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Abstract Background The cancer genome contains several driver mutations. However, in some cases, no known drivers have been identified; these remaining areas of unmet needs, leading to limited progress in cancer therapy. Whole-genome sequencing (WGS) can identify non-coding alterations associated with the disease. Consequently, exploration of non-coding regions using WGS and other omics data such as ChIP-sequencing (ChIP-seq) to discern novel alterations and mechanisms related to tumorigenesis have been attractive these days. Methods Integrated multi-omics analyses, including WGS, ChIP-seq, DNA methylation, and RNA-sequencing (RNA-seq), were conducted on samples from patients with non-clinically actionable genetic alterations (non-CAGAs) in lung adenocarcinoma (LUAD). Second-level cluster analysis was performed to reinforce the correlations associated with patient survival, as identified by RNA-seq. Subsequent differential gene expression analysis was performed to identify potential druggable targets. Results Differences in H3K27ac marks in non-CAGAs LUAD were found and confirmed by analyzing RNA-seq data, in which mastermind-like transcriptional coactivator 2 (MAML2) was suppressed. The down-regulated genes whose expression was correlated to MAML2 expression were associated with patient prognosis. WGS analysis revealed somatic mutations associated with the H3K27ac marks in the MAML2 region and high levels of DNA methylation in MAML2 were observed in tumor samples. The second-level cluster analysis enabled patient stratification and subsequent analyses identified potential therapeutic target genes and treatment options. Conclusions We overcome the persistent challenges of identifying alterations or driver mutations in coding regions related to tumorigenesis through a novel approach combining multi-omics data with clinical information to reveal the molecular mechanisms underlying non-CAGAs LUAD, stratify patients to improve patient prognosis, and identify potential therapeutic targets. This approach may be applicable to studies of other cancers with unmet needs.

Abstract Background Sarcomatoid carcinoma of the lung is a rare histological type of non-small cell lung cancer with a poor prognosis. We aimed to investigate the clinicopathological characteristics and prognostic factors of surgically resected sarcomatoid carcinoma of the lung. Methods We retrospectively reviewed 14999 patients who underwent surgical resection for non-small cell lung cancer accumulated by the Japanese Joint Committee of Lung Cancer Registry in 2010. Clinicopathological characteristics and survival were compared between the sarcomatoid carcinoma and other non-small cell cancer groups. The prognostic factors in the sarcomatoid carcinoma group were identified using a multivariate Cox proportional hazard model. Results Patients with sarcomatoid carcinoma comprised 1.4% of all patients. The sarcomatoid carcinoma group demonstrated a more aggressive pathology with presentation at more advanced stages, requiring more frequent extensive surgical resections. The sarcomatoid carcinoma group had remarkably poorer overall and recurrence-free survival than the other non-small cell lung cancer group. Adjuvant chemotherapy was associated with improved survival for pathological stage II–III sarcomatoid carcinoma cases rather than for pathological stage I disease. In the multivariate analysis, larger tumor size, lymphatic permeation, and no adjuvant chemotherapy were associated with the sarcomatoid carcinoma group’s overall and recurrence-free survival. Conclusions Surgically resected sarcomatoid carcinoma of the lung has a higher aggressive and metastatic potential and a worse prognosis than other non-small cell lung cancers. Adjuvant chemotherapy, which was associated with enhanced survival in patients with pathological stage II–III of the disease, could be considered for treating patients with pathological stage II–III sarcomatoid carcinoma of the lung.

Abstract Seafloor geodetic observation technology is a pivotal method for capturing plate boundary earthquakes and related phenomena. Currently, this task is performed by Global Navigation Satellite System-Acoustic ranging (GNSS-A), a technique that integrates GNSS observations with acoustic ranging. This method involves measuring the round-trip travel time of an acoustic signal between a surface platform and a preinstalled seafloor station, which is then converted into a distance for positioning. While the horizontal positioning accuracy can reach the centimeter level, the vertical accuracy tends to be loser. Recent research has revealed that equipment-dependent distortion of sound waves causes problems for accurate travel time identification, leading to a decrease in vertical component accuracy. In this paper, we conducted an analysis of distortion tendencies using water tank test data and quantitatively identified differences in distortion magnitude related to instruments, angle dependence, and directional dependence attributed to the sea surface platform. We proposed a new identification algorithm, Acoustic Ambiguity Reduction (AAR) method, using a catalog of distortions for each device, and improved the observational capabilities of the vertical component. Graphical Abstract

