In-depth proteomics reveals the characteristic developmental profiles of early lung adenocarcinoma with epidermal growth factor receptor mutation.

INTRODUCTION: Lung adenocarcinoma progresses stepwise from atypical adenomatous hyperplasia to adenocarcinoma in situ (AIS), followed by minimally invasive adenocarcinoma (MIA), and then obvious invasive adenocarcinoma. In this study, we examined the protein expression profiles of early and epidermal growth factor receptor (EGFR) mutation-positive lung adenocarcinomas. METHODS: Fifteen cases of small and EGFR mutation-positive adenocarcinomas were collected, including AIS, MIA, and small invasive adenocarcinoma (SIA). We examined their protein expression profiles by tandem mass tag (TMT)-labeling liquid chromatography-mass spectrometry (LC-MS/MS) and compared the results between AIS and MIA versus SIA. The differentially expressed proteins were then verified by Western blot analysis and immunohistochemistry (IHC). The clinicopathological implications of the proteins were also examined by IHC. RESULTS: A total of 4220 proteins were identified by LC-MS/MS analysis. Pathway analysis of the differentially expressed proteins revealed that pathways related to interferon α/ß signaling, glutamate and glutamine metabolism, and gluconeogenesis were upregulated in SIA relative to AIS. Among the 13 differentially expressed proteins, cellular retinoic acid binding protein 2 (CRABP2), delta(24)-sterol reductase (DHCR24), and adenylate kinase 4 (AK4) were expressed significantly more strongly in SIA than in AIS. Patients with high expression of CRABP2, DHCR24, and AK4 showed a significantly poorer outcome than those with low expression. CONCLUSION: In comparison with AIS, SIA shows differences in several different protein expression pathways. Furthermore, CRABP2, DHCR24, and AK4 are useful IHC markers for diagnosis of lung adenocarcinoma invasiveness and may be associated with malignant progression of AIS.

Diagnostic performance of RASSF1A and SHOX2 methylation combined with EGFR mutations for differentiation between small pulmonary nodules.

BACKGROUND AND AIM: Aberrant methylation of Ras association domain family 1, isoform A (RASSF1A), and short-stature homeobox gene 2 (SHOX2) promoters has been validated as a pair of valuable biomarkers for diagnosing early lung adenocarcinomas (LUADs). Epidermal growth factor receptor (EGFR) is the key driver mutation in lung carcinogenesis. This study aimed to investigate the aberrant promoter methylation of RASSF1A and SHOX2, and the genetic mutation of EGFR in 258 specimens of early LUADs. METHODS: We retrospectively selected 258 paraffin-embedded samples of pulmonary nodules measuring 2 cm or less in diameter and evaluated the diagnostic performance of individual biomarker assays and multiple panels between noninvasive (group 1) and invasive lesions (groups 2A and 2B). Then, we investigated the interaction between genetic and epigenetic alterations. RESULTS: The degree of RASSF1A and SHOX2 promoter methylation and EGFR mutation was significantly higher in invasive lesions than in noninvasive lesions. The three biomarkers distinguished between noninvasive and invasive lesions with reliable sensitivity and specificity: 60.9% sensitivity [95% confidence interval (CI) 52.41-68.78] and 80.0% specificity (95% CI 72.14-86.07). The novel panel biomarkers could further discriminate among three invasive pathological subtypes (area under the curve value > 0.6). The distribution of RASSF1A methylation and EGFR mutation was considerably exclusive in early LUAD (P = 0.002). CONCLUSION: DNA methylation of RASSF1A and SHOX2 is a pair of promising biomarkers, which may be used in combination with other driver alterations, such as EGFR mutation, to support the differential diagnosis of LUADs, especially for stage I.

Relationship between early lung adenocarcinoma and multiple driving genes based on artificial intelligence medical images of pulmonary nodules.

