Lung Biopsy Tissue Research Articles

Use of a pulmosphere model to evaluate drug antifibrotic responses in interstitial lung diseases

BackgroundInterstitial lung diseases (ILD) encompass a heterogenous group of diffuse parenchymal lung disorders characterized by variable degrees of inflammation and fibrosis. Pretherapeutic clinical testing models for such diseases can serve as a platform to test and develop effective therapeutic strategies. In this study, we developed patient derived 3D organoid model to recapitulate the disease process of ILDs. We characterized the inherent property of invasiveness in this model and tested for antifibrotic responses with an aim to develop a potential platform for personalized medicine in ILDs.MethodsIn this prospective study, 23 patients with ILD were recruited and underwent lung biopsy. 3D organoid-based models (pulmospheres) were developed from the lung biopsy tissues. Pulmonary functioning testing and other relevant clinical parameters were collected at the time of enrollment and follow up visits. The patient derived pulmospheres were compared to normal control pulmospheres obtained from 9 explant lung donor samples. These pulmospheres were characterized by their invasive capabilities and responsiveness to the antifibrotic drugs, pirfenidone and nintedanib.ResultsInvasiveness of the pulmospheres was measured by the zone of invasiveness percentage (ZOI%). The ILD pulmospheres (n = 23) had a higher ZOI% as compared to control pulmospheres (n = 9) (516.2 ± 115.6 versus 54.63 ± 19.6 respectively. ILD pulmospheres were responsive to pirfenidone in 12 of the 23 patients (52%) and responsive to nintedanib in all 23 patients (100%). Pirfenidone was noted to be selectively responsive in patients with connective tissue disease related ILD (CTD-ILD) at low doses. There was no correlation between the basal pulmosphere invasiveness, response to antifibrotics, and FVC change (Δ FVC).ConclusionsThe 3D pulmosphere model demonstrates invasiveness which is unique to each individual subject and is greater in ILD pulmospheres as compared to controls. This property can be utilized to test responses to drugs such as antifibrotics. The 3D pulmosphere model could serve as a platform for the development of personalized approaches to therapeutics and drug development in ILDs and potentially other chronic lung diseases.

Russian Registry of Idiopathic Pulmonary Fibrosis: Clinical Features, Treatment Management, and Outcomes.

This was a prospective multicenter, observational, non-interventional study. Patients' risk factors, demographics, clinical data, results of high-resolution computed tomography (HRCT) of the chest and pulmonary function testing, and lung tissue biopsy findings were analyzed. We also analyzed the exercise tolerance (6-min walking test) of patients, serological markers of systemic connective tissue diseases, treatment, clinical course, and outcomes of the disease. Multidisciplinary discussion (MDD) was used as needed. One thousand three hundred and fifty-three patients were included in the registry from 2016 to 2020. The mean age was 64.4 ± 10.7 years, most patients were active smokers or ex-smokers. Antifibrotic therapy was administered to 90 of 948 patients (9.5%). Since starting the registry in 2016, the incidences of IPF have increased and the time period from manifestation of the disease to making the diagnosis has shortened, the number of patients on antifibrotic therapy has increased and the number of patients taking systemic steroids decreased. The registry of patients with IPF was helpful to improve IPF diagnosis and to implement antifibrotic agents in clinical practice. Further analysis of the clinical course and prognostic markers of IPF in the Russian population is needed. An analysis of the long-term efficacy of antifibrotic therapy in this population is also important.

Swellable and Thermally Responsive Hydrogel/Shape Memory Polymer Foam Composites for Sealing Lung Biopsy Tracts.

