Lung Cancer Biopsies Research Articles

Proportion of biopsy specimens containing a tumor when compared to all biopsy specimens by transbronchial biopsy.

The lung cancer biopsy specimens obtained by endobronchial ultrasound-guide sheath (EBUS-GS) trans lung biopsy occasionally do not contain cancer cells. It is a problem that there is a possibility that they may not contain cancer cells. To investigate the proportion of biopsy specimens containing cancer cells in total biopsy specimens. Patients with lung cancer diagnosed by EBUS-GS were selected. The primary end point was the proportion of specimens containing tumors in the total specimens obtained by EBUS-GS. Twenty-six patients were investigated. The percentage of specimens containing cancer cells in the total specimens was 79.0%. The proportion of biopsy specimens containing cancer cell to all biopsy specimens by EBUS-GS was high, but not 100%.

Gene expression of programmed cell death ligand-1 (PDL-1) and vitamin D receptor (VDR) with the serum vitamin D3 in lung cancer

Abstract Background Lung cancer (LC) is one of the leading causes of death worldwide. Programmed cell death receptor 1 (PD-1) interacts with its ligand (PDL-1) on T cells inhibiting its functioning which may affect the patient's immunological response. Aim Investigate if there is a link between smoking and tissue expression of PDL-1 and vitamin D receptor (VDR) in lung cancer patients. In addition, the relation of vitamin D with smoking and these biochemical markers. Methods PDL-1 and VDR expressions were evaluated by real-time PCR in 54 lung cancer biopsy samples and 36 controls to prove this hypothesis. Vitamin D levels in the blood were measured using an ELISA. Results Expressions of PDL-1 were significantly upregulated in LC patients than in controls. The highest expression was in stage II and in squamous cell carcinoma followed by small cell carcinoma then adenocarcinoma. However, VDR expressions and vitamin D levels in serum were significantly downregulating in LC patients than in controls. There was a positive correlation between PDL-1expression and duration of smoking but not smoking index. Also, there is an inverse correlation between duration of smoking, smoking index, and VDR. Conclusion Expression of PDL-1 in LC was significantly upregulated and correlated with staging. Interestingly, our current study for the first time explained the role of duration of smoking on PDL-1 and VDR in the pathogenesis of LC. As PDL-1 expression increased with duration of smoking whereas VDR decreased, this novel findings may provide a possible link between the cumulative effect of smoking and the level of expressions of these biomarkers.

Preliminary Experience of Liquid Biopsy in Lung Cancer Compared to Conventional Assessment: Light and Shadows.

To assess the qualitative relationship between liquid biopsy and conventional tissue biopsy. As a secondary target, we evaluated the relationship between the liquid biopsy results and the T stage, N stage, M stage, and compared to grading. The Local Ethics Committee of the "Università degli Studi della Campania Luigi Vanvitelli", with the internal resolution number 24997/2020 of 12.11.2020, approved this spontaneous prospective study. According to the approved protocol, patients with lung cancer who underwent Fine-Needle Aspiration Cytology (FNAC), CT-guided biopsy, and liquid biopsy were enrolled. A Yates chi-square test was employed to analyze differences in percentage values of categorical variables. A p-value < 0.05 was considered statistically significant. Data analysis was performed using the Matlab Statistic Toolbox (The MathWorks, Inc., Natick, MA, USA). When a genetic mutation is present on the pathological examination, this was also detected on the liquid biopsy. ROS1 and PDL1 mutations were found in 2/29 patients, while EGFR Exon 21 was identified in a single patient. At liquid biopsy, 26 mutations were identified in the analyzed samples. The mutations with the highest prevalence rate in the study populations were: ALK (Ile1461Val), found in 28/29 patients (96.6%), EML4 (Lys398Arg), identified in 16/29 (55.2%) patients, ALK (Asp1529Glu), found in 14/29 (48.3%) patients, EGFR (Arg521Lys), found in 12/29 (41.4%) patients, ROS (Lys2228Gln), identified in 11/29 (37.9%) patients, ROS (Arg167Gln) and ROS (Ser2229Cys), identified in 10/29 (34.5%) patients, ALK (Lys1491Arg) and PIK3CA (Ile391Met), identified in 8/29 (27.6%) patients, ROS (Thr145Pro), identified in 6/29 (20.7%) patients, and ROS (Ser1109Leu), identified in 4/29 (13.8%) patients. No statistically significant differences can be observed in the mutation rate between the adenocarcinoma population and the squamous carcinoma population (p > 0.05, Yates chi-square test). We showed that, when a genetic mutation was detected in pathological examination, this was always detected by liquid biopsy, demonstrating a very high concordance rate of genomic testing between tissues and their corresponding mutations obtained by liquid biopsy, without cases of false-negative results. In addition, in our study, liquid biopsy highlighted 26 mutations, with the prevalence of ALK mutation in 96.6% of patients, supporting the idea that this approach could be an effective tool in cases with insufficient tumor tissue specimens or in cases where tissue specimens are not obtainable.

