Tsuyoshi Hata

Nuclear localization of beta-catenin involved in precancerous change in oral leukoplakia

BackgroundOral leukoplakia is a precancerous change developed in the oral mucosa, and the mechanism that oral leukoplakia becomes malignant through atypical epithelium is not known. Here we compared the β-catenin expression detected by immunohistochemical staining in the normal oral epithelium and in the oral leukoplakia with or without dysplasia.ResultsThe normal oral epithelium showed β-catenin expression only in the cell membrane, but not in the nuclei. In the oral leukoplakia without dysplasia, 7 out of 17 samples (41%) showed β-catenin expression in the cell membrane, and 5 samples (29%) showed expression in the nuclei. In the oral leukoplakia with dysplasia, nuclear expression of β-catenin was shown in 11 out of 12 samples (92%). Incidence of nuclear β-catenin expression was significantly different between dysplasia and normal oral epithelium (P < 0.01), and also between oral leukoplakia with dysplasia and those without dysplasia (P < 0.01). Wnt3 expression was detected in the epithelial cell membrane or cytoplasm in oral leukoplakia where nuclear expression of β-catenin was evident, but not in epithelial cells without nuclear expression of β-catenin.ConclusionThe components of canonical Wnt pathway, such as Wnt3, β-catenin, and cyclin D1, were detected, implying that this pathway is potentially involved in the progression of dysplasia in oral leukoplakia.

Desmoglein 3-specific CD4+ T cells induce pemphigus vulgaris and interface dermatitis in mice

Pemphigus vulgaris (PV) is a severe autoimmune disease involving blistering of the skin and mucous membranes. It is caused by autoantibodies against desmoglein 3 (Dsg3), an adhesion molecule critical for maintaining epithelial integrity in the skin, oral mucosa, and esophagus. Knowing the antigen targeted by the autoantibodies renders PV a valuable model of autoimmunity. Recently, a role for Dsg3-specific CD4+ T helper cells in autoantibody production was demonstrated in a mouse model of PV, but whether these cells exert cytotoxicity in the tissues is unclear. Here, we analyzed 3 Dsg3-specific TCRs using transgenic mice and retrovirus induction. Dsg3-specific transgenic (Dsg3H1) T cells underwent deletion in the presence of Dsg3 in vivo. Dsg3H1 T cells that developed in the absence of Dsg3 elicited a severe pemphigus-like phenotype when cotransferred into immunodeficient mice with B cells from Dsg3-/- mice. Strikingly, in addition to humoral responses, T cell infiltration of Dsg3-expressing tissues led to interface dermatitis, a distinct form of T cell-mediated autoimmunity that causes keratinocyte apoptosis and is seen in various inflammatory/autoimmune skin diseases, including paraneoplastic pemphigus. The use of retrovirally generated Dsg3-specific T cells revealed that interface dermatitis occurred in an IFN-γ- and TCR avidity-dependent manner. This model of autoimmunity demonstrates that T cells specific for a physiological skin-associated autoantigen are capable of inducing interface dermatitis and should provide a valuable tool for further exploring the immunopathophysiology of T cell-mediated skin diseases.

A combined anterolateral thigh flap, anteromedial thigh flap, and vascularized iliac bone graft for a full-thickness defect of the mental region.

A triple combined anterolateral thigh flap, anteromedial thigh flap, and vascularized iliac bone graft was used for reconstruction of a full-thickness defect of the mental region after wide resection of advanced tongue cancer. The distal end of the pedicle vessels of these double skin flaps, i.e., the lateral circumflex femoral system, was directly anastomosed in tandem to the pedicle of the iliac bone graft, which was enveloped by these flaps. The advantages of this iliac osteocutaneous flap are as follows: It can be transferred within a shorter surgical time because the flaps can be obtained in the supine position simultaneously with tumor resectioning; and its skin components can be separated from the bone because each component has its own pedicle vessels, has a longer vascular pedicle (> 10 cm), and has a thicker crest of the bone graft. This flap is believed to be suitable for reconstruction of large full-thickness defects in various sizes of the mandibular and maxillary regions of the face.

Tumour-suppressive function of SIRT4 in human colorectal cancer.

Background:SIRT4, which is localised in the mitochondria, is one of the least characterised members of the sirtuin family of nicotinamide adenine dinucleotide-dependent enzymes that play key roles in multiple cellular processes such as metabolism, stress response and longevity. There are only a few studies that have characterised its function and assessed its clinical significance in human cancers.Methods:We established colorectal cancer cell lines (SW480, HCT116, and HT29) overexpressing SIRT4 and investigated their effects on proliferation, migration and invasion, as well as E-cadherin expression, that negatively regulates tumour invasion and metastases. The associations between SIRT4 expression in colorectal cancer specimens and clinicopathological features including prognosis were assessed by immunohistochemistry.Results:SIRT4 upregulated E-cadherin expression and suppressed proliferation, migration and invasion through inhibition of glutamine metabolism in colorectal cancer cells. Moreover, SIRT4 expression in colorectal cancer decreased with the progression of invasion and metastasis, and a low expression level of SIRT4 was correlated with a worse prognosis.Conclusions:SIRT4 has a tumour-suppressive function and may serve as a novel therapeutic target in colorectal cancer.

