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BACKGROUND: T4b was previously defined as unresectable; however, it has been demonstrated that compartmental surgery can achieve treatment outcomes comparable to those of T4a cases. We frequently encounter posteriorly advanced oral squamous cell carcinoma (OSCC) in contact with the medial pterygoid muscle, and determining the appropriate extent of resection and managing subsequent recurrence in the masticator space remain challenging. Therefore, this study aimed to investigate the association between the tumor's spatial relationship with the medial pterygoid muscle and patient outcomes, providing insights to enhance diagnostic precision and guide therapeutic strategies. METHODS: This retrospective study included 50 patients with posteriorly advanced OSCC, excluding those classified as T4b. Preoperative magnetic resonance imaging classified lesions as either non-contact or contact type based on their relationship to the medial pterygoid muscle. Clinicopathological factors, overall survival rate (OS), and recurrence rates in the masticator space were compared between the two groups. RESULTS: No significant differences were observed in OS between the non-contact and contacttypes. Among non-contact types, no recurrence in the masticator space was observed, whereas contact-type tumors showed a recurrence rate of 22.6% (p = 0.04). Multivariate analysis revealed that lymphatic invasion was associated with an increased risk of primary recurrence, whereas postoperative radiotherapy was associated with a reduced risk among 31 patients with contact-type tumors. CONCLUSIONS: In summary, compartmental surgery with pterygomandibular space dissection should be considered for contact-type tumors near the medial pterygoid muscle, along with T4b cases. However, its indication must be assessed based on clinical findings. Postoperative radiotherapy is key to recurrence prevention, improving oncologic outcomes and long-term disease control.

BACKGROUND & AIMS: Long-term oral administration of zinc preparations to zinc-deficient patients has been shown to result in copper deficiency. Two patients who had zinc deficiency taste disorder and copper deficiency after upper gastrointestinal resection were treated with zinc acetate hydrate. METHODS: Two patients with abnormal taste were diagnosed as zinc deficiency taste disorder by an examination, and were treated with zinc replacement therapy. Case 1 (BMI 15.6 kg/mm2) was given 50 mg/day of zinc acetate hydrate for a year. On the other hand, case 2 (BMI 24.2 kg/mm2) started treatment with zinc acetate hydrate at a concentration of 100 mg/day, but due to pancytopenia, the dose was reduced to 25 mg/day, and the drug was administered for a total of 5 years. RESULTS: Both patients demonstrated an improvement in taste. In case 1, the serum zinc level reached 111 μg/dL after a year of zinc acetate hydrate treatment. There was no decrease in serum copper levels or blood cell counts. Case 2 had pancytopenia with neutrophils 320/mm3, Hb 7.1 g/dL, and platelets 164 × 109/L one year after zinc acetate hydrate administration. The patient was given subcutaneous G-CSF and a transfusion of concentrated red blood cells and was started on oral administration of pure cocoa 10 g/day. However, pancytopenia did not improve; thus, discontinuing and reducing zinc acetate hydrate increased serum copper and improvement of pancytopenia. Due to the history of spinal canal stenosis surgery, it was difficult to evaluate the neurologic abnormalities. CONCLUSION: Long-term administration of zinc supplements should be administered with caution to patients with zinc deficiency-related taste disorders following upper gastrointestinal resection, particularly those with poor nutritional status.

Objective Extranodal extension (ENE) is one of the major influencing factors for the oncological outcomes in oral squamous cell carcinoma (OSCC). We aimed to elucidate the clinical features predictive of ENE in OSCC. Materials and methods We conducted a retrospective analysis of patients with OSCC who underwent neck dissection (ND) with a confirmed pN + status. Cases in which the histopathological evaluation was compromised by preoperative chemotherapy or radiotherapy were excluded. Histopathological evaluation of extent of ENE category and grading of worst pattern of invasion (WPOI) was compared for available cases. Results Fifty-nine patients met the inclusion criteria for the study. Of these, 32/59 (54.2%) were ENE-positive. A higher incidence of ENE was observed in cases where ND was performed at a separate time from the primary tumor resection (odds ratio [OR] = 11.0, 95% confidence interval [95%CI] 2.23-54.5, P = 0.003). Additionally, a higher grade of WPOI (WPOI 4 or 5) was significantly associated with ENE occurrence (OR = 4.53, 95%CI 1.19-20.50, P = 0.026). A positive correlation between the WPOI grade and ENE extent was also identified (rho = 0.412, P <.001). Conclusion We demonstrated an association between WPOI and ENE in patients with OSCC. Objective Extranodal extension (ENE) is one of the major influencing factors for the oncological outcomes in oral squamous cell carcinoma (OSCC). We aimed to elucidate the clinical features predictive of ENE in OSCC. Materials and methods We conducted a retrospective analysis of patients with OSCC who underwent neck dissection (ND) with a confirmed pN + status. Cases in which the histopathological evaluation was compromised by preoperative chemotherapy or radiotherapy were excluded. Histopathological evaluation of extent of ENE category and grading of worst pattern of invasion (WPOI) was compared for available cases. Results Fifty-nine patients met the inclusion criteria for the study. Of these, 32/59 (54.2%) were ENE-positive. A higher incidence of ENE was observed in cases where ND was performed at a separate time from the primary tumor resection (odds ratio [OR] = 11.0, 95% confidence interval [95%CI] 2.23-54.5, P = 0.003). Additionally, a higher grade of WPOI (WPOI 4 or 5) was significantly associated with ENE occurrence (OR = 4.53, 95%CI 1.19-20.50, P = 0.026). A positive correlation between the WPOI grade and ENE extent was also identified (rho = 0.412, P <.001). Conclusion We demonstrated an association between WPOI and ENE in patients with OSCC.

Objectives: This study aimed to determine the salivary factors that influence salivary bacterial counts and the microbiome composition in children and young adults. Methods: This cross-sectional study included 382 patients who visited the dental clinic in Hiroshima University Hospital. All participants underwent a saliva test and were divided into high- and low-bacterial-count groups based on the median bacterial count. Salivary factors and clinical variables, including the total number of teeth and plaque control record, were analyzed to determine their association with salivary bacterial counts. Furthermore, a comprehensive analysis of microbiome diversity and composition was performed using 16S rRNA sequencing. Results: Univariate and multivariate analyses identified stimulated saliva volume (SSV) and plaque control record as independent factors influencing salivary bacterial counts. Principal coordinate analysis revealed a significant decrease in beta diversity in the high-bacterial-count group. LEfSe analysis revealed Prevotella, Veillonella, Megafaella, Selenomonas, and TM7X as the 5 most abundant bacteria. The relative abundance of the 35 KEGG pathways exhibited significant differences. Furthermore, Prevotella and Veillonella were strongly associated with 25 functional pathways. Conclusion: Oral hygiene instruction is necessary even for children and young adults with relatively adequate SSV to maintain a healthy oral microbiome. (c) 2025 The Authors. Published by Elsevier Inc. on behalf of FDI World Dental Federation. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/) Objectives: This study aimed to determine the salivary factors that influence salivary bacterial counts and the microbiome composition in children and young adults. Methods: This cross-sectional study included 382 patients who visited the dental clinic in Hiroshima University Hospital. All participants underwent a saliva test and were divided into high- and low-bacterial-count groups based on the median bacterial count. Salivary factors and clinical variables, including the total number of teeth and plaque control record, were analyzed to determine their association with salivary bacterial counts. Furthermore, a comprehensive analysis of microbiome diversity and composition was performed using 16S rRNA sequencing. Results: Univariate and multivariate analyses identified stimulated saliva volume (SSV) and plaque control record as independent factors influencing salivary bacterial counts. Principal coordinate analysis revealed a significant decrease in beta diversity in the high-bacterial-count group. LEfSe analysis revealed Prevotella, Veillonella, Megafaella, Selenomonas, and TM7X as the 5 most abundant bacteria. The relative abundance of the 35 KEGG pathways exhibited significant differences. Furthermore, Prevotella and Veillonella were strongly associated with 25 functional pathways. Conclusion: Oral hygiene instruction is necessary even for children and young adults with relatively adequate SSV to maintain a healthy oral microbiome. (c) 2025 The Authors. Published by Elsevier Inc. on behalf of FDI World Dental Federation. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

The role of inflammatory response by salivary gland cells is considered to be important in the pathogenesis of salivary gland chronic inflammation, as seen in Sjögren's syndrome patients. The primary salivary gland cell cultures are required to investigate such inflammatory responses, though primary cells exhibit a limited replicative short lifespan with only a few passages. An immortalized human minor salivary gland cell line, NSG cells, was established by transfection with human telomerase reverse transcriptase (hTERT) and SV40 large T antigen (SV40LT). The effects of IFN-γ, TNF-α, and IL-1β on chemokine expression in those cells were then examined. Following hTERT expression vector and SV40LT vector transfections into minor salivary gland cells with a non-viral method, real-time PCR was employed to examine the effects of IFN-γ, TNF-α, and IL-1β on chemokine mRNA expression. Additionally, ELISA was used to examine the effects of combinations of IFN-γ, TNF-α, and IL-1β on CXCL10 and CXCL1 protein expressions. NSG cell growth was found to continue for more than 100 population doublings and the cells constitutively expressed immortalized-related and salivary gland-associated genes. IFN-γ, TNF-α, and IL-1β each increased the examined chemokines by various levels. Both TNF-α and IL-1β separately increased IFN-γ-induced CXCL10 in the NSG cells, whereas IFN-γ decreased CXCL1 induced by TNF-α or IL-1β. An immortalized human minor salivary gland cell line was established by hTERT and SV40LT transfection. The examinations showed that IFN-γ, TNF-α, and IL-1β have important roles for development of salivary gland inflammation, such as Sjögren's syndrome.

