ObjectivesPulmonary sarcomatoid carcinomas are rare and generally aggressive tumors composed of carcinomatous and sarcomatous components; however, the evolution of sarcomatoid cancer has not been elucidated. Here, we aimed to evaluate the mutational profiles and phylogeny of sarcomatoid carcinomas using next generation sequencing and in-silico analysis to facilitate the development of novel therapies.MethodsFour patients who underwent surgery for sarcomatoid cancer were enrolled. Cancer cells were collected from carcinomatous and sarcomatous components in each tumor by laser capture microdissection. Next-generation sequencing was performed in each component, and the mutation profiles were compared. For further inference of phylogenies, phylogenetic and PyClone analyses were performed. Mismatch repair disturbance and programmed death ligand-1 (PD-L1) expression were also evaluated.ResultsComparative genetic analysis of different histological areas revealed that the separate components shared several common mutations, which showed relatively high cellular prevalence in the PyClone statistical inference. Phylogenetic analysis showed that the sarcomatous component had ramified from the carcinomatous component in the early phase of the evolution process and accumulated a number of mutations that were different from those of the carcinomatous component. Moreover, microsatellite instability was detected in a case of sarcomatoid cancer and PD-L1 was strongly positive (≥ 50%) in all sarcomatoid cancers.ConclusionsOur data suggest that sarcomatoid carcinoma evolves from a common ancestral clone, and its phylogenetic features may reflect high-grade malignancy in pulmonary sarcomatoid carcinoma. High tumor mutation burden and strong PD-L1 staining may provide a rationale for the use of targeted immunotherapies in pulmonary sarcomatoid carcinomas.

In cases of multiple lung cancers, individual tumors may represent either a primary lung cancer or both primary and metastatic lung cancers. In this study, we investigated the differences between clinical/histopathological and genomic diagnoses to determine whether they are primary or metastatic. 37 patients with multiple lung cancers were enrolled in this study. Tumor cells were selected from tissue samples using laser capture microdissection. DNA was extracted from those cells and subjected to targeted deep sequencing. In multicentric primary lung cancers, the driver mutation profile was mutually exclusive among the individual tumors, while it was consistent between metastasized tumors and the primary lesion. In 11 patients (29.7%), discrepancies were observed between genomic and clinical/histopathological diagnoses. For the lymph node metastatic lesions, the mutation profile was consistent with only one of the two primary lesions. In three of five cases with lymph node metastases, the lymph node metastatic route detected by genomic diagnosis differed from the clinical and/or pathological diagnoses. In conclusion, in patients with multiple primary lung cancers, cancer-specific mutations can serve as clonal markers, affording a more accurate understanding of the pathology of multiple lung cancers and their lymphatic metastases and thus improving both the treatment selection and outcome.

iventricular pacing (BVP) is currently indicated for the treatment of patients with medically refractory heart failure and electrical dyssynchrony, 1-4 but it is becoming increasingly clear that QRS duration is an inadequate predictor of the response to BVP therapy. [4][5][6] Because the improvement in mechanical dyssynchrony after BVP correlates with improvement in clinical status and reverse remodeling, [6][7][8][9][10][11][12] there are ongoing investigations to determine the most accurate and efficient method of detecting mechanical dyssynchrony. Echocardiography, including tissue Doppler imaging (TDI), is simple, easy and ideal for evaluating regional wall motion. [5][6][7][8][9][10][11][12][13] Furthermore, recent advances in TDI and strain Doppler imaging (SDI) have enhanced the ability to evaluate ventricular synchrony and regional myocardial function. [5][6][7][8][9][10][11][12][13][14][15][16] However, the utility and efficacy of TDI and SDI for evaluating ventricular synchrony and function and for predicting long-term clinical improvement in patients undergoing BVP have not been sufficiently determined. The purpose of this study was to clarify this point. Methods Study PopulationThis study included 17 patients with advanced heart failure and a wide QRS complex who received a pacemaker or an implantable cardioverter defibrillator (ICD) providing BVP. There were 12 men and 5 women, and their mean age was 66±9 years; 13 patients had idiopathic cardiomyopathy, 2 had ischemic heart disease, and the remaining 2 had valvular heart disease; 11 patients were in New York Heart Association (NYHA) functional class IV, and the remaining 6 were in class III despite maximal pharmacologic therapy at the time of pacemaker implantation. The QRS interval was >140 ms and the left ventricular (LV) ejection fraction determined by echocardiography was <40% in all the patients.Patients were divided into 2 groups according to their clinical status at the end of the follow-up period (23±7 months; range: 14-37 months): the responder group (n=12) and the nonresponder group (n=5; Table 1). The clinical status of each patient was determined at the end of the follow-up period by 2 cardiologists who did not have any information concerning the BVP status of the patients. A responder was defined as a patient who improved clinically to NYHA functional class I or II during the follow-up period and a nonresponder was one who did not. There was no significant difference between the 2 groups in the NYHA classification before the initiation of BVP (p=0.9; Background The purpose of this study was to determine the utility and efficacy of tissue Doppler imaging (TDI) and strain Doppler imaging (SDI) for evaluating ventricular synchrony and function, and for predicting the long-term clinical improvement in patients undergoing biventricular pacing (BVP). Methods and Results TDI and SDI were performed before and <1 month after initiating BVP in 17 patients with advanced heart failure. An intraventricular conduction delay between the left ventricu...

