Detection Of Early-stage Lung Cancer Research Articles

Optical Nanobiosensor Based on Surface-Enhanced Raman Spectroscopy and Catalytic Hairpin Assembly for Early-Stage Lung Cancer Detection via Blood Circular RNA.

Lung cancer has become the leading cause of cancer-related deaths globally. However, early detection of lung cancer remains challenging, resulting in poor outcomes for the patients. Herein, we developed an optical biosensor integrating surface-enhanced Raman spectroscopy (SERS) with a catalyzed hairpin assembly (CHA) to detect circular RNA (circRNA) associated with tumor formation and progression (circSATB2). The signals of the Raman reporter were considerably enhanced by generating abundant SERS "hot spots" with a core-shell nanoprobe and 2D SERS substrate with calibration capabilities. This approach enabled the sensitive (limit of detection: 0.766 fM) and reliable quantitative detection of the target circRNA. Further, we used the developed biosensor to detect the circRNA in human serum samples, revealing that patients with lung cancer had higher circRNA concentrations than healthy subjects. Moreover, we characterized the unique circRNA concentration profiles of the early stages (IA and IB) and subtypes (IA1, IA2, and IA3) of lung cancer. These results demonstrate the potential of the proposed optical sensing nanoplatform as a liquid biopsy and prognostic tool for the early screening of lung cancer.

Diagnosis, treatment, and prognosis of stage IB non-small cell lung cancer with visceral pleural invasion.

With the increasing implementation of early lung cancer screening and the increasing emphasis on physical examinations, the early-stage lung cancer detection rate continues to rise. Visceral pleural invasion (VPI), which denotes the tumor's breach of the elastic layer or reaching the surface of the visceral pleura, stands as a pivotal factor that impacts the prognosis of patients with non-small cell lung cancer (NSCLC) and directly influences the pathological staging of early-stage cases. According to the latest 9th edition of the TNM staging system for NSCLC, even when the tumor diameter is less than 3 cm, the final T stage remains T2a if VPI is present. There is considerable controversy within the guidelines regarding treatment options for stage IB NSCLC, especially among patients exhibiting VPI. Moreover, the precise determination of VPI is important in guiding treatment selection and prognostic evaluation in individuals with NSCLC. This article aims to provide a comprehensive review of the current status and advancements in studies pertaining to stage IB NSCLC accompanied by VPI.

An Observation and Analysis the role of Convolutional Neural Network towards Lung Cancer Prediction

Lung cancer is one of the most serious and prevalent diseases, causing many deaths each year. Though CT scan images are mostly used in the diagnosis of cancer, the assessment of scans is an error-prone and time-consuming task. Machine learning and AI-based models can identify and classify types of lung cancer quite accurately, which helps in the early-stage detection of lung cancer that can increase the survival rate. In this paper, Convolutional Neural Network is used to classify Adenocarcinoma, squamous cell carcinoma and normal case CT scan images from the Chest CT Scan Images Dataset using different combinations of hidden layers and parameters in CNN models. The proposed model was trained on 1000 CT Scan Images of cancerous and non-cancerous cells to find the best combination of parameters in CNN to predict lung cancer accurately. The proposed system recorded the highest accuracy of 92.79%. In addition to that, the paper addresses 192 observations made using the CNN model.

A novel three-biomarker signature (CA-62, CEA, CYFRA 21-1) and early-stage NSC lung cancer detection.

3038 Background: A double-blind clinical study was conducted on 304 clinically verified blood serum samples, including 141 non-small cell lung cancer (NSCLC), 133 healthy volunteers and 30 patients with chronic obstructive pulmonary disease (COPD). The objectives of the study were: The assessment of various cancer markers and their possible combinations for detection of early (I-II) stages of non-small cell lung cancer (NSCLC) and evaluation of the best cancer markers panel as a pre-screening tool for NSCLC before Low-Dose CT scan. Methods: Quantitative measurement of various tumor markers for serum samples was carried out using electrochemiluminescent immunoassays Elecsys CA-125, ELECSYS CA 19-9, ELECSYS CYFRA 21-1 and ELECSYS SCC (COBAS, Roche Diagnostics GmbH, Germany, EU), enzyme immunoassay CA15-3-ELISA-BEST, CEA-ELISA-BEST, NSE-ELISA-BEST (JSC "VECTOR-BEST", RF) and immunochemiluminescent analysis CLIA-CA-62 (LLC "JVS Diagnostics", RF). The results were compared to the literature data for other biomarkers and their panels. Results: Unlike other tumor markers, which are expressed proportionally to tumor size, the marker for epithelial carcinomas CA-62 is expressed from Stage I and demonstrated the highest diagnostic results in detecting early stages (I-II) of NSCLC. The biomarkers CYFRA 21-1 and SCC do not have sufficient sensitivity and specificity to diagnose asymptomatic lung cancer. However, the addition of cytokeratin 19 CYFRA 21-1 to glycoproteins CA-62 and CEA increases the specificity by eliminating false positive results, which significantly improves the diagnostic value of the cancer marker signature (CA-62, CEA and CYFRA 21-1): 100% Specificity, 93% Sensitivity, 100% PPV, 94% NPV and 94% test accuracy. Conclusions: The results of the study demonstrated that using the biomarkers signature (CA-62, CEA and CYFRA 21-1) allows increasing the specificity of CT scan for patients with suspicious abnormalities on the tomogram, improving the interpretation of visualized localized lesion, and improving the accuracy of differential diagnosis at detecting early stages of LC up to 94%. In the prospective, adding cancer markers panel signature (CA-62, CEA and CYFRA 21-1) to the current lung cancer risk assessment system as a pre-screening tool for LDCT may improve the quality of early-stage Lung cancer detection by significant increasing the sensitivity and by reducing the proportion of false positive results. [Table: see text]