Effects of preoperative needle biopsy for lung cancer on survival and recurrence: a systematic review and meta-analysis.

Needle biopsy (NB) is used for the diagnosis of lung cancer, but there is still controversy about its effect on the prognosis after surgery. We conducted this meta-analysis to compare the prognosis of lung cancer patients who underwent preoperative NB with that of those who did not. We systematically searched seven databases and Google Scholar for eligible studies. Recurrence-free survival (RFS) and overall survival (OS) were analyzed as primary outcome measures. Nine articles with a collective total of 13,541 patients (NB group, n = 4550; non-NB group, n = 8991) were included in our meta-analysis. OS [hazard ratio (HR) = 1.43 (0.96, 2.12), p = 0.08] and RFS (HR = 1.59 [1.25, 2.01], p = 0.0001) tended to be better in the non-NB group than in the NB group. Pleural recurrence (risk ratio (RR) = 2.40 [1.42, 4.07], p = 0.001) was significantly lower in the non-NB group than in the NB group. The recurrence analysis data did not reach significance, but the overall trend was better for the non-NB group. These findings demonstrate that NB is detrimental to the survival prognosis of lung cancer patients and increases the chance of pleural recurrence.

Assessment of Early Detection by Multi-omics-Based Liquid Biopsy in Lung Cancer: A Prospective Study (ASCEND-LUNG)

Assessment of Early Detection by Multi-omics-Based Liquid Biopsy in Lung Cancer: A Prospective Study (ASCEND-LUNG)

Utility of cell-free DNA from bronchial washing fluid in diagnosis and genomic determination for radiology-suspected pulmonary nodules.

Bronchial washing fluid (BWF) is a less-invasive specimen. Due to the limited sensitivity of BWF cellular component diagnosis, the aim of this study was to explore the potential role of BWF supernatant as a source of liquid biopsy of lung cancer. This prospective study enrolled 76 suspected and 5 progressed lung cancer patients. Transbronchial biopsy tissues, BWF supernatant (BWF_Sup) and BWF precipitant (BWF_Pre) were tested by a targeted panel of 1021 genes. BWF_Sup cell-free DNA (cfDNA) was superior to tissue biopsy and BWF_Pre in determining mutational allele frequency, tumour mutational burden, and chromosomal instability. Moreover, BWF_Sup and BWF_Pre achieved comparable efficacy to tissue samples in differentiating malignant and benign patients, but only BWF_Sup persisted differentiated performance after excluding 55 malignancies pathologically diagnosed by bronchoscopic biopsy. Among 67 malignant patients, 82.1% and 71.6% of tumour-derived mutations (TDMs) were detected in BWF_Sup and BWF_Pre, respectively, and the detectability of TDMs in BWF_Sup was independent of the cytological examination of BWF. BWF_Sup outperformed BWF_Pre in providing more subclonal information and thus might yield advantage in tracking drug-resistant markers. BWF_Sup cfDNA is a reliable medium for lung cancer diagnosis and genomic profiles and may provide important information for subsequent therapeutic regimens.

