disrupting gene function or altering gene expression, and in cancer, these may be associated with patient outcomes. CNV profiling can be valuable for understanding the biology of our immortalized cells and identifying genes that could serve as potential targets for diagnostic, prognostic, and therapeutic interventions. This study aimed to characterize previously immortalized NSCLC-derived cells, GL01, in comparison with an established lung adenocarcinoma (LUAD) cell line, A549, through whole-genome microarray-based copy number profiling. Methods: DNA was extracted from GL01 and A549 cells using a commercially-available silica-based DNA extraction kit. DNA extracts were quantified and normalized for microarray analysis. Whole-genome copy number profiling was done using the OncoScan CNV Plus Assay following the manufacturer’s protocols, and data was analyzed using the Chromosome Analysis Suite software. Functional analysis of genes identified to be involved in copy number aberrations was done using the PANTHER Classification System. Results: Copy number aberrations span 1,592,737,105 bp in GL01 and 1,715,708,552 bp in A549, with a high degree of concordance between the two. Largescale and focal copy number aberrations previously identified to be recurrent in various LUAD cohorts were present in both GL01 and A549. Focal copy number aberrations associated with previously described lung cancer-related genes involve the PDE4D gene in GL01 and the SKIL and CDKN2A/CDKN2B genes in both GL01 and A549. PANTHER Pathway analysis of genes positively correlated with mRNA expression showed that the ubiquitin proteasome pathway was significantly overrepresented in both GL01 (FDR p = 0.000074) and A549 (FDR p = 0.000075), with 20 genes involved. Additionally, the KRAS:p.G12C/S:c.34G>T/A somatic mutation variant was detected in both GL01 and A549. Conclusion This study provides a method for identifying potentially clinically-relevant genes associated with a sample’s copy number aberrations and the pathways they represent, providing personalized mechanistic, prognostic, and therapeutic insights into the cancer biology of our cells.
carcinoma, non-small cell lung ; adenocarcinoma of lung

BACKGROUND: Lung cancer prevails and induces high mortality around the world. This study provided real-world information on the evolution of clinicopathological profiles and survival outcomes of lung cancer, and provided survival information within stage I subtypes. METHODS: Patients pathologically confirmed with lung cancer between January 2009 and December 2018 were identified with complete clinicopathological information, molecular testing results, and follow-up data. Shifts in clinical characteristics were evaluated using χ2 tests. Overall survival (OS) was calculated through the Kaplan-Meier method. RESULTS: A total of 26,226 eligible lung cancer patients were included, among whom 62.55% were male and 52.89% were smokers. Non-smokers and elderly patients took increasingly larger proportions in the whole patient population. The proportion of adenocarcinoma increased from 51.63% to 71.80%, while that of squamous carcinoma decreased from 28.43% to 17.60%. Gene mutations including EGFR (52.14%), KRAS (12.14%), and ALK (8.12%) were observed. Female, younger, non-smoking, adenocarcinoma patients and those with mutated EGFR had better survival prognoses. Importantly, this study validated that early detection of early-stage lung cancer patients had contributed to pronounced survival benefits during the decade. Patients with stage I lung cancer, accounted for an increasingly considerable proportion, increasing from 15.28% to 40.25%, coinciding with the surgery rate increasing from 38.14% to 54.25%. Overall, period survival analyses found that 42.69% of patients survived 5 years, and stage I patients had a 5-year OS of 84.20%. Compared with that in 2009-2013, the prognosis of stage I patients in 2014-2018 was dramatically better, with 5-year OS increasing from 73.26% to 87.68%. Regarding the specific survival benefits among stage I patients, the 5-year survival rates were 95.28%, 93.25%, 82.08%, and 74.50% for stage IA1, IA2, IA3, and IB, respectively, far more promising than previous reports. CONCLUSIONS Crucial clinical and pathological changes have been observed in the past decade. Notably, the increased incidence of stage I lung cancer coincided with an improved prognosis, indicating actual benefits of early detection and management of lung cancer.
Humans ; Male ; Female ; Aged ; Lung Neoplasms/genetics* ; Adenocarcinoma/pathology* ; Prognosis ; Survival Rate ; Mutation ; ErbB Receptors/genetics* ; Neoplasm Staging ; Retrospective Studies

