Molecular Diagnosis Of Lung Cancer Research Articles

Molecular Diagnosis of Lung Cancer (LC) In the ERA of Cryobiopsy (CB). A Literature Review

Molecular Diagnosis of Lung Cancer (LC) In the ERA of Cryobiopsy (CB). A Literature Review

Transesophageal ultrasound-guided bronchoscopic Acquire TBNB versus Vizishot2 TBNA needles for neoplastic lesions: A retrospective study

BackgroundLung cancer is often diagnosed at an advanced stage; however, it has shown improved therapeutic efficacy with the introduction of molecularly targeted drugs and immune checkpoint inhibitors, necessitating accurate molecular diagnosis for effective treatment planning. Traditional sampling techniques, including endobronchial ultrasound-guided transbronchial needle aspiration, frequently require multiple biopsies to obtain sufficient tissues for multiplex testing, highlighting the need for more efficient methods. Therefore, we explored the diagnostic utility of endoscopic ultrasound with bronchoscope-guided fine-needle biopsy (EUS-B-FNB) versus fine-needle aspiration (EUS-B-FNA) in patients with lung cancer, focusing on tissue sample collection for molecular testing. The introduction of the Franseen needle in EUS-B-FNB, characterized by three beveled edges, allows for more tissue collection in cylinder form. MethodsWe retrospectively analyzed the data of 97 patients who underwent EUS-B-FNB or EUS-B-FNA at Hakodate Goryoukaku Hospital and evaluated diagnostic yields, safety, and nucleic acid concentrations using collected specimens. ResultsThe diagnostic yields of EUS-B-FNB and EUS-B-FNA were comparable (92.2% vs. 92.3%), with no significant differences in complications. However, EUS-B-FNB provided significantly higher DNA and RNA concentrations (DNA; 41.05 vs. 10.20 ng/mL; P < 0.0001, RNA; 36.80 vs. 11.80 ng/mL; P = 0.0009), essential for comprehensive molecular testing. ConclusionThis study highlights the potential of EUS-B-FNB for enhancing the molecular diagnosis of lung cancer by ensuring adequate tissue sample collection for multiplex testing, paving the way for personalized medicine. This technique is comparable in safety and efficacy to traditional methods while offering a substantial improvement in the quality of molecular diagnostics.

1187P Molecular diagnosis of lung cancer via ctDNA and ctRNA detection on bronchoscopic fluid specimens from 31 patients: A retrospective analysis

1187P Molecular diagnosis of lung cancer via ctDNA and ctRNA detection on bronchoscopic fluid specimens from 31 patients: A retrospective analysis

Molecular Profiling in Non-Squamous Non-Small Cell Lung Carcinoma: Towards a Switch to Next-Generation Sequencing Reflex Testing.

Molecular diagnosis of lung cancer is a constantly evolving field thanks to major advances in precision oncology. The wide range of actionable molecular alterations in non-squamous non-small cell lung carcinoma (NS-NSCLC) and the multiplicity of mechanisms of resistance to treatment resulted in the need for repeated testing to establish an accurate molecular diagnosis, as well as to track disease evolution over time. While assessing the increasing complexity of the molecular composition of tumors at baseline, as well as over time, has become increasingly challenging, the emergence and implementation of next-generation sequencing (NGS) testing has extensively facilitated molecular profiling in NS-NSCLC. In this review, we discuss recent developments in the molecular profiling of NS-NSCLC and how NGS addresses current needs, as well as how it can be implemented to address future challenges in the management of NS-NSCLC.

Cryobiopsy in Lung Cancer Diagnosis-A Literature Review.

Optimizing the diagnosis of lung cancer represents a challenge, as well as a necessity, for improving the low survival of these patients. Flexible bronchoscopy with forceps biopsy is one of the key diagnostic procedures used for lung tumors. The small sample size and crush artifacts are several factors that can often limit access to a complete diagnosis, therefore leading to the need of repeating the bronchoscopy procedure or other invasive diagnostic methods. The bronchoscopic cryobiopsy is a recent technique that proved its utility in the diagnosis of both endobronchial and peripheral lung tumors. In comparison with conventional forceps biopsy, studies report a higher diagnostic yield and a superior quality of the collected samples for both the histopathological and the molecular diagnosis of lung cancer. This method shows promising results in sampling lung tissue, alone, or in conjunction with fluoroscopy or radial endobronchial ultrasound (r-EBUS). With a good safety and cost-benefit profile, this novel method has the potential to improve the diagnosis, and therefore the management of lung cancer patients. The objective of this narrative review is to provide a comprehensive review of the recent data regarding the advantages of cryobiopsy and r-EBUS in lung cancer diagnosis.