conventional next generation sequencing analysis has provided important insights into cancer genetics. However, the detection of rare (low allele fraction) variants remains difficult because of the error-prone nucleotide changes derived from sequencing/pcR errors. to eliminate the false-positive variants and detect genuine rare variants, sequencing technology combined with molecular barcodes will be useful. Here, we used the newly developed dual-molecular barcode technology (ion AmpliSeq HD) to analyze somatic mutations in 24 samples (12 tumor tissues and 12 plasma) from 12 patients with biliary-pancreatic and non-small cell lung cancers. We compared the results between next generation sequencing analysis with or without molecular barcode technologies. the variant allele fraction (VAf) between non-molecular barcode and molecular barcode sequencing was correlated in plasma DnA (R 2 = 0.956) and tumor (R 2 = 0.935). Both methods successfully detected high VAF mutations, however, rare variants were only identified by molecular barcode sequencing and not by non-molecular barcode sequencing. Some of these rare variants in tumors were annotated as pathogenic, and therefore subclonal driver mutations could be observed. Furthermore, the very low VAF down to 0.17% were identified in cell free DNA in plasma. These results demonstrate that the dual molecular barcode sequencing technologies can sensitively detect rare somatic mutations, and will be important in the investigation of the clonal and subclonal architectures of tumor heterogeneity. Cancer acquires somatic mutations during the evolution of a tumor. Subclonal mutants are considered to be associated with drug resistance in various cancers, including non-small cell lung, breast and colorectal cancers 1,2. Cell free DNA (cfDNA) in plasma contains a small fraction of tumor DNA with tumor-derived mutations, which is called circulating tumor DNA. Plasma cfDNA is useful for monitoring tumor recurrence, estimating treatment effects and identifying drug-resistant mutations 3. However, only low levels of mutated alleles are present in the overall cfDNA circulating in blood. Therefore, the development of highly sensitive methods to detect rare variants is required. Various sensitive and accurate methods have been developed for the detection and quantification of mutated alleles in low abundance among high amounts of the wild-type allele 4. These methods are important for medical oncology, cancer research, infectious disease and microbial studies. To investigate the tumor heterogeneity and cfDNA in liquid biopsy, highly sensitive assays are necessary for detecting somatic mutations with low variant allele fraction (VAF). Droplet digital PCR, chip-based digital PCR and beads, emulsion, amplification, magnetics and flow cytometry (BEAMing) assays can sensitively detect rare mutations present at 0.1% VAF 4. Digital PCR and BEAMing have been applied for well-known pathogenic variants and detect several types of variants simultaneously; however, these may not be suitable for target...

Thymic epithelial tumors (TETs) are rare malignant mediastinal tumors that are difficult to diagnose and treat. The programmed death 1 (PD-1) receptor and its ligand (PD-L1) are expressed in various malignant tumors and have emerged as potential immunotherapeutic targets. However, the immunobiology of TETs is poorly understood. We evaluated PD-L1 expression and the presence of tumor-infiltrating lymphocytes (CD8 and CD3 expression) in surgical TET specimens from 39 patients via immunohistochemistry and determined their relation to clinicopathological parameters. Cases with membranous reactivity of the PD-L1 antibody in ≥1% of tumor cells were considered positive. Positive PD-L1 expression was observed in 53.9% of cases. Histologically, PD-L1 expression was positive in 2/6 type A, 2/6 type AB, 3/9 type B1, 4/4 type B2, 5/6 type B3, and 5/8 type C TET cases. Thus, the number of cases with PD-L1 expression and the percent expression of PD-L1 were significantly higher in more aggressive thymomas (type B2 or B3). CD3+ and CD8+ tumor-infiltrating lymphocytes were diffusely and abundantly distributed in all cases. These data suggest that a PD-1/PD-L1 blockade is a promising treatment for TETs, with more beneficial treatment effects for aggressive thymomas such as type B2 or B3.