Hiroyuki Tao

Prognostic potential of FOXP3 expression in non-small cell lung cancer cells combined with tumor-infiltrating regulatory T cells

Expression of the transcription factor FOXP3 characterizes regulatory T cells (Tregs) that engage in the maintenance of immunological self-tolerance and immune homeostasis. Intra-tumoral accumulation of Tregs is associated with unfavorable prognosis in several kinds of cancers. Recently, expression of FOXP3 and its association with prognosis have also been shown in some cancer cells in clinical studies. For non-small cell lung cancer (NSCLC), however, prognostic significance of tumor FOXP3 expression and its relationship with Tregs remain unknown. FOXP3 expression in cancer cells and tumor-infiltrating lymphocytes was examined by immunohistochemical staining of surgical specimens from 87 patients with NSCLC. Prognostic values of the tumor-infiltrating Treg count and tumor FOXP3 expression status were evaluated retrospectively. FOXP3-positive cancer cells were observed in 27 of 87 (31.0%) patients. There was no significant relationship between Treg count and tumor FOXP3 status. Increased Treg counts were associated with worse overall and relapse-free survival whereas the influence of tumor FOXP3 status on survival was not significant. However, when FOXP3-positive cancer cells were present, the relationship between Treg accumulation and worse prognosis was attenuated. In contrast, patients without tumor FOXP3 expression and high Treg count had significantly worse overall and relapse-free survival (hazard ratio: 3.118 and 3.325, p=0.028 and 0.024, respectively) than other groups. These results suggest that tumor FOXP3 expression has a better prognostic potential in NSCLC and that in combination with tumor-infiltrating Treg count the absence of tumor FOXP3 allows the selection of high-risk patients.

Sirolimus Ameliorated Post Lung Transplant Chylothorax in Lymphangioleiomyomatosis

We report a successful case of sirolimus treatment for chylous pleural and peritoneal effusions of lymphangioleiomyomatosis after lung transplantation. A 32-year-old woman underwent living donor lung transplantation. Persistent chylous pleural and peritoneal effusions were seen postoperatively. Pleurodesis by intrathoracic injection of OK-432, minomycin, and somatostatine analog failed to control chylous effusions. However, sirolimus treatment reduced the amount of chylous drainage and improved both chylous pleural and peritoneal effusions.

Development of a New Tissue-Engineered Sheet for Reconstruction of the Stomach

We have developed tissue-engineered digestive tracts composed of collagen scaffold and an inner silicon sheet and successfully used it to repair defects in parts of the esophagus, stomach, and small intestine. However, some improvements were demanded for clinical usage because the silicon sheet presented technical difficulties for suturing and endoscopic removal. New tissue-engineered sheet (New-sheet) was composed of a single-piece and reinforced collagen scaffold with biodegradable copolymer. One beagle dog was used to evaluate whether New-sheet could withstand suturing in comparison with native digestive tracts using a tensile tester. Seven beagle dogs had a 5-cm circular defect created in the stomach. New-sheet soaked with autologous peripheral blood or bone marrow aspirate was sutured to the gastric wall. Endoscopic, histological, and immunohistochemical assessment was performed to evaluate regeneration of the stomach up to 16 weeks. Tensile strength testing showed that the mucosal side of New-sheet had strength almost equivalent to the mucosa of the esophagus (P = 0.61). Endoscopically, regeneration of the mucosa started from the circumference after 4 weeks, but a small linear ulcer was still evident at 16 weeks. The regenerated stomach shrank by 60-80% of its original size and histologically showed villous mucosa and underlying dense connective tissue. Immunohistochemically, the regenerated area expressed alpha-smooth-muscle actin but was negative for basic calponin, irrespective of the source of soaked blood. New-sheet shows sufficient strength for suturing, no dehiscence, and better biocompatibility for clinical use, although further examination will be necessary to create a functional digestive tract.