We report a case of an industrial homeworker diagnosed with allergic contact dermatitis by UV-curing acrylic resin for crafts. Approximately 2 months after a female in her 40s started producing handicrafts using resin, itchy desquamative erythema and vesicles occurred on her eyelids and palms. The course of the symptoms suggested that her dermatitis was occupational origin. Interviews regarding the work environment indicated that her employer did not adequately explain the hazards of resin. An inappropriate work environment may have led to the development of allergy. For industrial homeworkers, managing chemical exposures is predicted to be challenging. However, the expansion of the handmade market and the spread of craft resin increase opportunities for exposure to resin. In order to prevent the spread of health problems, appropriate information should be provided regarding the hazards of resin.

Mesenchymal stem cells (MSCs) have gained significant attention in cell therapies due to their multipotency and immunomodulatory capacities. The transcriptional co-activators YAP/TAZ, central to the mechanotransduction system in MSCs, dominantly direct MSCs lineage commitment. However, their role in immunomodulation remains elusive. Accordingly, this present study aimed to investigate the role of mechanotransducer YAP/TAZ and their binding target transcriptional factor, TEAD, in the immunomodulatory capacities of human bone marrow-derived MSCs. Reducing YAP/TAZ activity by altering the matrix stiffness, disrupting the F-actin integrity with chemical inhibitors, or using siRNAs increased the expression of immunomodulatory genes, such as TSG-6 and IDO, upon TNF-α stimulation. Similarly, transfection of TEAD siRNA also increased the immunomodulatory capacities in MSCs. RNA-seq analysis and inhibition assays demonstrated that the immunomodulatory capacities caused by YAP/TAZ-TEAD axis disruption were due to the NF-κB signaling pathway activation. Then, we also evaluated the in vivo anti-inflammatory efficacy of MSCs in a dextran sulfate sodium (DSS)-induced mice colitis model. The administration of human MSCs transfected with TEAD siRNA, which exhibited enhanced immunomodulatory properties in vitro, significantly ameliorated inflammatory bowel disease symptoms, such as body weight loss and acute colon inflammation, in the DSS-induced mice colitis model. Our findings underscore the mechanosignaling YAP/TAZ-TEAD axis as a regulator of MSCs immunomodulation. Targeting these signaling pathways could herald promising MSCs-based therapies for immune disorders.

AIM: To retrospectively investigate the relationship between the CD4+ T-cell counts at baseline and the efficacy of the initial periodontal treatment of patients undergoing treatment for human immunodeficiency virus (HIV) infection using the periodontal inflamed surface area (PISA). MATERIALS AND METHODS: Thirty-three patients with chronic periodontitis who had undergone periodontal examination at baseline and after the initial periodontal treatment were enrolled. PISA was calculated from the periodontal probing depth and bleeding on probing, and the ratio of PISA after treatment to that at baseline (PISA response ratio) was calculated. Groups with a response ratio of <1 and ≥1 were defined as the improvement and the non-improvement groups, respectively. RESULTS: PISA after the initial periodontal treatment significantly decreased compared with that at baseline (p < .05). A weak negative correlation was found between the PISA response ratio and CD4+ T-cell counts at baseline (p < .05). The CD4+ T-cell counts at baseline were significantly higher in the improvement group than in the non-improvement group (p < .05). Multivariate analysis revealed that the CD4+ T-cell counts at baseline was an independent factor that affects the PISA (p < .05). CONCLUSIONS: The higher the CD4+ T-cell counts at baseline in patients undergoing treatment for HIV infection, the more effective the initial periodontal treatment.

PURPOSE: Chemotherapy-induced taste alteration is a side effect that can result in malnutrition and reduced quality of life in cancer patients. However, the underlying causes of this phenomenon remain unclear, and evidence-based treatments have not been established. This study focused on patients' subjective symptoms of taste alterations aimed to explore how the sensitivity to basic tastes changes due to anticancer agents and how alterations in one taste perception are associated with changes in other tastes during chemotherapy. METHODS: A cross-sectional questionnaire-based interview survey was conducted on 215 patients undergoing chemotherapy. The subjective sensitivity to each basic taste was assessed using a visual analog scale, and the incidence of taste alterations due to different chemotherapy regimens was calculated. Multivariate logistic regression analysis was performed to determine whether there were associations between changes in one taste sensitivity and changes in other taste sensitivities. RESULTS: Approximately half (49.5%) of the patients experienced chemotherapy-induced taste alterations. An analysis of subjective changes in basic tastes revealed that the salt and umami taste systems were more sensitive to chemotherapy than other taste systems. Patients with altered sensitivity to sweet taste were significantly more likely to report altered sensitivity to salt, bitter, and sour tastes. Moreover, umami-salt and bitter-sour taste sensitivities were significantly related to each other. CONCLUSION: This study suggests that changes in subjective sensitivities to one basic taste during chemotherapy may be accompanied by changes in other tastes in specific combinations. Considering taste associations in dietary guidance may help improve the nutritional status of cancer patients experiencing taste alterations due to chemotherapy.

The Hippo signaling pathway and its downstream effector YAP play a central role in cell proliferation. Dysregulation of the Hippo pathway triggers YAP hyperactivation, thereby inducing head and neck squamous cell carcinoma (HNSCC). Recently, we reported that EGFR promotes tyrosine phosphorylation of MOB1 and subsequent LATS1/2 inactivation, which are core components of the Hippo pathway, resulting in YAP activation. However, EGFR-targeted monotherapy has shown a low response rate in HNSCC patients. Given that YAP is activated in patient samples refractory to EGFR-targeted therapy, EGFR inhibitors may temporarily inactivate YAP, but intrinsic hyperactivation or acquired reactivation of YAP may confer resistance to EGFR inhibitors in HNSCC cells. The mechanism by which YAP is activated in HNSCC resistant to EGFR inhibitors remains unclear. Comprehensive transcriptional analysis revealed that AXL activates YAP through a novel mechanism: AXL heterodimerizes with EGFR, thereby activating YAP via the EGFR-LATS1/2 axis. The combination of AXL and EGFR inhibitors synergistically inactivates YAP and suppresses the viability of HNSCC and lung adenocarcinoma cells. In turn, LATS1/2 knockout and YAP hyperactivation confer resistance to the synergistic effects of these inhibitors. Our findings suggest that co-targeting both AXL and EGFR represent a promising therapeutic approach in patients with EGFR-altered cancers.

OBJECTIVES: The causes of hypogeusia include zinc deficiency, systemic illness, and consumption of drugs. Notably, patients with oral cavity diseases such as oral candidiasis and salivary gland hypofunction may present with risk factors that remain unreported. Hence, this study aimed to investigate the relationship between age, sex, smoking status, serum zinc concentration, oral candidiasis, saliva volume, and taste function in patients with hypogeusia. SUBJECTS AND METHODS: Overall, 335 participants who complained of taste abnormalities underwent a taste test. Based on the recognition threshold value, the participants were classified as normal individuals (recognition threshold of 1 and 2) and patients with hypogeusia (recognition threshold of ≥3). The clinical characteristics, including resting saliva volume (RSV) and stimulated saliva volume (SSV), were compared, and a multivariate logistic regression analysis focusing on RSV was performed. RESULTS: Patients with hypogeusia had a lower RSV than normal individuals for all tastes, but not for SSV. Based on the results of regression analysis, RSV was identified as an independent predictor of hypogeusia for salty and bitter tastes. Moreover, the proportion of patients with decreased RSV increased as the number of taste qualities exceeding the reference recognition threshold increased. Furthermore, a decrease in RSV was associated with an increase in the recognition threshold for salty and bitter tastes. CONCLUSIONS: Based on the results of the present study, moisturizing the oral cavity may be useful against hypogeusia.

Resin components, such as methyl methacrylate (MMA) can cause allergic contact dermatitis (ACD). Allergic reactions to resin are usually delayed. Only a few studies have reported dental resin allergy with acute symptoms. Here, a case of ACD with acute facial swelling after dental treatment using resin material is reported. A 55-year-old woman with a history of periungual inflammation when using gel nail polish had repeated episodes of facial swelling after dental treatment with resin material. The resin temporary crown was removed, and symptoms were alleviated with antihistamines and corticosteroids. With the suspicion of resin allergy, skin tests were performed. Patch testing revealed positive reactions to self-adhesive resin cement (primer and polymerized), self-curing acrylic resin (liquid and polymerized), 2-hydroxyethyl methacrylate (2-HEMA), and ethylene glycol dimethacrylate (EGDMA), whereas the prick test was negative for all allergens. Complement C4 and C1 inhibitor activity were reference values in the tests for hereditary angioedema. Based on these findings, the patient was diagnosed with ACD to 2-HEMA and EGDMA. Since diagnosis, no similar symptoms have been observed in subsequent dental treatment with non-resin materials. The use of dental resin materials may cause ACD with an acute reaction. This report alerts dentists who routinely use resin materials.