Abstract Background In an extensive genomic analysis of lung adenocarcinomas (LUADs), driver mutations have been recognized as potential targets for molecular therapy. However, there remain cases where target genes are not identified. Super-enhancers and structural variants are frequently identified in several hundred loci per case. Despite this, most cancer research has approached the analysis of these data sets separately, without merging and comparing the data, and there are no examples of integrated analysis in LUAD. Methods We performed an integrated analysis of super-enhancers and structural variants in a cohort of 174 LUAD cases that lacked clinically actionable genetic alterations. To achieve this, we conducted both WGS and H3K27Ac ChIP-seq analyses using samples with driver gene mutations and those without, allowing for a comprehensive investigation of the potential roles of super-enhancer in LUAD cases. Results We demonstrate that most genes situated in these overlapped regions were associated with known and previously unknown driver genes and aberrant expression resulting from the formation of super-enhancers accompanied by genomic structural abnormalities. Hi-C and long-read sequencing data further corroborated this insight. When we employed CRISPR-Cas9 to induce structural abnormalities that mimicked cases with outlier ERBB2 gene expression, we observed an elevation in ERBB2 expression. These abnormalities are associated with a higher risk of recurrence after surgery, irrespective of the presence or absence of driver mutations. Conclusions Our findings suggest that aberrant gene expression linked to structural polymorphisms can significantly impact personalized cancer treatment by facilitating the identification of driver mutations and prognostic factors, contributing to a more comprehensive understanding of LUAD pathogenesis.

Objective: Oncological feasibility of segmentectomy for internal non–small cell lung cancer (NSCLC) has not been assessed adequately. We assessed the oncological feasibility of segmentectomy for inner-located NSCLC by investigating surgical margins and patient prognosis after undergoing the procedure. Methods: Of the 3555 patients who underwent resection for lung cancer between 2013 and 2019 at our institution, 659 patients who underwent segmentectomy for clinical stage 0 to stage1A NSCLC were included in this study. Patients were separated into 2 groups according to whether the tumor was in the inner or outer third of the lung area. Clinical characteristics and prognoses were retrospectively compared between the groups. Results: Of the included 659 cases, 183 (27.8%) were inner-located, and 476 (72.2%) had outer-located NSCLC. The surgical margin was significantly shorter in the inner-located group than in the outer group (median, 16 vs 25 mm; P

Abstract DNA methylation is an epigenetic modification that results in dynamic changes during ontogenesis and cell differentiation. DNA methylation patterns regulate gene expression and have been widely researched. While tools for DNA methylation analysis have been developed, most of them have focused on intergroup comparative analysis within a dataset; therefore, it is difficult to conduct cross-dataset studies, such as rare disease studies or cross-institutional studies. This study describes a novel method for DNA methylation analysis, namely, methPLIER, which enables interdataset comparative analyses. methPLIER combines Pathway Level Information Extractor (PLIER), which is a non-negative matrix factorization (NMF) method, with regularization by a knowledge matrix and transfer learning. methPLIER can be used to perform intersample and interdataset comparative analysis based on latent feature matrices, which are obtained via matrix factorization of large-scale data, and factor-loading matrices, which are obtained through matrix factorization of the data to be analyzed. We used methPLIER to analyze a lung cancer dataset and confirmed that the data decomposition reflected sample characteristics for recurrence-free survival. Moreover, methPLIER can analyze data obtained via different preprocessing methods, thereby reducing distributional bias among datasets due to preprocessing. Furthermore, methPLIER can be employed for comparative analyses of methylation data obtained from different platforms, thereby reducing bias in data distribution due to platform differences. methPLIER is expected to facilitate cross-sectional DNA methylation data analysis and enhance DNA methylation data resources.

Abstract Inflammatory factors in the peripheral blood, such as the C-reactive protein level and neutrophil-to-lymphocyte ratio (NLR), are prognostic markers in multiple types of cancer, including non-small cell lung cancer (NSCLC). However, the association between inflammatory factors and prognosis based on histological types has not been adequately reported. In addition, the relationship between these factors and the immune condition of the tumor microenvironment (TME) is unclear. In this single center, retrospective study, we first investigated the relationship between preoperative inflammatory markers and clinical outcomes in 176 patients with NSCLC who underwent surgery. Lung adenocarcinoma (LUAD) showed no significant prognostic marker, whereas for lung squamous cell carcinoma (LUSC), a multivariate analysis showed that a high NLR was significantly associated with postoperative recurrence. In LUSC patients, the median time of postoperative recurrence-free survival in patients with a low NLR was longer than that in patients with a high NLR. We then compared the tumor-infiltrating lymphocyte (TIL) profile with inflammatory markers in peripheral blood and found that the NLR was negatively correlated with the frequencies of T cells and B cells in LUSC tissues. Thus, the NLR is a useful predictive biomarker for postoperative recurrence and may reflect the immune condition of the TME in LUSC.

Abstract The mechanism underlying the development of tumors, particularly at early stages, still remains mostly elusive. Here, we report whole-genome long and short read sequencing analysis of 76 lung cancers, focusing on very early-stage lung adenocarcinomas such as adenocarcinoma in situ (AIS) and minimally invasive adenocarcinoma. The obtained data is further integrated with bulk and spatial transcriptomic data and epigenomic data. These analyses reveal key events in lung carcinogenesis. Minimal somatic mutations in pivotal driver mutations and essential proliferative factors are the only detectable somatic mutations in the very early-stage of AIS. These initial events are followed by copy number changes and global DNA hypomethylation. Particularly, drastic changes are initiated at the later AIS stage, i.e., in Noguchi type B tumors, wherein cancer cells are exposed to the surrounding microenvironment. This study sheds light on the pathogenesis of lung adenocarcinoma from integrated pathological and molecular viewpoints.