European guidelines for the surgical management of pure ground-glass opacities and part-solid nodules: Task Force of the European Association of Cardio-Thoracic Surgery and the European Society of Thoracic Surgeons.

Pulmonary nodules are often encountered by respiratory physicians, radiologists, and thoracic surgeons alike. The European Society of Thoracic Surgery (ESTS) and European Association of Cardiothoracic Surgery (EACTS) established a multidisciplinary collaboration of clinicians with expertise in managing pulmonary nodules with the aim of producing the first joint comprehensive review of the scientific literature, with a specific focus on the management of pure ground glass opacities and part-solid nodules. The scope of the document has been defined by the EACTS and ESTS governing bodies and focuses on six areas of main interest agreed upon by the Task Force. These include: management of solitary and multiple pure ground glass nodules, solitary part-solid nodules, identifying the non-palpable lesion, the role of minimally invasive surgery and the decision making behind sub-lobar versus lobar resection. The literature revealed that with the increasing use of incidental CT scans and CT lung cancer screening programs, the detection of early-stage lung cancer is going to rise with a higher number of potential cancers presenting on the ground glass and part-solid nodule spectrum. Given that the gold standard for improved survival is surgical resection, there is an urgent need for comprehensive characterisation of these nodules and guidelines that are directed towards their surgical management. It is recommended that standard decision-making tools are used to determine the risk of malignancy and thus guide referral for surgical management and decisions for surgical resection are made in a multidisciplinary setting with equitable consideration given to radiological characteristics, the evolution of the lesion, presence of solid component, patient fitness and co-morbidities. Given the recent surge in robust level I data comparing sublobar and lobar resection with the release of JCOG0802 and CALGB140503 data, a global overview of an individual case must be adopted into clinical practice. This set of recommendations are based on the available literature; however, close collaboration in the design and conduct of randomized controlled trials is still of utmost importance to answer further questions in this rapidly evolving field.

Accurate detection of early-stage lung cancer using a panel of circulating cell-free DNA methylation biomarkers

BackgroundLung cancer remains the leading cause of cancer mortality worldwide. Early detection of lung cancer helps improve treatment and survival. Numerous aberrant DNA methylations have been reported in early-stage lung cancer. Here, we sought to identify novel DNA methylation biomarkers that could potentially be used for noninvasive early diagnosis of lung cancers.MethodsThis prospective-specimen collection and retrospective-blinded-evaluation trial enrolled a total of 317 participants (198 tissues and 119 plasmas) comprising healthy controls, patients with lung cancer and benign disease between January 2020 and December 2021. Tissue and plasma samples were subjected to targeted bisulfite sequencing with a lung cancer specific panel targeting 9,307 differential methylation regions (DMRs). DMRs associated with lung cancer were identified by comparing the methylation profiles of tissue samples from patients with lung cancer and benign disease. Markers were selected with minimum redundancy and maximum relevance algorithm. A prediction model for lung cancer diagnosis was built through logistic regression algorithm and validated independently in tissue samples. Furthermore, the performance of this developed model was evaluated in a set of plasma cell-free DNA (cfDNA) samples.ResultsWe identified 7 DMRs corresponding to 7 differentially methylated genes (DMGs) including HOXB4, HOXA7, HOXD8, ITGA4, ZNF808, PTGER4, and B3GNTL1 that were highly associated with lung cancer by comparing the methylation profiles of lung cancer and benign nodule tissue. Based on the 7-DMR biomarker panel, we developed a new diagnostic model in tissue samples, termed “7-DMR model”, to distinguish lung cancers from benign diseases, achieving AUCs of 0.97 (95%CI: 0.93-1.00)/0.96 (0.92-1.00), sensitivities of 0.89 (0.82–0.95)/0.92 (0.86–0.98), specificities of 0.94 (0.89–0.99)/1.00 (1.00–1.00), and accuracies of 0.90 (0.84–0.96)/0.94 (0.89–0.99) in the discovery cohort (n = 96) and the independent validation cohort (n = 81), respectively. Furthermore, the 7-DMR model was applied to noninvasive discrimination of lung cancers and non-lung cancers including benign lung diseases and healthy controls in an independent validation cohort of plasma samples (n = 106), yielding an AUC of 0.94 (0.86-1.00), sensitivity of 0.81 (0.73–0.88), specificity of 0.98 (0.95-1.00), and accuracy of 0.93 (0.89–0.98).ConclusionThe 7 novel DMRs could be promising methylation biomarkers that merits further development as a noninvasive test for early detection of lung cancer.Graphical abstract