Novel Robotic-Assisted Cryobiopsy for Peripheral Pulmonary Lesions

PurposeTissue acquisition in lung cancer is vital for multiple reasons. Primary reasons reported for molecular testing failure in lung cancer biopsy specimens include insufficient amount of tumor cells provided and inadequate tissue quality. Robotic bronchoscopy is a new tool enabling peripheral pulmonary lesion sampling; however, diagnostic yield remains imperfect possibly due to the location of nodules adjacent to or outside of the airway. The 1.1-mm cryoprobe is a novel diagnostic tool and accesses tissue in a 360-degree manner, thus potentially sampling eccentric/adjacent lesions. This study examines the diagnostic yield of the cryoprobe compared to standard needle aspiration and forceps biopsy. It additionally evaluates yield for molecular markers in cases of lung cancer.MethodsThis is a retrospective analysis of 112 patients with 120 peripheral pulmonary lesions biopsied via robotic bronchoscopy using needle aspirate, forceps, and cryobiopsy.ResultsThe overall diagnostic yield was 90%. Nearly 18% of diagnoses were made exclusively from the cryobiopsy sample. Molecular analysis was adequate on all cryobiopsy samples sent. Digital imaging software confirmed an increase in quantity and quality of samples taken via cryobiopsy compared to needle aspirate and traditional forceps biopsy.ConclusionUsing the 1.1-mm cryoprobe to biopsy PPN combined with the Ion robotic bronchoscopy system is safe, feasible, and provides more diagnostic tissue than needle aspirates or traditional forceps biopsies. The combination of cryobiopsy with robotic-assisted bronchoscopy increased diagnostic yield, likely due to its 360-degree tissue acquisition which is beneficial when targeting extraluminal lesions adjacent to the airway.

910P Assessment of early detection by multi-omics-based liquid biopsy in lung cancer: A prospective study (ASCEND-LUNG)

Liquid biopsy offers an optimal approach for the early detection of lung cancer that is the leading cause of cancer mortality worldwide. Here, we report the results of AssesSment of early-deteCtion basEd oN liquiD biopsy in LUNG cancer (ASCEND-LUNG, NCT04817046), a prospective study designed to develop early detection models for lung cancer based on multi-omics assays including cell-free DNA (cfDNA) methylation, mutation, and tumor proteins.

EP16.02-001 Integration of Primer Exchange Reaction and DNAzyme for Electrochemical Detection of PD-L1-Expressing Exosomes in Lung Cancer

Lung cancer is the most common malignant diseases and the leading cause of cancer-related deaths worldwide. To date, PD-L1-expressing exosomes, containing abundant tumor-related biological substances, have attracted great interest as a kind of tumor-specific biomarkers in liquid biopsy of lung cancer. Also,exosome PD-L1 has been reported to contribute to tumour immune evasion, and the potential role of its level in predicting immunotherapeutic efficacy has raised concerns but remains unclear.

P2.10-01 Transcriptional Diversity of Emerging Cell Populations in Refractory Small Cell Lung Cancer Biopsies and Xenografts

Small cell lung cancer (SCLC) is a high-grade neuroendocrine lung carcinoma notable for early dissemination and rapid, albeit transient, responses to standard of care (SOC) treatment [platinum-based chemotherapy plus immune checkpoint blockade (ICB)] that are rapidly undone by refractory relapses. Using single cell profiling, we previously demonstrated that a major factor regulating resistance is an increase in plasticity and transcriptional intratumoral heterogeneity (ITH) in response to treatment.

Abstract 5978: Real time pathologic diagnosis of digital biopsy during lung cancer surgery using confocal laser endomicroscopy

