Yoshimasa Maniwa

A metabolomic approach to lung cancer

Lung cancer is one of the most common cancers in the world, but no good clinical markers that can be used to diagnose the disease at an early stage and predict its prognosis have been found. Therefore, the discovery of novel clinical markers is required. In this study, metabolomic analysis of lung cancer patients was performed using gas chromatography mass spectrometry. Serum samples from 29 healthy volunteers and 33 lung cancer patients with adenocarcinoma (n=12), squamous cell carcinoma (n=11), or small cell carcinoma (n=10) ranging from stage I to stage IV disease and lung tissue samples from 7 lung cancer patients including the tumor tissue and its surrounding normal tissue were used. A total of 58 metabolites (57 individual metabolites) were detected in serum, and 71 metabolites were detected in the lung tissue. The levels of 23 of the 58 serum metabolites were significantly changed in all lung cancer patients compared with healthy volunteers, and the levels of 48 of the 71 metabolites were significantly changed in the tumor tissue compared with the non-tumor tissue. Partial least squares discriminant analysis, which is a form of multiple classification analysis, was performed using the serum sample data, and metabolites that had characteristic alterations in each histological subtype and disease stage were determined. Our results demonstrate that changes in metabolite pattern are useful for assessing the clinical characteristics of lung cancer. Our results will hopefully lead to the establishment of novel diagnostic tools.

The alternative Ctf18-Dcc1-Ctf8-replication factor C complex required for sister chromatid cohesion loads proliferating cell nuclear antigen onto DNA

The linkage of sister chromatids after DNA replication ensures the faithful inheritance of chromosomes by daughter cells. In budding yeast, the establishment of sister chromatid cohesion requires Ctf8, Dcc1, and Ctf18, a homologue of the p140 subunit of the replication factor C (RFC). In this report we demonstrate that in 293T cells, Flag-tagged Ctf18 forms a seven-subunit cohesion-RFC complex comprised of Ctf18, Dcc1, Ctf8, RFCp40, RFCp38, RFCp37, and RFCp36 (Ctf18-RFC). We demonstrate that a stoichiometric heteroheptameric Ctf18-RFC complex can be assembled by coexpressing the seven proteins in baculovirus-infected insect cells. In addition, the two other stable subcomplexes were formed, which include a pentameric complex comprised of Ctf18, RFCp40, RFCp38, RFCp37, and RFCp36 and a dimeric Dcc1-Ctf8. Both the five- and seven-subunit Ctf18-RFC complexes bind to single-stranded and primed DNAs and possess weak ATPase activity that is stimulated by the addition of primed DNA and proliferating cell nuclear antigen (PCNA). These complexes catalyzed the ATP-dependent loading of PCNA onto primed and gapped DNA but not onto double-stranded nicked or single-stranded circular DNAs. Consistent with these observations, both Ctf18-RFC complexes substituted for the replicative RFC in the PCNA-dependent DNA polymerase δ-catalyzed DNA replication reaction. These results support a model in which sister chromatid cohesion is linked to DNA replication.

Differentiation of Malignant and Benign Pulmonary Nodules with Quantitative First-Pass 320–Detector Row Perfusion CT versus FDG PET/CT

PURPOSE To prospectively compare the capability of quantitative first-pass perfusion 320-detector row computed tomography (CT) (ie, area-detector CT) with that of combined positron emission tomography and CT (PET/CT) for differentiation between malignant and benign pulmonary nodules. MATERIALS AND METHODS This prospective study was approved by the institutional review board, and written informed consent was obtained from 50 consecutive patients with 76 pulmonary nodules. All patients underwent dynamic area-detector CT, PET/CT, and microbacterial and/or histopathologic examinations. All pulmonary nodules were divided into three groups: malignant nodules (n = 43), benign nodules with low biologic activity (n = 6), and benign nodules with high biologic activity (n = 27). For each dynamic area-detector CT data set, the perfusion derived by using the maximum slope model (PF(MS)), extraction fraction derived by using the Patlak plot model (EF(PP)), and blood volume derived by using the Patlak plot model (BV(PP)) were calculated. These parameters were statistically compared among the three nodule groups. Receiver operating characteristic (ROC) analyses were used to compare the diagnostic capability of the CT and PET/CT indexes. Finally, the sensitivity, specificity, and accuracy of each index were compared by using the McNemar test. RESULTS All indexes in the malignant nodule group were significantly different from those in the low-biologic-activity benign nodule group (P < .05). Areas under the ROC curve for PF(MS) and EF(PP) were significantly larger than those for BV(PP) (P < .05) and maximal standard uptake value (SUV(max)) (P < .05). The specificity and accuracy of PF(MS) and EF(PP) were significantly higher than those of BV(PP) and SUV(max) (P < .05). CONCLUSION Dynamic first-pass area-detector perfusion CT has the potential to be more specific and accurate than PET/CT for differentiating malignant from benign pulmonary nodules. SUPPLEMENTAL MATERIAL http://radiology.rsna.org/lookup/suppl/doi:10.1148/radiol.10100245/-/DC1.