Abstract 5434: Machine-learning-based epigenetic detection of early-stage lung cancers using the EpiCheck liquid biopsy platform

Abstract Alterations in DNA methylation are one of the earliest, most common signatures of cancer development, making them ideal biomarkers for early detection. Methylation profiling of plasma cell-free DNA (cfDNA) has significant potential to expand the use of liquid biopsies to cancer screening. The EpiCheck platform combines methylation-sensitive restriction endonuclease (MSRE) digestion with whole genome sequencing to comprehensively map genome-wide DNA methylation changes with high fidelity. In this proof-of-concept study, MSRE-NGS was used to interrogate a liquid biopsy atlas focused on early-stage lung cancers to i) identify informative NGS methylation biomarkers in plasma, and ii) develop a machine learning method to discriminate between cancer and high-risk, non-cancer controls. The EpiCheck lung cancer atlas was constructed using biospecimens from academic (UBC, Vanderbilt, Cleveland Clinic) and commercial biobanks, and from a prospective multi-center study (NCT04968548). Plasma samples were collected from 93 high-risk primary lung cancer patients (62% stage I) and 90 high-risk individuals without cancer. Extracted DNA was digested with methylation-sensitive endonucleases and sequenced at an average depth of 600x. Methylation levels of ~6 million genomic loci were rank ordered using Student's t-test. Gene set enrichment analysis (GSEA) was performed on the top ranking 1000 differentially methylated loci. A logistic regression classifier with Lasso regularization was trained on 100,000 loci, and performance was examined by mean AUC using 5-fold cross-validation. A total of 23,130 loci exhibited significant differential methylation patterns between cancers and controls (p<0.01, FDR corrected). Of these, 20,159 and 2,971 lung cancer loci were hypermethylated and hypomethylated, respectively. Biological characterization using GSEA identified enrichments in transcriptional regulation and developmental control. In particular, loci were enriched for Polycomb Repressive Complex regulated genes, suggesting a possible connection to abnormal epigenetic regulation via histone modification in lung cancer. Construction of a machine learning logistic regression model based on the five training folds utilized 224 loci on average, and achieved a mean cross-validation AUC of 0.93 when distinguishing plasma cancer cases vs controls. Our findings demonstrate that the MSRE-NGS EpiCheck platform identified putative biomarkers within the plasma methylome for detecting early-stage lung cancer. A machine learning model trained on methylation targets performed with high accuracy in discriminating lung cancer patients from high-risk healthy individuals. Additional studies are required for defining the strength of the approach and for validating its use in non-invasive lung cancer screening. Citation Format: Dvir Netanely, Stephen Lam, Anna McGuire, Stephen Deppen, Eric Grogan, Fabien Maldonado, Michael Gieske, Joseph Seaman, Kimberly Rieger-Christ, Satish Kalanjeri, Luis Herrera, Nichole Tanner, Garrett B. Sherwood, Orna Savin, Shacade Danan, Sarah Zaouch, Nimrod Axelrad, Revital Knirsh, Ofir Shliefer, Keren Manor, Radha Duttagupta, Aharona Shuali, Peter J. Mazzone, Gerard A. Silvestri, Catherine A. Schnabel, Danny Frumkin, Adam Wasserstrom. Machine-learning-based epigenetic detection of early-stage lung cancers using the EpiCheck liquid biopsy platform. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5434.