Ghost cell odontogenic carcinoma (GCOC) is an extremely rare intraosseous malignant odontogenic tumor with prominent ghost cell keratinization and dentinoid formation. Here, we present the first case of GCOC arising in dentinogenic ghost cell tumor (DGCT), peripheral. The patient was a man in his 60s with an exophytic mass in the anterior part of lower gingiva. The resected tumor measured 4.5 cm in maximum diameter. Histologically, the nonencapsulated tumor proliferated in the gingiva without bone invasion. It was predominantly composed of ameloblastoma-like nests and islands of basaloid cells with ghost cells and dentinoid in the mature connective tissue, suggesting DGCT, peripheral. As minor components, sheets of atypical basaloid cells and ameloblastic carcinoma-like nests with pleomorphism and high proliferative activity (Ki-67 labeling index up to 40%) consistent with malignancy were identified. CTNNB1 mutation and β-catenin nuclear translocation were observed in both benign and malignant components. Final diagnosis was GCOC arising in DGCT, peripheral. GCOC shows similar histological features to DGCT. In this unique case without invasion, the cytological atypia and high proliferative activity supports the diagnosis of malignant transformation from DGCT.

Multiple myeloma is a hematologic malignancy characterized by neoplastic proliferation of monoclonal plasma cells. The mass formed within the soft tissue is particularly called extramedullary disease in multiple myeloma. This disease rarely occurs in the head and neck regions. Notably, submandibular extramedullary lesions have not been reported. Therefore, we report a case of multiple myeloma diagnosed with extramedullary disease in the submandibular region. A 70-year-old woman was referred to our clinic due to painless swelling on the left side of the submandibular region. We confirmed the presence of a relatively well-circumscribed subglobular tumor on the left side of the submandibular region on contrast-enhanced computed tomography and magnetic resonance imaging. The tumor was resected surgically, including the circumferential tissue, with the patient under general anesthesia. The tumor was histologically diagnosed as a plasma cell neoplasm. Positron emission tomographycomputed tomography showed an osteolytic lesion in the sacral region. A bone marrow examination diagnosed multiple myeloma of IgG-lambda-type, ISS II stage. The patient has been receiving continuous chemotherapy for multiple myeloma for 1 year after operation and has no recurrence in the submandibular region. Data Availability Statement: The data presented in this study are available on request from the corresponding author. Publicly available datasets were analyzed in this study.

Abstract Background Dysgeusia is a relatively early symptom of zinc deficiency, and zinc replacement is effective in treating dysgeusia. The administration of zinc acetate hydrate (ZAH) was approved in 2017 for patients with hypozincemia in Japan. This retrospective study was conducted to explore the efficacy and safety of ZAH administration in patients with hypozincemia-induced dysgeusia. Methods Patients with hypozincemia-induced dysgeusia who visited our hospital from May 2013 to December 2019 were included in this study. ZAH (zinc content; 50 mg/day) was administered to 42 patients for 24 weeks. The taste test was performed using the filter paper disk method, and the total cognitive thresholds of the left and right chorda tympani regions were used. Changes in taste function, serum zinc and copper levels, and copper/zinc ratio were analyzed. A total of 28 patients who received polaprezinc (PPZ, zinc content; 34 mg/day) for 24 weeks, who were prescribed until ZAH was approved, were registered as controls. Results Serum zinc levels at 12 and 24 weeks after ZAH or PPZ administration were higher than those before administration. These levels were significantly higher in the ZAH-treated group than in the PPZ-treated group. However, serum copper levels did not significantly change before and after administration. In the taste test, the taste thresholds for the acidity and salty at 12 and 24 weeks after ZAH administration were significantly decreased compared to before administration. In contrast, in the PPZ group, the taste thresholds for the acidity and salty were significantly decreased 24 weeks after administration. Conclusions ZAH (50 mg/day) administration was effective in improving the gustatory sensitivity of patients with dysgeusia and hypozincemia 12 weeks after administration without affecting the serum copper level. ZAH was also more effective than PPZ.

The presence of a supernumerary tooth is one of the most common dental anomalies, and surgical treatment is often required to address this anomaly. Moreover, it may lead to malocclusion, and long-term follow-up is important to monitor its status. A 4-year-and-11-month-old boy was referred to our hospital for dental caries treatment. At 5 years and 5 months of age, a radiographic examination showed a supernumerary tooth (first supernumerary tooth) near the permanent maxillary left central incisor, and it was extracted 6 months later. Eighteen months after the extraction of the first supernumerary tooth, a new supernumerary tooth (second supernumerary tooth) was detected in the same region, which was extracted when the patient was aged seven years and seven months. Seven months later, another supernumerary tooth (third supernumerary tooth) was detected and extracted immediately. However, the permanent maxillary left central incisor did not erupt spontaneously even after 6 months. Therefore, surgical exposure was performed, and the central incisor erupted into the oral cavity. This report describes our experience with this patient with three metachronous supernumerary teeth and their management until the eruption of the permanent tooth. This report highlights the importance of long-term follow-up after supernumerary tooth extraction until the permanent teeth in that region have erupted completely.

Tumor angiogenesis is essential for tumor progression. The inhibition of tumor angiogenesis is a promising therapy for tumors. Bovine lactoferrin (bLF) has been reported as an anti-tumor agent. However, bLF effects on tumor angiogenesis are not well demonstrated. This study evaluated the inhibitory effects of bLF on tumor angiogenesis in vivo and in vitro. Herein, tumor endothelial cells (TECs) and normal endothelial cells (NECs) were used. Proliferation, migration, tube formation assays, RT-PCR, flow cytometry, Western blotting, siRNA experiments and immunoprecipitation were conducted to clarify the mechanisms of bLF-induced effects. CD-31 immunoexpression was examined in tumor tissues of oral squamous cell carcinoma mouse models with or without Liposomal bLF (LbLF)-administration. We confirmed that bLF inhibited proliferation/migration/tube formation and increased apoptosis in TECs but not NECs. TNF receptor-associated factor 6 (TRAF6), p-p65, hypoxia inducible factor-α (HIF-1α) and vascular endothelial growth factor (VEGF) were highly expressed in TECs. In TECs, bLF markedly downregulated VEGF-A, VEGF receptor (VEGFR) and HIF-1α via the inhibition of p-p65 through binding with TRAF6. Since NECs slightly expressed p-p65, bLF-TRAF-6 binding could not induce detectable changes. Moreover, orally administrated LbLF decreased CD31-positive microvascular density only in TECs. Hence, bLF specifically suppressed tumor angiogenesis through p-p65 inhibition by binding to TRAF6 and suppressing HIF-1α activation followed by VEGF/VEGFR down-regulation. Collectively, bLF can be an anti-angiogenic agent for tumors.

Genetic alterations and dysregulation of signaling pathways are indispensable for the initiation and progression of cancer. Understanding the genetic, molecular, and signaling diversities in cancer patients has driven a dynamic change in cancer therapy. Patients can select a suitable molecularly targeted therapy or immune checkpoint inhibitor based on the driver gene alterations determined by sequencing of cancer tissue. This "precision medicine" approach requires detailed elucidation of the mechanisms connecting genetic alterations of driver genes and aberrant downstream signaling pathways. The regulatory mechanisms of the Hippo pathway and Yes-associated protein/transcriptional co-activator with PDZ binding motif (YAP/TAZ) that have central roles in cancer cell proliferation are not fully understood, reflecting their recent discovery. Nevertheless, emerging evidence has shown that various genetic alterations dysregulate the Hippo pathway and hyperactivate YAP/TAZ in cancers, including head and neck squamous cell carcinoma (HNSCC). Here, we summarize the latest evidence linking genetic alterations and the Hippo pathway in HNSCC, with the aim of contributing to the continued development of precision medicine.