Objective: Segmentectomy as a parenchymal-sparing surgical approach has been recommended over lobectomy in select patients with early-stage non–small cell lung cancer. This study aimed to address 3 aspects of segmentectomy (“patient indication”; “segmentectomy approaches”; “lymph node assessment”) where there is limited clinical guidance. Methods: A modified Delphi approach comprising 3 anonymous surveys and 2 expert discussions was used to establish consensus on the aforementioned topics among 15 thoracic surgeons (2 Steering Committee; 2 Task Force; 11 Voting Experts) from Asia who have extensive segmentectomy experience. Statements were developed by the Steering Committee and Task Force based on their clinical experience, published literature (rounds 1-3), and comments received from Voting Experts through surveys (rounds 2-3). Voting Experts indicated their agreement with each statement on a 5-point Likert scale. Consensus was defined as ≥70% of Voting Experts selecting either “Agree”/“Strongly Agree” or “Disagree”/“Strongly Disagree.” Results: Consensus from the 11 Voting Experts was reached on 36 statements (11 “patient indication” statements; 19 “segmentation approaches” statements; 6 “lymph node assessment” statements). In rounds 1, 2, and 3, consensus was reached on 48%, 81%, and 100% of drafted statements, respectively. Conclusions: A recent phase 3 trial reported significantly improved 5-year overall survival rates for segmentectomy compared with lobectomy, proposing thoracic surgeons to consider segmentectomy as a surgical option in suitable patients. This consensus serves as a guidance to thoracic surgeons considering segmentectomy in patients with early non–small cell lung cancer, outlining key principles that surgeons should consider in surgical decision-making.

Abstract Lung adenocarcinoma is the most common type of lung cancer. Known risk variants explain only a small fraction of lung adenocarcinoma heritability. Here, we conducted a two-stage genome-wide association study of lung adenocarcinoma of East Asian ancestry (21,658 cases and 150,676 controls; 54.5% never-smokers) and identified 12 novel susceptibility variants, bringing the total number to 28 at 25 independent loci. Transcriptome-wide association analyses together with colocalization studies using a Taiwanese lung expression quantitative trait loci dataset (n = 115) identified novel candidate genes, including FADS1 at 11q12 and ELF5 at 11p13. In a multi-ancestry meta-analysis of East Asian and European studies, four loci were identified at 2p11, 4q32, 16q23, and 18q12. At the same time, most of our findings in East Asian populations showed no evidence of association in European populations. In our studies drawn from East Asian populations, a polygenic risk score based on the 25 loci had a stronger association in never-smokers vs. individuals with a history of smoking (Pinteraction = 0.0058). These findings provide new insights into the etiology of lung adenocarcinoma in individuals from East Asian populations, which could be important in developing translational applications.

Abstract Thoracic SMARCA4‐deficient undifferentiated tumors (SMARCA4‐UT) have a poor prognosis and are often diagnosed at an inoperable advanced stage. Herein, we report a case of SMARCA4‐UT diagnosed by endobronchial ultrasound‐guided transbronchial cryobiopsy (EBUS‐cryo). The patient was a 42‐year‐old man with a history of smoking. Chest computed tomography revealed a right upper lobe nodule and an enlarged #11s lymph node. Core tissues could not be obtained by EBUS‐guided transbronchial needle aspiration (EBUS‐TBNA) for diagnosis and mediastinal staging; hence, EBUS‐guided intranodal forceps biopsy (EBUS‐IFB) was performed. However, a detailed diagnosis beyond poorly differentiated carcinoma could not be obtained. Subsequent EBUS‐cryo provided sufficient specimens for immunohistochemical and molecular evaluation and SMARCA4‐UT was definitively diagnosed. Thus, EBUS‐cryo could be of additional diagnostic value for uncommon tumors, such as SMARCA4‐UT, conjointly with EBUS‐IFB as well as EBUS‐TBNA.