Lung tissue biopsies can result in a leakage of blood (hemothorax) and air (pneumothorax) from the biopsy tract, which threatens the patient with a collapsed lung and other complications. We have developed a lung biopsy tract sealant based on a thiol-ene-crosslinked PEG hydrogel and polyurethane shape memory polymer (SMP) foam composite. After insertion into biopsy tracts, the PEG hydrogel component contributes to sealing through water-driven swelling, whereas the SMP foam contributes to sealing via thermal actuation. The gelation kinetics, swelling properties, and rheological properties of various hydrogel formulations were studied to determine the optimal formulation for composite fabrication. Composites were then fabricated via vacuum infiltration of the PEG hydrogel precursors into the SMP foam followed by thermal curing. After drying, the composites were crimped to enable insertion into biopsy tracts. Characterization revealed that the composites exhibited a slight delay in shape recovery compared to control SMP foams. However, the composites were still able to recover their shape in a matter of minutes. Cytocompatibility testing showed that leachable byproducts can be easily removed by washing and washed composites were not cytotoxic to mouse lung fibroblasts (L929s). Benchtop testing demonstrated that the composites can be easily deployed through a cannula, and the working time for deployment after exposure to water was 2 min. Furthermore, testing in an in vitro lung model demonstrated that the composites were able to effectively seal a lung biopsy tract and prevent air leakage. Collectively, these results show that the PEG hydrogel/SMP foam composites have the potential to be used as lung biopsy tract sealants to prevent pneumothorax post-lung biopsy.

A Millimeter-Scale Soft Robot for Tissue Biopsy Procedures.

While interest in soft robotics as surgical tools has grown due to their inherently safe interactions with the body, their feasibility is limited in the amount of force that can be transmitted during procedures. This is especially apparent in minimally invasive procedures where millimeter-scale devices are necessary for reaching the desired surgical site, such as in interventional bronchoscopy. To leverage the benefits of soft robotics in minimally invasive surgery, a soft robot with integrated tip steering, stabilization, and needle deployment capabilities is proposed for lung tissue biopsy procedures. Design, fabrication, and modeling of the force transmission of this soft robotic platform allows for integration into a system with a diameter of 3.5 mm. Characterizations of the soft robot are performed to analyze bending angle, force transmission, and expansion during needle deployment. In-vitro experiments of both the needle deployment mechanism and fully integrated soft robot validate the proposed workflow and capabilities in a simulated surgical setting.

Differences in interleukin-6 and interleukin-17 expression in covid-19 post-mortem lung tissue biopsy compared with noncovid- 19

Differences in interleukin-6 and interleukin-17 expression in covid-19 post-mortem lung tissue biopsy compared with noncovid- 19

Phenotypic and Genotypic Appraisal of Pulmonary Mucinous Adenocarcinoma: A Retrospective Study

Introduction: Lung cancer is the second most common cancer diagnosed in the world. According to GLOBOCAN 2020, it is approximately 11.4% of all malignancies and accounts for one in five cancer deaths worldwide. Several driver gene mutations are known in lung adenocarcinoma including Epidermal Growth Factor Receptor (EGFR), Kirsten rat sarcoma viral oncogene homolog(KRAS), v-raf murine sarcoma viral oncogene homolog B1(BRAF), Human epidermal growth factor receptor2 (HER 2), anaplastic lymphoma kinase (ALK), Ros Protooncogene 1, Receptor Tyrosine Kinase (ROS1) and rearranged during transfection (RET). Among these, KRAS mutation has been classically defined as bearing a negative prognostic factor. They are described to have unfavourable survival rates as compared with KRAS wild- type tumours. However, its real significance remains controversial due to heterogeneity amongst studies. Aim: The aim of this study was to assess the histomorphology, immunophenotype, EGFR and KRAS mutation profile in invasive mucinous and mixed mucinous/non mucinous type lung carcinoma. Materials and Methods: This retrospective study was done at a tertiary cancer centre in the state of Kerala, South India to identify the frequency of EGFR and KRAS mutations in invasive mucinous or mixed mucinous/non mucinous type lung carcinoma diagnosed during the period June 2017 to March 2019. Immunohistochemical assays for CK7, CK20, TTF-1, Napsin-A, CDX-2 and p63 were done on all cases. EGFR mutation analysis for exons 18, 19, 20, 21 were done using ARMS PCR and KRAS mutation analysis for codons 12, 13, 61, 146 by Sanger sequencer. Descriptive statistics was used wherever applicable. Results: Out of a total of 290 patients of pulmonary adenocarcinoma during the two year study period, twelve cases (4.1%) were diagnosed as invasive mucinous or mixed mucinous/non mucinous types based on histomorphology and immunophenotype, on both lung tissue biopsies as well as material from metastatic sites. Of the twelve, nine (75%) were pure mucinous and three (25%) were mixed mucinous and non mucinous. All the twelve cases were subjected to molecular testing for both EGFR and KRAS. Three cases (25%) showed KRAS mutations in codon 12. EGFR mutation (TKI resistant T790M mutation) was found only in a single case which had a histology of mixed mucinous and non mucinous carcinoma and a coexistent mutation in KRAS (c12 G>C) gene. Conclusions: Our study corroborates the described fact that pulmonary mucinous carcinoma differ from the conventional adenocarcinoma in terms of its histology, immunoprofile and molecular profile. KRAS mutation appears to be the main pathogenetic factor in this variant. In the current day practice, there is no demonstrable effective therapy for KRAS mutant lung adenocarcinoma and more trials are needed to identify newer mutations that can modify treatment outcomes.