Bidimensional analysis of desmoglein 1 distribution on the outermost corneocytes provides the structural and functional information of the stratum corneum

BACKGROUND The stratum corneum (SC) plays an important role in cutaneous barrier function. Recent clarification of the pathophysiology of several keratoses has suggested that adhesive molecules contribute not only to SC construction but also to SC barrier function. OBJECTIVE The purpose of this study is to clarify how the distribution of adhesion molecules on corneocytes contributes to the construction of the SC and the overall organization and function of the cutaneous barrier. METHODS To investigate the distribution of desmoglein 1 (Dsg1), which may be a main component of corneodesmosomes (CDSs) in the SC, we used a bidimensional observation method using tape-stripped corneocytes and several immunohistochemical techniques to demonstrate the distribution of Dsg1 and to deduce internal events in the SC. RESULTS Immunofluorescence labeling showed that Dsg1 distributed on corneocytes of the outermost SC with a characteristic pattern at the periphery, or over the entire surface, and differences in this distribution pattern correlated with the transepidermal water loss (TEWL). Furthermore, electron microscopic analysis showed that (1) Dsg1 was localized on CDSs involved in adhesion, and (2) CDSs on the periphery of corneocytes contributed to the generation of the characteristic basket-weave structure. CONCLUSION We explored the distribution pattern of Dsg1 in the SC via a non-invasive investigation tool. Our findings indicate the significance of adhesion molecules in the formation and function of the SC, and suggest that adhesion molecules are one of the important elements in barrier formation in addition to corneocytes, which act as bricks, and intercellular lipids, which act as mortar.

Ectopic expression of epidermal antigens renders the lung a target organ in paraneoplastic pemphigus.

Paraneoplastic pemphigus (PNP) is an autoimmune disease of the skin and mucous membranes that can involve fatal lung complications. IgG autoantibodies target the cell adhesion molecules desmoglein (Dsg)3 and plakins, but the nature and targets of infiltrating T cells are poorly characterized. Moreover, the lung involvement in this skin Ag-specific autoimmune condition represents a paradox. To mimic autoimmunity in PNP, we grafted wild-type skin onto Dsg3−/− mice, which resulted in graft rejection and generation of anti-Dsg3 IgG and Dsg3-specific T cells. Transfer of splenocytes from these mice into Rag2−/− mice induced a combination of suprabasilar acantholysis and interface dermatitis, a histology unique to PNP. Furthermore, the recipient mice showed prominent bronchial inflammation of CD4+ and CD8+ T cells with high mortality. Intriguingly, ectopic Dsg3 expression was observed in the lungs of PNP mice, mirroring the observation that squamous metaplasia is often found in the lungs of PNP patients. Dsg3 and other epidermal Ags were ectopically expressed in the lungs after pulmonary injuries by naphthalene, which was sufficient for recruitment of Dsg3-specific CD4+ T cells. These findings demonstrate that squamous metaplasia after pulmonary epithelial injury may play a crucial role in redirecting the skin-specific autoimmune reaction to the lungs in PNP.

Mycosis fungoides with involvement of the oral mucosa

Mycosis fungoides (MF) is a T-cell lymphoma that predominantly affects the skin. Involvement of the oral tissues in MF is rare. A case of MF with involvement of the maxillary gingiva and cervical lymph nodes is presented. Electron-beam radiation with 40 Gy and systemic chemotherapy with vincristine and interferon gamma produced almost complete remission of the oral lesion while the persistent pain was relieved.

MRI in carcinomatous encephalitis

Abstract We report a rare case of miliary brain metastases presenting with symptoms similar to encephalitis (“carcinomatous encephalitis”). Contrast-enhanced MRI demonstrated miliary metastases more distinctly than other imaging methods and reproduced the pathological features.

Heterotopic salivary gland adenocarcinoma in the cervical region

A case of heterotopic salivary gland adenocarcinoma (HSGA) in the right cervical region is presented. The carcinoma cells were positive for alpha-amylase, carcinoembryonic antigen, epithelial membrane antigen, cytokeratin as well as for expression of human salivary alpha-amylase messenger ribonucleic acid. The possibility of HSGA should be considered when an adenocarcinoma producing human salivary alpha-amylase is diagnosed away from sites where major and minor salivary glands normally are found.

MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer

The immune checkpoint ligand PD-L1 and the transmembrane mucin MUC1 are upregulated in triple-negative breast cancer (TNBC), where they contribute to its aggressive pathogenesis. Here, we report that genetic or pharmacological targeting of the oncogenic MUC1 subunit MUC1-C is sufficient to suppress PD-L1 expression in TNBC cells. Mechanistic investigations showed that MUC1-C acted to elevate PD-L1 transcription by recruitment of MYC and NF-κB p65 to the PD-L1 promoter. In an immunocompetent model of TNBC in which Eo771/MUC1-C cells were engrafted into MUC1 transgenic mice, we showed that targeting MUC1-C associated with PD-L1 suppression, increases in tumor-infiltrating CD8+ T cells and tumor cell killing. MUC1 expression in TNBCs also correlated inversely with CD8, CD69, and GZMB, and downregulation of these markers associated with decreased survival. Taken together, our findings show how MUC1 contributes to immune escape in TNBC, and they offer a rationale to target MUC1-C as a novel immunotherapeutic approach for TNBC treatment.Significance: These findings show how upregulation of the transmembrane mucin MUC1 contributes to immune escape in an aggressive form of breast cancer, with potential implications for a novel immunotherapeutic approach. Cancer Res; 78(1); 205-15. ©2017 AACR.