182P Impact of SARS CoV 2 outbreak in the molecular diagnosis of advanced NSCLC: A retrospective comparative cohort study

Background: The SARS CoV 2 coronavirus pandemic has rocked health care systems to the core. Concurrent circulation of COVID 19 has led to service disruptions and delays in the standard procedures related to lung cancer (LC) diagnose and may negatively impact in the management, care and therapeutic patient intervention. We aimed to determine COVID 19 impact in the molecular diagnosis of LC at our institution.

Ambient mass spectrometry for the molecular diagnosis of lung cancer.

Lung cancer is one of the most common malignancies and the leading cause of cancer-related death worldwide. Among the technologies suitable for the rapid and accurate molecular diagnosis of lung cancer, ambient mass spectrometry (AMS) has gained increasing interest as it allows the direct profiling of molecular information from various biological samples (e.g., tissue, serum, urine and sputum) in real-time and with minimal or no sample pretreatment. This minireview summarizes the applications of AMS in lung cancer studies (including tissue molecular identification, the discovery of potential biomarkers, and surgical margin assessment), and discusses the challenges and perspectives of AMS in the clinical precision molecular diagnosis of lung cancer.

Template-ready PCR method for detection of human telomerase reverse transcriptase mRNA in sputum

Human telomerase reverse transcriptase (hTERT) mRNA in tissue is a biomarker of lung cancer, but hTERT mRNA in sputum had not been successfully detected with conventional reverse transcription PCR methods. Here, we developed a novel PCR protocol: Template-Ready PCR (TRPCR), to detect sputum hTERT mRNA, in which probes serve as templates of amplification. While free probes and dsDNA were removed in template preparation through aspiration and restriction digestion, probes that formed into heterocomplex with target RNA remained intact for PCR amplification. By fishing out the heterocomplex and amplifying the probes, TRPCR achieved sensitivity higher than reverse transcription-quantitative PCR (RT-qPCR). ROC curve of sputum hTERT mRNA by TRPCR assay showed the discrimination in high sensitivity and specificity between patients with lung cancer and lung cancer-free donors at the PCR Ct cutoff of 33. We further validated this approach through TRPCR assay of sputum from 858 lung cancer patients and 480 non-malignant pulmonary disease patients. 722 (84.2%) cases from 858 with lung cancer patients were detected as positive, whereas 461 (96.0%) cases from 480 non-malignant pulmonary disease patients were detected as negative, suggesting that TRPCR assay of sputum hTERT mRNA can serve as a non-invasive molecular diagnosis of lung cancer.

Identification of Proteomic Features To Distinguish Benign Pulmonary Nodules from Lung Adenocarcinoma.

We hypothesized that distinct protein expression features of benign and malignant pulmonary nodules may reveal novel candidate biomarkers for the early detection of lung cancer. We performed proteome profiling by liquid chromatography-tandem mass spectrometry to characterize 34 resected benign lung nodules, 24 untreated lung adenocarcinomas (ADCs), and biopsies of bronchial epithelium. Group comparisons identified 65 proteins that differentiate nodules from ADCs and normal bronchial epithelium and 66 proteins that differentiate ADCs from nodules and normal bronchial epithelium. We developed a multiplexed parallel reaction monitoring (PRM) assay to quantify a subset of 43 of these candidate biomarkers in an independent cohort of 20 benign nodules, 21 ADCs, and 20 normal bronchial biopsies. PRM analyses confirmed significant nodule-specific abundance of 10 proteins including ALOX5, ALOX5AP, CCL19, CILP1, COL5A2, ITGB2, ITGAX, PTPRE, S100A12, and SLC2A3 and significant ADC-specific abundance of CEACAM6, CRABP2, LAD1, PLOD2, and TMEM110-MUSTN1. Immunohistochemistry analyses for seven selected proteins performed on an independent set of tissue microarrays confirmed nodule-specific expression of ALOX5, ALOX5AP, ITGAX, and SLC2A3 and cancer-specific expression of CEACAM6. These studies illustrate the value of global and targeted proteomics in a systematic process to identify and qualify candidate biomarkers for noninvasive molecular diagnosis of lung cancer.

Biomarker Testing in Lung Carcinoma Cytology Specimens: A Perspective From Members of the Pulmonary Pathology Society.