Lung adenocarcinoma is one of the most common cancers in the world, and accurate diagnosis of lung nodules is an important factor in reducing its mortality. In the diagnosis of pulmonary nodules, artificial intelligence (AI) assisted diagnosis technology has been rapidly developed, so testing its effectiveness is conducive to promoting its important role in clinical practice. This paper introduces the background of early lung adenocarcinoma and lung nodule AI medical imaging, and then makes academic research on early lung adenocarcinoma and AI medical imaging, and finally summarizes the biological information. In the experimental part, the relationship analysis of 4 driver genes in group X and group Y showed that there were more abnormal invasive lung adenocarcinoma genes, and the maximum uptake value and uptake function of metabolic value were also higher. However, there was no significant correlation between mutations in the four driver genes and metabolic values, and the average accuracy of AI-based medical images was 3.88% higher than that of traditional images.

The Diagnostic Potential of SHOX2 and RASSF1A DNA Methylation in Early Lung Adenocarcinoma.

Objective: Methylation of the promoters of SHOX2 and RASSF1A are potentially informative biomarkers for the diagnosis of early lung adenocarcinoma (LUAD). Abnormal methylation of SHOX2 and RASSF1A promoters may promote the occurrence and facilitate the progression of LUAD. Materials and Methods: We selected 54 patients with early LUAD and 31 patients with benign lung nodules as a NJDT cohort and evaluated their DNA methylation and mRNA sequencing levels. The DNA methylation sequencing, mRNA sequencing, and clinical data for patients with LUAD were obtained from The Cancer Genome Atlas, and served as a TCGA cohort. We evaluated the diagnostic potential of a SHOX2 and RASSF1A combined promoter methylation assay for detection of early LUAD in the NJDT cohort. Then we explored the promoter methylation levels of SHOX2 and RASSF1A and their gene expression between normal and tumor samples at different stages in both cohorts. Pathways enriched between tumor and normal samples of methylation-positive patients in the NJDT cohort were analyzed. Results: In the NJDT cohort, the sensitivity of the combined promoter methylation assay on tumor samples was 74.07%, the sensitivity on paired tumor and paracancerous samples was 77.78%, and the specificities in both contexts were 100%. The combined promoter methylation-positive patients had clinicopathologic features including older age, larger tumors, deeper invasion, and higher Ki-67 expression. In both cohorts, SHOX2 expression increased and RASSF1A expression decreased in tumor samples. The promoter methylation level of SHOX2 and RASSF1A was significantly higher in tumor samples at stage I-II than that in normal samples. The promoter methylation levels of these two genes were both negative associated with their expression in early tumor samples. In the NJDT cohort, methylation-positive patients of both individual SHOX2 and RASSF1A assays exhibited upregulation of folate acid metabolism and nucleotide metabolism in tumor samples. The SHOX2 methylation-positive and RASSF1A methylation-positive patients showed the downregulation of pathways related to cell proliferation and apoptosis and pathways involved in DNA repair, cell growth and cell adhesion, respectively. Conclusion: The combined promoter methylation assay for SHOX2 and RASSF1A can be used for screening and diagnosis of early LUAD, with good sensitivity and specificity. The promoter methylation levels of SHOX2 and RASSF1A were associated with their abnormal mRNA expression, and affected DNA instability, cell proliferation, apoptosis and tumor microenvironment in patients with LUAD.

Case Report: Primary Bilateral Minimally Invasive Adenocarcinoma of the Lungs in an 11-Year-Old Child: A Rare Case.

There are many types of benign and malignant tissue, but primary lung tumor is very rare in children and often remains undiagnosed until after distant metastasis has occurred. Few cases of early lung adenocarcinoma in children have been reported. However, this case concerns an 11-year-old child with primary bilateral minimally invasive adenocarcinoma. As far as we know, this is the youngest reported case of its type.

Decreased IL-6 and NK Cells in Early-Stage Lung Adenocarcinoma Presenting as Ground-Glass Opacity.