Automated Particle Analysis Using Field-Emission Scanning Electron Microscopy (FE-SEM) and Energy Dispersive X-Ray Spectroscopy (EDS) to Characterize Inhaled Particulate Matter (PM) in Biopsied Lung Tissue

Abstract The role of inhaled particulate matter in the development of chronic lung disease in military veterans remains ill-defined and controversial. Characterizing the nature of such inhaled particulates in surgical lung biopsy tissue is an important step in evaluating the association between exposure and illness. We have developed a protocol for automated in-situ analysis of inorganic inhaled particulate matter using field-emission scanning electron microscopy with energy dispersive X-ray spectroscopy. Over 120,000 particles of both a geological and manufactured nature were analyzed and characterized according to their chemistry and morphology. The most abundant particles were clays, feldspars, silica, titanium dioxides, and metals and were by and large less than 2.5 µm in the greatest dimension. Automating the process allows for efficient in-situ analysis of thousands of particles each day and the development of large datasets.

The performance of detecting Mycobacterium tuberculosis complex in lung biopsy tissue by metagenomic next-generation sequencing

BackgroundTuberculosis (TB) is a chronic infectious disease caused by the Mycobacterium tuberculosis complex (MTBC), which is the leading cause of death from infectious diseases. The rapid and accurate microbiological detection of the MTBC is crucial for the diagnosis and treatment of TB. Metagenomic next-generation sequencing (mNGS) has been shown to be a promising and satisfying application of detection in infectious diseases. However, relevant research about the difference in MTBC detection by mNGS between bronchoalveolar lavage fluid (BALF) and lung biopsy tissue specimens remains scarce.MethodsWe used mNGS to detect pathogens in BALF and lung biopsy tissue obtained by CT-guide percutaneous lung puncture (CPLP) or radial endobronchial ultrasound transbronchial lung biopsy (R-EBUS-TBLB) from 443 hospitalized patients in mainland China suspected of pulmonary infections between May 1, 2019 and October 31, 2021. Aim to evaluate the diagnostic performance of mNGS for detecting MTBC and explore differences in the microbial composition in the 2 specimen types.ResultsAmong the 443 patients, 46 patients finally were diagnosed with TB, of which 36 patients were detected as MTBC positive by mNGS (8.93%). Striking differences were noticed in the higher detection efficiency of lung biopsy tissue compared with BALF (P = 0.004). There were no significant differences between the 2 specimen types in the relative abundance among the 27 pathogens detected by mNGS from the 36 patients.ConclusionsThis study demonstrates that mNGS could offer an effective detection method of MTBC in BALF or lung tissue biopsy samples in patients suspected of TB infections. When it comes to the situations that BALF samples have limited value to catch pathogens for special lesion sites or the patients have contraindications to bronchoalveolar lavage (BAL) procedures, lung biopsy tissue is an optional specimen for MTBC detection by mNGS. However, whether lung tissue-mNGS is superior to BALF-mNGS in patients with MTBC infection requires further prospective multicenter randomized controlled studies with more cases.