The advent of targeted therapy in lung cancer has heralded a paradigm shift in the practice of cytopathology with the need for accurately subtyping lung carcinoma, as well as providing adequate material for molecular studies, to help guide clinical and therapeutic decisions. The variety and versatility of cytologic-specimen preparations offer significant advantages to molecular testing; however, they frequently remain underused. Therefore, evaluating the utility and adequacy of cytologic specimens is critical, not only from a lung cancer diagnosis standpoint but also for the myriad ancillary studies that are necessary to provide appropriate clinical management. A large fraction of lung cancers are diagnosed by aspiration or exfoliative cytology specimens, and thus, optimizing strategies to triage and best use the tissue for diagnosis and biomarker studies forms a critical component of lung cancer management. This review focuses on the opportunities and challenges of using cytologic specimens for molecular diagnosis of lung cancer and the role of cytopathology in the molecular era.

Evaluation of lung flute in sputum samples for molecular analysis of lung cancer

BackgroundMolecular analysis of sputum provides a promising approach for lung cancer diagnosis, yet is limited by the difficulty in collecting the specimens from individuals who can’t spontaneously expectorate sputum. Lung Flute is a small self-powered audio device that can induce sputum by generating sound waves and vibrating in the airways of the lungs. Here we propose to evaluate the usefulness of Lung Flute for sputum sampling to assist diagnosis of lung cancer.MethodsForty-three stage I lung cancer patients and 47 cancer-free individuals who couldn’t spontaneously cough sputum were instructed to use Lung Flute for sputum sampling. Expressions of two microRNAs, miRs-31 and 210, were determined in the specimens by qRT-PCR. The results were compared with sputum cytology.ResultsSputum was easily collected from 39 of 43 (90.7%) lung cancer patients and 42 of 47 (89.4%) controls with volume ranges from 1 to 5 ml (median, 2.6 ml). The specimens had less than 4% oral squamous cells, indicating that sputum was obtained from low respiratory tract. Expressions of miRs-31 and 210 in sputum were considerably higher in cancer patients than cancer-free individuals (8.990 vs. 4.514; 0.6847 vs. 0.3317; all P <0.001). Combined use of the two miRNAs produced a significantly higher sensitivity (61.5% vs. 35.9%, P = 0.002) and a slightly lower specificity (90.5% vs. 95.2%, p = 0.03) compared with cytology for lung cancer diagnosis.ConclusionLung Flute could potentially be useful in convenient and efficient collection of sputum for molecular diagnosis of lung cancer.

Aberrant anaplastic lymphoma kinase expression in high-grade pulmonary neuroendocrine carcinoma

AimsIn lung cancer with anaplastic lymphoma kinase (ALK) gene rearrangement, accurate diagnosis is essential to identify patients who can be treated with a specific kinase inhibitor. Sensitive ALK immunostaining can...

Mise en place d’un secteur de pathologie moléculaire en oncologie au sein d’un laboratoire d’anatomie pathologique (LPCE, CHU de Nice)

The advent of targeted therapies and personalized medicine in oncology has led in France to the settlement and organisation of a network of hospital molecular genetic platforms under the impetus of the National Cancer Institute (INCa). These platforms are, according to the concerned sites, integrated or not in pathology laboratories. The development of molecular biology methods, the choice of the procedures, the establishment of sample workflow, the quality control and the selection of the genomic alterations to be detected on each platform, have been left to the discretion of the different laboratories. Based on calls for project made by the INCa, hospital molecular genetic platforms were able to adapt their activity according to the assigned budgets. While the presence of some genomic alterations (i.e. KRAS gene mutations in metastatic colon adenocarcinoma or EGFR gene mutations in lung adenocarcinomas), may lead to administration of targeted therapies under the Marketing Authorization Application (MAA), others are associated with therapeutic clinical trials. However, increasing number of MAA for new molecules targeting genomic alterations is likely in the near future. In this context, it is necessary to quickly adapt the organisation of work of the hospital pathology laboratories performing molecular biology tests in order to meet the growing demand of oncologists in the field of targeted therapies. The purpose of this article is to describe the different steps of the settlement of a molecular genetic platform in an academic pathology laboratory (LPCE, CHU de Nice) and to show the experience of this laboratory specifically oriented on the support of the morphological and molecular diagnosis of lung cancer, thyroid cancer and malignant melanoma.

The morphological and molecular diagnosis of lung cancer.

