Since the introduction of video-assisted thoracoscopic surgery, the demand for its use in resecting small pulmonary nodules has increased. In parallel, the development of high-resolution computed ...tomography has led to an increase in the detection of the early lung cancers appearing as nodules with ground-glass opacity. Several techniques to resect these small lesions have been devised, the most familiar of which is the use of a computed tomography-guided percutaneous hook wire. We recently developed virtual-assisted lung mapping to achieve safer and more reliable resection of these lesions. Virtual-assisted lung mapping is carried out using three-dimensional computed tomography, bronchoscopy, and fluoroscopically guided navigation to mark the lung surface with dye. A prospective study showed that this technique was safe and had a high success rate. Multiple marking around the targeted lesion allows for a sufficient surgical margin at resection. We recently introduced an electromagnetic navigation bronchoscopy system to confirm the sites to be marked by virtual-assisted lung mapping in the operating room prior to video-assisted thoracoscopic surgery. We are now studying a method to enable minimally invasive, safe, and reliable resection of lesions located deep in the lung parenchyma.
The integration of metabolomics and transcriptomics can provide precise information on gene-to-metabolite networks for identifying the function of unknown genes unless there has been a ...post-transcriptional modification. Here, we report a comprehensive analysis of the metabolome and transcriptome of Arabidopsis thaliana over-expressing the PAP1 gene encoding an MYB transcription factor, for the identification of novel gene functions involved in flavonoid biosynthesis. For metabolome analysis, we performed flavonoid-targeted analysis by high-performance liquid chromatography-mass spectrometry and non-targeted analysis by Fourier-transform ion-cyclotron mass spectrometry with an ultrahigh-resolution capacity. This combined analysis revealed the specific accumulation of cyanidin and quercetin derivatives, and identified eight novel anthocyanins from an array of putative 1800 metabolites in PAP1 over-expressing plants. The transcriptome analysis of 22 810 genes on a DNA microarray revealed the induction of 38 genes by ectopic PAP1 over-expression. In addition to well-known genes involved in anthocyanin production, several genes with unidentified functions or annotated with putative functions, encoding putative glycosyltransferase, acyltransferase, glutathione S-transferase, sugar transporters and transcription factors, were induced by PAP1. Two putative glycosyltransferase genes (At5g17050 and At4g14090) induced by PAP1 expression were confirmed to encode flavonoid 3-O-glucosyltransferase and anthocyanin 5-O-glucosyltransferase, respectively, from the enzymatic activity of their recombinant proteins in vitro and results of the analysis of anthocyanins in the respective T-DNA-inserted mutants. The functional genomics approach through the integration of metabolomics and transcriptomics presented here provides an innovative means of identifying novel gene functions involved in plant metabolism.
The interaction of immune cells and cancer cells shapes the immunosuppressive tumor microenvironment. For successful cancer immunotherapy, comprehensive knowledge of antitumor immunity as a dynamic ...spatiotemporal process is required for each individual patient. To this end, we developed an immunogram for the cancer-immunity cycle by using next-generation sequencing.
Whole exome sequencing and RNA sequencing were performed in 20 patients with NSCLC (12 with adenocarcinoma, seven with squamous cell carcinoma, and one with large cell neuroendocrine carcinoma). Mutated neoantigens and cancer germline antigens expressed in the tumor were assessed for predicted binding to patients’ human leukocyte antigen molecules. The expression of genes related to cancer immunity was assessed and normalized to construct a radar chart composed of eight axes reflecting seven steps in the cancer-immunity cycle.
Three immunogram patterns were observed in patients with lung cancer: T-cell–rich, T-cell–poor, and intermediate. The T-cell–rich pattern was characterized by gene signatures of abundant T cells, regulatory T cells, myeloid-derived suppressor cells, checkpoint molecules, and immune-inhibitory molecules in the tumor, suggesting the presence of antitumor immunity dampened by an immunosuppressive microenvironment. The T-cell–poor phenotype reflected lack of antitumor immunity, inadequate dendritic cell activation, and insufficient antigen presentation in the tumor. Immunograms for both the patients with adenocarcinoma and the patients with nonadenocarcinoma tumors included both T-cell–rich and T-cell–poor phenotypes, suggesting that histologic type does not necessarily reflect the cancer immunity status of the tumor.
The patient-specific landscape of the tumor microenvironment can be appreciated by using immunograms as integrated biomarkers, which may thus become a valuable resource for optimal personalized immunotherapy.