Abstract 6510: REM-DREAMing: Low-cost digital microfluidic analysis of DNA methylation heterogeneity for enhanced, liquid biopsy-based detection of early-stage lung cancer

Abstract We report the results of a study exploring the ability to exploit molecular heterogeneity in DNA methylation for improving the performance of liquid biopsy-based screening for early-stage (I & II) non-small cell lung cancer (NSCLC). Annual low-dose CT (LDCT) screening is currently recommended for adults aged 50 or older who are at high-risk of developing lung cancer. While this approach has resulted in improvements in survival, the false positive rate of lung nodules detection by LDCT remains over 95%, leading to unnecessary invasive follow-up procedures and further points to the need for new, complementary methods to improve diagnostics performance and reduce patient risk. DNA methylation biomarkers have demonstrated considerable potential as tumor-specific biomarkers for blood-based detection of early-stage NSCLC. Nonetheless, cell-free DNA (cfDNA) assessment techniques, such as methylation-specific PCR (MSP) or bisulfite sequencing, have limited sensitivity to assess epigenetic heterogeneity of rare epiallelic variants in a cost-effective manner. Here we reported a new platform, named REM-DREAMing (Ratiometric-Encoded Multiplex Discrimination of Rare EpiAlleles by Melt), which provides a simple, low-cost solution for multiplexed assessment of loci-specific DNA methylation heterogeneity at single molecule sensitivity. The microfluidic nanoarray contains four independent but identical 10,040 nanowell modules. Methylation biomarkers are differentiated by a ratiometric fluorescence scheme and the assessment of individual epiallele species of each locus are achieved through digitization in the nanoarray and precise high-resolution melt (HRM) analysis. In this study, we explore the potential utility of REM-DREAMing as a complementary assay for improving LDCT screening of NSCLC by testing a cohort of 48 clinical samples (28 cancer and 20 control samples) of low-volume liquid biopsy specimens from patients with CT-scan indeterminant pulmonary nodules. A machine learning algorithm incorporating logistic regression models with leave-one-out cross validation was developed to identify a proper methylation density threshold of each biomarker in the panel. Evaluation of the receiver operation characteristic (ROC) curve yielded an area under the curve (AUC) of 0.97 (95% CI, 0.94-1) with 93% sensitivity at 95% specificity for the REM-DREAMing assay, compared with 93% sensitivity at 62% specificity achieved using a traditional, MSP-based approach. These results suggest that the assessment of intermolecular epigenetic heterogeneity can provide superior clinical performance for cfDNA methylation and noninvasive detection of early-stage NSCLC, in particular. Our results warrant further investigation in a larger sample cohort to validate its utility for improving routine screening of NSCLC in high-risk populations. Citation Format: Yang Zhao, Christine M. O’Keefe, James G. Herman, Thomas R. Pisanic, Tza-Huei Wang. REM-DREAMing: Low-cost digital microfluidic analysis of DNA methylation heterogeneity for enhanced, liquid biopsy-based detection of early-stage lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6510.

Abstract 3330: Development of a genome-wide multiomic atlas of early-stage lung cancer enables identification of novel methylation biomarkers for disease detection beyond TCGA

Abstract Lung cancer is the leading cause of cancer mortality, mainly due to diagnosis at advanced stages. Early detection reduces mortality and screening by CT is recommended for a high-risk population, however, uptake is low. Biomarkers for early-stage detection may enhance screening. Liquid biopsy and blood circulating tumor DNA (ctDNA) are established plasma surrogates of tumor tissue but their use in early detection remains challenging due to the high noise level of current techniques. We constructed a novel multiomic atlas through whole genome sequencing (WGS) to map all genetic and epigenetic changes associated with early-stage lung cancer as a tool to identify markers. Biospecimens from 48 early-stage lung cancer cases (58% stage I) and 29 cancer-free controls, all high-risk by USPSTF, were acquired from academic (UBC, Vanderbilt, Cleveland Clinic) and commercial biobanks. Cases had tumor and normal lung tissue, whole blood (WB) and plasma samples. Controls had WB and plasma samples. EpiCheck sequencing (ECS) which combines methylation-sensitive restriction endonuclease digestion to detect differential methylation with WGS for multiomic analyses was performed on each sample. Extracted DNA was digested, underwent standard library preparation and sequenced at an average depth of 600x for cfDNA and 80x for tissue and WB. A subset also underwent WG bisulfite sequencing (BS) and standard WGS for comparison. Data analysis was performed using customized software. ECS outperformed BS. Mapping rate was 99.6%, 99.7% and 85.7% and unique mapping rate was 94.1%, 94.3% and 81.4% for WGS, ECS, and BS samples, respectively. Copy number integrity showed Pearson correlations of 0.9 for ECS and 0.67 for BS. Somatic mutation analysis in tissue identified a subset of cases with relatively high ctDNA shedding in plasma that were associated with larger tumors, older age and squamous cell carcinoma histology. This subset was further used to identify tumor derived plasma-based markers and assess fragmentation with high confidence. Shorter ctDNA fragments were observed in some, but not all cases. Additional analysis of WB and plasma identified host derived blood-based markers using methylation and copy number. This orthogonal approach enabled detection of small, low shedding tumors. A discovery panel of 87 markers showed 100% sensitivity and 97% specificity to discriminate cases vs controls. Only 11% of these markers are represented in the TCGA data. ECS generates high integrity sequencing data superior to BS, enabling genome-wide multiomic analyses (methylation, mutation, copy number, fragmentomics), with methylation signatures beyond the scope of TCGA. Results, which require validation, underscore the potential of the EpiCheck lung cancer atlas as a development platform of novel blood biomarkers for early-stage lung cancer detection. Citation Format: Revital Knirsh, Stephen Lam, Anna McGuire, Peter J. Mazzone, Stephen Deppen, Eric Grogan, Fabien Maldonado, Orna Savin, Shacade Danan, Sarah Zaouch, Nimrod Axelrad, Dvir Netanely, Aharona Shuali, Catherine A. Schnabel, Adam Wasserstrom, Danny Frumkin. Development of a genome-wide multiomic atlas of early-stage lung cancer enables identification of novel methylation biomarkers for disease detection beyond TCGA [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3330.