多彩な細胞像を示し、異型に乏しい低悪性度腫瘍の多い唾液腺腫瘍では、(1)臨床的特徴の把握、(2)複数箇所からの採取とギムザ染色の併用、(3)悪性の危険度、臨床的対応を主眼に分類されたミラノシステムの利用、(4)代表的な腫瘍型の定型像とピットフォールの理解、(5)補助診断の実施、といった悪性の見落としを防ぐ方策が必要である。(著者抄録)

Non-sebaceous lymphadenoma is a rare benign salivary gland tumor comprised of non-sebaceous epithelial cells and lymphoid tissue. Although its clinicopathological features have been described, its histogenesis and genetic background have not yet been elucidated. MAML2 rearrangement and the resultant CRTC1/3-MAML2 fusion gene are well-known specific genetic changes in mucoepidermoid carcinoma. Here, we present a case of lymphoepithelial tumor characterized by histomorphology of the non-sebaceous lymphadenoma and CRTC1-MAML2 fusion gene. The patient was an 83-year-old woman with an 8-year history of a solid, well-circumscribed tumor in the parotid gland. Histologically, the tumor was surrounded by thin fibrous connective tissue and was composed of tubular-cystic and solid nests of epithelial cells equally distributed in the lymphoid tissue. The histological features were suggestive of non-sebaceous lymphadenoma. Although the histomorphology was not consistent with mucoepidermoid carcinoma, a diagnosis of non-sebaceous lymphadenoma-like mucoepidermoid carcinoma was made based on the presence of the CRTC1-MAML2 fusion gene. The histological features alone could not establish the diagnosis, and ancillary molecular analysis was required.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by inflammatory bone destruction in which tumor necrosis factor alpha (TNF-α) plays a key role. Bovine lactoferrin (bLF) is a multifunctional protein with anti-inflammatory and immunomodulatory properties. This study aimed to clarify the inhibitory effects of bLF on the pathological progression of RA. The mannan-induced arthritis model in SKG mice (genetic RA model) was used. Orally applied liposomal bLF (LbLF) markedly reduced ankle joint swelling and bone destruction. Histologically, pannus formation and osteoclastic bone destruction were prevented in the LbLF-treated animals. Moreover, orally administered LbLF improved the balance between Th17 cells and regulatory T cells isolated from the spleen of mannan-treated SKG mice. In an in vitro study, the anti-inflammatory effects of bLF on TNF-α-induced TNF-α production and downstream signaling pathways were analyzed in human synovial fibroblasts from RA patients (RASFs). bLF suppressed TNF-α production from RASFs by inhibiting the nuclear factor kappa B and mitogen-activated protein kinase pathways. The intracellular accumulation of bLF in RASFs increased in an applied bLF dose-dependent manner. Knockdown of the lipoprotein receptor-related protein-1 (LRP1) siRNA gene reduced bLF expression in RASFs, indicating that exogenously applied bLF was mainly internalized through LRP-1. Immunoprecipitated proteins with anti-TNF receptor-associated factor 2 (TRAF2; an adapter protein/ubiquitin ligase) included bLF, indicating that bLF binds directly to the TRAF2-TRADD-RIP complex. This indicates that LbLF may effectively prevent the pathological progression of RA by suppressing TNF-α production by binding to the TRAF2-TRADD-RIP complex from the RASFs in the pannus. Therefore, supplemental administration of LbLF may have a beneficial effect on preventive/therapeutic reagents for RA.

The Hippo pathway is frequently dysregulated in cancer, leading to the unrestrained activity of its downstream targets, YAP/TAZ, and aberrant tumor growth. However, the precise mechanisms leading to YAP/TAZ activation in most cancers is still poorly understood. Analysis of large tissue collections revealed YAP activation in most head and neck squamous cell carcinoma (HNSCC), but only 29.8% of HNSCC cases present genetic alterations in the FAT1 tumor suppressor gene that may underlie persistent YAP signaling. EGFR is overexpressed in HNSCC and many other cancers, but whether EGFR controls YAP activation is still poorly understood. Here, we discover that EGFR activates YAP/TAZ in HNSCC cells, but independently of its typical signaling targets, including PI3K. Mechanistically, we find that EGFR promotes the phosphorylation of MOB1, a core Hippo pathway component, and the inactivation of LATS1/2 independently of MST1/2. Transcriptomic analysis reveals that erlotinib, a clinical EGFR inhibitor, inactivates YAP/TAZ. Remarkably, loss of LATS1/2, resulting in aberrant YAP/TAZ activity, confers erlotinib resistance on HNSCC and lung cancer cells. Our findings suggest that EGFR-YAP/TAZ signaling plays a growth-promoting role in cancers harboring EGFR alterations, and that inhibition of YAP/TAZ in combination with EGFR might be beneficial to prevent treatment resistance and cancer recurrence.

The diagnosis of ameloblastic carcinoma is based on a combination of cytological features of malignancy and histological pattern of an ameloblastoma. We report a very rare case of ameloblastic carcinoma arising from a pre-existing ameloblastoma of the maxilla with challenging diagnostic aspects due to atypical histology. A 74-year-old female with a swelling in the right maxillary region was referred to our hospital in March 2015. Intraoral examination revealed a 45 x 30 mm sized swelling extending from the right upper first premolar to the maxillary tuberosity. Radiographic examination revealed a tumor mass with heterogeneous enhancement from the right maxillary alveolar area to the maxillary sinus, invading the pterygopalatine fossa under the cranial base, with bone destruction of the alveolar bone, floor of the maxillary sinus, and pterygoid plate. Incisional biopsy results revealed round or short spindle cells with clear cytoplasm, with low mitotic activity, cellular atypism, and Ki-67 labeling index. A low grade malignant epithelial tumor was suspected. Tumor resection with hemimaxillectomy and partial mandibulectomy was performed in April 2015. The surgical specimen was composed of diffusely distributed spindle cells and clusters of round cells with clear cytoplasm, similar to the biopsy findings. Although cellular atypism was scant, Ki-67 labeling index was high in most sections. Ameloblastoma was detected in part of the tumor and a definitive diagnosis of ameloblastic carcinoma was made. The patient died of metastatic colon cancer 3 years and 10 months post-operatively; there was no evidence of recurrence and metastasis of ameloblastic carcinoma in that duration. (C) 2021 Asian AOMS, ASOMP, JSOP, JSOMS, JSOM, and JAMI. Published by Elsevier Ltd. All rights reserved.

症例1は75歳女性で、歯科治療を目的に近在歯科医院を受診し、口蓋の腫瘤について当科紹介受診となった。生検によりMALTリンパ腫の病理組織学的診断を得て、諸検査よりシェーグレン症候群の合併例と診断した。病変は口腔内に限局していたため外科的切除を先行し、術後化学療法を施行、治療終了後5年間で局所再発や転移は認めていない。症例2は70歳女性で、口腔内に18F-FDGの異常集積を認め当科紹介となり、MALTリンパ腫の病理組織学的診断を得た。R-CVP療法を6コース施行され、その後8年、再発は認めなかった。症例3は62歳女性で、左側口底部に無痛性腫瘤を自覚して当科を紹介され、生検にてMALTリンパ腫との確定診断を得た。外科的切除+術後化学療法を行い、その後7年、再発は認めていない。症例4は70歳女性で、左側軟口蓋部の腫瘤を自覚したため当科受診となり、生検にてMALTリンパ腫の確定診断を得て、リツキシマブ単独療法を行った。3年後、再発は認めていない。

Cetuximab, a monoclonal antibody targeting EGFR, is a standard of care for the treatment for locally advanced or metastatic head and neck squamous cell carcinoma (HNSCC). However, despite overexpression of EGFR in over 90% of HNSCC lesions, most HNSCC patients fail to respond to cetuximab treatment. In addition, there are no available biomarkers to predict sensitivity or resistance to cetuximab in the clinic. Here, we sought to advance precision medicine approaches for HNSCC by identifying PI3K-mTOR signaling-network-specific cetuximab resistance mechanisms. We first analyzed the frequency of genomic alterations in genes involved in the PI3K-mTOR signaling circuitry in the HNSCC TCGA dataset. Experimentally, we took advantage of CRISPR/Cas9 genome editing approaches to systematically explore the contribution of genomic alterations in each tumor suppressor gene (TSG) controlling the PI3K-mTOR pathway to cetuximab resistance in HNSCC cases that do not exhibit PIK3CA mutations. Remarkably, we found that many HNSCC cases exhibit pathway-specific gene copy number loss of multiple TSGs that normally restrain PI3K-mTOR signaling. Among them, we found that both engineered and endogenous PTEN gene deletions can mediate resistance to cetuximab. Our findings suggest that PTEN gene copy number loss, which is highly prevalent in HNSCC, may result in sustained PI3K/mTOR signaling independent of EGFR, thereby representing a promising mechanistic biomarker predictive of cetuximab resistance in this cancer type. Further prospective studies are needed to investigate the impact of PTEN loss on cetuximab efficacy in the clinic.

Cetuximab, a monoclonal antibody targeting EGFR, is a standard of care for the treatment for locally advanced or metastatic head and neck squamous cell carcinoma (HNSCC). However, despite overexpression of EGFR in over 90% of HNSCC lesions, most HNSCC patients fail to respond to cetuximab treatment. In addition, there are no available biomarkers to predict sensitivity or resistance to cetuximab in the clinic. Here, we sought to advance precision medicine approaches for HNSCC by identifying PI3K-mTOR signaling-network-specific cetuximab resistance mechanisms. We first analyzed the frequency of genomic alterations in genes involved in the PI3K-mTOR signaling circuitry in the HNSCC TCGA dataset. Experimentally, we took advantage of CRISPR/Cas9 genome editing approaches to systematically explore the contribution of genomic alterations in each tumor suppressor gene (TSG) controlling the PI3K-mTOR pathway to cetuximab resistance in HNSCC cases that do not exhibit PIK3CA mutations. Remarkably, we found that many HNSCC cases exhibit pathway-specific gene copy number loss of multiple TSGs that normally restrain PI3K-mTOR signaling. Among them, we found that both engineered and endogenous PTEN gene deletions can mediate resistance to cetuximab. Our findings suggest that PTEN gene copy number loss, which is highly prevalent in HNSCC, may result in sustained PI3K/mTOR signaling independent of EGFR, thereby representing a promising mechanistic biomarker predictive of cetuximab resistance in this cancer type. Further prospective studies are needed to investigate the impact of PTEN loss on cetuximab efficacy in the clinic.