In Germany, lung cancer causes more deaths than any other malignant disease. Its main etiology is smoking, but other risk factors need to be considered as well. The morphological, molecular and biological phenotype is complex and should no longer be just categorized as either small-cell or non-small cell lung cancer. This review article is based on the authors' longstanding involvement in the scientific investigation and diagnostic evaluation of lung cancer, including contributions to the current WHO classification and collaboration in the new interdisciplinary classification of adenocarcinoma. The relevant literature was selectively reviewed. Lung cancer is morphologically classified into four main subtypes-small-cell carcinoma, squamous-cell carcinoma, adenocarcinoma, and large-cell carcinoma. Genetic and molecular analyses have revealed distinct differences within subtypes; in particular, adenocarcinomas can be further subdivided. Complex techniques of genomic analysis are now available, but clinicopathological data are still the most important determinants of prognosis and are clearly better for this purpose than molecular classification alone. Nonetheless, the assessment of specific molecular markers is becoming increasingly important. The morphological and molecular classification of lung cancer is undergoing a re-evaluation which will lead to more accurate assessment of individual prognoses and to improved prediction of the response to specific treatment regimens.

Functionalization of upconverted luminescent NaYF4 : Yb/Er nanocrystals by folic acid–chitosan conjugates for targeted lung cancer cell imaging

We demonstrate a robust approach to functionalize the upconverted luminescent NaYF4:Yb/Er nanocrystals with folic acid (FA)–chitosan conjugates for specific targeting and imaging of lung cancer cells. In this approach, the amine groups of FA–chitosan conjugates covalently reacted with the carboxyl groups located on the surface of dimercaptosuccinic acid (DMSA)-modified NaYF4 : Yb/Er nanoparticles. The produced FA–chitosan-coated NaYF4 : Yb/Er nanoparticles were characterized by means of transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) and photoluminescence (PL) spectroscopy. Additionally, a cell viability assay showed that those FA–chitosan-coated fluorescence nanocrystals were fully biocompatible. Moreover, confocal fluorescence images illustrated that the functionalized-NaYF4 : Yb/Er nanoparticles with FA–chitosan conjugates were specific in targeting lung cancer cell lines (H460) rather than human lung fibroblast (HLF) cells. Consequently, the surface chemistry and functional design of the upconverted luminescence nanocrystals using biocompatible FA–chitosan conjugates might provide a powerful nanoplatform for the detection and molecular diagnosis of lung cancer.

A novel strategy for surface modification of superparamagnetic iron oxide nanoparticles for lung cancer imaging

Superparamagnetic iron oxide (SPIO) nanoparticles are widely used in magnetic resonance imaging (MRI) as versatile ultra-sensitive nanoprobes for cellular and molecular imaging of cancer. In this study, we report a one-step procedure for the surface functionalization of SPIO nanoparticles with a lung cancer-targeting peptide. The hydrophobic surfactants on the as-synthesized SPIO are displaced by the peptide containing a poly(ethylene glycol)-tethered cysteine residue through ligand exchange. The resulting SPIO particles are biocompatible and demonstrate high T(2) relaxivity. The nanoprobes are specific in targeting α(v)β(6)-expressing lung cancer cells as demonstrated by MR imaging and Prussian blue staining. This facile surface chemistry and the functional design of the proposed SPIO system may provide a powerful nanoplatform for the molecular diagnosis of lung cancer.

Combined detection of p53, p16, Rb, and EGFR mutations in lung cancer by suspension microarra

Mutations of some contributing factors (p53, p16, Rb, and EGFR) are believed to affect diagnosis and drug resistance of lung cancer. We evaluated the efficacy of a multimarker panel for molecular diagnosis of lung cancer, using a high-throughput suspension microarray. One hundred and twenty-five lung cancer specimens and 30 tumor-free lung tissue samples were assayed by multiplex polymerase chain reaction with specific probes designed to detect hot-spot mutations in p53, p16, Rb, and EGFR. The mutation rates of p53, p16, Rb, or EGFR in the lung cancer specimens were 36.8, 15.2, 11.2, and 18.4%, respectively. Inclusion of four markers elevated sensitivity to 68.0%. The specificity and accuracy of four-marker detection were 90.0 and 72.3%, and the mutation rates of this panel in stage I, stage II and stage III disease were 62.2, 65.9 and 75.0%, respectively. Mutation at p16 occurred more frequently in non-small cell lung cancer (19.3%) than in small cell lung cancer (5.4%); while the mutation rate of Rb was 32.4% in small cell lung cancer versus 2.3% in non-small cell lung cancer. We conclude that simultaneous detection of p53, Rb, p16, and EGFR in a suspension microarray facilitates rapid diagnosis of lung cancer.