Abstract Purpose: Portable hand held confocal laser endomicroscopy is a novel, noninvasive technology that provides real time imaging during biopsy procedure and could be used to perform rapid histopathological diagnosis. However, its ability of determining whether the ex vivo samples obtained during lung cancer surgery or biopsy are tumor or non tumor is yet to be evaluated. Methods: Real time imaging of 75 surgical samples was performed. In preanalytic process, the samples were stained with fluorescent dye. Benign and malignant histologic structures were classified by cell density, nuclear size variability, and nuclear arrangement, which are compared with HE images from the same section. Results: We defined the confocal laser endomicroscopy atypia classification from the findings of the cells and made a comprehensive atlas of digital biopsy images for pleural surface, bronchus, and lung parenchyma. A total of 3,734 digital images were correlated with 75 frozen lung specimens. Three pathologists assessed digital biopsy images for defining cancer versus non cancer. Overall accuracy was 92.2%, The sensitivity for malignancy was 100% and the specificity was 84.4%. Both inter observer (k = 0.51) and intra observer (k = 0.52) agreement confirmed moderate agreement. Conclusion: Portable confocal laser endomicroscopy was useful to differentiate malignant from benign, to assess the histology of pleural surface and bronchial margin and might be substitute for rapid on site evaluation of pathologic diagnosis during surgical or biopsy procedure. Citation Format: Tae-Jung Kim, Young Jo Sa, Sung Ah Kim, Kyungmin Hwang. Real time pathologic diagnosis of digital biopsy during lung cancer surgery using confocal laser endomicroscopy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5978.

Abstract 2233: Pilot study of liquid biopsy of lung cancer circulating tumor cells (CTC) captured by a novel carbon nano-tube (CNT) differential adhesion platform

Abstract Introduction: Diagnosing and profiling lung cancers (LC) is challenging across the disease spectrum. Early-stage LC are small and hard to distinguish from benign nodules. Patients are often infirm, increasing biopsy risks. Monitoring for LC progression, recurrence or second primaries are further challenges. CTCs recovered from blood sample is non-invasive, easily repeatable across the clinical course of disease treatment and progression. Unlike cell-free nucleic acid profiles, CTC is mutation agnostic. Aims/Methods: Pilot test of a Carbon-nanotube (CNT) platform on capturing suspected LC cells across a range of stages/cell types.51 “normal controls” recruited to balance cancer: benign cases and to test specificity. Study period 1/2020 - 8/2021Procedure details per Loeian M Lab on a Chip 2019;11:1899. 16 ml of whole blood was lysed, nucleated cells cultured for 48 hours on a CNT coated surface. Cells were fixed, stained with CK7/8/18/pancytokeratinAE1/2 epithelial markers (CTC), CD45(WBC), nuclear DAPI control. A sample is considered CTC positive if 1 or more CD45-/CK+ cells are found. Results: 142 subjects were recruited from four clinical sites. 18 samples were non-analyzable due to shipping and preanalytical error.43 of 56 cancers had pathologic staging: 30% I, 16% II, 35% III, 19% IV.CTCs were found in 77% of LC samples (sensitivity), specificity 94%, Positive Predictive Value 92% and Negative Predictive Value 83%.Even in Stages I/II patients, 75% were found to have CTCs. The number of CTC found varied by patient, (range 0-31). Conclusion: CNT adherence enables antigen independent capture of live, viable LC CTCs from whole blood with high specificity and PPV. Planned inclusion of additional CTC markers should improve the detection of other classes of CTCs. High specificity and PPV identify this as a useful “rule-in” test to pursue downstream testing to confirm early LC diagnosis or follow-up surveillance for recurrence. Table 1. Performance of SS-CNT in capturing CTCs Test Result Tissue Positive Cancer Tissue Negative Cancer or Control Total CNT Test + True Pos (TP) 43 False Pos (FP) 4 47 CNT Test - False Neg (FN) 13 True Neg (TN) 64 77 56 68 124 Test Performance 1 or more CTC as positive Sensitivity TP/(TP + FN) 76.80% Specificity TN/(TN + FP) 94.10% Positive Predictive Value PPV TP/(TP + FP) Rule-in 91.50% Negative Predictive Value NPV TN/(TN + FN) Rule- out 83.10% Citation Format: Rex Chin-Wei Yung, Connor Tyree, Ashlee Colbert, Ganesh Krishna, Rakesh Mehta, Jessica Moja, Daniel Fox, Jeff Allard, John Park, Mark Connelly. Pilot study of liquid biopsy of lung cancer circulating tumor cells (CTC) captured by a novel carbon nano-tube (CNT) differential adhesion platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2233.