Objective: We aimed to visualize complicated patterns of lymph node metastases in surgically resected non–small cell lung cancer by applying a data mining technique. Methods: In this retrospective study, 783 patients underwent lobectomy or pneumonectomy with systematic mediastinal lymph node dissection for non–small cell lung cancer between January 2010 and December 2018. Surgically resected lymph nodes were classified according to the International Association for the Study of Lung Cancer lymph node map. Network analysis generated patterns of lymph node metastases from stations 1 to 14, and the degree of connection between 2 lymph node stations was assessed. Results: The median number of lymph nodes examined per patient was 20, and the pathological N category was pN0 in 428 cases, pN1 in 132, pN2 in 221, and pN3 in 2. N1 lymph node stations had strong associations with superior mediastinal lymph node stations for patients with primary tumors in the upper lobes and with station 7 for the lower lobes. There was also a connection from the N1 lymph node stations to superior mediastinal lymph node stations in the lower lobes. In the right middle lobe, an even distribution from station 12m toward stations 2R, 4R, and 7 was noted. We released an interactive web application to visualize these data: http://www.canexapp.com. Conclusions: Lymph node metastasis patterns differed according to the lobe bearing the tumor. Our results support the need for clinical trials to further investigate selective mediastinal lymph node dissection.

Abstract Cryobiopsy is advantageous for collecting larger specimens with minimum crushing compared to forceps biopsy and transbronchial needle aspiration (TBNA), but it has not been widely used for mediastinal tumors. In this report, a leiomyoma of the thoracic esophagus was diagnosed with endobronchial ultrasound‐guided transbronchial mediastinal cryobiopsy (EBUS‐cryo). An asymptomatic 49‐year‐old woman had a 2.6‐cm sized submucosal tumor (SMT) of the esophagus adjacent to the trachea and left main bronchus. EBUS‐TBNA and EBUS‐guided intranodal forceps biopsy were performed, followed by EBUS‐cryo. The biopsy forceps could not be inserted into the tumor, but the cryoprobe was smoothly inserted. EBUS‐TBNA could not obtain enough spindle‐shaped tumor cells for immunohistochemical staining, but EBUS‐cryo provided sufficient specimens for diagnosing the leiomyoma. Adding EBUS‐cryo to EBUS‐TBNA has recently been reported to achieve high diagnostic yields for lymphomas, uncommon tumors, and benign diseases. EBUS‐cryo seems a valid diagnostic option for esophageal SMTs that are difficult to diagnose with needles and forceps.

Introduction: To determine the rate of deteriorating activities of daily living (ADL) and to investigate predictive factors in elderly patients undergoing surgery for NSCLC. Methods: Patients with NSCLC aged 75 years or older who underwent curative surgical resection were evaluated using the Tokyo Metropolitan Institute of Gerontology Index of Competence Instrumental ADL (TMIG-IADL) and the Japanese version of EuroQol 5-dimensions 5-level (EQ-5D-5L) quality-of-life scale administered at baseline and at 6 months postoperative. The primary end point was the rate of living patients without substantial deterioration of TMIG-IADL, defined as a decline greater than or equal to three points. Multivariable logistic regression was performed to determine risk factors for deteriorating ADL. Results: Between May 2019 and May 2020, 876 of the 986 screened patients enrolled from 47 institutions were eligible and included in the analysis. TMIG-IADL and EQ-5D-5L scores were obtained from 96.0% and 92.6% of the patients, respectively. At 6 months postoperative, 745 patients (85.1%, 95% confidence interval: 82.5%–87.3%) reported no significant ADL deterioration, and 96 of 841 patients (11.4%) with postoperative score data reported significant deterioration. The social domain was the most frequently affected activity. In multivariable analysis, poor performance status, low G8 geriatric screening score, segmentectomy (versus wedge resection), and surgery lasting less than 3 hours were associated with deteriorating ADL. Worsening EQ-5D-5L scores by minimally important difference or more were observed in 22.1% of the patients. Changes in TMIG-IADL and EQ-5D-5L scores were poorly correlated. Conclusions: Approximately 15% of elderly patients with NSCLC experienced significant ADL deterioration at 6 months postoperative.

Abstract The GNSS-A technique is an observation method that can detect seafloor crustal deformations with centimeter-level positioning accuracy. The GNSS-A seafloor geodetic observation array operated by the Japan Coast Guard (SGO-A) has been constructed near the Japanese Islands along the Nankai Trough and the Japan Trench. This observation array has detected several earthquakes’ displacements and episodic slow crustal deformation. To compare the detection results of SGO-A with other observation networks and expand the SGO-A coverage area, it is necessary to correctly understand its detection capability. In this paper, numerical simulations and statistical verifications were used to assess the capabilities of the present GNSS-A system using a manned vessel (observation frequency: 4–6 times/year, positioning accuracy: standard deviation = 1.5 cm) to detect (1) secular deformation only, (2) a transient slip event only and (3) secular deformation and a transient event together. We verified these results with appropriate thresholds and found the following features: When it is known that there is no transient event, the 95% confidence level (CL) for the estimation of secular crustal deformation rate with 4-year observation is about 0.5–0.8 cm/year; when the deformation rate is known, a signal of about 3.0 cm can be detected by observations of about 4 times before and after the transient event. When the deformation rate and the transient event are detected together, to keep the false positive low (about 0.05), the false negative becomes high (about 0.7–0.2 for detecting a signal of 4.5–6.0 cm). The determined rate and event variations are approximately 1.8 cm/year (95%CL) and 1.5 cm (standard deviation), respectively. We also examined the detection capability for higher observation frequency and positioning accuracy, to examine how the detection capability improves by technological advancements in the future. Additionally, we calculated the spatial range of event detectability using the determined values of detection sensitivity. Obtained results show that each seafloor site can detect a slip event of