A Comparative Review of the Detection of Early-stage Lung Cancer by Exosomal and Free Nucleic Acids and Standard Screening Methods

Lung cancer (LC), with its high incidence and less effective treatment strategies, is often not detected until it is in an advanced stage, which contributes to its high mortality rate. Hence, screening the DNA and RNA content in exosomes offers a promising method for its early diagnosis. New technologies for the early detection of LC are absolutely necessary to improve patient outcomes. Such an approach could be the exploitation of exosomes and their content. Exosomes contain DNA, different RNA species, proteins, ceramides, and cholesterol. They can transport their oncogenetic cargo derived from tumor cells to healthy cells over a range of distances to help propagate genetic information that could contribute to the initiation of cancer. This review provides an overview of the involvement of exosomes and their DNA/RNA together with cell-free DNA, summarizes potential biomarkers and describes the application of technologies for the characterization of exosomes with their possible tumor markers. Current radiological and scanning methods for the early detection of LC are also described and compared with nucleic acid analysis. The strengths and weaknesses of both approaches are considered.

Combinatorial Blood Platelets-Derived circRNA and mRNA Signature for Early-Stage Lung Cancer Detection

Despite the diversity of liquid biopsy transcriptomic repertoire, numerous studies often exploit only a single RNA type signature for diagnostic biomarker potential. This frequently results in insufficient sensitivity and specificity necessary to reach diagnostic utility. Combinatorial biomarker approaches may offer a more reliable diagnosis. Here, we investigated the synergistic contributions of circRNA and mRNA signatures derived from blood platelets as biomarkers for lung cancer detection. We developed a comprehensive bioinformatics pipeline permitting an analysis of platelet-circRNA and mRNA derived from non-cancer individuals and lung cancer patients. An optimal selected signature is then used to generate the predictive classification model using machine learning algorithm. Using an individual signature of 21 circRNA and 28 mRNA, the predictive models reached an area under the curve (AUC) of 0.88 and 0.81, respectively. Importantly, combinatorial analysis including both types of RNAs resulted in an 8-target signature (6 mRNA and 2 circRNA), enhancing the differentiation of lung cancer from controls (AUC of 0.92). Additionally, we identified five biomarkers potentially specific for early-stage detection of lung cancer. Our proof-of-concept study presents the first multi-analyte-based approach for the analysis of platelets-derived biomarkers, providing a potential combinatorial diagnostic signature for lung cancer detection.

Serum proteomics profiling identifies a preliminary signature for the diagnosis of early-stage lung cancer.

Lung cancer is the most common cause of death from cancer worldwide, largely due to late diagnosis. Thus, there is an urgent need to develop new approaches to improve the detection of early-stage lung cancer, which would greatly improve patient survival. The quantitative protein expression profiles of microvesicles isolated from the sera from 46 lung cancer patients and 41 high-risk non-cancer subjects were obtained using a mass spectrometry method based on a peptide library matching approach. We identified 33 differentially expressed proteins that allow discriminating the two groups. We also built a machine learning model based on serum protein expression profiles that can correctly classify the majority of lung cancer cases and that highlighted a decrease in the levels of Arysulfatase A (ARSA) as the most discriminating factor found in tumors. Our study identified a preliminary, non-invasive protein signature able to discriminate with high specificity and selectivity early-stage lung cancer patients from high-risk healthy subjects. These results provide the basis for future validation studies for the development of a non-invasive diagnostic tool for lung cancer.