Prostatic Abscess – A Rare Causative Organism

A 60-year-old male with diabetes presented with fatigue, urinary frequency, and suprapubic pain for 2 weeks. Patient resides in southern California and denied other symptoms. Laboratory workup showed leukocytosis (16.4 bil/L) and hyperglycemia (463 mg/dL) and urine culture grew 50,000 cfu/mL mixed flora with negative blood cultures. After no clinical improvement with piperacillin/tazobactam for presumed UTI, computed tomography demonstrated a multi-loculated prostatic abscess with regional cystitis (Fig. 1). Chest imaging showed cavitary right upper lobe opacities and miliary pulmonary nodules (Fig. 2). Percutaneous drainage of the prostatic abscess and lung tissue biopsies were obtained (Fig. 3). Figure 2Computed tomography of the chest. (A) Cavitary right upper lobe lesion with background of extensive miliary pulmonary nodules. View Large Image Figure Viewer Download Hi-res image Figure 3Pathology results of lung tissue biopsies. (A) Coccidioides spherule (thick-walled spherule with endospores). View Large Image Figure Viewer Download Hi-res image

Correlation between cytology Oil Red O staining and lung biopsy specimens: utility of the lipid-laden macrophage index

Oil Red O staining is used for enumeration of the lipid-laden macrophage index (LLMI) as a surrogate for aspiration. As part of quality improvement efforts aimed at optimizing resource utilization, the utility of this stain in current cytopathology practice was re-evaluated. The objective of this study was to explore the clinical utility of Oil Red O staining in bronchoalveolar lavage (BAL) samples by correlating the LLMI with findings in concurrent histologic tissue samples. Lung transbronchial biopsy specimens that suggested aspiration that were submitted with concurrent BAL cytology samples were retrieved. Lung tissue biopsies were reviewed for the presence of foamy alveolar macrophages (graded as 0, 1+, and 2+), foreign material, and giant cells. The concurrent BAL was reviewed with consensus determination of the LLMI. A total of 53 cases were identified. On histology, 13 cases (24.5%) were found to have no foamy alveolar macrophages, 23 cases (43.4%) were found to have 1+ foamy alveolar macrophages, and 17 cases (32.1%) were found to have 2+ foamy alveolar macrophages. Six cases (11.3%) were found to have foreign material, and 10 cases (18.9%) were found to have multinucleated giant cells. The average LLMI score was 16, with 44 (83.0%) in the low range (LLMI <40) and 9 (17.0%) in the intermediate range (LLMI of 40-90). None of the cases in our study had an LLMI that exceeded the cutoff value for which aspiration would be suspected. We found no correlation of the LLMI with lung biopsies that suggested aspiration.

Causal Path of COPD Progression-Associated Genes in Different Biological Samples

Chronic obstructive pulmonary disease (COPD) is a progressive inflammatory disease with pulmonary and extra-pulmonary complications. Due to the disease’s systemic nature, many investigations investigated the genetic alterations in various biological samples. We aimed to infer causal genes in COPD’s pathogenesis in different biological samples using elastic-net logistic regression and the Structural Equation Model. Samples of small airway epithelial cells, bronchoalveolar lavage macrophages, lung tissue biopsy, sputum, and blood samples were selected (135, 70, 235, 143, and 226 samples, respectively). Elastic-net Logistic Regression analysis was implemented to identify the most important genes involved in COPD progression. Thirty-three candidate genes were identified as essential factors in the pathogenesis of COPD and regulation of lung function. Recognized candidate genes in small airway epithelial (SAE) cells have the highest area under the ROC curve (AUC = 97%, SD = 3.9%). Our analysis indicates that macrophages and epithelial cells are more influential in COPD progression at the transcriptome level.