Abstract Purpose We aimed to examine the characteristics of imaging findings of adenocarcinoma in situ (AIS) and minimally invasive adenocarcinoma (MIA) in the lungs of smokers compared with those of non-smokers. Materials and methods We included seven cases of AIS and 20 cases of MIA in lungs of smokers (pack-years ≥ 20) and the same number of cases of AIS and MIA in lungs of non-smokers (pack-years = 0). We compared the diameter of the entire lesion and solid component measured on computed tomography (CT) images, pathological size and invasive component diameter measured from pathological specimens, and CT values of the entire lesion and ground-glass opacity (GGO) portions between the smoker and non-smoker groups. Results The diameters of AIS and MIA on CT images and pathological specimens of the smoker group were significantly larger than those of the non-smoker group (p = 0.036 and 0.008, respectively), whereas there was no significant difference in the diameter of the solid component on CT images or invasive component of pathological specimens between the two groups. Additionally, mean CT values of the entire lesion and GGO component of the lesions in the smoker group were significantly lower than those in the non-smoker group (p = 0.036 and 0.040, respectively). Conclusion AIS and MIA in smoker’s lung tended to have larger lesion diameter and lower internal CT values compared with lesions in non-smoker’s lung. This study calls an attention on smoking status in CT-based diagnosis for early stage adenocarcinoma.

Abstract Primary squamous cell carcinoma (SqCC) often occurs in the trachea and bronchi. Recently, intraluminal bronchoscopic treatment (IBT) has emerged as an option for curative treatment, not just surgery, in patients with central early‐stage SqCC (CES‐SqCC). However, patients that can be cured by IBT are limited. We report a rare case of CES‐SqCC that was surgically confirmed to be cured by IBT alone. A 72‐year‐old woman had a nodular bronchial tumor at the bifurcation of right upper and intermediate bronchi. For histological diagnosis, the tumor was resected and incinerated using high‐frequency snare (HFS). Obtained specimens were diagnosed as SqCC; wedge resection of the bifurcation was performed to remove the residual lesion. However, no malignant findings were found in the excised specimens. Some patients with CES‐SqCC may be cured by aggressive diagnostic bronchoscopy. The risk of postoperative complications cannot be ignored because the surgery requires bronchoplasty in patients with CES‐SqCC. For patients with CES‐SqCC, surgical resection may be avoided by detailed assessment of residual lesion with radial probe endobronchial ultrasonography (RP‐EBUS).

Abstract Investigations of the co- and postseismic processes of the 2011 Tohoku-oki earthquake provide essential information on the seismic cycle in the Japan Trench. Although almost all of the source region lies beneath the seafloor, recent seafloor geophysical instruments have enabled to detect the near-field signals of both the coseismic rupture and the postseismic stress relaxation phenomena. Annual-scale seafloor geodesy contributed to refining the postseismic deformation models, specifically to the incorporation of viscoelastic effects. However, because of the insufficiency in the spatial coverage and observation period of seafloor geodetic observations, no consensus on crustal deformation models has been reached, especially on the along-strike extent of the main rupture, even for the coseismic process. To decompose the postseismic transient processes in and around the source region, i.e., viscoelastic relaxation and afterslip, long-term postseismic geodetic observations on the seafloor play an essential role. Here, from decadal seafloor geodetic data, we provide empirical evidence for offshore aseismic afterslip on the rupture edges that had almost decayed within 2–3 year. The afterslip regions are considered to have stopped the north–south rupture propagation due to their velocity strengthening frictional properties. In the southern source region (~ 37° N), despite not being resolved by coseismic geodetic data, shallow tsunamigenic slip near the trench is inferred from postseismic seafloor geodesy as a subsequent viscoelastic deformation causing persistent seafloor subsidence at a geodetic site off-Fukushima. After a decade from the earthquake, the long-term viscoelastic relaxation process in the oceanic asthenosphere is currently in progress and is still dominant not only in the rupture area, but also in the off-Fukushima region.