Prostate cancer (PCa) innervation contributes to the progression of PCa. However, the precise impact of innervation on PCa cells is still poorly understood. By focusing on muscarinic receptors, which are activated by the nerve-derived neurotransmitter acetylcholine, we show that muscarinic receptors 1 and 3 (m1 and m3) are highly expressed in PCa clinical specimens compared with all other cancer types, and that amplification or gain of their corresponding encoding genes (CHRM1 and CHRM3, respectively) represent a worse prognostic factor for PCa progression free survival. Moreover, m1 and m3 gene gain or amplification is frequent in castration-resistant PCa (CRPC) compared with hormone-sensitive PCa (HSPC) specimens. This was reflected in HSPC-derived cells, which show aberrantly high expression of m1 and m3 under androgen deprivation mimicking castration and androgen receptor inhibition. We also show that pharmacological activation of m1 and m3 signaling is sufficient to induce the castration-resistant growth of PCa cells. Mechanistically, we found that m1 and m3 stimulation induces YAP activation through FAK, whose encoding gene, PTK2 is frequently amplified in CRPC cases. Pharmacological inhibition of FAK and knockdown of YAP abolished m1 and m3-induced castration-resistant growth of PCa cells. Our findings provide novel therapeutic opportunities for muscarinic-signal-driven CRPC progression by targeting the FAK-YAP signaling axis.

Prostate cancer (PCa) innervation contributes to the progression of PCa. However, the precise impact of innervation on PCa cells is still poorly understood. By focusing on muscarinic receptors, which are activated by the nerve-derived neurotransmitter acetylcholine, we show that muscarinic receptors 1 and 3 (m1 and m3) are highly expressed in PCa clinical specimens compared with all other cancer types, and that amplification or gain of their corresponding encoding genes (CHRM1 and CHRM3, respectively) represent a worse prognostic factor for PCa progression free survival. Moreover, m1 and m3 gene gain or amplification is frequent in castration-resistant PCa (CRPC) compared with hormone-sensitive PCa (HSPC) specimens. This was reflected in HSPC-derived cells, which show aberrantly high expression of m1 and m3 under androgen deprivation mimicking castration and androgen receptor inhibition. We also show that pharmacological activation of m1 and m3 signaling is sufficient to induce the castration-resistant growth of PCa cells. Mechanistically, we found that m1 and m3 stimulation induces YAP activation through FAK, whose encoding gene, PTK2 is frequently amplified in CRPC cases. Pharmacological inhibition of FAK and knockdown of YAP abolished m1 and m3-induced castration-resistant growth of PCa cells. Our findings provide novel therapeutic opportunities for muscarinic-signal-driven CRPC progression by targeting the FAK-YAP signaling axis.

Liposarcoma (LS) is the most common soft-tissue sarcoma. Dedifferentiated liposarcoma (DDLS) shows more aggressive biological behavior than that of well-differentiated liposarcoma (WDLS), so advanced therapeutic agents based on molecular mechanism are urgently needed. Here we show that tissue inhibitors of metalloproteinases (TIMPs) from TIMP-1 to TIMP-4 are differently expressed and regulate yes-associated protein (YAP)/transcriptional co-activator with PDZ binding motif (TAZ) in LS. Database analysis showed high TIMP-1 expression in DDLS patients correlating with poor prognosis, but high TIMP-4 expression in WDLS patients with better prognosis. Stable TIMP-1 knockdown inactivated YAP/TAZ and inhibited proliferation, colony formation and migration in DDLS cells, which was rescued by a constitutive active YAP. However, stable overexpression of TIMP-1 showed the opposite in WDLS cells. Stable TIMP-4 knockdown activated YAP/TAZ and promoted proliferation and migration in WDLS cells, which was suppressed by YAP/TAZ inhibitor (verteporfin) or knockdown of YAP/TAZ. Recombinant TIMP-4 showed opposite results in DDLS cells. These results indicate that dedifferentiation in LS shifts the expression of TIMPs from type 4 to type 1, inducing more aggressive behavior and poor prognosis through YAP/TAZ activation, which can be prognostic markers and therapeutic targets for LS patients.

Liposarcoma (LS) is the most common soft-tissue sarcoma. Dedifferentiated liposarcoma (DDLS) shows more aggressive biological behavior than that of well-differentiated liposarcoma (WDLS), so advanced therapeutic agents based on molecular mechanism are urgently needed. Here we show that tissue inhibitors of metalloproteinases (TIMPs) from TIMP-1 to TIMP-4 are differently expressed and regulate yes-associated protein (YAP)/transcriptional co-activator with PDZ binding motif (TAZ) in LS. Database analysis showed high TIMP-1 expression in DDLS patients correlating with poor prognosis, but high TIMP-4 expression in WDLS patients with better prognosis. Stable TIMP-1 knockdown inactivated YAP/TAZ and inhibited proliferation, colony formation and migration in DDLS cells, which was rescued by a constitutive active YAP. However, stable overexpression of TIMP-1 showed the opposite in WDLS cells. Stable TIMP-4 knockdown activated YAP/TAZ and promoted proliferation and migration in WDLS cells, which was suppressed by YAP/TAZ inhibitor (verteporfin) or knockdown of YAP/TAZ. Recombinant TIMP-4 showed opposite results in DDLS cells. These results indicate that dedifferentiation in LS shifts the expression of TIMPs from type 4 to type 1, inducing more aggressive behavior and poor prognosis through YAP/TAZ activation, which can be prognostic markers and therapeutic targets for LS patients.

Metformin may reduce the progression of head and neck squamous cell carcinoma (HNSCC); however, whether metformin acts by altering the host metabolism or targets cancer-initiating cells remains poorly understood. This gap in knowledge has prevented the stratification of patient populations who are most likely to benefit from metformin treatment. Here, we explored whether metformin acts directly on HNSCC cells to inhibit aberrant cell growth. To investigate the tumor cell autonomous effects of metformin, we engineered representative HPV- and HPV+ HNSCC cells harboring typical genetic alternations to express the yeast mitochondrial NADH dehydrogenase (NDI1) protein, which is insensitive to metformin. NDI1 expression rescued the inhibitory effects of metformin on mitochondrial complex I, abolished the ability of metformin to activate AMP-activated protein kinase, and inhibited mTOR signaling both in vitro and in vivo, and was sufficient to render metformin ineffective to prevent HNSCC tumor growth. This experimental system provided an opportunity to identify metformin-regulated transcriptional programs linked to cancer cell growth inhibition in the tumor microenvironment. Remarkably, computational analysis of the metformin-induced transcriptome revealed that metformin downregulated gene expression signatures associated with cancer stemness and epithelial-mesenchymal transition, concomitant with increased expression of squamous differentiation genes. These findings support that metformin may act directly on cancer-initiating cells to prevent their progression to HNSCC, which may inform the selection of patients at risk of developing HNSCC in future early-stage clinical trials. SIGNIFICANCE

Metformin may reduce the progression of head and neck squamous cell carcinoma (HNSCC); however, whether metformin acts by altering the host metabolism or targets cancer-initiating cells remains poorly understood. This gap in knowledge has prevented the stratification of patient populations who are most likely to benefit from metformin treatment. Here, we explored whether metformin acts directly on HNSCC cells to inhibit aberrant cell growth. To investigate the tumor cell autonomous effects of metformin, we engineered representative HPV- and HPV+ HNSCC cells harboring typical genetic alternations to express the yeast mitochondrial NADH dehydrogenase (NDI1) protein, which is insensitive to metformin. NDI1 expression rescued the inhibitory effects of metformin on mitochondrial complex I, abolished the ability of metformin to activate AMP-activated protein kinase, and inhibited mTOR signaling both in vitro and in vivo, and was sufficient to render metformin ineffective to prevent HNSCC tumor growth. This experimental system provided an opportunity to identify metformin-regulated transcriptional programs linked to cancer cell growth inhibition in the tumor microenvironment. Remarkably, computational analysis of the metformin-induced transcriptome revealed that metformin downregulated gene expression signatures associated with cancer stemness and epithelial-mesenchymal transition, concomitant with increased expression of squamous differentiation genes. These findings support that metformin may act directly on cancer-initiating cells to prevent their progression to HNSCC, which may inform the selection of patients at risk of developing HNSCC in future early-stage clinical trials. SIGNIFICANCE: Metformin's ability to directly target HNSCC-initiating cells instead of exerting cancer preventive activity based solely on its systemic effects may inform the selection of patients in future precision prevention trials.

