Non-invasive detection of orthotopic human lung tumors by microRNA expression profiling of mouse exhaled breath condensates and exhaled extracellular vesicles.

Aim: The lung is the second most frequent site of metastatic dissemination. Early detection is key to improving survival. Given that the lung interfaces with the external environment, the collection of exhaled breath condensate (EBC) provides the opportunity to obtain biological material including exhaled miRNAs that originate from the lung. Methods: In this proof-of-principal study, we used the highly metastatic MDA-MB-231 subline 3475 breast cancer cell line (LM-3475) to establish an orthotopic lung tumor-bearing mouse model and investigate non-invasive detection of lung tumors by analysis of exhaled miRNAs. We initially conducted miRNA NGS and qPCR validation analyses on condensates collected from unrestrained animals and identified significant miRNA expression differences between the condensates of lung tumor-bearing and control mice. To focus our purification of EBC and evaluate the origin of these differentially expressed miRNAs, we developed a system to collect EBC directly from the nose and mouth of our mice. Results: Using nanoparticle distribution analyses, TEM, and ONi super-resolution nanoimaging, we determined that human tumor EVs could be increasingly detected in mouse EBC during the progression of secondary lung tumors. Using our customizable EV-CATCHER assay, we purified human tumor EVs from mouse EBC and demonstrated that the bulk of differentially expressed exhaled miRNAs originate from lung tumors, which could be detected by qPCR within 1 to 2 weeks after tail vein injection of the metastatic cells. Conclusion: This study is the first of its kind and demonstrates that lung tumor EVs are exhaled in mice and provide non-invasive biomarkers for detection of lung tumors.

A clinical nomogram for predicting tumor regression grade in esophageal squamous-cell carcinoma treated with immune neoadjuvant immunotherapy.

Background: There are various treatment options for esophageal squamous cell cancer. including surgery, peri-operative chemotherapy, and radiation. More recently, neoadjuvant immunotherapy has also been shown improve outcomes. In this study, we addressed the question, "Can we predict which patients with esophageal squamous cell cancer will benefit from neoadjuvant immunotherapy?". Methods: All patients with thoracic esophageal squamous-cell carcinoma (T2N+M0-T3-4N0/+M0) (according to the eighth edition of the National Comprehensive Cancer Network guidelines) who underwent immune neoadjuvant immunochemotherapy with programmed cell death protein 1 (PD-1) combined with paclitaxel plus cisplatin or nedaplatin in the Affiliated Cancer Hospital of Zhengzhou University, China, between November 2019 and August 2021 were included in this study. All patients underwent surgical resection. We developed a response [tumor regression grade (TRG)] prediction model using the least absolute shrinkage and selection operator (LASSO) regression incorporating factors associated with response. The accuracy of the prediction model was then validated. Results: We included 79 patients who underwent neoadjuvant immunotherapy combined with chemotherapy, aged 48-78 years (62.05±6.67), including 21 males and 58 females. There were five cases of immune-related pneumonia, of which three cases were diagnosed as immune-related pneumonia during the perioperative period, and one case of immune-related thyroid dysfunction changes. After LASSO regression, the factors that were independently associated with TRG were clinical T stage before neoadjuvant therapy, clinical N stage before neoadjuvant therapy, albumin level difference from before to after neoadjuvant therapy, white blood cell (WBC) count before neoadjuvant therapy, and T stage before surgery. We constructed a prediction model, plotted the nomogram, and verified its accuracy. Its Brier score was 0.13, its calibration slope was 0.98, and its C-index was 0.90 (95% CI: 0.82-0.97). Conclusions: Our prediction model can predict the likelihood of TRG in patients with esophageal squamous cell cancer after immunotherapy combined with neoadjuvant chemotherapy. Using this prediction model, we plan to conduct a subsequent neoadjuvant radiotherapy in patients with of TRG 2-3 patients with neoadjuvant radiotherapy.

Esophageal cancer: the rise of adenocarcinoma over squamous cell carcinoma in the Asian belt.

Esophageal cancer is a common cancer worldwide with a high associated mortality rate. Amongst the two most frequent subsets of disease, squamous cell carcinoma (SCC) and adenocarcinoma (AC), there has been an epidemiologic shift towards adenocarcinoma over the last few decades. However, squamous cell carcinoma still predominates worldwide. Within Western countries, obesity has been associated with an increase in esophageal AC. A striking report from the World Health Organization (WHO) stated that worldwide obesity has tripled since 1975. In 2016, the WHO reported that greater than 1.9 billion adults are overweight and over 650 million were obese. In this review our goal is to analyze the esophageal cancer trends of China, which is the largest contributor among the esophageal cancer "Asian belt." Our intent is to evaluate whether there is a correlation between the rise in esophageal adenocarcinoma and obesity in this esophageal cancer "hotspot." With further analysis, the high-risk populations that are identified can be targeted to implement preventative strategies. This focus will aid in decreasing the burden of esophageal cancer at the global health level by addressing preventative strategies, such as screening endoscopy and lifestyle modifications. For example, WHO developed a global action plan on physical activity in response to the rise in obesity worldwide. Prevention is key to decreasing the rate of esophageal adenocarcinoma as majority of cases are diagnosed in the late stages leading to high mortality rates.