Detection of early-stage lung cancer in sputum using automated flow cytometry and machine learning

BackgroundLow-dose spiral computed tomography (LDCT) may not lead to a clear treatment path when small to intermediate-sized lung nodules are identified. We have combined flow cytometry and machine learning to develop a sputum-based test (CyPath Lung) that can assist physicians in decision-making in such cases.MethodsSingle cell suspensions prepared from induced sputum samples collected over three consecutive days were labeled with a viability dye to exclude dead cells, antibodies to distinguish cell types, and a porphyrin to label cancer-associated cells. The labeled cell suspension was run on a flow cytometer and the data collected. An analysis pipeline combining automated flow cytometry data processing with machine learning was developed to distinguish cancer from non-cancer samples from 150 patients at high risk of whom 28 had lung cancer. Flow data and patient features were evaluated to identify predictors of lung cancer. Random training and test sets were chosen to evaluate predictive variables iteratively until a robust model was identified. The final model was tested on a second, independent group of 32 samples, including six samples from patients diagnosed with lung cancer.ResultsAutomated analysis combined with machine learning resulted in a predictive model that achieved an area under the ROC curve (AUC) of 0.89 (95% CI 0.83–0.89). The sensitivity and specificity were 82% and 88%, respectively, and the negative and positive predictive values 96% and 61%, respectively. Importantly, the test was 92% sensitive and 87% specific in cases when nodules were < 20 mm (AUC of 0.94; 95% CI 0.89–0.99). Testing of the model on an independent second set of samples showed an AUC of 0.85 (95% CI 0.71–0.98) with an 83% sensitivity, 77% specificity, 95% negative predictive value and 45% positive predictive value. The model is robust to differences in sample processing and disease state.ConclusionCyPath Lung correctly classifies samples as cancer or non-cancer with high accuracy, including from participants at different disease stages and with nodules < 20 mm in diameter. This test is intended for use after lung cancer screening to improve early-stage lung cancer diagnosis.Trial registration ClinicalTrials.gov ID: NCT03457415; March 7, 2018

Abstract PR006: Screening under-served populations by mobile low dose computerized tomography (LDCT) scans results in stage shift with potential cures: Time for a change in health policy?

Abstract Background: More than 60% of under-served patients with lung cancer first present with advanced/metastatic disease. Randomized trials have shown that LDCT of chest reduces lung cancer mortality but under-served populations have been excluded from these studies because of financial, geographic, insurance and access problems. We have tested the first mobile LDCT units and demonstrated improved access. We report a shift to earlier stage at diagnosis and potential cures in a large under-served population that includes African American, Hispanic/LatinX and Native American subjects. Methods: We developed two coaches fitted with portable 32 slice low-dose CT scanners to screen uninsured and under-served heavy smokers for lung cancer in an IRB-approved program ("The Oncologist", 2019). All films were reviewed by a central panel (using LUNG RADS protocol). Medicare patients were excluded because their insurance covers LDCT lung screening (but it should be noted that the elderly population is at increased risk for lung cancer, leading to a negative case selection bias). Results: We initially screened 1200 uninsured/under-insured subjects, mean age 61 years (range 55-64), with average pack year history of 47.8 (range 30- &amp;gt;150); 61% male; 18% Black, 3% Hispanic/LatinX, &amp;lt;1% Native American; 78% rural. We found 97 pts with LUNG RADS 4 (very high risk) lesions, 30 lung cancers (2.5%), including 18 at stage I-III treated with curative intent (60%); &amp;gt;50% had intercurrent cardiovascular disease and COPD seen on LDCT. We have additionally screened 574 cases, including 51 Native Americans (in a structured assessment of one tribe). Of the Native Americans, 5 had LUNG RADS 3-4 (high risk) lesions, but no cancers have been identified to date. For the total series of 1774 subjects with age-related negative case selection bias, 42 lung cancers have been identified (2.4%), 26 of which were treated with curative intent (62%) via surgery, radiation, or chemo-radiation. Only one patient of 18 treated with curative intent in the first series of 30 lung cancers has relapsed to date (median follow up 3 years, with 5 without evidence of disease beyond 3.5 years). These data confirm a shift to detection of early-stage lung cancer in this large under-served population, with an apparent potential for cure. Conclusion: Mobile LDCT yields a higher screening rate for under-served patients than prior hallmark trials, with a consequent shift to early-stage detection of lung cancer, with sustained treatment-induced complete remissions beyond 4 years. This approach could be applied to improve national lung cancer survival in the under-served. Citation Format: Derek Raghavan, Darcy Doege, Mellisa Wheeler, Kia Dungan, John Doty III, Glenn Hickman, Kathryn Mileham, Daniel Carrizosa. Screening under-served populations by mobile low dose computerized tomography (LDCT) scans results in stage shift with potential cures: Time for a change in health policy? [abstract]. In: Proceedings of the AACR Special Conference: Precision Prevention, Early Detection, and Interception of Cancer; 2022 Nov 17-19; Austin, TX. Philadelphia (PA): AACR; Can Prev Res 2023;16(1 Suppl): Abstract nr PR006.