Dental infection is risk for preterm birth (PTB) through unclear mechanisms. We established a dental infection-induced PTB mouse model, in which Porphyromonas gingivalis (P.g.) induced PTB by 2 days. We analysed pathogenic factors contributing to PTB and their effects on trophoblasts in vitro. TNF-α, IL-8, and COX-2 were upregulated in P.g.-infected placenta. Galectin-3 (Gal-3), an immune regulator, was significantly upregulated in placenta, amniotic fluid, and serum. In vitro, P.g.-lipopolysaccharide (P.g.-LPS) increased TNF-α and Gal-3 in trophoblasts via NF-κB/MAPK signalling. Gal-3 inhibition significantly downregulated P.g.-LPS-induced TNF-α production. TNF-α upregulated Gal-3. Gal-3 also increased cytokines and Gal-3 through NF-κB/MAPK signalling. Moreover, Gal-3 suppressed CD-66a expression at the maternal-foetal interface. Co-stimulation with Gal-3 and P.g.-LPS upregulated cytokine levels, while Gal-3 plus Aggregatibacter actinomycetemcomitans (A.a.)-or Escherichia coli (E. coli)-LPS treatment downregulated them, indicating the critical role of Gal-3 especially in P.g. dental infection-induced PTB. P.g.-dental infection induced PTB, which was associated with Gal-3-dependent cytokine production. New therapies and/or diagnostic systems targeting Gal-3 may reduce PTB.

Dental infection is risk for preterm birth (PTB) through unclear mechanisms. We established a dental infection-induced PTB mouse model, in which Porphyromonas gingivalis (P.g.) induced PTB by 2 days. We analysed pathogenic factors contributing to PTB and their effects on trophoblasts in vitro. TNF-α, IL-8, and COX-2 were upregulated in P.g.-infected placenta. Galectin-3 (Gal-3), an immune regulator, was significantly upregulated in placenta, amniotic fluid, and serum. In vitro, P.g.-lipopolysaccharide (P.g.-LPS) increased TNF-α and Gal-3 in trophoblasts via NF-κB/MAPK signalling. Gal-3 inhibition significantly downregulated P.g.-LPS-induced TNF-α production. TNF-α upregulated Gal-3. Gal-3 also increased cytokines and Gal-3 through NF-κB/MAPK signalling. Moreover, Gal-3 suppressed CD-66a expression at the maternal-foetal interface. Co-stimulation with Gal-3 and P.g.-LPS upregulated cytokine levels, while Gal-3 plus Aggregatibacter actinomycetemcomitans (A.a.)-or Escherichia coli (E. coli)-LPS treatment downregulated them, indicating the critical role of Gal-3 especially in P.g. dental infection-induced PTB. P.g.-dental infection induced PTB, which was associated with Gal-3-dependent cytokine production. New therapies and/or diagnostic systems targeting Gal-3 may reduce PTB.

Cleft lip and palate is the most common congenital anomaly in the orofacial region. Autogenous iliac bone graft, in general, has been employed for closing the bone defect at the alveolar cleft. However, such iliac bone graft provides patients with substantial surgical and psychological invasions. Consequently, development of a less invasive method has been highly anticipated. Stem cells from human exfoliated deciduous teeth (SHED) are a major candidate for playing a significant role in tissue engineering and regenerative medicine. The aim of this study was to elucidate the nature of bone regeneration by SHED as compared to that of human dental pulp stem cells (hDPSCs) and bone marrow mesenchymal stem cells (hBMSCs). The stems cells derived from pulp tissues and bone marrow were transplanted with a polylactic-coglycolic acid barrier membrane as a scaffold, for use in bone regeneration in an artificial bone defect of 4 mm in diameter in the calvaria of immunodeficient mice. Three-dimensional analysis using micro CT and histological evaluation were performed. Degree of bone regeneration with SHED relative to the bone defect was almost equivalent to that with hDPSCs and hBMSCs 12 weeks after transplantation. The ratio of new bone formation relative to the pre-created bone defect was not significantly different among groups with SHED, hDPSCs and hBMSCs. In addition, as a result of histological evaluation, SHED produced the largest osteoid and widely distributed collagen fibers compared to hDPSCs and hBMSCs groups. Thus, SHED transplantation exerted bone regeneration ability sufficient for the repair of bone defect. The present study has demonstrated that SHED is one of the best candidate as a cell source for the reconstruction of alveolar cleft due to the bone regeneration ability with less surgical invasion.

Cleft lip and palate is the most common congenital anomaly in the orofacial region. Autogenous iliac bone graft, in general, has been employed for closing the bone defect at the alveolar cleft. However, such iliac bone graft provides patients with substantial surgical and psychological invasions. Consequently, development of a less invasive method has been highly anticipated. Stem cells from human exfoliated deciduous teeth (SHED) are a major candidate for playing a significant role in tissue engineering and regenerative medicine. The aim of this study was to elucidate the nature of bone regeneration by SHED as compared to that of human dental pulp stem cells (hDPSCs) and bone marrow mesenchymal stem cells (hBMSCs). The stems cells derived from pulp tissues and bone marrow were transplanted with a polylactic-coglycolic acid barrier membrane as a scaffold, for use in bone regeneration in an artificial bone defect of 4 mm in diameter in the calvaria of immunodeficient mice. Three-dimensional analysis using micro CT and histological evaluation were performed. Degree of bone regeneration with SHED relative to the bone defect was almost equivalent to that with hDPSCs and hBMSCs 12 weeks after transplantation. The ratio of new bone formation relative to the pre-created bone defect was not significantly different among groups with SHED, hDPSCs and hBMSCs. In addition, as a result of histological evaluation, SHED produced the largest osteoid and widely distributed collagen fibers compared to hDPSCs and hBMSCs groups. Thus, SHED transplantation exerted bone regeneration ability sufficient for the repair of bone defect. The present study has demonstrated that SHED is one of the best candidate as a cell source for the reconstruction of alveolar cleft due to the bone regeneration ability with less surgical invasion.

Tissue inhibitor of metalloproteinase-1 (TIMP-1), a member of the TIMP family (TIMP-1 to 4), is highly expressed in various types of cancer and forms a complex with its receptor CD63 and Integrin β1. However, the precise oncogenic mechanism of TIMP-1 remains unclear. Yes-associated protein (YAP) and transcriptional co-activator with PDZ binding motif (TAZ) are transcription co-activators enhancing the transcription of specific genes related to cell proliferation. But the mechanism of aberrant YAP/TAZ activation in cancer is not fully understood. Here, we showed that TIMP-1 activates YAP/TAZ as novel downstream targets to promote cell proliferation. The TIMP-1-CD63-Integrin β1 axis activates Src and promotes RhoA-mediated F-actin assembly, leading to LATS1/2 inactivation. This results in under-phosphorylation, protein stabilization and nuclear translocation of YAP/TAZ (YAP/TAZ activation) CTGF production and cell proliferation. Furthermore, the TIMP-1-YAP/TAZ axis is aberrantly activated in various types of cancer cells or tissues. TIMP-1 knockdown inhibits cell proliferation through YAP/TAZ inactivation in cancer cells. This study found that TIMP-1 accelerates cell proliferation through YAP/TAZ activation in cancer, and suggests the TIMP-1-YAP/TAZ axis may be a novel potential drug target for cancer patients.

Tissue inhibitor of metalloproteinase-1 (TIMP-1), a member of the TIMP family (TIMP-1 to 4), is highly expressed in various types of cancer and forms a complex with its receptor CD63 and Integrin β1. However, the precise oncogenic mechanism of TIMP-1 remains unclear. Yes-associated protein (YAP) and transcriptional co-activator with PDZ binding motif (TAZ) are transcription co-activators enhancing the transcription of specific genes related to cell proliferation. But the mechanism of aberrant YAP/TAZ activation in cancer is not fully understood. Here, we showed that TIMP-1 activates YAP/TAZ as novel downstream targets to promote cell proliferation. The TIMP-1-CD63-Integrin β1 axis activates Src and promotes RhoA-mediated F-actin assembly, leading to LATS1/2 inactivation. This results in under-phosphorylation, protein stabilization and nuclear translocation of YAP/TAZ (YAP/TAZ activation) CTGF production and cell proliferation. Furthermore, the TIMP-1-YAP/TAZ axis is aberrantly activated in various types of cancer cells or tissues. TIMP-1 knockdown inhibits cell proliferation through YAP/TAZ inactivation in cancer cells. This study found that TIMP-1 accelerates cell proliferation through YAP/TAZ activation in cancer, and suggests the TIMP-1-YAP/TAZ axis may be a novel potential drug target for cancer patients.

Central mucoepidermoid carcinoma (MEC) poses a diagnostic challenge because of its rarity and histological overlap with glandular odontogenic cyst (GOC). In MEC of both salivary glands and jaws, MAML2 arrangement has been well known as the specific gene alteration. We report a case of central MEC arising from GOC diagnosed by MAML2 fusion gene. A 57-year-old male presented a multilocular cystic lesion in left molar region of the mandible. Histopathologically, multiple cysts lined by thin cuboidal or non-keratinized squamous epithelium with small duct-like structures, mucous cells and ciliated cells were present. It was diagnosed as GOC. The recurrent lesion after nine years showed the proliferation of many cystic and solid nests composed of epidermoid, mucous and intermediated cells. Nested PCR revealed CRTC3-MAML2 fusion gene in the recurrent lesion, but not in the primary one. Similarly, MAML-2 rearrangement by FISH analysis was positive in the recurrent lesion, while negative for the primary one, thus confirming the diagnosis of central MEC arising from GOC. Analysis of MAML2 rearrangement can be used as a supportive evidence to distinguish central MEC from GOC.