Objective: To evaluate efficacy of S-1 (tegafur/gimeracil/oteracil), an active novel fluoropyrimidine, as compared to UFT (tegafur/uracil) as a postoperative adjuvant therapy in patients with node-negative non–small cell lung cancer (NSCLC). Methods: Eligible patients had undergone complete resection of p-stage I (T1 with tumor diameter >2 cm or T2-N0M0 by 5th edition Union for International Cancer Control TNM) NSCLC, and were randomized to receive oral UFT 250 mg/m2/day for 2 years (Arm A) or oral S-1 80 mg/m2/day for 2 weeks with a 1-week rest period, for 1 year (Arm B). The primary end point was relapse-free survival (RFS), with 80% power and a one-sided type I error of 0.05. Results: From November 2008 to December 2013, 963 patients were enrolled (Arm A: 482, Arm B: 481). Toxicities (hematologic/nonhematologic) of grade 3 or more were observed in 15.9 (1.5/14.7)% in Arm A, and in 14.9 (3.6/12.1)% in Arm B, respectively. At data cut-off in December 2018, the hazard ratio for RFS was 1.06 (95% confidence interval, 0.82-1.36), showing no superiority of S-1 over UFT. The hazard ratio of overall survival (OS) was 1.10 (95% confidence interval, 0.81-1.50). The 5-year RFS/OS were 79.4%/88.8% in Arm A and 79.5%/89.7% in Arm B, respectively. The original NSCLC accounted for 58%/53%, respectively, of the Arm A/Arm B OS events. Secondary malignancies were observed in 85 (17.8%) and 84 (17.8%) individuals in Arm A and Arm B, respectively. Conclusions: S-1 was not superior to UFT as postoperative adjuvant therapy in node-negative NSCLC. Future investigation should incorporate identification of high-risk populations for recurrence.

Abstract The tumor microenvironment strongly influences clinical outcomes of immunotherapy. By transfecting genes of relevant cytokines into tumor cells, we sought to manipulate the microenvironment so as to elicit activation of T helper type 1 (Th1) responses and the maturation of dendritic cells (DCs). Using a synthetic vehicle, the efficiency of in vivo transfection of GFP‐cDNA into tumor cells was about 7.5% by intratumoral injection and about 11.5% by intravenous injection. Survival was significantly improved by both intratumoral and intravenous injection of the plasmid containing cDNA of interferon‐gamma, followed by intratumoral injection of DCs presenting the tumor antigens. Also, tumor growth was inhibited by these treatments. A more significant effect on survival and tumor growth inhibition was observed following injection of the plasmid containing cDNA of CD40 ligand, which is a potent inducer of DC‐maturation. Furthermore, the co‐injection of both IFNγ‐ and CD40 ligand‐encoding cDNA‐plasmids, followed by DC treatment, gave rise to further marked and enhancement, including 100% survival and more than 50% complete remission. This treatment regimen elicited significant increases in mature DCs and types of cells contributing to Th1 responses, and significant decreases in immune suppressor cells in the tumor. In the spleen, the treatment significantly increased activities of tumor‐specific killer and natural killer cells, but no alteration was observed in mature DCs or suppressor cells. These results indicate that transfection of these cytokine genes into tumor cells significantly alter the tumor microenvironment and improve the therapeutic results of DC‐based immunotherapy.

Here, we report a rare case of lung adenocarcinoma with intrapulmonary metastases that have “spread through air spaces” (STAS) by means of the alveoli and bronchioles. The peripheral intrapulmonary metastases were exhibiting pure ground-glass nodules along the bronchioles on computed tomography. The primary pathologic diagnosis was micropapillary adenocarcinoma with prominent tumor STAS. Histopathologic examination revealed that the cancer cells in the bronchioles around the primary tumor revealed micropapillary clusters on the mucosal surface or in the air spaces and reached peripheral intrapulmonary metastatic nodules. Notably, no vascular and stromal invasion was observed. The pathologic findings suggest that cancer cells are viable in the airspace of the bronchioles and alveoli and may support the significance of STAS as a pattern of airborne metastasis.

The Global Navigation Satellite System-Acoustic ranging combination technique (GNSS-A) has enabled us to measure seafloor crustal deformation in the precision of centimeters, leading to numerous discoveries of subseafloor tectonic phenomena. The moving observation conducted by our research group allows us to measure both the horizontal and vertical absolute positions of a reference point on the seafloor. However, the observation frequency of our GNSS-A observation system is still insufficient to observe short-term phenomena. This paper focused on the possibility to reduce the observation time per a seafloor site by shrinking the seafloor transponder array size and the survey line radius, which were empirically defined to be equal to the seafloor site depth in the early research. We evaluated the effects of changing these sizes on the GNSS-A positioning accuracy by conducting a series of numerical experiments. The results of the numerical experiments indicated that for a seafloor site with a depth of 3,000 m, the positioning accuracy is rapidly degraded as the transponder array size and the survey line radius are reduced to less than 3,000 m. Additional experiments done for transponder array sizes and survey line radii around 2,000–4,000 m revealed that shrinking the survey line radius has a dominant effect on the decrease in positioning accuracy. Thus, shrinking the transponder array size and the survey line radius is not a suitable option for reducing observation time, and the empirically defined observation configurations are concluded to be quite optimal when regarding both the positioning accuracy and the observation time.