Plasma extracellular vesicle phenotyping for the differentiation of early-stage lung cancer and benign lung diseases.

The development of a minimally invasive technique for early-stage lung cancer detection is crucial to reducing mortality. Phenotyping of tumor-associated extracellular vesicles (EVs) has the potential for early-stage lung cancer detection, yet remains challenging due to the lack of sensitive, integrated techniques that can accurately detect rare tumor-associated EV populations in blood. Here, we integrated gold core-silver shell nanoparticles and nanoscopic mixing in a microfluidic assay for sensitive phenotypic analysis of EVs directly in plasma without EV pre-isolation. The assay enabled multiplex detection of lung cancer-associated markers PTX3 and THBS1 and canonical EV marker CD63 by surface-enhanced Raman spectroscopy, providing a squared correlation coefficient of 0.97 in the range of 103-107 EVs mL-1 and a limit of detection of 19 EVs mL-1. Significantly, our machine learning-based nanostrategy provided 92.3% sensitivity and 100% specificity in differentiating early-stage lung cancer from benign lung diseases, superior to the CT scan-based lung cancer diagnosis (92.3% sensitivity and 71.4% specificity). Overall, our integrated nanostrategy achieved an AUC value of 0.978 in differentiating between early-stage lung cancer patients (n = 28) and controls consisting of patients with benign lung diseases (n = 23) and healthy controls (n = 26), which showed remarkable diagnostic performance and great clinical potential for detecting the early occurrence of lung cancer.

LCSCNet: A multi-level approach for lung cancer stage classification using 3D dense convolutional neural networks with concurrent squeeze-and-excitation module

Lung cancer, the deadliest disease worldwide, poses a massive threat to humankind. Various researchers have designed Computer-Aided-Diagnosis systems for the early-stage detection of lung cancer. However, patients are primarily diagnosed in advanced stages when treatment becomes complicated and dependent on multiple factors like size, nature, location of the tumor, and proper cancer staging. TNM (Tumor, Node, and Metastasis) staging provides all this information. This study aims to develop a novel and efficient approach to classify lung cancer stages based on TNM standards. We propose a multi-level 3D deep convolutional neural network, LCSCNet (Lung Cancer Stage Classification Network). The proposed network architecture consists of three similar classifier networks to classify three labels, T, N, and M-labels. First, we pre-process the data, in which the CT images are augmented, and the label files are processed to get the corresponding TNM labels. For the classification network, we implement a dense convolutional neural network with a concurrent squeeze & excitation module and asymmetric convolutions for classifying each label separately. The overall stage is determined by combining all three labels. The concurrent squeeze & excitation module helps the network focus on the essential information of the image, due to which the classification performance is enhanced. The asymmetric convolutions are introduced to reduce the computation complexity of the network. Two publicly available datasets are used for this study. We achieved average accuracies of 96.23% for T-Stage, 97.63% for N-Stage, and 96.92% for M-Stage classification. Furthermore, an overall stage classification accuracy of 97% is achieved.

Multidimensional Cell-Free DNA Fragmentomic Assay for Detection of Early-Stage Lung Cancer.