Cardiac Sarcoidosis Managed by Biventricular Pacing: A Case Report

Background: Cardiac sarcoidosis (CS) is a rare pathology that affects the middle-aged population. Its diagnosis, as well as its treatment, can be quite challenging, especially dealing with device management. We hereby discuss a case of CS, managed by a biventricular pacemaker in a 43-year-old man. Case Report: A 43-year-old male presented to the emergency department with a 15-day history of chronic cough and worsening dyspnea. On examination, he was found to be bradycardic in the context of a third-degree heart block on ECG as well as bilateral hilar lymphadenopathy on chest radiography. Based on these findings, cardiac sarcoidosis was suspected. A temporary right internal jugular transvenous pacemaker was implanted in the right ventricle and empiric corticosteroid management was commenced. Lung tissue biopsies were obtained via bronchoscopy and were positive for granulomas consistent with a diagnosis of sarcoidosis. Even though the patient showed a partial positive response to corticosteroid treatment, he required a permanent pacemaker. A dual-chamber implantable cardioverter-defibrillator (ICD) was implanted and the patient was discharged on prednisone. His follow-up plan consisted of serial echocardiography and consideration of a new coronary sinus lead in the event of left ventricular dysfunction in the future. Conclusion: Cardiac rhythm management in the context of CS is often difficult, with decisions that need to be made between a pacemaker, a defibrillator, or a cardiac resynchronization device. The idea that biventricular pacing has a preventative role against heart failure in patients with normal left ventricular ejection fraction and CS deserves more attention and discussion.

Utilizing Natural Language Processing (NLP) to identify breast cancer associated-lung metastases from pathology reports to delineate characteristics and challenges of this common site of breast cancer recurrence.

e13592 Background: NLP (artificial intelligence) can automate the identification of records in large datasets. The purpose of this study was to evaluate the feasibility of NLP to identify breast cancer-associated lung metastases to understand clinical characteristics and challenges posed by this common site of breast cancer recurrence. Methods: Patients with pathologically confirmed breast cancer associated-lung metastases seen at a large academic center between 3/2012-5/2019 were identified using NLP of institutional pathology reports, with an IRB approved protocol. Chart review was performed to confirm breast cancer associated-lung metastases and determine clinical and pathological features. Results: Using NLP, 32 patients with pathology reports denoting breast cancer associated-lung metastases were identified, with pathologic confirmation of lung biopsy tissue in the majority of cases (24), and pleural fluid specimens (8) on the remainder. Ten of 32 (31%) were HR+/HER2-, 3/32 (9.3%) HER2+, and 19/32 (59%) TNBC. The majority were invasive ductal carcinoma (21/26) with the remainder invasive lobular carcinoma (2/26) or mixed histology (3/26). Median age at lung metastasis diagnosis was 62 years (range 31-88). The median time to development of lung metastasis following primary breast cancer was 5.6 years (range 0-24.8 years). Fifty six percent of lung metastases were detected on imaging and 44% by symptoms including dyspnea, cough, or pain. Tumor tissue genotyping results on the lung metastases were available for 8 patients showing PI3KCA (5), TP53 (3), SMARCA4 (2), ERBB2 (1), FGFR3 (1), ATM (1), CDK4 (1), MYC (1), and ESR1 (1). Treatment after diagnosis of lung metastases included hormone therapy (61%), chemotherapy (84%), lung irradiation (26%), and surgical resection of lung metastases (6%). Lung metastases were associated with considerable morbidity including pleural effusion (15%), dyspnea (6%), pneumothorax (3%), hemothorax (3%), and atelectasis (3%). Patients diagnosed with lung metastases had brain (32%), bone (35%), renal (6%), skin (3%) and adrenal (3%) metastases during disease course. Conclusions: NLP can help identify organ specific metastases from pathology reports, such as breast cancer associated-lung metastases, which can then facilitate observational, translational, and clinical research to characterize and address challenges posed by this common site of breast cancer recurrence. This cohort of patients highlights the morbidity of breast cancer associated-lung metastases and potential role of NLP for disease characterization and clinical research. (Support from ASCO Medical Student Rotation for Underrepresented Populations Award.)

Extracellular Vesicle-Based Bronchoalveolar Lavage Fluid Liquid Biopsy for EGFR Mutation Testing in Advanced Non-Squamous NSCLC.