BACKGROUND: Since the popularization of computed tomography (CT) technology, the detection rate of pulmonary ground glass nodules (GGNs) with imaging follow-up as the main management method has increased significantly. The purpose of this study is to quantitatively analyze the changes of pulmonary GGNs during the follow-up process with three-dimensional reconstruction technology, explore the natural progression of pulmonary GGNs, and provide effective basis for clinical guidance for patients to conduct reasonable management of nodules. METHODS: A total of 115 cases of pulmonary GGNs with regular follow-up in the Combined Outpatient Department of Zhoushan Hospital from March 2015 to November 2022 were enrolled. Quantitative imaging features of nodules were extracted by semi-automatic segmentation of 3D Slicer software to evaluate the growth of nodules and clinical intervention during follow-up. RESULTS: The average baseline age of the patients was (56.9±10.1) yr. The mean follow-up time was (48.8±18.9) months. The two-dimensional diameter of baseline CT scan was (7.9±2.9) mm, and the maximum three-dimensional diameter was (10.1±3.4) mm. The two-dimensional diameter of the last CT scan was (9.9±4.7) mm, and the maximum three-dimensional diameter was (11.4±5.1) mm. A total of 27 cases (23.5%) showed an increase during follow-up, with a median volume doubling time of 822 days and a median mass doubling time of 1,007 days. 32 cases were surgically resected, including 6 cases of invasive adenocarcinoma (IAC), 16 cases of minimally invasive adenocarcinoma (MIA), 8 cases of adenocarcinoma in situ (AIS) and 2 cases of atypical adenomatous hyperplasia (AAH). Five nodules underwent surgical intervention due to the progression of two-dimensional diameter, which was pathologically confirmed as pre-invasive lesions, but their three-dimensional maximum diameter showed no significant change. Nodular morphology, lobulated sign, spiculated sign and vacuole signs all promoted the growth of nodules in univariate analysis. There were significant differences in age, baseline diameter, mean CT value, median CT value, 10% and 90% percentile CT number between the growth group and the stable group (P<0.05). Multivariate Logistic regression analysis showed that age and average CT value were risk factors for nodule growth (P<0.05). Receiver-operating characteristic (ROC) curve analysis results indicated that the age ≥63 years old, the baseline three-dimensional maximum diameter ≥9.2 mm, and the average CT value ≥-507.8 HU were more likely to accelerate the growth of GGNs. The maximum three-dimensional diameter ≥14.4 mm and the average CT value ≥-495.7 HU may be a higher malignant probability. CONCLUSIONS GGNs show an inert growth process, and the use of three-dimensional measurements during follow-up is of greater significance. For persistent glass grinding nodules ≥63 years old, the baseline three-dimensional maximum diameter ≥9.2 mm, and the average CT value ≥-507.8 HU are more likely to increase. However, most nodules still have good prognosis after progression, and long-term follow-up is safe.
Humans ; Middle Aged ; Lung Neoplasms/pathology* ; Imaging, Three-Dimensional ; Neoplasm Invasiveness ; Retrospective Studies ; Multiple Pulmonary Nodules/pathology* ; Adenocarcinoma/pathology*

Lung cancer is the most common in incidence and mortality worldwide. With the development of next generation sequencing (NGS) detection technology, more and more patients with rare anaplastic lymphoma kinase (ALK) fusion mutations were detected. A case of advanced lung adenocarcinoma with rare COX7A2L-ALK (C2:A20) fusion detected by NGS was reported in Peking Union Medical College Hospital, and all cases with rare ALK fusion mutations were searched from medical datebase from January 1, 2014 to March 31, 2021, to investigate the treatment of rare ALK fusion mutations with ALK inhibitors. The best response of the patient was assessed as partial response (PR) with Ceritinib treatment. By literature review, 22 cases of rare ALK fusion were reported in 19 articles. Combined with this case, 23 cases were analyzed. The objective response rate (ORR) was 82.6% (19/23) and disease control rate (DCR) was 95.7% (22/23) for rare ALK fusions patients treated with ALK inhibitors. Lung adenocarcinoma patients with rare ALK fusion could benefit from ALK inhibitors.
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Humans ; Anaplastic Lymphoma Kinase/genetics* ; Lung Neoplasms/diagnosis* ; Crizotinib ; Adenocarcinoma of Lung/genetics* ; Protein Kinase Inhibitors/pharmacology* ; Oncogene Proteins, Fusion/genetics*

BACKGROUND: Lung cancer is one of the most common malignant tumors in the world. The accuracy of intraoperative frozen section (FS) in the diagnosis of lung adenocarcinoma infiltration cannot fully meet the clinical needs. The aim of this study is to explore the possibility of improving the diagnostic efficiency of FS in lung adenocarcinoma by using the original multi-spectral intelligent analyzer. METHODS: Patients with pulmonary nodules who underwent surgery in the Department of Thoracic Surgery, Beijing Friendship Hospital, Capital Medical University from January 2021 to December 2022 were included in the study. The multispectral information of pulmonary nodule tissues and surrounding normal tissues were collected. A neural network model was established and the accuracy of the neural network diagnostic model was verified clinically. RESULTS: A total of 223 samples were collected in this study, 156 samples of primary lung adenocarcinoma were finally included, and a total of 1,560 sets of multispectral data were collected. The area under the curve (AUC) of spectral diagnosis in the test set (10% of the first 116 cases) of the neural network model was 0.955 (95%CI: 0.909-1.000, P<0.05), and the diagnostic accuracy was 95.69%. In the clinical validation group (the last 40 cases), the accuracy of spectral diagnosis and FS diagnosis were both 67.50% (27/40), and the AUC of the combination of the two was 0.949 (95%CI: 0.878-1.000, P<0.05), and the accuracy was 95.00% (38/40). CONCLUSIONS The accuracy of the original multi-spectral intelligent analyzer in the diagnosis of lung invasive adenocarcinoma and non-invasive adenocarcinoma is equivalent to that of FS. The application of the original multi-spectral intelligent analyzer in the diagnosis of FS can improve the diagnostic accuracy and reduce the complexity of intraoperative lung cancer surgery plan.
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Humans ; Lung Neoplasms/surgery* ; Adenocarcinoma of Lung/surgery* ; Adenocarcinoma/surgery* ; Hospitals ; Multiple Pulmonary Nodules