Central mucoepidermoid carcinoma (MEC) poses a diagnostic challenge because of its rarity and histological overlap with glandular odontogenic cyst (GOC). In MEC of both salivary glands and jaws, MAML2 arrangement has been well known as the specific gene alteration. We report a case of central MEC arising from GOC diagnosed by MAML2 fusion gene. A 57-year-old male presented a multilocular cystic lesion in left molar region of the mandible. Histopathologically, multiple cysts lined by thin cuboidal or non-keratinized squamous epithelium with small duct-like structures, mucous cells and ciliated cells were present. It was diagnosed as GOC. The recurrent lesion after nine years showed the proliferation of many cystic and solid nests composed of epidermoid, mucous and intermediated cells. Nested PCR revealed CRTC3-MAML2 fusion gene in the recurrent lesion, but not in the primary one. Similarly, MAML-2 rearrangement by FISH analysis was positive in the recurrent lesion, while negative for the primary one, thus confirming the diagnosis of central MEC arising from GOC. Analysis of MAML2 rearrangement can be used as a supportive evidence to distinguish central MEC from GOC.

Primordial odontogenic tumor (POT) is a rare lesion in the jaw which has been included as a new entity of benign mixed epithelial and mesenchymal odontogenic tumour in the latest World Health Organization (WHO) classification (2017). Only seven cases have been reported. It typically occurs in the posterior mandible. We report an additional case of POT in the maxilla of an 8-year-old girl presenting with an asymptomatic buccal enlargement. A well-defined, unilocular, radiolucent lesion was observed radiographically. Histologically, the tumor was mostly composed of loose fibrous connective tissue resembling dental papilla and a single layer of columnar epithelium covering the periphery of the tumor. In part, cords or nests of epithelium were present in the mesenchyme close to the periphery. Nestin, a marker of odontogenic ectomesenchyme, was positive in the mesenchymal tumor cells. We finally diagnosed the lesion as POT considering the possibility of other odontogenic tumors like ameloblastic fibroma or developing odontoma as a differential diagnosis. The patient shows no recurrence after 16 months. This case is the first report from Japan using this novel diagnosis POT after it was recognized and defined in the latest WHO classification.

Primordial odontogenic tumor (POT) is a rare lesion in the jaw which has been included as a new entity of benign mixed epithelial and mesenchymal odontogenic tumour in the latest World Health Organization (WHO) classification (2017). Only seven cases have been reported. It typically occurs in the posterior mandible. We report an additional case of POT in the maxilla of an 8-year-old girl presenting with an asymptomatic buccal enlargement. A well-defined, unilocular, radiolucent lesion was observed radiographically. Histologically, the tumor was mostly composed of loose fibrous connective tissue resembling dental papilla and a single layer of columnar epithelium covering the periphery of the tumor. In part, cords or nests of epithelium were present in the mesenchyme close to the periphery. Nestin, a marker of odontogenic ectomesenchyme, was positive in the mesenchymal tumor cells. We finally diagnosed the lesion as POT considering the possibility of other odontogenic tumors like ameloblastic fibroma or developing odontoma as a differential diagnosis. The patient shows no recurrence after 16 months. This case is the first report from Japan using this novel diagnosis POT after it was recognized and defined in the latest WHO classification.

症例は17歳男性で、左側下顎臼歯部から骨体部の透過病変を指摘された。デンタル、パノラマX線およびCT画像で、左側下顎小臼歯から下顎骨体部におよぶ広範で境界明瞭なX線透過像を認めた。臨床および画像所見からエナメル上皮腫と臨床診断し、生検を行った。病理組織診断にて充実型/多嚢胞型エナメル上皮腫の診断を得た。全身麻酔下に摘出開窓術を施行した。骨形成は良好であったが処置後1年8ヵ月で進学のため転院となり、外来受診は中断された。その後、左側頬部のび漫性腫脹を自覚するも放置した。38歳時、左側頬部および口腔内の腫瘤形成を認め、既往歴よりエナメル上皮腫の再発を疑った。パノラマX線にて、左側下顎犬歯根当部から下顎頭に及ぶ広範な骨吸収像を認めた。エナメル上皮腫再発と臨床診断し、生検にてエナメル上皮腫の確定診断後、腫瘍切除術、左側下顎半側切除及び腓骨遊離皮弁再建術を行った。手術1年後、再発所見を認めないことを確認し、咀嚼機能改善のため顎骨支持型補綴装置作製目的に二次手術を施行した。現在術後約5年経過し、腫瘍再発は認めない。左側下顎エナメル上皮腫(再発)と診断した。

症例は17歳男性で、左側下顎臼歯部から骨体部の透過病変を指摘された。デンタル、パノラマX線およびCT画像で、左側下顎小臼歯から下顎骨体部におよぶ広範で境界明瞭なX線透過像を認めた。臨床および画像所見からエナメル上皮腫と臨床診断し、生検を行った。病理組織診断にて充実型/多嚢胞型エナメル上皮腫の診断を得た。全身麻酔下に摘出開窓術を施行した。骨形成は良好であったが処置後1年8ヵ月で進学のため転院となり、外来受診は中断された。その後、左側頬部のび漫性腫脹を自覚するも放置した。38歳時、左側頬部および口腔内の腫瘤形成を認め、既往歴よりエナメル上皮腫の再発を疑った。パノラマX線にて、左側下顎犬歯根当部から下顎頭に及ぶ広範な骨吸収像を認めた。エナメル上皮腫再発と臨床診断し、生検にてエナメル上皮腫の確定診断後、腫瘍切除術、左側下顎半側切除及び腓骨遊離皮弁再建術を行った。手術1年後、再発所見を認めないことを確認し、咀嚼機能改善のため顎骨支持型補綴装置作製目的に二次手術を施行した。現在術後約5年経過し、腫瘍再発は認めない。左側下顎エナメル上皮腫(再発)と診断した。

Ameloblastin (AMBN), the most abundant non-amelogenin enamel matrix protein, plays a role in ameloblast differentiation. Previously, we found that AMBN promoted osteogenic differentiation via the interaction between CD63 and integrin β1, leading to the inactivation of Src; however, how AMBN affects the malignant behavior of osteosarcoma is still unclear. Osteosarcoma affects the bone and is associated with poor prognosis because of the high rate of pulmonary metastases and drug resistance. Here we demonstrated that stable overexpression of AMBN induced apoptosis and suppressed colony formation and cell migration via the inactivation of Src-Stat3 pathway in human osteosarcoma cells. Moreover, AMBN induced chemosensitivity to doxorubicin. Thus, AMBN induced a tumor suppressive phenotype and chemosensitivity to doxorubicin via the AMBN-Src-Stat3 axis in osteosarcoma. Indeed, immunohistochemical expression of AMBN was significantly correlated with better outcome of osteosarcoma patients. Our findings suggest that AMBN can be a new prognostic marker and therapeutic target for osteosarcoma combined with conventional doxorubicin treatment.

Ameloblastin (AMBN), the most abundant non-amelogenin enamel matrix protein, plays a role in ameloblast differentiation. Previously, we found that AMBN promoted osteogenic differentiation via the interaction between CD63 and integrin β1, leading to the inactivation of Src; however, how AMBN affects the malignant behavior of osteosarcoma is still unclear. Osteosarcoma affects the bone and is associated with poor prognosis because of the high rate of pulmonary metastases and drug resistance. Here we demonstrated that stable overexpression of AMBN induced apoptosis and suppressed colony formation and cell migration via the inactivation of Src-Stat3 pathway in human osteosarcoma cells. Moreover, AMBN induced chemosensitivity to doxorubicin. Thus, AMBN induced a tumor suppressive phenotype and chemosensitivity to doxorubicin via the AMBN-Src-Stat3 axis in osteosarcoma. Indeed, immunohistochemical expression of AMBN was significantly correlated with better outcome of osteosarcoma patients. Our findings suggest that AMBN can be a new prognostic marker and therapeutic target for osteosarcoma combined with conventional doxorubicin treatment.

The aim of this study was to examine the effect of 16 amino acids of the N-terminal region of human ameloblastin (16N-AMBN) synthetic peptide, on the proliferation and differentiation of MC3T3-E1 cells and bone regeneration. While 16N-AMBN did not affect the proliferation, it induced mRNA expression of type I collagen, alkaline phosphatase (ALP), bone sialoprotein, and osteocalcin. 16N-AMBN also stimulated ALP activity and promoted mineralized nodule formation. On the other hand, these activities were inhibited by anti-16N-AMBN antibody. Treatment of rat calvarial bone defects with 16N-AMBN resulted in almost complete healing compared to that of the control treatments. These findings suggest that 16N-AMBN may be applicable for regeneration therapy of bone defects.

The aim of this study was to examine the effect of 16 amino acids of the N-terminal region of human ameloblastin (16N-AMBN) synthetic peptide, on the proliferation and differentiation of MC3T3-E1 cells and bone regeneration. While 16N-AMBN did not affect the proliferation, it induced mRNA expression of type I collagen, alkaline phosphatase (ALP), bone sialoprotein, and osteocalcin. 16N-AMBN also stimulated ALP activity and promoted mineralized nodule formation. On the other hand, these activities were inhibited by anti-16N-AMBN antibody. Treatment of rat calvarial bone defects with 16N-AMBN resulted in almost complete healing compared to that of the control treatments. These findings suggest that 16N-AMBN may be applicable for regeneration therapy of bone defects.