Simple SummaryTissue biopsy is the gold standard for molecular genotyping in lung cancer. However, obtaining tumor tissue is challenging due to its invasiveness, inadequate amount of tissue, or complications. To overcome the limitations of tissue biopsy, plasma liquid biopsy using cfDNA has been investigated extensively; however, its low sensitivity limits the clinical application. Therefore, we used the tumor-specific DNA of extracellular vesicles (EVs) in bronchoalveolar lavage fluid (BALF) as DNA source for EGFR genotyping. As a result, we demonstrated that EV-based BALF EGFR testing in advanced lung NSCLC is a highly accurate rapid method overcoming low sensitivity of plasma cfDNA-based EGFR genotyping. It can be used as an adjuvant or alternative method for lung biopsy in cases where obtaining an adequate amount of tissue is difficult.To overcome the limitations of the tissue biopsy and plasma cfDNA liquid biopsy, we performed the EV-based BALF liquid biopsy of 224 newly diagnosed stage III-IV NSCLC patients and compared it with tissue genotyping and 110 plasma liquid biopsies. Isolation of EVs from BALF was performed by ultracentrifugation. EGFR genotyping was performed through peptide nucleic acid clamping-assisted fluorescence melting curve analysis. Compared with tissue-based genotyping, BALF liquid biopsy demonstrated a sensitivity, specificity, and concordance rates of 97.8%, 96.9%, and 97.7%, respectively. The performance of BALF liquid biopsy was almost identical to that of standard tissue-based genotyping. In contrast, plasma cfDNA-based liquid biopsy (n = 110) demonstrated sensitivity, specificity, and concordance rates of 48.5%, 86.3%, and 63.6%, respectively. The mean turn-around time of BALF liquid biopsy was significantly shorter (2.6 days) than that of tissue-based genotyping (13.9 days; p < 0.001). Therefore, the use of EV-based BALF shortens the time for confirmation of EGFR mutation status for starting EGFR-TKI treatment and can hence potentially improve clinical outcomes. As a result, we suggest that EV-based BALF EGFR testing in advanced lung NSCLC is a highly accurate rapid method and can be used as an alternative method for lung tissue biopsy.

Expression of CD91 in extracellular vesicles: A potential biomarker for the diagnosis of non-small cell lung cancer

Lung cancer is the leading cause of death; by the time it is diagnosed, the patient is usually in late-stage grade IV. Late-stage lung cancer is mainly associated with metastasis in the liver, brain, and lymphoid tissues; as a result, a localized lung cancer treatment remains meaningless. Early diagnosis of non-small cell lung cancer (NSCLC) may be curable or will improve the survival rate. Although with advanced developments to screen high-risk patients by LDCT scan, false-positive rates and limited resolution necessitated the development of advanced diagnostic techniques for NSCLC. Extracellular vesicles (EVs) released from cells freely circulate in the blood and contain various transmembrane proteins, and they may be a non-invasive biomarker for cancer diagnosis and prognosis. Current studies predict that the CD91 marker in EVs may be a potential biomarker of NSCLC; however, the expression of CD91 in lung cancer tissues is not fully known. Here, this study determines the differential expression of CD91 in lung cancer cells and in circulating EVs in blood as a potential biomarker of NSCLC patients. Our results confirmed the expression of CD91 in NCI–H1975 cultured cells and NSCLC lung biopsy tissues. Furthermore, biophysical characterization of EVs from lung cancer cells determines the substantial expression of CD91, which the Transmission Electron Microscope confirms. Thus, this study suggests EVs containing CD91 could be an asset to studying the development of diagnostic and prognostic biomarkers in NSCLC disease.

Cryptococcal antigen testing of lung tissue homogenate improves pulmonary cryptococcosis diagnosis: Two case reports

BACKGROUNDPulmonary cryptococcosis (PC) is an opportunistic infectious disease of the respiratory system. Lung tissue biopsies, culture of respiratory samples (e.g., sputum, lung tissue, pleural fluid, and bronchoalveolar lavage fluid), and cryptococcal antigen (CrAg) testing are helpful for a definitive diagnosis. However, these tests are sometimes falsely negative. PC is often misdiagnosed or underdiagnosed owing to the absence of obvert symptoms, poor imaging specificity, and false-negative laboratory tests.CASE SUMMARYWe report two female patients who underwent computed tomography-guided percutaneous needle pulmonary biopsy of a lung nodule for a confirmed diagnosis. In both patients, the CrAg test on the lung biopsy tissue homogenate was positive, while the serum CrAg test was negative. Combined with the lung tissue pathology, we made the diagnosis of PC. Antifungal therapy was effective in both patients.CONCLUSIONGiven the findings of our cases and the literature review, lung tissue homogenate CrAg testing can be helpful in improving the diagnosis of PC.