Paraffin-based molecular diagnosis of lung cancer reproduces morphologic and molecular subtypes of lung cancer

7579 Background: Molecular profiling of lung cancer by gene expression analyses has documented potential to guide therapy. However, quality ‘fresh’ tissue for RNA analyses is generally unavailable in clinical practice. We introduce a (q)RT-PCR assay and analytic method for profiling lung cancers from clinically obtained formalin-fixed paraffin-embedded tissues. Methods: Approximately 1,000 DNA microarrays were analyzed to select genes distinguishing the major histological variants (adenocarcinoma, small cell carcinoma, etc) and previously described molecular subtypes of lung cancer. Based on gene expression, a classifier of 62 genes was constructed to assign each sample to its morphologic tumor type, as well as to risk stratify by molecular subtypes. A real-time qRT-PCR assay was developed to evaluate the expression of the 62 genes from paraffin-embedded samples. This assay was used to profile RNA extracted from a cohort of surgically treated lung cancer patients. Samples were procured as fresh frozen and formalin-fixed, paraffin-embedded and were archived between 1–15 years. Results: Fifty-eight of 62 genes passed performance criteria and were used for analyses. Normalized gene expression was used for sample classification. The cohort contained a broad spectrum of tumors in proportions consistent with clinical practice. Gene amplification was successful in 139 of 142 (98%) lung cancer samples. Matched frozen-paraffin and replicate paraffin samples had mean correlations of approximately 80%. Linear discriminant analysis of gene expression data agreed with morphologic classification by light microscopy in &gt;99% of cases. More importantly, the method successfully re-identified molecular subtypes of lung cancer for the first time through the use of a paraffin-based assay. Clinical outcomes previously associated with molecular tumor subtypes, including differential survival and metastatic patterns, were again seen in our cohort. Conclusions: We describe for the first time a clinically meaningful and robust molecular diagnosis of a clinical cohort of lung cancer patients which is complementary to morphologic cancer diagnosis. This assay is easily implemented using specimens routinely collected in current patient care. No significant financial relationships to disclose.

Hypermethylation of p16INK4a in Chinese lung cancer patients: biological and clinical implications.

Promoter hypermethylation of the p16INK4a gene was investigated in 111 cases of tumor tissue, as well as in 136 circulating plasma and 95 sputum samples from Chinese patients with primary lung cancer, using a modified protocol of semi-nested methylation-specific-PCR (MSP). The results showed hypermethylated p16 sequence in 80.2% of tumor tissues and frequencies of 75.7 and 74.7% in plasma and sputum specimens, respectively. Among the patients, 50 cases of matched plasma, sputum and tumor tissue from the same individual were analyzed. Of these, hypermethylation of the p16 promoter was detected in 84.0% of the tumor tissues, with frequencies of 72.0 and 76.0% in the corresponding plasma and sputum, respectively. Notably, only patients whose tumor tissue showed hypermethylation of p16 exhibited the same aberrant methylation in their sputum and/or plasma. Hypermethylation of p16 in sputum and plasma samples may provide a more sensitive approach to molecular diagnosis of lung cancer than relying on conventional cytological analysis. Our data show that a combination of cytological analysis of sputum and examination of p16 hypermethylation in sputum and plasma identified 92.0% (46/50) of the lung cancer patients studied, offering an effective means of early detection of lung cancer.

Recent advances in the molecular diagnosis of lung cancer.

Despite extensive effort in improvement of diagnosis and treatment of patients with lung cancer in past three decades, the overall survival of patients with the disease remains dismal. Because the development of lung cancer takes a few decades, early diagnosis of the disease or identification of truly high-risk populations may provide us opportunity to successfully cure or prevent the disease. Recent advances in understanding biological basis of lung tumorigenesis provide new tools for detecting malignant cells or the process of malignant transformation and progression. Along with identification of molecular abnormalities in the early lung tumorigenesis, advanced molecular analytic technologies have been emerged, which may facilitate development of rapid and effective methods for early diagnosis and risk assessment. Here, I discuss recent progresses in understanding of early molecular abnormalities in lung cancer, efforts of translating laboratory findings to clinical tests, and prospective of biomarkers in lung cancer diagnosis and risk assessment.