Only a small fraction of tumor‐infiltrating lymphocytes can specifically recognize and attack cancer cells in PD‐1/PD‐L1 blockade therapy. Here, we investigate approaches to expand the ...neoantigen‐specific CD8+ T cells to overcome the difficulties in treating PD‐1/PD‐L1 blockade‐resistant tumors. Mutation‐associated neoepitopes of murine nonsmall cell lung cancer ASB‐XIV were estimated by whole‐exome and RNA sequencing and predicted by MHC‐I binding affinity (FPKM >1) in silico. Using ASB‐XIV‐specific CD8+ T cells, we screened a panel of 257 neoepitope peptides derived from ASB‐XIV missense and indel mutations. Mutated Phf3 peptide (mPhf3) was successfully identified as an immunogenic neoepitope. Prophylactic mPhf3‐DC vaccination inhibited ASB‐XIV tumor growth through CD8+ T cell‐mediated antitumor immunity. Combining the mPhf3‐DC vaccine and anti‐PD‐1 treatment elicited robust antitumor activity through the induction of mPhf3‐specific CD8+ T cells in the tumor microenvironment. Furthermore, the adoptive transfer of mPhf3‐specific CD8+ T cells eradicated ASB‐XIV tumors. Likewise, the combination of mutated Cdt1 peptide (mCdt1)‐DC vaccine and anti‐PD‐1 treatment or adoptive transfer of mCdt1‐specific CD8+ T cells also led to significant regression of PD‐1 blockade‐resistant murine gastric YTN16 tumors. In conclusion, a novel immunogenic neoepitope of ASB‐XIV was identified for immunotherapy targeting neoantigens. Identification of immunogenic neoantigens can extend the therapeutic strategies by increasing the frequency of neoantigen‐specific T cells, even for PD‐1/PD‐L1 blockade‐resistant tumors.
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Only a small fraction of tumor‐infiltrating lymphocytes can specifically recognize and attack cancer cells during PD1/PD‐L1 blockade therapy. Here, the authors identified a novel immunogenic tumor mutation‐associated neoepitope in two murine models for lung and gastric cancer. The combination of anti‐PD‐1 treatment and neoantigen vaccination or adoptive transfer of neoantigen‐specific CD8+ T cells led to significant regression of PD‐1 blockade‐resistant lung and gastric tumors. The identification of neoantigens could help monitor antitumor immunity dynamics and expand potential therapeutic strategies by increasing the frequency of neoantigen‐specific T cells, even for PD‐1/PD‐L1 blockade‐resistant solid tumors.
Virtual-assisted lung mapping is a preoperative bronchoscopic multi-spot dye-marking technique. This study aimed to examine the efficacy of virtual-assisted lung mapping for obtaining sufficient ...surgical margins in sublobar lung resection.
The multicenter, prospective, single-arm study was conducted from September 2016 to July 2017 in 19 registered centers. Patients who required sublobar lung resection and careful determination of resection margins underwent virtual-assisted lung mapping followed by thoracoscopic surgery. Successful resection was defined as resection of the lesion with margins greater than the lesion diameter or 2 cm using the preoperatively planned resection without additional resection. We defined the primary goal of the study as achieving successful resection in 95% of lesions.
The resection of 203 lesions (average diameter, 9.6 ± 5.3 mm) was intended in 153 patients. The lesions included pure and mixed ground-glass nodules (75 35.9% and 36 17.2%, respectively), solid nodules (91 43.5%), and others (7 3.3%). Surgical procedures included wedge resection (131, 71.2%), segmentectomy (51, 27.7%), and others (2, 1.1%). Successful resection was achieved in 178 lesions (87.8% 95% confidence interval, 82.4-91.9%), and virtual-assisted lung mapping markings successfully aided in the identification of 190 lesions (93.6% 95% confidence interval, 89.3-96.5%). Multivariable analysis showed that the most significant factor affecting resection success was the depth of the necessary resection margin (P = .0072).
This study showed that virtual-assisted lung mapping has reasonable efficacy, although the successful resection rate did not reach the primary goal. The depth of the required margin was the most significant factor leading to resection failure.
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Adenosine‐to‐inosine (A‐to‐I) microRNA editing is associated with tumor phenotypes in various cancer types. Recent analyses of The Cancer Genome Atlas (TCGA) dataset have shown several microRNAs that ...undergo A‐to‐I editing in human cancers, some of which have been reported to be associated with prognosis. Herein, we examined published small RNA deep sequencing data of 74 cases of lung adenocarcinoma (AD) and the corresponding normal counterpart (NC) specimen in silico in order to identify A‐to‐I microRNA editing events. Editing levels of miR‐379‐5p, miR‐99a‐5p, and miR‐497‐5p were lower in AD than in NC and, in a large number of cases, the editing level of miR‐200b‐3p was higher in AD than in NC. Difference in the editing level between AD and NC was largest for miR‐99a‐5p. Then, we examined the editing level of miR‐99a‐5p in 50 surgically resected lung adenocarcinoma cases at our institution by a conventional sequence‐based method, and its association with clinical outcomes. The editing level of miR‐99a‐5p was significantly lower in 19 cases of AD (38%) than in corresponding NC. These cases showed a shorter overall survival as assessed using the log‐rank test (P = .047). This trend was consistent with previous analyses of TCGA dataset. The altered editing level of microRNAs in lung adenocarcinoma could serve as a potential biomarker.