Performance of rapid on-site evaluation of touch imprints of lung tissue biopsies for the diagnosis of pulmonary cryptococcosis in patients without HIV infection.

The diagnosis of pulmonary cryptococcosis depends on serum testing, histopathology and mycological culture; there are few studies on touch imprints of lung tissue biopsies for the diagnosis of pulmonary cryptococcosis in patients without HIV infection. The purpose of the current study was to investigate the accuracy and timeliness of on-site touch imprint cytology in the diagnosis of pulmonary cryptococcosis during CT-guided percutaneous lung biopsy. We retrospectively analysed the diagnosis and treatment of 56 patients with final proof of pulmonary cryptococcosis through histopathology and culture or surgical resection from September 2015 to February 2021. Diagnostic methods and treatment and the turnaround time for diagnosis were analysed. The sensitivity of rapid on-site evaluation was 89.3%, and the sensitivity of serology, histopathology and mycological culture was 53.6%, 91.1% and 61.5%, respectively, compared with the final diagnosis. The average turnaround time to diagnose pulmonary cryptococcosis by on-site touch imprint cytology was 8.3±0.9min, which was significantly faster than serum testing, histopathology and mycological culture. On-site touch imprint cytology showed good sensitivity and timeliness in the diagnosis of pulmonary cryptococcosis. In addition, it contributed to the triage of biopsies based on the preliminary diagnosis. On-site touch imprint cytology should be applied and promoted in the diagnosis of pulmonary cryptococcosis during biopsy.

Diagnostic performance of the metagenomic next-generation sequencing in lung biopsy tissues in patients suspected of having a local pulmonary infection

PurposeThis study aims to evaluate the diagnostic application and performance of the metagenomic next-generation sequencing (mNGS) in patients suspected of local pulmonary infection by comparing it to the traditional pathogen detection methods in lung tissue specimens obtained by a computerized tomography-guided biopsy (CT-guided biopsy).MethodsWe retrospectively reviewed patients, admitted to the First Affiliated Hospital of Wenzhou Medical University, China from May 2018 to December 2020, who were suspected of local pulmonary infection. All cases received a CT-guided lung biopsy, tissue samples were sent both for conventional examinations (CE) and mNGS tests. The sensitivity and specificity of the two diagnostic approaches were compared.Results106 patients enrolled, 76 patients were diagnosed with a pulmonary infection. Among 49 patients with identified pathogens, CE confirmed pathogenic infections in 32 cases. Mycobacterium spp. and fungi accounted for 37.5% (12/32) and 28.1% (9/32), respectively, with bacteria 34.4% (11/32). The mNGS examination detected extra pathogenic microorganisms in 22 patients that were consistent with the patients' clinical and radiographic pictures. The sensitivity of mNGS was 53.9% vs. 42.1% for the CE, while the specificity was 56.7% versus 96.7%. For detection rate, mNGS was significantly superior to CE in bacterial (96.3% vs. 40.7%, p < 0.05), and mixed infections (100% vs. 50%, p < 0.05), but inferior to CE in fungal (60% vs. 90%, p > 0.05) and Mycobacterium spp. infections (66.7% vs. 100%, p > 0.05) with no significant difference. Among 31 cases diagnosed with lung abscess, the diagnostic performance of the detection rate was 67.7% (21/31) in favour of mNGS compared to 29.0% (9/31) for CE (p < 0.05). Most polymicrobial infections were induced by anaerobic species that coexisted with Streptococcus constellatus. And Klebsiella pneumoniae was the most common isolated monomicrobial infection.ConclusionsThe most commonly detected causative pathogens for local pulmonary infections were bacteria, Mycobacterium spp. and fungi. Compared with the CE, the advantages of mNGS in the pathogens detection lie in the discovery of bacterial and mixed infections, as well as in the detection of lung abscess. Conversely, mNGS is not good enough to be recommendable for the detection of Mycobacterium spp. and fungi.