Replacement of the Left Main Bronchus With a Tissue-Engineered Prosthesis in a Canine Model

BACKGROUND Stenosis of the left main bronchus caused by inflammatory diseases and neoplasms is a serious clinical problem because it can cause obstructive pneumonia and may require pneumonectomy. As an alternative to various treatments currently available, including balloon dilatation, stenting, and bronchoplasty, we propose the use of a prosthesis developed based on the concept of in situ tissue engineering for replacement of the left main bronchus. METHODS The main frame of the tissue-engineered prosthesis is a polypropylene mesh tube, 12 to 15 mm in inner diameter and 30 mm in length, with reinforcing rings. Collagen extracted from porcine skin is conjugated to this frame. A consecutive series of 8 beagle dogs underwent replacement of the left main bronchus with this tissue-engineered prosthesis. RESULTS All dogs survived the postoperative period with no morbidity except 1, which required intravenous administration of antibiotic for a week for pneumonia and recovered. Three dogs were euthanized for examination at 3 and 4 months after bronchus replacement, and the other five were monitored for more than 1 year. In two dogs, histologic examination revealed that the luminal surface was completely covered with ciliated columnar epithelium or nonciliated squamous epithelium. Exposure of the polypropylene mesh to various degrees was observed in 6 dogs, but the prosthesis remained stable and no adverse effects such as infection, sputum retention, or dehiscence were observed. CONCLUSIONS These long-term results suggest that our tissue-engineered prosthesis is applicable for replacement of the left main bronchus.

NONINVASIVE ASSESSMENT OF PULMONARY EMPHYSEMA USING DYNAMIC CONTRAST-ENHANCED MAGNETIC RESONANCE IMAGING

Emphysema tends to be complicated by diffuse abnormalities in the pulmonary peripheral microvasculature. The aim of this study was to evaluate whether dynamic contrast-enhanced magnetic resonance imaging (MRI) could provide a valid assessment of pulmonary blood flow as an indicator of the severity of emphysema. To do this, the authors compared MRI data with the pathological findings in lung tissue. Dynamic contrast-enhanced MRI is a noninvasive method and can be used to repeatedly monitor clinicopathological severity. Using MRI clear pulmonary vascular information can be obtained easily, and the relative pulmonary blood flow in the lung parenchyma can be quantified.

Prognostic impact of lymphovascular invasion compared with that of visceral pleural invasion in patients with pN0 non–small-cell lung cancer and a tumor diameter of 2 cm or smaller

BACKGROUND Both visceral pleural invasion (VPI) and lymphovascular invasion (LVI) have been shown to be adverse prognostic factors for early-stage non-small-cell lung cancer (NSCLC). Positive VPI upstages the T category of tumors ≤ 2 cm (T1a) to T2a, whereas LVI is not adapted as a descriptor for the Tumor, Node, Metastasis classification system. This study was conducted to evaluate the prognostic impacts of VPI and LVI in patients with pN0 NSCLC and a tumor diameter of ≤ 2 cm. METHODS We reviewed records of a total of 142 patients with pN0 NSCLC and a tumor diameter of ≤ 2 cm, who underwent lobectomy with hilar and mediastinal lymph node dissection between January 2001 and December 2009. We conducted univariate and multivariate analyses to evaluate the impact of VPI, LVI, and other clinicopathologic factors on survival. RESULTS Visceral pleural invasion and LVI were diagnosed as positive in 18 (12.7%) and 22 (15.5%) patients, respectively. Male sex, squamous cell carcinoma, positive VPI, and positive LVI were risk factors for overall survival. Squamous cell carcinoma, positive VPI, and positive LVI were risk factors for relapse-free survival. In multivariate analysis, squamous cell carcinoma and positive LVI were independent risk factors for overall survival, and positive LVI was an independent risk factor for relapse-free survival. CONCLUSIONS Positive LVI was more important than VPI as a prognostic factor in patients with pN0 NSCLC and a tumor diameter of ≤ 2 cm. Adjuvant chemotherapy should be considered for such patients, to improve the treatment outcomes.

FIBROBLAST GROWTH FACTOR-2 PROMOTES RECOVERY OF PULMONARY FUNCTION IN A CANINE MODELS OF ELASTASE-INDUCED EMPHYSEMA

Bronchoscopic lung volume reduction (BLVR) for severe emphysema is less invasive than lung volume reduction surgery. Fibroblast growth factor-2 (FGF-2) has been reported to enhance fibrogenesis and angiogenesis. The aim of this study was to investigate the feasibility of BLVR with the FGF-2, and ability to reduce lung volume and promote recovery of lung function. The BLVR based on FGF-2 is less invasive than surgical procedures, and can be performed repeatedly if the effectiveness of volume reduction is inadequate. This simple bronchoscopic approach allows selective reduction in the volume of the emphysematous parenchyma, and intratracheal administration of FGF-2 induces an increase in pulmonary blood flow, thus allowing recovery of pulmonary function.