Global Navigation Satellite System–Acoustic ranging combined seafloor geodetic technique (GNSS-A) has extended the geodetic observation network into the ocean. The key issue for analyzing the GNSS-A data is how to correct the effect of sound speed variation in the seawater. We constructed a generalized observation equation and developed a method to directly extract the gradient sound speed structure by introducing appropriate statistical properties in the observation equation, especially the data correlation term. In the proposed scheme, we calculate the posterior probability based on the empirical Bayes approach using the Akaike’s Bayesian Information Criterion for model selection. This approach enabled us to suppress the overfitting of sound speed variables and thus to extract simpler sound speed field and stable seafloor positions from the GNSS-A dataset. The proposed procedure is implemented in the Python-based software “GARPOS” (GNSS-Acoustic Ranging combined POsitioning Solver).

This article presents supplementary data for the research article by Yotsukura et al. entitled “Prognostic impact of cancer-associated active fibroblasts and invasive architectural patterns on early-stage lung adenocarcinoma” [1], which presented the postoperative prognosis for early-stage lung adenocarcinoma categorized according to histological findings. We included data of 1,032 resected cases of lung adenocarcinoma, which consisted of pathological stage IA invasive cancer and adenocarcinoma in situ resected at National Cancer Center Hospital, Tokyo, Japan, between 2007 and 2012. A pathological review was performed to assess total tumor size, size of invasion, histological subtype, lymphovascular invasion, and presence of cancer-associated active fibroblast (CAF). Tumor recurrence and overall survival were retrospectively recorded. Of the included cases, 166 (16.1%), and 866 (83.9%) were adenocarcinoma in situ and pathological stage IA, respectively. Pathological stage IA adenocarcinoma was further classified based on the histologial subtype and the presence of CAF. This data set may be useful for analyzing the postoperative prognosis of early-stage lung adenocarcinoma, in combination with detailed pathological findings including size of invasion, histological subtype, and presence of CAF.

The Global Navigation Satellite System-Acoustic ranging combination technique (GNSS-A) is a recently developed technology to precisely detect seafloor crustal deformation. This method can also estimate km-scale underwater sound speed structure (SSS) as a by-product of monitoring seafloor crustal deformation. This paper evaluates the validity of the spatial gradient and its temporal variation of the SSS estimated by GNSS-A observations off the Kii channel before and after Kuroshio meandering. According to the comparison of the JCOPE2M reanalysis data and the in situ observation data, the GNSS-A estimated SSS has local structures that are not reproduced in this reanalysis but were detected by in situ data. In addition, we investigate the effect of observation time on the stability of SSS estimation. The results suggest that the sufficient time required for stable estimation depends on the spatial coverage of observation data, which depends on the depth of the site. As a result, the time resolution was derived to be about one hour at a site whose depth is 1500 m.

The Global Navigation Satellite System-Acoustic ranging combination technique (GNSS-A) is a seafloor geodetic technique that enables precise global seafloor positioning to detect subseafloor geophysical phenomena. The technique requires a sea surface observation platform that combines GNSS positioning and acoustic ranging. Currently, a survey vessel is used as the platform, which entails substantial financial and human resources costs, which makes increasing observation frequency difficult. It is possible to detect long-term average seafloor movement at the centimeter level, but it is difficult to detect short-term variation due to the insufficiency of observation frequency. The terrestrial GNSS observation network has detected temporal changes in crustal deformation fields. These precise observations provide useful information on the megathrust seismogenic zone. To detect such phenomena on the seafloor, the temporal resolution of the GNSS-A observation needs to be improved. Advances in vessel equipment technology are crucial for increasing observation frequency. In this paper, we review the historical development of the Japan Coast Guard’s GNSS-A observation system, focusing on technological developments of on-board equipment installed on a sea surface platform, and explain how such improvements have increased observation frequency over time. In the present, ranging frequency has improved from 40–60 s to 15–20 s by the introduction of the multiple ranging system, resulting in more frequent observations up to five times per year for an individual site.

Abstract Crustal deformation data obtained by geodetic observation networks are foundations in the fields of geodesy and seismology. These data are essential for understanding plate motion and earthquake sources and for simulating earthquake and tsunami scenarios. Although relatively scarce, seafloor geodetic data are particularly important for monitoring the behaviour of undersea interplate boundary regions. Since the mid-1990s, we have been developing the combined Global Navigation Satellite System-Acoustic ranging (GNSS-A) technique for realizing seafloor geodesy. This technique allows us to collect time series of seafloor crustal deformation. Our published data can be used to investigate several seismological phenomena along the subduction zones around Japan, namely the Nankai Trough, Sagami Trough and Japan Trench. These regions are globally important places in geodesy and seismology and are also suitable for comparison with other geophysical datasets. Our intention is for these data to promote further understanding of megathrust zones.

EGFR mutations in lung adenocarcinoma are more frequent in East Asians compared to other populations. Here, the authors carry out a genome-wide association study in EGFRmutant cancers and identify loci that are associated with risk of developing this molecular subtype of cancer.