Development of a novel miR-3648-related gene signature as a prognostic biomarker in esophageal adenocarcinoma.

BACKGROUND: Esophageal adenocarcinoma (EA) is a typical immunogenic malignant tumor with a dismal 5-year survival rate lower than 20%. Although miRNA-3648 (miR-3648) is expressed abnormally in EA, its impact on the tumor immune microenvironment remains unknown. In this study, we sought to identify immune-related genes (IRGs) that are targeted by miR-3648 and develop an EA multigene signature. METHODS: The gene expression data of 87 EA tumor samples and 67 normal tissue samples from The Cancer Genome Atlas (TCGA) database and the Genotype-Tissue Expression (GTEx) database were downloaded, respectively. Weighted gene co-expression network analysis (WGCNA), the CIBERSORT algorithm, and Cox regression analysis were applied to identify IRGs and to construct a prognostic signature and nomogram. RESULTS: MiR-3648 was expectedly highly expressed in EA tumor tissues (P=2.6e-8), and related to the infiltration of activated natural killer cells (NK cells) and activated CD4 T lymphocytes (CD4 cells). A total of 70 miR-3648-targeted genes related to immune cell infiltration were identified. Among them, 4 genes (C10orf55, DLL4, PANX2, and NKAIN1) were closely related to overall survival (OS), and were thus selected to construct a 4-gene risk score (RS). The RS had a superior capability to predict OS [area under the curve (AUC) =0.740 for 1 year; AUC =0.717 for 3 years; AUC =0.622 for 5 years]. A higher score was indicative of a poorer prognosis than a lower score [hazard ratio (HR) =2.71; 95% confidence interval (CI): 1.45-5.09; P=0.002]. Furthermore, the nomogram formed by combining the RS and the TNM classification of malignant tumors (TNM stage) improved the accuracy of survival prediction [Harrell's concordance index (C-index) =0.698]. CONCLUSIONS: MiR-3648 may play a critical role in EA pathogenesis. The novel 4-gene signature may serve as a prognostic tool to manage patients with EA.

A volatile biomarker in breath predicts lung cancer and pulmonary nodules.

BACKGROUND: previous studies have reported volatile organic compounds (VOCs) in the breath as apparent biomarkers of lung cancer. We tested the hypothesis that a robust breath VOC biomarker of lung cancer should also predict pulmonary nodules in chest CT images. METHODS: Biomarker discovery study (unblinded): 301 subjects were screened for lung cancer with low dose chest CT (LDCT), and donated duplicate samples of alveolar breath for analysis with gas chromatography mass spectrometry (GC MS). Monte Carlo analysis of breath chromatograms revealed a mass ion as a biomarker that identified biopsy-proven lung cancer as well as suspicious pulmonary nodules on LDCT. The biomarker was termed Mass Abnormalities in Gaseous Ions with Imaging Correlates (MAGIIC). The chemical structure of MAGIIC was tentatively identified from the NIST library of mass spectra; the best-fit compounds included C4 and C5 alkane derivatives that were consistent with metabolic products of oxidative stress. Blinded validation of MAGIIC: the abundance of the MAGIIC biomarker was determined in a different group of 161 subjects undergoing screening with LDCT. They donated duplicate alveolar breath VOC samples that were analyzed at two independent laboratories. The study was blinded and monitored with Good Clinical Practice. The abundance of MAGIIC in breath predicted biopsy-proven lung cancer with 84% accuracy, sensitivity = 75.4% and specificity = 85.0%. MAGIIC also predicted pulmonary nodules in LDCT with 80.5% accuracy, sensitivity = 80.1% and specificity = 75.0%. Breath MAGIIC abundance was not significantly affected by tobacco smoking history. CONCLUSIONS: in a blinded study, breath VOC MAGIIC accurately predicted lung cancer confirmed on a tissue biopsy, as well as suspicious pulmonary nodules observed on LDCT. MAGIIC may have been a product of oxidative stress and it could potentially be employed as an ancillary to LDCT to predict the likelihood that a pulmonary nodule is malignant.

Bronchoscopy: Diagnostic and Therapeutic for Non-Small Cell Lung Cancer.