Density of Tumor-Infiltrating FOXP3 + T Cells as a Response Marker for Induction Chemoradiotherapy and a Potential Prognostic Factor in Patients Treated with Trimodality Therapy for Locally Advanced Non-Small Cell Lung Cancer

PURPOSE To examine the relationship between the density of tumor-infiltrating T cell subpopulations and the pathological response to induction chemoradiotherapy (CRT) in patients with locally advanced NSCLC, and to assess the impact of T cell density on patient prognosis. METHODS A total of 64 patients with c-stages IIA-IIIB NSCLC who underwent induction CRT followed by R0 surgery were enrolled. Tumor-infiltrating T cells expressing either FOXP3 or CD8 were detected by immunohistochemical staining. RESULTS Mean numbers of tumor-infiltrating FOXP3+ T cells were 39.9 for patients with minor pathological responses (n = 9), 18.4 for those with major pathological responses (n = 25), and 12.9 for those with complete pathological responses (n = 30; P <0.001). The number of CD8+ T cells was not associated with pathological responses. Patients with lower FOXP3+ T cell densities showed better survival, although the difference was not statistically significant. CONCLUSION Our study demonstrated that the density of tumor-infiltrating FOXP3+ T cells indicated the degree of response for induction CRT and prognosis in patients treated with trimodality therapy for locally advanced NSCLC, suggesting that FOXP3+ T cells may be target for adjunct immunotherapy.

Bronchial mucoepidermoid carcinoma with recurrent hemoptysis in a pregnant woman: Report of a case

Mucoepidermoid carcinoma of the tracheobronchial tree, which is thought to arise from the minor salivary gland tissue of the proximal airway, is an extremely rare neoplasm. We herein report a case of a 31-year-old pregnant woman in her 36th week, who complained of recurrent hemoptysis from bronchial mucoepidermoid carcinoma. As massive bleeding from the tumor was seen during a caesarean section, it was necessary to add bronchial artery embolization. The tumor was successfully removed by a sleeve resection of the left main bronchus after the embolization. The patient is doing well without any signs of recurrence at 3 years after undergoing the operation.

Doublecortin and CaM kinase-like-1 expression in pathological stage I non-small cell lung cancer.

PurposeDoublecortin and CaM kinase-like-1 (DCLK1) regulates microtubule polymerization in migrating neurons. Recently, DCLK1 has been reported to act as an intestinal tumor stem cell marker and has been shown to be expressed in cancer cells and in the stroma of breast, colon, pancreatic, and prostate cancers. Here, we studied DCLK1 expression in non-small cell lung cancer (NSCLC) by immunohistochemistry in association with clinicopathological factors and patient prognosis.MethodsDCLK1 expression was analyzed by immunohistochemical staining of surgical specimens from 232 patients with pathological stage I NSCLC, including 187 adenocarcinomas. Relationships between the expression status of DCLK1 and clinicopathological factors were examined. The impact of DCLK1 expression status and other clinicopathological factors on survival was evaluated by univariate and multivariate analyses.ResultsThirty-three (14.2%) of 232 patients had DCLK1-positive cancer cells. DCLK1 was also expressed in the tumor stroma in most of the specimens and was significantly associated with DCLK1 expression in cancer cells. DCLK1-positive cancer cells were more common in non-adenocarcinoma tissues (44.4%) than in adenocarcinoma tissues (7.0%). Moreover, positive DCLK1 expression in cancer cells and stromal cells was correlated with a worse prognosis. Histological analyses revealed that the presence of DCLK1-positive cancer cells was an independent risk factor for poor prognosis in adenocarcinoma, but was not significantly associated with prognosis in non-adenocarcinoma.ConclusionsDCLK1 expression was observed in tumor cells in patients with pathological stage I NSCLC and was correlated with adverse prognosis, especially in patients with adenocarcinoma. DCLK1 may be a potential new therapeutic target.