Abstract 1393: A Novel platform providing automated, consistent and gentle cell isolation from tissues for downstream applications

Abstract Tumor biologists have long studied tumor tissue to understand cancer progression and to ultimately treat the disease. The gold standard would be to analyze tumor and immune cells from the same sample, providing purified cells that are interacting within the tumor. Many tissue homogenization techniques have been developed to release cells for downstream analysis, however, an automated and gentle process of purifying multiple cell types from the same processed tissue has remained elusive. The industry-standard workflows, such as centrifugation, have presented long-standing performance issues in terms of low cell recovery and inefficient removal of molecular debris, which often leads to sequencing results that are difficult to interpret. To ensure the quality of sequencing data from tissues, upstream processes need to improve to get the most from the precious patient tissue. Applied Cells has developed an integrated system based on multiple physics principles to achieve cell wash, concentration, and target cell isolation, with high recovery at an unprecedented flow rate. This platform combines tunable, acoustic cell purification and in-flow, column-free immuno-magnetic separation technologies, enabling automation of the entire cell sample preparation workflow for genomics and transcriptomics analysis. By harnessing the power and precision of acoustic cell manipulation, the MARS System can process a variety of samples including dissociated solid tissue in enzymatic buffer with high cell recovery and superior molecular debris removal compared to standard methods. For instance, when purifying leukocytes from a lung cancer biopsy tissue samples, MARS can remove 99.50% debris and 91% of dead cells in one single step, which is supreme than the conventional centrifuge cell wash while having >90% leukocyte recovery at 1mL/min flow rate. The in-flow microfluidics design drastically reduces residual DNA and RNA fragments that are difficult to remove using centrifuge wash methods. Furthermore, the system enables parallel cell purification and concentration for high throughput sample processing. Tumor or TIL cells can be further enriched by means of positive magnetic selection for genomic or cell biology applications. This novel platform allows improved tumor and TIL recovery in a gentle, consistent manner while having minimal interference with downstream applications, all from a single source starting material. Citation Format: Liping Yu. A Novel platform providing automated, consistent and gentle cell isolation from tissues for downstream applications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1393.

Abstract 1099: Resistance to anti-EGFR therapy is transferred horizontally between lung cancer cells through caveolin-associated intercellular trafficking

Abstract Despite initial benefit from anti-EGFR therapy, patients with EGFR-mutated lung cancer (LC) inevitably relapse. Knowing that the majority of the relapsing cancer cells do not carry the resistance mutation, we hypothesized that resistance to anti-EGFR therapy can be transferred horizontally between LC cells, through intercellular communication. Using a cell culture system that allows communication of secreted molecules and extracellular vesicles, we performed indirect co-culture of Erlotinib-sensitive LC HCC827 cells (S cells) and Erlotinib-resistant LC H1975 cells (R cells, carrying the EGFR T790M resistance mutation), where S cells were treated with conditioned medium from R cells under selective pressure of Erlotinib (EGFR inhibitor). In vivo, immunodeficient mice (Rag2-/-Il2rg-/-) were engrafted with S cells in a single flank (n=12), or with S and R cells in opposite flanks (n=20) and treated with Erlotinib three times a week. Our results showed that resistance to anti-EGFR therapy can be transferred in vitro from R to S cells. Through digital PCR we provide evidence that transfer of resistance is neither due to cell contamination, nor transfer of DNA molecules with the T790M mutation. We also showed that resistance to anti-EGFR therapy can be transferred in vivo from R to S cells, originating “de novo” resistant tumors (SR tumors) and thereby decreasing progression-free survival of the animals. Using DNA fingerprinting we demonstrated that SR tumors do not contain R cells, ruling out cell migration as an explanation for the growth of the SR tumors. Moreover, the acquired resistance phenotype is stable over several in vivo passages of the tumor. We have analyzed the SR tumors with NGS and did not detect any known resistance mutations to anti-EGFR therapy. Through RNA-Seq and pathway analysis we found that SR tumors show significant changes in genes involved in intracellular trafficking, as evidenced by increased expression of CAV1 and CAV2. This increment of caveolins expression was validated in the SR tumors by real-time PCR and immunohistochemistry. CAV1 and CAV2 expression analysis was also performed in human LC biopsies, showing that in a subset of EGFR-mutated LC there is an increase in caveolins expression from the untreated primary to the anti-EGFR-treated relapsing tumor. Our results show that the anti-EGFR resistance phenotype can be transferred from R to S cells both in vivo and in vitro, and that it results in reprogramming of the expression profile of SR cells. Our findings point out a key role for caveolin-associated intercellular trafficking in the acquisition of resistance to targeted therapy in cancer. This challenges the current paradigm of cell division and selection as the sole mechanism underlying transmission of mutation-driven therapy resistance, and reveals an unforeseen level of plasticity in cancer. PhD Scholarship SFRH/BD/115099/2016 Citation Format: Susana Junqueira Neto, Ana Rita Oliveira, Joana Fernandes Marques, Gabriela Fernandes, Venceslau Hespanhol, Conceição Souto Moura, Sónia Almeida Melo, José Luís Costa, José Carlos Machado. Resistance to anti-EGFR therapy is transferred horizontally between lung cancer cells through caveolin-associated intercellular trafficking [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1099.