PURPOSE:The image noise and image quality of a prototype ultra-high-resolution computed tomography (U-HRCT) scanner was evaluated and compared with those of conventional high-resolution CT (C-HRCT) scanners. MATERIALS AND METHODS:This study was approved by the institutional review board. A U-HRCT scanner prototype with 0.25 mm x 4 rows and operating at 120 mAs was used. The C-HRCT images were obtained using a 0.5 mm x 16 or 0.5 mm x 64 detector-row CT scanner operating at 150 mAs. Images from both scanners were reconstructed at 0.1-mm intervals; the slice thickness was 0.25 mm for the U-HRCT scanner and 0.5 mm for the C-HRCT scanners. For both scanners, the display field of view was 80 mm. The image noise of each scanner was evaluated using a phantom. U-HRCT and C-HRCT images of 53 images selected from 37 lung nodules were then observed and graded using a 5-point score by 10 board-certified thoracic radiologists. The images were presented to the observers randomly and in a blinded manner. RESULTS:The image noise for U-HRCT (100.87 ± 0.51 Hounsfield units [HU]) was greater than that for C-HRCT (40.41 ± 0.52 HU; P < .0001). The image quality of U-HRCT was graded as superior to that of C-HRCT (P < .0001) for all of the following parameters that were examined: margins of subsolid and solid nodules, edges of solid components and pulmonary vessels in subsolid nodules, air bronchograms, pleural indentations, margins of pulmonary vessels, edges of bronchi, and interlobar fissures. CONCLUSION:Despite a larger image noise, the prototype U-HRCT scanner had a significantly better image quality than the C-HRCT scanners.

The aim of this study was to clarify the significance of DNA methylation alterations at precancerous stages of lung adenocarcinoma. Using single-CpG resolution Infinium array, genome-wide DNA methylation analysis was performed in 36 samples of normal lung tissue obtained from patients without any primary lung tumor, 145 samples of non-cancerous lung tissue (N) obtained from patients with lung adenocarcinomas, and 145 samples of tumorous tissue (T). Stepwise progression of DNA methylation alterations from normal lung tissue to non-cancerous lung tissue obtained from patients with lung adenocarcinomas, and then tumorous tissue samples, was observed at 3,270 CpG sites, suggesting that non-cancerous lung tissue obtained from patients with lung adenocarcinomas was at precancerous stages with DNA methylation alterations. At CpG sites of 2,083 genes, DNA methylation status in samples of non-cancerous lung tissue obtained from patients with lung adenocarcinomas was significantly correlated with recurrence after establishment of lung adenocarcinomas. Among such recurrence-related genes, 28 genes are normally unmethylated (average β-values based on Infinium assay in normal lung tissue samples was less than 0.2) and their DNA hypermethylation at precancerous stages was strengthened during progression to lung adenocarcinomas (Δβ(T-N)>0.1). Among these 28 genes, we focused on 6 for which implications in transcription regulation, apoptosis or cell adhesion had been reported. DNA hypermethylation of the ADCY5, EVX1, GFRA1, PDE9A, and TBX20 genes resulted in reduced mRNA expression in tumorous tissue samples. 5-Aza-2'-deoxycytidine treatment of lung cancer cell lines restored the mRNA expression levels of these 5 genes. Reduced mRNA expression in tumorous tissue samples was significantly correlated with tumor aggressiveness. These data suggest that DNA methylation alterations at precancerous stages determine tumor aggressiveness and outcome through silencing of specific genes.

The nature of microRNA (miRNA) dysfunction in carcinogenesis remains controversial because of the complex connection between miRNA structural diversity and biological processes. Here, we found that oncofetal IGF2BP3 regulates the selective production of a subset of 3'-isoforms (3'-isomiRs), including miR-21-5p and Let-7 family, which induces significant changes in their cellular seed occupancy and structural components, establishing a cancer-specific gene expression profile. The D-score, reflecting dominant production of a representative miR-21-5p+C (a 3'-isomiR), discriminated between clinical early-stage lung adenocarcinoma (LUAD) cases with low and high recurrence risks, and was associated with molecular features of cell cycle progression, epithelial-mesenchymal transition pressure, and immune evasion. We found that IGF2BP3 controls the production of miR-21-5p+C by directing the nuclear Drosha complex to select the cleavage site. IGF2BP3 was also involved in the production of 3'-isomiRs of miR-425-5p and miR-454-3p. IGF2BP3-regulated these three miRNAs are suggested to be associated with the regulation of p53, TGF-β, and TNF pathways in LUAD. Knockdown of IGF2BP3 also induced a selective upregulation of Let-7 3'-isomiRs, leading to increased cellular Let-7 seed occupancy and broad repression of its target genes encoding cell cycle regulators. The D-score is an index that reflects this cellular situation. Our results suggest that the aberrant regulation of miRNA structural diversity is a critical component for controlling cellular networks, thus supporting the establishment of a malignant gene expression profile in early stage LUAD. [ABSTRACT FROM AUTHOR]