We compared the editing levels of miR‐99a‐5p between cancerous tissues and corresponding normal tissues from 50 cases of lung adenocarcinoma at our institution. Nineteen cases with significant loss of editing in adenocarcinoma showed a shorter overall survival (P = .047). The altered editing level of microRNAs in lung adenocarcinoma could serve as a potential biomarker.
YAP1, the main Hippo pathway effector, is a potent oncogene and is overexpressed in non‐small‐cell lung cancer (NSCLC); however, the YAP1 expression pattern in small‐cell lung cancer (SCLC) has not ...yet been elucidated in detail. We report that the loss of YAP1 is a special feature of high‐grade neuroendocrine lung tumors. A hierarchical cluster analysis of 15 high‐grade neuroendocrine tumor cell lines containing 14 SCLC cell lines that depended on the genes of Hippo pathway molecules and neuroendocrine markers clearly classified these lines into two groups: the YAP1‐negative and neuroendocrine marker‐positive group (n = 11), and the YAP1‐positive and neuroendocrine marker‐negative group (n = 4). Among the 41 NSCLC cell lines examined, the loss of YAP1 was only observed in one cell line showing the strong expression of neuroendocrine markers. Immunostaining for YAP1, using the sections of 189 NSCLC, 41 SCLC, and 30 large cell neuroendocrine carcinoma (LCNEC) cases, revealed that the loss of YAP1 was common in SCLC (40/41, 98%) and LCNEC (18/30, 60%), but was rare in NSCLC (6/189, 3%). Among the SCLC and LCNEC cases tested, the loss of YAP1 correlated with the expression of neuroendocrine markers, and a survival analysis revealed that YAP1‐negative cases were more chemosensitive than YAP1‐positive cases. Chemosensitivity test for cisplatin using YAP1‐positive/YAP1‐negative SCLC cell lines also showed compatible results. YAP1‐sh‐mediated knockdown induced the neuroendocrine marker RAB3a, which suggested the possible involvement of YAP1 in the regulation of neuroendocrine differentiation. Thus, we showed that the loss of YAP1 has potential as a clinical marker for predicting neuroendocrine features and chemosensitivity.
YAP1 is possibly involved in the regulation of neuroendocrine differentiation of lung tumors. Loss of YAP1 correlated with strong expression of neuroendocrine markers. Loss of YAP1 is a predictor of chemothensitivity in high‐grade neuroendocrine tumors.
Cancer-associated fibroblasts (CAFs) interact closely with cancer cells, supporting their growth and invasion. To investigate the role of microRNA-21 (miR-21) in lung adenocarcinoma, and especially ...in its CAF component, in situ hybridisation was applied to samples from 89 invasive lung adenocarcinoma cases. MiR-21 expression was observed in both cancer cells and CAFs. When the patients were stratified by expression, miR-21 levels in CAFs (n = 9), but not in cancer cells (n = 21), were inversely correlated with patient survival; patients with miR-21
CAFs exhibited lower survival than those with miR-21
CAFs. The underlying mechanism was investigated in vitro. Conditioned medium (CM) from A549 lung cancer cells increased miR-21 expression in MRC-5 and IMR-90 lung fibroblasts through the transforming growth factor-β pathway, and induced CAF-like morphology and migratory capacity. MiR-21 up-regulation in lung fibroblasts induced a novel CAF-secreted protein, calumenin, as well as known CAF markers (periostin, α-smooth muscle actin, and podoplanin). Moreover, CM from the lung fibroblasts increased A549 cell proliferation in a calumenin-dependent manner. Thus, miR-21 expression in lung fibroblasts may trigger fibroblast trans-differentiation into CAFs, supporting cancer progression. Therefore, CAF miR-21 represents a pivotal prognostic marker for this scar-forming cancer of the lungs.
Background:The aim of this study was to categorize the conduction patterns between the right atrium (RA) and the superior vena cava (SVC), and to determine the ideal procedure for SVC isolation using ...a novel high-resolution mapping system.Methods and Results:RA-SVC conduction was evaluated using the RHYTHMIA system in 113 patients (age 62.8±11.5 years, paroxysmal: 67) with atrial fibrillation (AF) after pulmonary vein (PV) isolation. In 56 patients, a line of conduction block was found to run obliquely just above the sinus node (Block group). The remaining 57 patients did not have block (Non-block group). Non-PV foci were spontaneous or provoked with isoproterenol after electrical cardioversion of pacing-induced AF. In 43 patients with SVC foci (Block group: 22, Non-Block group: 21), SVC was isolated by radiofrequency applications delivered along the line connecting the open ends of the block line (Block group) or by conventional methods (Non-block group). The Block group required fewer radiofrequency deliveries for SVC isolation than the Non-Block group (4.2±0.9 vs. 10.2±2.8 times; P<0.0001). The isolated SVC area was larger in the Block group (15.7±3.7 vs. 10.5±3.1 cm2; P<0.0001).Conclusions:We found that approximately half of patients with AF had a diagonal line of block at the RA-SVC junction that could be utilized to isolate the SVC with fewer radiofrequency deliveries.
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