N stage disease in patients with non-small cell lung cancer: efficacy of quantitative and qualitative assessment with STIR turbo spin-echo imaging, diffusion-weighted MR imaging, and fluorodeoxyglucose PET/CT.

PURPOSE To prospectively compare the diagnostic capability of short inversion time inversion-recovery (STIR) turbo spin-echo (SE) imaging, diffusion-weighted (DW) magnetic resonance (MR) imaging, and fluorodeoxyglucose (FDG) combined positron emission tomography (PET) and computed tomography (CT) in N stage assessment in patients with non-small cell lung cancer (NSCLC). MATERIALS AND METHODS This prospective study was approved by the institutional review board, and written informed consent was obtained from all patients. A total of 250 consecutive patients with NSCLC (136 men; mean age, 73 years; 114 women; mean age, 72 years) prospectively underwent pretherapeutic STIR turbo SE imaging, DW MR imaging, and FDG PET/CT, as well as surgical and pathologic examinations (N0 disease, n = 157; N1 disease, n = 72; N2 disease, n = 16; N3 disease, n = 5). Lymph node-to-saline ratio (LSR), lymph node-to-muscle ratio (LMR), apparent diffusion coefficient (ADC), maximal standardized uptake value (SUV(max)), and visual scoring were assessed for 135 metastatic lymph nodes and 135 randomly selected nonmetastatic lymph nodes. Receiver operating characteristic curve analysis was used to determine feasible threshold values. Diagnostic capabilities for N stage assessment were compared with the McNemar test on a per-patient basis. RESULTS When feasible, threshold values were used for quantitative assessment; sensitivity and accuracy of LSR and LMR (sensitivity, 82.8%; accuracy, 86.8%) proved to be significantly higher than those of ADC (sensitivity: 74.2%, P = .01; accuracy: 84.4%, P = .04) and SUV(max) (sensitivity: 74.2%, P = .01). For qualitative assessment, sensitivity of STIR turbo SE imaging (77.4%) was significantly higher than that of DW MR imaging (71.0%, P = .03) and FDG PET/CT (69.9%, P = .02). CONCLUSION Quantitative and qualitative assessments of N stage disease in patients with NSCLC obtained with STIR turbo SE MR imaging are more sensitive and/or more accurate than those obtained with DW MR imaging and FDG PET/CT. SUPPLEMENTAL MATERIAL http://radiology.rsna.org/lookup/suppl/doi:10.1148/radiol.11110281/-/DC1.

Role of pleural lavage cytology before resection for primary lung carcinoma.

OBJECTIVE To investigate the role of pleural lavage cytology (PLC) in resection for primary lung carcinoma. SUMMARY BACKGROUND DATA The prognostic significance of PLC before manipulation is still controversial. METHODS Cytology of pleural lavage immediately after thoracotomy but before any manipulation of the lung was examined in 500 consecutive patients with lung cancer with no pleural effusion who underwent pulmonary resections. Eighteen patients who already had pleural dissemination were excluded from this study. RESULTS Eighteen of 482 patients (3.7%) had positive cytologic findings. The positivity of PLC was significantly correlated with histology, extension of tumor to pleura, and presence of lymphatic permeation or vascular involvement by tumor. Positive lavage findings were seen only in adenocarcinoma. Because 6.3% of the patients with adenocarcinoma had positive cytologic findings, it is vital to perform PLC before curative resections for lung cancer, especially adenocarcinoma. The 5-year survival rates of the patients having negative and positive lavage findings were 52.9% and 14.6%, respectively. The prognosis of the patients with positive lavage findings was as poor as that of the patients with stage IIIB disease and that of the patients with malignant effusion. CONCLUSIONS Positive findings on PLC indicate exfoliation of cancer cells into the pleural cavity, which is an essential prognostic factor. In addition, we should regard positive cytologic findings as a subclinical malignant pleural effusion that is pathologic stage T4.

Dynamic MRI, dynamic multidetector-row computed tomography (MDCT), and coregistered 2-[fluorine-18]-fluoro-2-deoxy-D-glucose–positron emission tomography (FDG-PET)/CT: Comparative study of capability for management of pulmonary nodules

To compare the nodule management capabilities of dynamic MRI, dynamic multidetector‐row computed tomography (MDCT) and coregistered positron emission tomography (PET)/CT.

Postoperative radiotherapy is effective for thymic carcinoma but not for thymoma in stage II and III thymic epithelial tumors: the Japanese Association for Research on the Thymus Database Study.

The efficacy of postoperative radiotherapy (PORT) for thymic epithelial tumors is still controversial. Using the Japanese Association for Research on the Thymus (JART) database, this study was aimed at clarifying the efficacy of PORT for Masaoka stage II and III thymic carcinoma and thymoma.