BACKGROUND: Lung ground-glass opacities (GGOs) are an early manifestation of lung adenocarcinoma. It is of great value to study the changes in the immune microenvironment of GGO to elucidate the occurrence and evolution of early lung adenocarcinoma. Although the changes of IL-6 and NK cells in lung adenocarcinoma have caught global attention, we have little appreciation for how IL-6 and NK cells in the lung GGO affect the progression of early lung adenocarcinoma. METHODS: We analyzed the RNA sequencing data of surgical specimens from 21 patients with GGO-featured primary lung adenocarcinoma and verified the changes in the expression of IL-6 and other important immune molecules in the TCGA and GEO databases. Next, we used flow cytometry to detect the protein expression levels of important Th1/Th2 cytokines in GGO and normal lung tissues and the changes in the composition ratio of tumor infiltrating lymphocytes (TILs). Then, we analyzed the effect of IL-6 on NK cells through organoid culture and immunofluorescence. Finally, we explored the changes of related molecules and pathway might be involved. RESULTS: IL-6 may play an important role in the tumor microenvironment of early lung adenocarcinoma. Further research confirmed that the decrease of IL-6 in GGO tissue is consistent with the changes in NK cells, and there seems to be a correlation between these two phenomena. CONCLUSION: The IL-6 expression status and NK cell levels of early lung adenocarcinoma as GGO are significantly reduced, and the stimulation of IL-6 can up-regulate or activate NK cells in GGO, providing new insights into the diagnosis and pathogenesis of early lung cancer.

Signature identification of relapse-related overall survival of early lung adenocarcinoma after radical surgery.

BACKGROUND: The widespread use of low-dose chest CT screening has improved the detection of early lung adenocarcinoma. Radical surgery is the best treatment strategy for patients with early lung adenocarcinoma; however, some patients present with postoperative recurrence and poor prognosis. Through this study, we hope to establish a model that can identify patients that are prone to recurrence and have poor prognosis after surgery for early lung adenocarcinoma. MATERIALS AND METHODS: We screened prognostic and relapse-related genes using The Cancer Genome Atlas (TCGA) database and the GSE50081 dataset from the Gene Expression Omnibus (GEO) database. The GSE30219 dataset was used to further screen target genes and construct a risk prognosis signature. Time-dependent ROC analysis, calibration degree analysis, and DCA were used to evaluate the reliability of the model. We validated the TCGA dataset, GSE50081, and GSE30219 internally. External validation was conducted in the GSE31210 dataset. RESULTS: A novel four-gene signature (INPP5B, FOSL2, CDCA3, RASAL2) was established to predict relapse-related survival outcomes in patients with early lung adenocarcinoma after surgery. The discovery of these genes may reveal the molecular mechanism of recurrence and poor prognosis of early lung adenocarcinoma. In addition, ROC analysis, calibration analysis and DCA were used to verify the genetic signature internally and externally. Our results showed that our gene signature had a good predictive ability for recurrence and prognosis. CONCLUSIONS: We established a four-gene signature and predictive model to predict the recurrence and corresponding survival rates in patients with early lung adenocarcinoma after surgery. These may be helpful for reforumulating post-operative consolidation treatment strategies.

Neutrophil-Lymphocyte Ratio as a Predictive Factor of Growing Malignant Pulmonary Ground-glass Opacity.

BACKGROUND: The object of the study was to elucidate the relationship between the neutrophil-lymphocyte ratio (NLR) and the growth of pulmonary ground-glass opacity (GGO) in stage IA lung adenocarcinoma. METHODS: All patients with GGO following surgical procedures were enrolled, with the time of follow-up and the variation tendency of GGO recorded. Meanwhile, laboratory parameters, age, gender, smoking history, histology, tumor size, and stage were recorded. Logistic regression was used to evaluate the value of NLR and the cutoff value was calculated by SPSS 22.0. RESULTS: In the whole cohort, 30 cases of growing GGO and 43 cases of stable GGO undergoing surgical procedures were diagnosed as lung adenocarcinoma. There was significant statistical difference between the two groups. Multivariable analysis showed that NLR could predict the GGOs with growth (odds ratio 5.198, 95% confidence interval (95%CI: 1.583-14.581, P=0.002). Receiver operating characteristics analysis for NLR showed the optimal cutoff value of 2.38, with a sensitivity of 60.0% and specificity of 81.4%. CONCLUSION: Our study demonstrated that the NLR appeared to have value as a promising clinical predictor of GGOs with growth. Further studies are needed to confirm this conclusion.