The bronchoscope has gone through much advancement from its origin as a thin metal tube. It has become a highly sophisticated tool for clinicians. Both rigid and the flexible bronchoscopes are invaluable in the diagnosis and treatment of non-small cell lung cancer. Treatment of this disease process hinges on accurate diagnosis and lymph node staging. Technologies, such as endobronchial ultrasound, navigational bronchoscopy, and autofluorescence, have improved efficacy of endobronchial diagnosis and sample collection. If a patient is not a candidate for surgery and has a complication from a centrally located mass, the bronchoscope has been used to deliver palliative therapies.

Epithelioid inflammatory myofibroblastic sarcoma: a case report.

Inflammatory myofibroblastic tumor (IMT) of the lung is a rare malignancy with few cases reported in the literature. Histologically, it is composed by spindle cells and an infiltrate of inflammatory cells. Children and young, non-smoking adults constitute the majority of cases, the clinical behavior ranges from a benign entity to a malignant process with rapid recurrence and metastatic progression. We present a case of epithelioid inflammatory myofibroblastic sarcoma (EIMS) of the pleura, a malignant variant of IMT, which was initially treated with debulking surgical resection followed by systemic chemotherapy. The tumor was found to have an anaplastic lymphoma kinase (ALK) gene rearrangement. An ALK directed tyrosine kinase inhibitor was used with an impressive response, the patient remains in remission nearly 1 year after presentation. The pathogenesis, pathologic findings, clinical behavior and imaging of pulmonary EIMS are discussed.

Blinded Validation of Breath Biomarkers of Lung Cancer, a Potential Ancillary to Chest CT Screening.

BACKGROUND: Breath volatile organic compounds (VOCs) have been reported as biomarkers of lung cancer, but it is not known if biomarkers identified in one group can identify disease in a separate independent cohort. Also, it is not known if combining breath biomarkers with chest CT has the potential to improve the sensitivity and specificity of lung cancer screening. METHODS: Model-building phase (unblinded): Breath VOCs were analyzed with gas chromatography mass spectrometry in 82 asymptomatic smokers having screening chest CT, 84 symptomatic high-risk subjects with a tissue diagnosis, 100 without a tissue diagnosis, and 35 healthy subjects. Multiple Monte Carlo simulations identified breath VOC mass ions with greater than random diagnostic accuracy for lung cancer, and these were combined in a multivariate predictive algorithm. Model-testing phase (blinded validation): We analyzed breath VOCs in an independent cohort of similar subjects (n = 70, 51, 75 and 19 respectively). The algorithm predicted discriminant function (DF) values in blinded replicate breath VOC samples analyzed independently at two laboratories (A and B). Outcome modeling: We modeled the expected effects of combining breath biomarkers with chest CT on the sensitivity and specificity of lung cancer screening. RESULTS: Unblinded model-building phase. The algorithm identified lung cancer with sensitivity 74.0%, specificity 70.7% and C-statistic 0.78. Blinded model-testing phase: The algorithm identified lung cancer at Laboratory A with sensitivity 68.0%, specificity 68.4%, C-statistic 0.71; and at Laboratory B with sensitivity 70.1%, specificity 68.0%, C-statistic 0.70, with linear correlation between replicates (r = 0.88). In a projected outcome model, breath biomarkers increased the sensitivity, specificity, and positive and negative predictive values of chest CT for lung cancer when the tests were combined in series or parallel. CONCLUSIONS: Breath VOC mass ion biomarkers identified lung cancer in a separate independent cohort, in a blinded replicated study. Combining breath biomarkers with chest CT could potentially improve the sensitivity and specificity of lung cancer screening. TRIAL REGISTRATION: ClinicalTrials.gov NCT00639067.

An immersive surgery training system with live streaming capability.

Providing real-time, interactive immersive surgical training has been a key research area in telemedicine. Earlier approaches have mainly adopted videotaped training that can only show imagery from a fixed view point. Recent advances on commodity 3D imaging have enabled a new paradigm for immersive surgical training by acquiring nearly complete 3D reconstructions of actual surgical procedures. However, unlike 2D videotaping that can easily stream data in real-time, by far 3D imaging based solutions require pre-capturing and processing the data; surgical trainings using the data have to be conducted offline after the acquisition. In this paper, we present a new real-time immersive 3D surgical training system. Our solution builds upon the recent multi-Kinect based surgical training system [1] that can acquire and display high delity 3D surgical procedures using only a small number of Microsoft Kinect sensors. We build on top of the system a client-server model for real-time streaming. On the server front, we efficiently fuse multiple Kinect data acquired from different viewpoints and compress and then stream the data to the client. On the client front, we build an interactive space-time navigator to allow remote users (e.g., trainees) to witness the surgical procedure in real-time as if they were present in the room.