症例は7歳女児で、左側の頸部腫脹を主訴とした。パノラマX線で両側下顎臼歯部から下顎枝に及ぶ多房性X線透過病変を認め、左下7歯胚の欠損を認めた。CTでは皮質骨の菲薄化と内部に隔壁形成を認めた。血清アルカリホスファターゼ以外に異常所見は認めなかった。両側下顎骨良性病変(ケルビズム疑い)の臨床診断のもと、生検を行った。細胞異型など悪性細胞は認めず、全身麻酔下に両側下顎骨病変の摘出掻爬術を行った。病理組織学的診断は中心性巨細胞病変であった。初回の外科手術から1年6ヵ月で左側下顎病変に再発を認め、摘出掻爬術を再度施行した。

Background

Background: A number of studies have revealed a link between chronic periodontitis and cardiovascular disease in obese patients. However, there is little information about the influence of periodontitis-associated bacteria, Porphyromonas gingivalis (Pg), on pathogenesis of atherosclerosis in obesity.Methods: In vivo experiment: C57BL/6J mice were fed with a high-fat diet (HFD) or normal chow diet (CD), as a control. Pg was infected from the pulp chamber. At 6 weeks post-infection, histological and immunohistochemical analysis of aortal tissues was performed. In vitro experiment: hTERT-immortalized human umbilical vein endothelial cells (HuhT1) were used to assess the effect of Pg/Pg-LPS on free fatty acid (FFA) induced endothelial cells apoptosis and regulation of cytokine gene expression.Results: Weaker staining of CD31 and increased numbers of TUNEL positive cells in aortal tissue of HFD mice indicated endothelial injury. Pg infection exacerbated the endothelial injury. Immunohistochemically, Pg was detected deep in the smooth muscle of the aorta, and the number of Pg cells in the aortal wall was higher in HFD mice than in CD mice. Moreover, in vitro, FFA treatment induced apoptosis in HuhT1 cells and exposure to Pg-LPS increased this effect. In addition, Pg and Pg-LPS both attenuated cytokine production in HuhT1 cells stimulated by palmitate.Conclusions: Dental infection of Pg may contribute to pathogenesis of atherosclerosis by accelerating FFA-induced endothelial injury.

症例は41歳女性で、8年前に前医にて左側下顎臼歯部のX線透過病変を指摘されていた。今回、同部違和感で前医を受診し、パノラマX線で同部に不透過像を含む透過像、右側下顎智歯部に歯牙様不透過像を認め紹介受診した。前医による8年間のパノラマX線では、初診時に右側下顎智歯部に類円形、歯冠大の境界不明瞭で中央に不透過像を含んだ病変を認め、中央の不透過像は年々増大して歯根様を呈し、左側下顎臼歯部に歯冠大で類円形の透過性病変を認め、徐々に中央部に不透過像の増大が認められた。当院のデンタルX線・CBCTでは、右側下顎智歯部の病変周囲に一層の透過体を有する歯根様硬組織塊を認め、左側下顎臼歯部の病変の中央に不透過像が混在し、第一大臼歯近心根の歯槽硬線と連続した像を認め、病変下端は下顎管に近接していた。両側下顎骨性異形成症と診断し、局所麻酔下に両側下顎部病変を摘出した。病理診断は限局性骨性異形成(右側

症例は41歳女性で、8年前に前医にて左側下顎臼歯部のX線透過病変を指摘されていた。今回、同部違和感で前医を受診し、パノラマX線で同部に不透過像を含む透過像、右側下顎智歯部に歯牙様不透過像を認め紹介受診した。前医による8年間のパノラマX線では、初診時に右側下顎智歯部に類円形、歯冠大の境界不明瞭で中央に不透過像を含んだ病変を認め、中央の不透過像は年々増大して歯根様を呈し、左側下顎臼歯部に歯冠大で類円形の透過性病変を認め、徐々に中央部に不透過像の増大が認められた。当院のデンタルX線・CBCTでは、右側下顎智歯部の病変周囲に一層の透過体を有する歯根様硬組織塊を認め、左側下顎臼歯部の病変の中央に不透過像が混在し、第一大臼歯近心根の歯槽硬線と連続した像を認め、病変下端は下顎管に近接していた。両側下顎骨性異形成症と診断し、局所麻酔下に両側下顎部病変を摘出した。病理診断は限局性骨性異形成(右側:成熟期、左側:中期)であった。

Background: Metastasis to regional lymph nodes via lymphatic vessels plays a key role in cancer progression. Tumor lymphangiogenesis is known to promote lymphatic metastasis, and vascular endothelial growth factor C (VEGF-C) is a critical activator of tumor lymphangiogenesis during the process of metastasis. We previously identified periostin as an invasion-and angiogenesis-promoting factor in head and neck squamous cell carcinoma (HNSCC). In this study, we discovered a novel role for periostin in tumor lymphangiogenesis. Methods and Findings: Periostin overexpression upregulated VEGF-C mRNA expression in HNSCC cells. By using conditioned media from periostin-overexpressing HNSCC cells, we examined tube formation of lymphatic endothelial cells. Conditioned media from periostin-overexpressing cells promoted tube formation. To know the correlation between periostin and VEGF-C, we compared Periostin expression with VEGF-C expression in 54 HNSCC cases by immunohistochemistry. Periostin expression was correlated well with VEGF-C expression in HNSCC cases. Moreover, correlation between periostin and VEGF-C secretion was observed in serum from HNSCC patients. Interestingly, periostin itself promoted tube formation of lymphatic endothelial cells independently of VEGF-C. Periostin-promoted lymphangiogenesis was mediated by Src and Akt activity. Indeed possible correlation between periostin and lymphatic status in periostin-overexpressing xenograft tumors and HNSCC cases was observed. Conclusions: Our findings suggest that periostin itself as well as periostin-induced upregulation of VEGF-C may promote lymphangiogenesis. We suggest that periostin may be a marker for prediction of malignant behaviors in HNSCC and a potential target for future therapeutic intervention to obstruct tumoral lymphatic invasion and lymphangiogenesis in HNSCC patients.

Background

Survivin belongs to the inhibitors of apoptosis (IAP) gene family and inhibits apoptosis. Besides its role as IAP, Survivin recently appears to function as a subunit of the chromosomal passenger complex (CPC) for regulating cell division with other CPC proteins including Aurora-B and INCENP. Nuclear Survivin is suspected to control cell division, whereas cytoplasmic Survivin is considered cytoprotective. Although there are several studies on Survivin expression and its function as inhibition of apoptosis, there is no study on Survivin function as a CPC and its correlation with other CPC proteins in head and neck squamous cell carcinoma (HNSCC). Here, therefore, we examined nuclear Survivin expression and its functional correlation with Aurora-B in HNSCC. High expression of Survivin was well correlated with Aurora-B expression in nuclear fraction of HNSCC cell lines and tissues. Moreover, nuclear Survivin expression was significantly correlated with Ki-67 and Aurora-B expression by immunohistochemistry. Notably, HNSCC cases with nuclear Survivin and Aurora-B expression exhibited marked malignant behaviors. Interestingly, both Survivin and Aurora-B knockdown inhibited cell growth and tumorsphere formation. Overall suggest that nuclear Survivin may be involved in tumor progression together with Aurora-B, and that Survivin and Aurora-B can be useful diagnostic markers and therapeutic targets. (C) 2010 Elsevier Ltd. All rights reserved.

Survivin belongs to the inhibitors of apoptosis (IAP) gene family and inhibits apoptosis. Besides its role as IAP, Survivin recently appears to function as a subunit of the chromosomal passenger complex (CPC) for regulating cell division with other CPC proteins including Aurora-B and INCENP. Nuclear Survivin is suspected to control cell division, whereas cytoplasmic Survivin is considered cytoprotective. Although there are several studies on Survivin expression and its function as inhibition of apoptosis, there is no study on Survivin function as a CPC and its correlation with other CPC proteins in head and neck squamous cell carcinoma (HNSCC). Here, therefore, we examined nuclear Survivin expression and its functional correlation with Aurora-B in HNSCC. High expression of Survivin was well correlated with Aurora-B expression in nuclear fraction of HNSCC cell lines and tissues. Moreover, nuclear Survivin expression was significantly correlated with Ki-67 and Aurora-B expression by immunohistochemistry. Notably, HNSCC cases with nuclear Survivin and Aurora-B expression exhibited marked malignant behaviors. Interestingly, both Survivin and Aurora-B knockdown inhibited cell growth and tumorsphere formation. Overall suggest that nuclear Survivin may be involved in tumor progression together with Aurora-B, and that Survivin and Aurora-B can be useful diagnostic markers and therapeutic targets. (C) 2010 Elsevier Ltd. All rights reserved.

日本臨床口腔病理学会奨励賞（実験病理部門） 受賞 / encouragement award

日本臨床口腔病理学会奨励賞（実験病理部門） 受賞

Poster Award

Poster Award

2013 IADR/Unilever Hatton Divisional Award

2013 IADR/Unilever Hatton Divisional Award

Encouragement Award

Encouragement Award