Landscape and dynamics of single tumor and immune cells in early and advanced-stage lung adenocarcinoma.

BACKGROUND: Lung adenocarcinoma (LUAD) patients with different American Joint Committee on Cancer stages have different overall 5-year survival rates. The tumor microenvironment (TME) and intra-tumor heterogeneity (ITH) have been shown to play a crucial role in the occurrence and development of tumors. However, the TME and ITH in different lesions of LUAD have not been extensively explored. METHODS: We present a 204,157-cell catalog of the TME transcriptome in 29 lung samples to systematically explore the TME and ITH in the different stages of LUAD. Traditional RNA sequencing data and complete clinical information were downloaded from publicly available databases. RESULTS: Based on these high-quality cells, we constructed a single-cell network underlying cellular and molecular features of normal lung, early LUAD, and advanced LUAD cells. In contrast with early malignant cells, we noticed that advanced malignant cells had a remarkably more complex TME and higher ITH level. We also found that compared with other immune cells, more differences in CD8+/CTL T cells, regulatory T cells, and follicular B cells were evident between early and advanced LUAD. Additionally, cell-cell communication analyses, revealed great diversity between different lesions of LUAD at the single-cell level. Flow cytometry and qRT-PCR were used to validate our results. CONCLUSION: Our results revealed the cellular diversity and molecular complexity of cell lineages in different stages of LUAD. We believe our research, which serves as a basic framework and valuable resource, can facilitate exploration of the pathogenesis of LUAD and identify novel therapeutic targets in the future.

The application and value of three-dimensional reconstruction technology in the differential diagnosis and surgical treatment of ground-glass opacity / 中华胸心血管外科杂志

Objective To explore the application and value of three-dimensional reconstruction technology in diagnosis and surgical therapy of ground-glass opacity.Methods We analyzed 188 cases with GGO during October 2012 and October 2014 in Shanghai First People’s Hospital affiliated to Shanghai Jiao tong University School of Medicine respectively.Several relative parameters based on three-dimensional reconstruction were selected to differentiate GGO pathology .By ROC curve anal-ysis, optimal cut-off points were determined.Volume-doubling time(VDT) was calculated among 104 cases which had been following up at least for three months and compared among AAH, MIA and IAC.Results Cases’ mean age was 56.66 ± 9.673years(ranged from 27 to 82 years) with 67 males and 121 females.The diameter(P<0.001), total volume(TV)(P<0.001), the maximum CT attenuation(MAX)(P<0.001) and standard deviation of distribution of CT attenuation within the whole GGO(STD)(P=0.015) of malignant group were significant larger than those of benign ones except for average CT at-tenuation(AVG)(P=0.094).The AUC were just 0.64-0.80 when differentiated benign group from AIS, benign group from MIA and AAH from MIA in comparison to 0.70-0.95 when it comes to differentiating benign group and AAH from IAC .Ad-ditionally, the optimal cut-off points were 13.5-15.5 mm in diameter, 400-600 mm3 in TV, -110 -20 Hu in MAX and 120-160 in STD.The mean VDT was 865.00 ±111.33, 464.67 ±44.40 and 238.36 ±76.71 for AAH, MIA and IAC re-spectively(AAH/MIA P<0.001, MIA/IAC P=0.003).Conclusion The diameter, TV, MAX and STD of GGO based on three-dimensional reconstruction could discriminate GGO availably .Combined utilization of HRCT and three-dimensional re-construction has a valuable significance in diagnosis and treatment of GGO .