Blood-based liquid biopsy: Insights into early detection and clinical management of lung cancer.

Currently, early detection of lung cancer relies on the characterisation of images generated from computed tomography (CT). However, lung tissue biopsy, a highly invasive surgical procedure, is required to confirm CT-derived diagnostic results with very high false-positive rates. Hence, a non-invasive or minimally invasive biomarkers is essential to complement the existing low-dose CT (LDCT) for early detection, improve responses to a certain treatment, predict cancer recurrence, and to evaluate prognosis. In the past decade, liquid biopsies (e.g., blood) have been demonstrated to be highly effective for lung cancer biomarker discovery. In this review, the roles of emerging liquid biopsy-derived biomarkers such as circulating nucleic acids, circulating tumour cells (CTCs), long non-coding RNA (lncRNA), and microRNA (miRNA), as well as exosomes, have been highlighted. The advantages and limitations of these blood-based minimally invasive biomarkers have been discussed. Furthermore, the current progress of the identified biomarkers for clinical management of lung cancer has been summarised. Finally, a potential strategy for the early detection of lung cancer, using a combination of LDCT scans and well-validated biomarkers, has been discussed.

Analysis of the Single-Cell Heterogeneity of Adenocarcinoma Cell Lines and the Investigation of Intratumor Heterogeneity Reveals the Expression of Transmembrane Protein 45A (TMEM45A) in Lung Adenocarcinoma Cancer Patients.

Simple SummaryNon-small cell lung cancer (NSCLC) is one of the main causes of cancer-related deaths worldwide. Intratumoral heterogeneity (ITH) is responsible for the majority of difficulties encountered in the treatment of lung-cancer patients. Therefore, the heterogeneity of NSCLC cell lines and primary lung adenocarcinoma was investigated by single-cell mass cytometry (CyTOF). Human NSCLC adenocarcinoma cells A549, H1975, and H1650 were studied at single-cell resolution for the expression pattern of 13 markers: GLUT1, MCT4, CA9, TMEM45A, CD66, CD274, CD24, CD326, pan-keratin, TRA-1-60, galectin-3, galectin-1, and EGFR. The intra- and inter-cell-line heterogeneity of A549, H1975, and H1650 cells were demonstrated through hypoxic modeling. Additionally, human primary lung adenocarcinoma, and non-involved healthy lung tissue were homogenized to prepare a single-cell suspension for CyTOF analysis. The single-cell heterogeneity was confirmed using unsupervised viSNE and FlowSOM analysis. Our results also show, for the first time, that TMEM45A is expressed in lung adenocarcinoma. Intratumoral heterogeneity (ITH) is responsible for the majority of difficulties encountered in the treatment of lung-cancer patients. Therefore, the heterogeneity of NSCLC cell lines and primary lung adenocarcinoma was investigated by single-cell mass cytometry (CyTOF). First, we studied the single-cell heterogeneity of frequent NSCLC adenocarcinoma models, such as A549, H1975, and H1650. The intra- and inter-cell-line single-cell heterogeneity is represented in the expression patterns of 13 markers—namely GLUT1, MCT4, CA9, TMEM45A, CD66, CD274 (PD-L1), CD24, CD326 (EpCAM), pan-keratin, TRA-1-60, galectin-3, galectin-1, and EGFR. The qRT-PCR and CyTOF analyses revealed that a hypoxic microenvironment and altered metabolism may influence cell-line heterogeneity. Additionally, human primary lung adenocarcinoma and non-involved healthy lung tissue biopsies were homogenized to prepare a single-cell suspension for CyTOF analysis. The CyTOF showed the ITH of human primary lung adenocarcinoma for 14 markers; particularly, the higher expressions of GLUT1, MCT4, CA9, TMEM45A, and CD66 were associated with the lung-tumor tissue. Our single-cell results are the first to demonstrate TMEM45A expression in human lung adenocarcinoma, which was verified by immunohistochemistry.