Elimination of socioeconomic and racial disparities related to lung cancer: closing the gap at a high volume community cancer center.

BACKGROUND: Healthcare disparities have afflicted the healthcare industry for decades and there have been many campaigns in recent years to identify and eliminate disparities. The purpose of this study was to identify disparities in the lung cancer population of a single community cancer center and to report the results in accordance with industry goals. METHODS: This was a retrospective cohort study of data on non-small cell lung cancer patients recorded in the Christiana Care Tumor Registry (CCTR) in Delaware. Gender, age, race, socioeconomic status and insurance status were used as potential variables in identifying disparities. RESULTS: We found no significant disparities between sexes, race or patients who were classified as having socioeconomic status 1-3. There was a lower survival rate associated with having the poorest socioeconomic status and in patients who used Medicare. Uninsured patients had the best survival outcomes and patients with Medicare had the poorest survival outcomes. CONCLUSION: Although we have closed the gap on sex and racial disparities, there remains a difference in survival outcomes across socioeconomic classes and insurance types.

A portable immersive surgery training system using RGB-D sensors.

Surgical training plays an important role in assisting residents to develop critical skills. Providing effective surgical training, however, remains as a challenging task. Existing videotaped training instructions can only show imagery from a fixed viewpoint that lacks both depth perception and interactivity. We present a new portable immersive surgical training system that is capable of acquiring and displaying high fidelity 3D reconstructions of actual surgical procedures. Our solution utilizes a set of Microsoft Kinect sensors to simultaneously recover the participants, the surgical environment, and the surgical scene itself. We then develop a space-time navigator to allow the trainees to witness and explore a prior procedure as if they were there. Preliminary feedback from residents shows that our system is much more effective than conventional videotaped system.

G-rich oligonucleotides alter cell cycle progression and induce apoptosis specifically in OE19 esophageal tumor cells.

Short synthetic oligonucleotides (ODNs) can be used to block cellular processes involved in cell growth and proliferation. Often acting as aptamers, these molecules interact with critical proteins that regulate the induction of apoptosis or necrosis. We have used a specialized class of ODNs that contain a monomeric sequence of guanosine to induce apoptosis specifically in the malignant esophageal cell line, OE19, in cell culture, and in a NODscid mouse model. OE19 cells were grown in culture and treated with a stable G-rich oligonucleotide (GRO). Cells were processed and apoptosis was measured by FACS analyses, caspase activity, and Hoescht staining. Circular dichroism (CD) was used to define the structure and stability of various GROs. The GRO works by first inducing retardation in the progression of the cell cycle and then by creating a sub-G1 population of apoptotic cells. The reaction is dose dependent, and appears to rely on the capacity of the G-rich ODN to adopt a G-quartet conformation. Apoptosis was measured by determining caspase 3/7 levels and by staining for nuclear fragmentation using the Hoechst dye. Importantly, nonmalignant esophageal cells or normal human lung fibroblasts are not impeded in their cell cycle progression when incubated with the G-rich ODNs. These results suggest that a selective killing of esophageal tumor cells is directed by G-rich ODNs. Selective killing was demonstrated in the unique activity of the GRO compared to other ODNs of different sequences as well as the response of oncogenic cells compared to nononcogenic cells.

Virtual bronchoscopy: clinical applications and limitations.

Virtual bronchoscopy (VB) is a term that describes a variety of software-based three-dimensional visualizations created from noninvasive medical imaging methods such as CT or MRI scans, with the goal of creating results similar to minimally invasive bronchoscopy procedures of the trachea and upper airways. This technique offers a detailed, noninvasive view of the airways, with reduced risk of infection or perforation, and facilitates preoperative planning for airway interventions that would otherwise not be possible. It is unlikely that VB will replace fiberoptic bronchoscopy; however, as VB techniques become more sophisticated and as sufficient computing and imaging power become more readily available, noninvasive visualization of the airways will play an important and useful role in the evaluation of airway diseases in well defined clinical situations.

Epidemiology of adenocarcinoma of the esophagogastric junction.

The incidence of adenocarcinoma of the esophagogastric junction has increased rapidly in the later half of the twentieth century in the United States and most western countries. Although squamous cell carcinoma of the esophagus used to predominate, adenocarcinoma of the distal esophagus and esophagogastric junction now accounts for more than half of new diagnoses in western countries. There also has been a shift from the development of distal gastric cancers to ones in a more proximal location. These recent epidemiologic shifts have led to controversy regarding the etiology and treatment of adenocarcinoma of the esophagogastric junction. Uncertainty still exists with regards to nomenclature and classification, risk factors, treatment, and screening and surveillance of esophagogastric adenocarcinoma. This article examines the epidemiology and etiologies of adenocarcinoma of the esophagogastric junction.