A predictive model based on liquid biopsy for non-small cell lung cancer to assess patient’s prognosis: Development and application

Background and objectiveImproving ability to predict the prognosis of patients with progressive lung cancer is an important task in the era of precision medicine. Here, a predictive model based on liquid biopsy for non-small cell lung cancer (NSCLC) was established to improve prognosis prediction in patients with progressive NSCLC. MethodsClinical data and blood samples of 500 eligible patients were collected and screened from the electronic case database and blood sample center of Hwa Mei Hospital, University of Chinese Academy of Sciences and Ningbo Institute of Life and Health Industry, University of Chinese Academy of Sciences. Patients were randomly assigned to training set (300 cases) and validation set (200 cases) in a ratio of 3:2 by random number method. Baseline levels of the two datasets were compared. Progression-free survival (PFS) analysis was performed on the training set using Kaplan-Meier method. The independent prognostic factors affecting patients’ PFS were determined by multivariate Cox regression analysis. The prognosis predictive model of patients was constructed by using the nomogram. Calibration curve and C-index were used to evaluate the accuracy of the prognosis predictive model in both internal and external validations. ResultsIn training set, the age distribution of patients was 59.00 (46.00, 71.00) years, including 137 (45.7 %) females and 163 (54.3 %) males, 198 cases (66.0 %) with Eastern Cooperative Oncology Group (ECOG) score 0–1, and 102 cases (34.0 %) with ECOG score 2. In verification set, the age distribution of patients was 60.00 (48.25, 73.00) years, including 92 females (46.0 %) and 108 males (54.0 %), 130 cases (65.0%) with ECOG score 0–1, and 70 cases (35.0 %) with ECOG score 2. Patients in training set showed PFS differences stratified by gene mutation type (p < 0.0001), differentiation degree (p < 0.0001), circulating tumor cell (CTC) content (p = 0.00026), and brain metastasis (p < 0.0001). Besides, multivariate Cox regression analysis indicated that gene mutation type, differentiation degree, CTC content (p = 0.002), and brain metastasis (p = 0.005) are independent prognostic factors for PFS. These factors were included in the nomogram parameters, and both internally validated calibration curve (C-index = 0.672) and externally validated calibration curve (C-index = 0.657), showing good predictive performance of the model. ConclusionThe predictive model has a good predictive ability for prognosis of patients with progressive NSCLC. Notably, the differentiation degree and CTC content are both impact factors for PFS of patients, and the performance of these indicators in predicting the survival of patients with progressive NSCLC needs to be clarified in the future.

