Lung cancer is a highly heterogeneous disease. Cancer cells and cells within the tumor microenvironment together determine disease progression, as well as response to or escape from treatment. To map ...the cell type-specific transcriptome landscape of cancer cells and their tumor microenvironment in advanced non-small cell lung cancer (NSCLC), we analyze 42 tissue biopsy samples from stage III/IV NSCLC patients by single cell RNA sequencing and present the large scale, single cell resolution profiles of advanced NSCLCs. In addition to cell types described in previous single cell studies of early stage lung cancer, we are able to identify rare cell types in tumors such as follicular dendritic cells and T helper 17 cells. Tumors from different patients display large heterogeneity in cellular composition, chromosomal structure, developmental trajectory, intercellular signaling network and phenotype dominance. Our study also reveals a correlation of tumor heterogeneity with tumor associated neutrophils, which might help to shed light on their function in NSCLC.
Lung cancer is one of the leading causes of cancer-related death worldwide. Cytology plays an important role in the initial evaluation and diagnosis of patients with lung cancer. However, due to the ...subjectivity of cytopathologists and the region-dependent diagnostic levels, the low consistency of liquid-based cytological diagnosis results in certain proportions of misdiagnoses and missed diagnoses. In this study, we performed a weakly supervised deep learning method for the classification of benign and malignant cells in lung cytological images through a deep convolutional neural network (DCNN). A total of 404 cases of lung cancer cells in effusion cytology specimens from Shanghai Pulmonary Hospital were investigated, in which 266, 78, and 60 cases were used as the training, validation and test sets, respectively. The proposed method was evaluated on 60 whole-slide images (WSIs) of lung cancer pleural effusion specimens. This study showed that the method had an accuracy, sensitivity, and specificity respectively of 91.67%, 87.50% and 94.44% in classifying malignant and benign lesions (or normal). The area under the receiver operating characteristic (ROC) curve (AUC) was 0.9526 (95% confidence interval (CI): 0.9019-9.9909). In contrast, the average accuracies of senior and junior cytopathologists were 98.34% and 83.34%, respectively. The proposed deep learning method will be useful and may assist pathologists with different levels of experience in the diagnosis of cancer cells on cytological pleural effusion images in the future.
Abstract
Background
Malignant transformation and progression of cancer are driven by the co-evolution of cancer cells and their dysregulated tumor microenvironment (TME). Recent studies on ...immunotherapy demonstrate the efficacy in reverting the anti-tumoral function of T cells, highlighting the therapeutic potential in targeting certain cell types in TME. However, the functions of other immune cell types remain largely unexplored.
Results
We conduct a single-cell RNA-seq analysis of cells isolated from tumor tissue samples of non-small cell lung cancer (NSCLC) patients, and identify subtypes of tumor-infiltrated B cells and their diverse functions in the progression of NSCLC. Flow cytometry and immunohistochemistry experiments on two independent cohorts confirm the co-existence of the two major subtypes of B cells, namely the naïve-like and plasma-like B cells. The naïve-like B cells are decreased in advanced NSCLC, and their lower level is associated with poor prognosis. Co-culture of isolated naïve-like B cells from NSCLC patients with two lung cancer cell lines demonstrate that the naïve-like B cells suppress the growth of lung cancer cells by secreting four factors negatively regulating the cell growth. We also demonstrate that the plasma-like B cells inhibit cancer cell growth in the early stage of NSCLC, but promote cell growth in the advanced stage of NSCLC. The roles of the plasma-like B cell produced immunoglobulins, and their interacting proteins in the progression of NSCLC are further validated by proteomics data.
Conclusion
Our analysis reveals versatile functions of tumor-infiltrating B cells and their potential clinical implications in NSCLC.
There are few studies on excimer laser (308 nm) atherectomy in the treatment of infrapopliteal artery disease. The purpose of this retrospective clinical study was to assess the efficacy and safety ...of excimer laser atherectomy (ELA) in combination with adjuvant drug-coated balloon angioplasty (DCB) compared to DCB for infrapopliteal arterial revascularization in patients with ischemic diabetic foot. From September 2018 to February 2019, a total of 79 patients with diabetic foot were treated for infrapopliteal arterial revascularization at Tianjin First Central Hospital (Tianjin, China). In this project, 35 patients were treated with ELA combined with DCB angioplasty, and 44 patients were treated with DCB angioplasty. The patients’ baseline characteristics were similar between the 2 groups. The primary efficacy endpoints through 24 months were clinically driven target lesion revascularization (CD-TLR), wound healing rate, major amputation rate, and target vessel patency rate. The primary safety endpoint through 24 months was all-cause mortality. The primary efficacy results at 24 months of ELA + DCB versus DCB were CD-TLR of 14.3% versus 34.1% (
p
= 0.044), wound healing rate of 88.6% versus 65.9% (
p
= 0.019), target vessel patency rate of 80.0% versus 52.3% (
p
= 0.010), and major amputations rate of 5.7% versus 22.7% (
p
= 0.036). The safety signal at 24 months of all-cause mortality rate was 2.9% for ELA + DCB group and 4.5% for DCB group (
p
= 0.957). ELA combined with DCB angioplasty is more effective than DCB in the treatment of infrapopliteal artery disease in patients with ischemic diabetic foot, which can improve the wound healing rate and target vessel patency rate. There was no statistical difference in the safety results between the two groups.