Value of diffusion-weighted MR imaging using various parameters for assessment and characterization of solitary pulmonary nodules

OBJECTIVES To determine the appropriate parameters and evaluation method for characterizing solitary pulmonary nodules (SPNs) using quantitative parameters of diffusion-weighted imaging (DWI). METHODS Thirty-two subjects with 36 SPNs underwent DWI with seven different b values (0, 50, 100, 150, 300, 500, and 1000s/mm(2)). Five quantitative parameters were obtained from the region of interest drawn over each SPN: apparent diffusion coefficients (ADCs), true diffusion coefficients (DCs), and perfusion fractions (PFs), and signal-intensity ratios between lesion and spinal cord from DWI (b values: 1000 [LSR1000] and 500 [LSR500)]). All quantitative parameters and the diagnostic capabilities were statistically compared. RESULTS SPNs were diagnosed as follow: malignant (n=27) and benign (n=9). Parameter comparisons for malignant and benign showed both LSRs differed significantly (p<0.05). Applying feasible threshold values showed LSR500 specificity (88.9% [8/9]) and accuracy (77.8% [28/36]) were significantly higher than ADC, DC, and PF specificity and accuracy (p<0.05). LSR1000 accuracy (72.2% [26/36]) was significantly higher than DC accuracy, and its specificity (88.9% [8/9]) was significantly higher than ADC, DC, and PF specificities (p<0.05). CONCLUSIONS For quantitative differentiation of SPNs, LSR evaluation was more useful and practical than ADC, DC, and PF, and choice of b values showed little impact for the differentiation.

Oncogenic phosphatase Wip1 is a novel prognostic marker for lung adenocarcinoma patient survival.

DNA damage response pathways are important for maintaining genomic stability. The oncogenic phosphatase Wip1 plays a crucial role in DNA damage response by inhibiting several cell cycle proteins, including p53. Although Wip1 gene amplification has been reported in various primary tumors, including lung cancer, its biological significance for survival of primary lung tumor patients remains unclear. We investigated the expression of Wip1 in cancer epithelial cells immunohistochemically in 84 consecutive resected cases of lung adenocarcinoma. Increased Wip1 expression was observed in 54 (64.3%) of the 84 cases. Wip1 expression was found to be correlated significantly with two clinicopathological factors: γ‐H2AX expression, and invasion to the pulmonary vein. A univariate analysis and log–rank test indicated a significant association between Wip1 expression and lower overall survival rate (P = 0.019 and P = 0.0099, respectively). A multivariate analysis also indicated a statistically significant association between increased Wip1 expression and lower overall survival rate (hazard ratio, 4.3; P = 0.026). The Ki67 index level was higher in the Wip1‐positive group than in the negative group (P < 0.04, Mann–Whitney U‐test). Moreover, in a subgroup analysis of only stage I patients, increased Wip1 expression was also significantly associated with a lower overall survival rate (P = 0.023, log–rank test). These results indicate that the increased expression of Wip1 in cancer epithelial cells has significant value for tumor progression and the clinical prognosis of patients with primary lung adenocarcinoma. (Cancer Sci, 2011; 102: 1101–1106)

Avian-Type Receptor-Binding Ability Can Increase Influenza Virus Pathogenicity in Macaques

ABSTRACT The first influenza pandemic of the 21st century was caused by novel H1N1 viruses that emerged in early 2009. An Asp-to-Gly change at position 222 of the receptor-binding protein hemagglutinin (HA) correlates with more-severe infections in humans. The amino acid at position 222 of HA contributes to receptor-binding specificity with Asp (typically found in human influenza viruses) and Gly (typically found in avian and classic H1N1 swine influenza viruses), conferring binding to human- and avian-type receptors, respectively. Here, we asked whether binding to avian-type receptors enhances influenza virus pathogenicity. We tested two 2009 pandemic H1N1 viruses possessing HA-222G (isolated from severe cases) and two viruses that possessed HA-222D. In glycan arrays, viruses possessing HA-222D preferentially bound to human-type receptors, while those encoding HA-222G bound to both avian- and human-type receptors. This difference in receptor binding correlated with efficient infection of viruses possessing HA-222G, compared to those possessing HA-222D, in human lung tissue, including alveolar type II pneumocytes, which express avian-type receptors. In a nonhuman primate model, infection with one of the viruses possessing HA-222G caused lung damage more severe than did infection with a virus encoding HA-222D, although these pathological differences were not observed for the other virus pair with either HA-222G or HA-222D. These data demonstrate that the acquisition of avian-type receptor-binding specificity may result in more-efficient infection of human alveolar type II pneumocytes and thus more-severe lung damage. Collectively, these findings suggest a new mechanism by which influenza viruses may become more pathogenic in mammals, including humans.