Molecular profiling of dedicated lung cancer biopsy tissue sample collected at time of diagnostic bronchoscopy.

e20587 Background: The combined use of molecular biomarker testing and targeted precision therapeutics has led to improved survival in lung cancer. Broadening access to national guideline recommended comprehensive molecular testing requires overcoming the challenges of inadequate tissue biopsies, which can lead to the need for additional procedures and ultimately, delays in initiation of care. We show Percepta Genomic Atlas identifies key molecular alterations in transbronchial needle aspirate (TBNA) and transbronchial biopsy (TBB) samples of lung or lymph node collected during the initial diagnostic bronchoscopy. Methods: Percepta Genomic Atlas combines the whole exome TruSeq RNA Exome and targeted AmpliSeq Focus DNA assays (Illumina) for a comprehensive gene panel including ALK, RET, ROS1, NTRK1/3, MET, EGFR, BRAF, KRAS and HER2. TruSight Oncology 500 DNA and AmpliSeq Focus RNA assays (Illumina) were used as reference assays. DNA and RNA were extracted from samples with the AllPrep Micro kit (Qiagen) and analyzed by Percepta Genomic Atlas and reference assays. 94 biopsy samples (73 TBNA and 21 TBB from 71 patients undergoing a diagnostic bronchoscopy for suspected lung cancer were collected into RNAprotect (Qiagen) under an IRB approved protocol. Local molecular testing results from FFPE biopsy samples taken during the same bronchoscopy procedure were collected. Results: RNA and DNA in sufficient amounts to run the Percepta Genomic Atlas and reference assays was obtained from 85 of 94 lung biopsies from 63 of 71 patients. Percepta Genomic Atlas identified pathogenic variants in 29 bronchoscopy biopsy samples from 23 patients including single nucleotide variants in EGFR, KRAS, BRAF and PIK3CA, an EGFR exon 19 deletion/insertion and copy number amplications in AR, EGFR, CDK4, CCND1, MYC, MYCN and PIK3CA. No fusions were identified. This results in a 100% sensitivity for detecting pathogenic alterations when compared to reference assay results. When comparing Percepta Genomic Atlas results to local molecular testing of bronchoscopy samples performed as part of routine clinical care, we found 100% concordance with 7 of 7 alterations detected in 7 patients, including mutations in EGFR, KRAS and BRAF. In a further 4 patients, Percepta Genomic Atlas and local multi-gene NGS testing results agreed, with neither assay identifying guideline recommended alterations. Conclusions: Using a combination of whole exome RNA and targeted DNA sequencing, Percepta Genomic Atlas detects the clinically actionable mutations in patients with non-small cell lung cancer using fresh tissue specimens collected during bronchoscopic tissue sampling, with high concordance to standard of care testing. By initiating broad molecular testing at the time of the bronchoscopy, Percepta Genomic Atlas may provide timelier results for patients with lung cancer.

The utilization of artificial intelligence to produce clinically relevant scores on PD-L1 immunostained non-small cell lung cancer biopsies.

e14576 Background: Immunotherapies targeting complex programmed death-ligand 1 (PD-L1) interactions have been groundbreaking in the treatment of non-small cell lung cancer (NSCLC), offering new strategies for patients who are likely to respond. PD-L1 binds to the checkpoint PD-1 to regulate T cells, which has an important role in boosting the immune response. Identifying patients who may benefit from PD-L1 focused care is dependent on the interpretation of the drug’s associated companion diagnostic (CDx) immunohistochemistry (IHC) assay. PD-L1 IHC assays stratify NSCLC patients using a Tumor Proportion Score (TPS), a manual scoring paradigm which must be applied by a clinical pathologist. Application of the score is complex as it requires the reader to visually exclude stromal and tumor-infiltrating immune cells across a full sample, retaining PD-L1 expression information for true tumor cells only. Methods: In this study, we utilized the PD-L1 [28-8] immunohistochemistry (IHC) assay to stain 10 NCSLC specimens. Stained slides were then imaged using the AT2 scanner (Leica Biosystems, Buffalo Grove, IL) scanner and analyzed using the Visiopharm Image Analysis platform. Results: The intricacy of TPS application is time consuming for the pathologist and introduces opportunity for inter- and intra-reader variability. Our novel image analysis approach utilizes artificial intelligence (AI) to automatically denote tumor nests and exclude tumor-infiltrating immune cells. The remaining true tumor cells may then be quantified for PD-L1 expression across the full biopsy, producing a TPS for each sample. Conclusions: This method allows for great accuracy and time-efficiency in providing a TPS than traditional methods, which may result in improved patient care.