To depict the genomic landscape of Chinese early-stage lung squamous cell carcinoma (LUSC) and investigate its correlation with tumor mutation burden (TMB), PD-L1 expression, and immune infiltrates.
...Whole-exome sequencing was performed on 189 surgically resected LUSC. TMB was defined as the sum of nonsynonymous single nucleotide and indel variants. CD8
tumor-infiltrating lymphocyte (TIL) density and PD-L1 expression were evaluated by immunohistochemistry. Six immune infiltrates were estimated using an online database.
The median TMB was 9.43 mutations per megabase. Positive PD-L1 expression and CD8
TILs density were identified in 24.3% and 78.8%. PIK3CA amplification was associated with significantly higher TMB (P = 0.036). Frequent genetic alterations had no impact on PD-L1 expression but PIK3CA amplification and KEAP1 mutation were independently associated with significantly lower CD8
TIL density (P < 0.001, P = 0.005, respectively). Low TMB and high CD8
TIL density were independently associated with longer disease-free survival (DFS) while none of them could individually predict the overall survival (OS). Combination of TMB and PD-L1 expression or TMB and CD8
TIL density could stratify total populations into two groups with distinct prognosis. Classifying tumor-immune microenvironment based on PD-L1 expression and CD8
TIL density showed discrepant genomic alterations but similar TMB, clinical features, and OS. Notably, patients with different smoking status had distinct prognostic factors.
The combination of TMB, PD-L1 expression, immune infiltrates, and smoking status showed the feasibility to subgroup stratification in Chinese patients with early-stage LUSC, which might be helpful for future design of personalized immunotherapy trials in LUSC.
Mesothelioma, with various clinical manifestations, radiological features, and histomorphological types, can be divided into epithelioid, sarcomatoid, and biphasic types, according to their ...histomorphological characteristics. There is a rare growth pattern of pleural mesothelioma: diffuse intrapulmonary mesothelioma (DIM), with a distinctive pattern of predominantly intrapulmonary growth, has no or minimal pleural involvement, and simulates interstitial lung disease(ILD) clinically and radiologically. A 59-year-old man presented to the hospital with recurrent pleural effusions for 4 years and a history of asbestos exposure. Computed tomography (CT) showed bilateral pure ground-glass opacity lesions, and the tumor cells showed a lepidic growth pattern pathologically. Immunohistochemical staining was positive for CK, WT-1, calretinin, D2-40, CK5/6, and Claudin4, while TTF-1, CEA, EMA, CK7, CK20, and other epithelial markers were negative. BAP1 loss its expression, and MTAP was positive in cytoplasm. CDKN2A was negative tested by Fluorescence in situ hybridization (FISH). The final diagnosis was DIM. In conclusion, we should recognize this rare disease to avoid misdiagnosis and delayed treatment.
Our aim was to validate and analyze the prognostic impact of the novel International Association for the Study of Lung Cancer (IASLC) Pathology Committee grading system for invasive pulmonary ...adenocarcinomas (IPAs) in Chinese patients and to evaluate its utility in predicting a survival benefit from adjuvant chemotherapy (ACT). In this multicenter, retrospective, cohort study, we included 926 Chinese patients with completely resected stage I IPAs and classified them into three groups (Grade 1, n = 119; Grade 2, n = 431; Grade 3, n = 376) according to the new grading system proposed by the IASLC. Recurrence-free survival (RFS) and overall survival (OS) were estimated by the Kaplan-Meier method, and prognostic factors were assessed using univariable and multivariable Cox proportional hazards models. All included cohorts were well stratified in terms of RFS and OS by the novel grading system. Furthermore, the proposed grading system was found to be independently associated with recurrence and death in the multivariable analysis. Among patients with stage IB IPA (N = 490), the proposed grading system identified patients who could benefit from ACT but who were undergraded by the adenocarcinoma (ADC) classification. The novel grading system not only demonstrated prognostic significance in stage I IPA in a multicenter Chinese cohort but also offered clinical value for directing therapeutic decisions regarding adjuvant chemotherapy.