Rationale: Cell-free DNA (cfDNA) analysis holds promise for early detection of lung cancer and benefits patients with higher survival. However, the detection sensitivity of previous cfDNA-based studies was still low to suffice for clinical use, especially for early-stage tumors. Objectives: Establish an accurate and affordable approach for early-stage lung cancer detection by integrating cfDNA fragmentomics and machine learning models. Methods: This study included 350 participants without cancer and 432 participants with cancer. The participants' plasma cfDNA samples were profiled by whole-genome sequencing. Multiple cfDNA features and machine learning models were compared in the training cohort to achieve an optimal model. Model performance was evaluated in three validation cohorts. Measurements and Main Results: A stacked ensemble model integrating five cfDNA features and five machine learning algorithms constructed in the training cohort (cancer: 113; healthy: 113) outperformed all the models built on individual feature-algorithm combinations. This integrated model yielded superior sensitivities of 91.4% at 95.7% specificity for cohort validation I (area under the curve [AUC], 0.984), 84.7% at 98.6% specificity for validation II (AUC, 0.987), and 92.5% at 94.2% specificity for additional validation (AUC, 0.974), respectively. The model's high performance remained consistent when sequencing depth was down to 0.5× (AUC, 0.966-0.971). Furthermore, our model is sensitive to identifying early pathological features (83.2% sensitivity for stage I, 85.0% sensitivity for <1 cm tumor at the 0.66 cutoff). Conclusions: We have established a stacked ensemble model using cfDNA fragmentomics features and achieved superior sensitivity for detecting early-stage lung cancer, which could promote early diagnosis and benefit more patients.

10P Detection of early-stage lung cancer using 5-hydroxymethylcytosine signatures in circulating cell-free DNA

10P Detection of early-stage lung cancer using 5-hydroxymethylcytosine signatures in circulating cell-free DNA

The role of low-dose computed tomography in lung cancer screening

Introduction: Lung cancer is a serious public health problem in Hungary, but currently there is no nationwide screening program for the early detection of the disease. Several technological developments have been carried out recently to improve the effectiveness of lung cancer screening. Low-dose computed tomography (LDCT) is one of these technologies. Objective: The objective of this study is to provide an overview on guidelines and recommendations related to the application of LDCT as a novel lung cancer screening modality and to summarize the scientific literature and screening practices of other countries. Method: We performed a targeted literature review to collect information about LDCT in lung cancer screening. We searched in publicly available databases for guidelines and recommendations as well as scientific publications on screening and early detection of lung cancer. Results: In our literature search, we identified 16 guidelines and recommendations for lung cancer screening and LDCT. Regarding the efficacy of LDCT lung cancer screening, 10 foreign randomized controlled trials and 2 Hungarian trials were reviewed. Information on screening practices of 10 European countries were identified. Discussion: Evidences suggest that LDCT screening improves the detection of lung cancer, especially at early stages, and reduces cancer-specific mortality. Conclusion: In summary, in the high-risk population, LDCT can be considered an effective screening modality for the early-stage detection of lung cancer and for reducing lung cancer mortality. The ongoing Hungarian and foreign pilot programs may provide futher evidence for the implementation of a nationwide LDCT lung cancer screening program.

Abstract 727: Detection of early-stage lung cancer using a liquid biopsy test based on extracellular vesicle proteins

Abstract We have recently conducted a study to evaluate the use of proteins associated with extracellular vesicles (EVs) isolated using an alternating current electrokinetic (ACE) system for the detection of early stage (I &amp; II) non-small cell lung cancer (NSCLC). Most lung cancers are diagnosed at advanced stages when treatments are less effective, pointing to the need for developing non-invasive screening strategies such as liquid biopsies. We tested the feasibility of detecting the presence of early stage (I-II) lung cancer using protein biomarkers carried by EVs in patient blood. Using an ACE microelectrode array, EVs were purified from NSCLC patient (N = 62; Stage I = 39, Stage II = 23) or control plasma (N = 390) and tested for the presence of cancer biomarker proteins with bead-based multiplex immunoassay kits. A machine learning algorithm based on recursive feature elimination with cross-validation was developed to identify the most important features for differentiation between cases and controls. The algorithm identified 10 protein biomarkers that were able to distinguish between the NSCLC and control patient samples with a receiver-operator characteristic (AUC) of 0.986 (95% CI: 0.976 - 0.996). Sensitivity was 66.1% at 99.2% specificity, rising to 79.0% and 93.6% sensitivity at 98.2% and 95.1% specificity, respectively. When further stratified, Stage I sensitivity was 53.8% and stage II sensitivity was 86.7% at the 99.2% specificity level. These results suggest that it is possible to detect early-stage NSCLC when treatments may be more effective, that EVs may carry the earliest signals of cancer development, and that a test based on blood-based EV-protein biomarkers can yield high sensitivity and specificity. Our results warrant investigation in larger datasets to validate the current observations with the potential application of this methodology for routine screening of at-risk populations. Citation Format: Juan P. Hinestrosa, Jean M. Lewis, Gregor Schroder, Heath I. Balcer, Razelle Kurzrock, Rajaram Krishnan. Detection of early-stage lung cancer using a liquid biopsy test based on extracellular vesicle proteins [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 727.