The pattern of association between histopathological vs clinical and radiological findings of lung cancer biopsy in patients of lung cancer

Background: Lung cancer is one of the most deadly tumours known. It is accurately found by many radiographic testing methods occasionally initiated for an unrelated ailment. In light of new histology guided therapeutic modalities and lung cancer genetic categorization, histological characterisation of lung cancer has risen in prominence. Aim: To link histology findings with clinical and radiographic features. Methods: This prospective investigation followed 40 patients with suspected lung cancer for a year, looking at clinical, radiological, and histological features. The research covered a clinical history, smoking habits, full physical examination of the respiratory system, chest roentgenogram, computed tomography of the thorax, fiberoptic bronchoscopy, and others. Results: Patients were aged 56.7 years with 80.2% male and 19.8% female. The most frequent symptom was cough 84.6%. Lesion 85.5% followed by collapse consolidation 35.26% were the most frequent radiological results. Squamous cell carcinoma most typically showed as a hilar mass 54.4%, adenocarcinoma as a peripheral mass 67.4%. Squamous cell carcinoma 48% was the most frequent form, followed by small cell carcinoma 13% and adenocarcinoma 2.98% . Conclusion: In order to confirm a clinical or radiological diagnosis of lung cancer, endobronchial lung biopsy and histopathology are both extremely necessary tests to do. Keywords: Lung Cancer, Radiological Patterns, Histopathological Types

Notch-mediated lactate metabolism regulates MDSC development through the Hes1/MCT2/c-Jun axis.

Myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs) play critical roles in tumorigenesis. However, the mechanisms underlying MDSC and TAM development and function remain unclear. In this study, we find that myeloid-specific activation of Notch/RBP-J signaling downregulates lactate transporter MCT2 transcription via its downstream molecule Hes1, leading to reduced intracellular lactate levels, blunted granulocytic MDSC (G-MDSC) differentiation, and enhanced TAM maturation. We identify c-Jun as a novel intracellular sensor of lactate in myeloid cells using liquid-chromatography-mass spectrometry (LC-MS) followed by CRISPR-Cas9-mediated gene disruption. Meanwhile, lactate interacts with c-Jun to protect from FBW7 ubiquitin-ligase-mediated degradation. Activation of Notch signaling and blockade of lactate import repress tumor progression by remodeling myeloid development. Consistently, the relationship between the Notch-MCT2/lactate-c-Jun axis in myeloid cells and tumorigenesis is also confirmed in clinical lung cancer biopsies. Taken together, our current study shows that lactate metabolism regulated by activated Notch signaling might participate in MDSC differentiation and TAM maturation.

Classification of subtypes including LCNEC in lung cancer biopsy slides using convolutional neural network from scratch

Identifying the lung carcinoma subtype in small biopsy specimens is an important part of determining a suitable treatment plan but is often challenging without the help of special and/or immunohistochemical stains. Pathology image analysis that tackles this issue would be helpful for diagnoses and subtyping of lung carcinoma. In this study, we developed AI models to classify multinomial patterns of lung carcinoma; ADC, LCNEC, SCC, SCLC, and non-neoplastic lung tissue based on convolutional neural networks (CNN or ConvNet). Four CNNs that were pre-trained using transfer learning and one CNN built from scratch were used to classify patch images from pathology whole-slide images (WSIs). We first evaluated the diagnostic performance of each model in the test sets. The Xception model and the CNN built from scratch both achieved the highest performance with a macro average AUC of 0.90. The CNN built from scratch model obtained a macro average AUC of 0.97 on the dataset of four classes excluding LCNEC, and 0.95 on the dataset of three subtypes of lung carcinomas; NSCLC, SCLC, and non-tumor, respectively. Of particular note is that the relatively simple CNN built from scratch may be an approach for pathological image analysis.