Visceral pleural invasion (VPI) is a critical component in the staging of peripheral non-small cell lung carcinoma (NSCLC). We aim to investigate whether dual-block elastic stain increases visceral ...pleural invasion positivity compared with single-block elastic stain. We further analyze the potential predictors of visceral pleural invasion. 8419 peripheral NSCLC patients (including 6008 patients with tumor size≤3 cm in stage I) were divided into a cohort using one paraffin block (single-block group, n = 5184) and a cohort using dual paraffin blocks (dual-block group, n = 3235) for elastic stain. The VPI-positive rate demonstrated by the dual-block elastic stains group was significantly higher than that of the single-block elastic stain group (17.7% (573/3235) versus 9.1% (474/5184), respectively, P < .001). The presence of visceral pleural invasion in T1 (≤3 cm) patients detected by single- and dual-block elastic stain was 6.3% (235/3730) and 12.0% (273/2278), respectively (P < .001). 5.7% of T1 patients (stage IA) were additionally upstaged to T2a (stage IB) by dual-block elastic stain. However, the incidence of visceral pleural invasion in pT2a patients showed no significant difference between the single-block group and the dual-block group (16.8% vs. 17.1%, P = .916). Lymphovascular invasion, lymph node metastasis, dedifferentiated carcinomas, the presence of spread through airspaces (STAS) and a poorly differentiated adenocarcinomatous growth pattern could be significant predictors of visceral pleural invasion (P < .001). Our results indicate that using dual-block elastic stain identifies more visceral pleural invasion positive T1 NSCLC patients who are upstaged to T2a, and who could benefit from optimal management post-operatively. The application of dual-block elastic stain is an efficient and practical method to detect visceral pleural invasion status.
•Droplet digital PCR (ddPCR) showed greatly improved sensitivity over real-time PCR for detecting Mycobacterium tuberculosis (MTB) in formalin-fixed and paraffin-embedded samples.•ddPCR is a ...sensitive approach for the detection of MTB in both pulmonary and extrapulmonary samples.•The ddPCR method could be used as an additional tool for the diagnosis of tuberculosis from pathological samples.
Droplet digital PCR (ddPCR) is a technology that has higher sensitivity than real-time PCR for the identification of trace DNA. However, the use of ddPCR for the detection of Mycobacterium tuberculosis DNA in pathological samples has not been fully studied.
A total of 65 formalin-fixed and paraffin-embedded (FFPE) specimens were included in this study. Twenty samples with definite results for tuberculosis (TB) were used to establish the ddPCR system for TB detection. ddPCR was then conducted to detect TB DNA in the 45 patients who were classified as ‘possible TB’ (real-time PCR results in the gray area, Ziehl–Neelsen staining-negative, and hematoxylin and eosin staining showing morphology suspicious for TB). The clinical treatment and disease outcomes were followed to assess the accuracy of ddPCR in the detection of TB DNA.
Among the 45 possible TB samples, 26 were ddPCR-positive, 12 were ddPCR-negative, and seven were in the gray area. ddPCR improved the positive rate of 57.8% (26/45) for the samples that were in the gray area by real-time PCR. Moreover, several patients received anti-TB therapy, and the effective ratio of therapy for the ddPCR-positive, ddPCR-negative, and ddPCR-gray area cases was 61.9% (13/21), 50.0% (2/4), and 33.3% (1/3), respectively.
ddPCR is more sensitive for detecting mild TB via FFPE samples than real-time PCR. The ddPCR method is of additional value in the diagnosis of TB from pathological samples.
Brain metastasis is a major cause for cancer death in patients with lung cancer. Sirtuin 1 (SIRT1) and hsa-miR-217 has been identified to mediate the development of non-small cell lung cancer ...(NSCLC).
We performed this study to investigate the roles of hsa-miR-217, its target SIRT1, as well as P53/KAI1 axis in the brain metastasis from NSCLC.
This is a cell culture study.
Human pulmonary adenocarcinoma brain metastasis cell line PC-14/B were incubated and treated with constructed lentiviral plasmids expressing miR-217 and/or SIRT1. BEAS-2B cell line was used as a control. The targeted regulation of miR-217 to SIRT1 was examined using dual luciferase reporter assay. Cell proliferation, migration, invasion and the expression of relate proteins were detected to examine the effect of miR-217/SIRT1 expression on metastasis.
PC-14/B cells expressed higher SIRT1 and lower p53 and KAI1 compared with BEAS-2B control cells (P<0.05). SIRT1 was a direct target of miR-217. MiR-217 expression suppressed PC-14/B cell invasion (P=0.004), migration (P=0.001) and proliferation (P<0.05), whereas SIRT1 overexpression reversed all processes. SIRT1 expression inhibited P53, KAI1/CD82, matrix metalloproteinase (MMP)-9 and β-catenin but upregulated E-cadherin protein. MiR-217 overexpression induced reverse changes.
Hsa-miR-217 and its target SIRT1 acted as metastasis suppressor and promoter gene in NSCLC, respectively. The hsa-miR-217/SIRT1/P53/KAI1 metastasis regulatory pathway showed novel and crucial roles in brain metastasis from NSCLC. This axis might be a potential target for the treatment of brain metastasis of lung cancer.
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