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1.
Hum Genome Var ; 11(1): 22, 2024 May 16.
Article in English | MEDLINE | ID: mdl-38755192

ABSTRACT

Neurofibromatosis type 1 (NF1) is an autosomal dominant nevus disease characterized by multiple manifestations, primarily café-au-lait macules and neurofibromas. Here, we present the case of an NF1 patient with 47,XYY mosaicism whose diagnosis was prompted by café-au-lait macules on the skin and mandibular neurofibromas. Targeted next-generation sequencing of the patient's blood sample revealed a novel frameshift mutation in NF1 (NM_000267.3:c.6832dupA:p.Thr2278Asnfs*8) that is considered a pathogenic variant.

2.
Cranio ; : 1-11, 2022 May 04.
Article in English | MEDLINE | ID: mdl-35506653

ABSTRACT

OBJECTIVE: To evaluate the relationship between the changes in condylar volume and maxillofacial skeletal morphology according to sex as well as the relationship between condylar volume reduction and skeletal relapse in patients who underwent orthognathic surgery. METHODS: Ninety-five patients were categorized into skeletal Class III, Class II, and facial asymmetry groups. Computed tomography scans taken preoperatively and at 1 year postoperatively were used for quantitative measurement. RESULTS: Postoperative condylar volume was reduced in both the Class II group and the deviated side of the asymmetry group. Both female and Class II deformity were significant predictors of postoperative reduction in the condylar volume. There was a significant correlation between skeletal relapse and postoperative change in condylar volume in the Class II group. CONCLUSION: Postoperative condylar resorption may be associated with preoperative maxillofacial skeletal morphology and sex and also with skeletal relapse in the Class II group.

3.
J Craniofac Surg ; 33(2): e135-e138, 2022.
Article in English | MEDLINE | ID: mdl-34456281

ABSTRACT

PURPOSE: To retrospectively evaluate skeletal stability after Le Fort I maxillary impaction surgery and mandibular autorotation without bilateral sagittal split osteotomy (BSSO) in high-angle class II patients. MATERIALS AND METHODS: Seven female high-angle class II patients who underwent maxillary impaction surgery and mandibular autorotation without bilateral sagittal split osteotomy were included in this study. Surgical changes and relapse were measured on lateral cephalograms taken preoperatively and at 1 month, 6 months and 1 year postoperatively. RESULTS: The horizontal movement of the maxilla at point A was 5.8 ±â€Š3.3 mm backward, and the upward movement at the posterior nasal spine was 3.3 ±â€Š1.4 mm. The mean horizontal change at point A during the 1-year follow-up period was 0.1 ±â€Š0.2 mm, and the vertical change at posterior nasal spine was 0.2 ±â€Š1.3 mm, which were not statistically significant. The horizontal surgical change at point B was 4.0 ±â€Š1.8 mm forward and the vertical surgical change at point B was 4.7 ±â€Š1.8 mm upward. Postoperative relapse was 10.9% and 13.7% in the horizontal and vertical directions, respectively. CONCLUSIONS: Le Fort I maxillary impaction surgery with mandibular autorotation may be 1 of the suitable procedures for high-angle class II patients.


Subject(s)
Malocclusion, Angle Class III , Malocclusion, Angle Class II , Tooth, Impacted , Cephalometry/methods , Female , Follow-Up Studies , Humans , Malocclusion, Angle Class II/diagnostic imaging , Malocclusion, Angle Class II/surgery , Malocclusion, Angle Class III/surgery , Mandible/diagnostic imaging , Mandible/surgery , Maxilla/diagnostic imaging , Maxilla/surgery , Osteotomy, Le Fort/methods , Recurrence , Retrospective Studies
4.
Article in English | MEDLINE | ID: mdl-34413003

ABSTRACT

OBJECTIVE: Adenoid cystic carcinoma (AdCC) is a rare, indolent salivary gland tumor that is reported to be driven by fusion genes. However, MYB/MYBL1-NFIB fusions have been detected in <60% of all AdCC cases and the oncogenic driver mutations in approximately 40% of AdCC remain unknown. Our aim was to identify novel gene fusions in AdCC. STUDY DESIGN: We investigated 20 AdCC cases using a targeted RNA sequencing panel to identify gene fusions and performed quantitative real-time reverse transcription polymerase chain reaction to assess MYB, MYBL1, and NFIB expression levels. RESULTS: A total of 36 fusion transcripts in 15 cases were detected and validated by Sanger sequencing. The MYB-NFIB and MYBL1-NFIB fusion genes were detected in 9 and 3 cases, respectively, in a mutually exclusive manner. Furthermore, novel gene fusions, namely, NFIB-EPB41L2, MAP7-NFIB, NFIB-MCMDC2, MYBL1-C8orf34, C8orf34-NFIB, and NFIB-CASC20, were identified. Among them, NFIB-EPB41L2 and NFIB-MCMDC2 are thought to activate MYB and MYBL1 expression, respectively, through the insertion of a genomic segment in proximity to MYB and MYBL1 genes, respectively. CONCLUSION: Six novel gene fusions other than MYB/MYBL1-NFIB were identified. The detection of novel fusion genes and investigation of the molecular mechanism will contribute to the development of novel molecular targeted therapies for this disease.


Subject(s)
Carcinoma, Adenoid Cystic , Salivary Gland Neoplasms , Carcinoma, Adenoid Cystic/genetics , High-Throughput Nucleotide Sequencing , Humans , Oncogene Proteins, Fusion/genetics , Proto-Oncogene Proteins , Salivary Gland Neoplasms/genetics , Sequence Analysis, RNA , Trans-Activators/genetics
5.
Oncogene ; 40(31): 5013-5025, 2021 08.
Article in English | MEDLINE | ID: mdl-34183772

ABSTRACT

Accumulation of mutant p53 (mutp53) is crucial for its oncogenic gain of function activity. DNAJA1, a member of J-domain containing proteins or heat shock protein 40, is shown to prevent unfolded mutp53 from proteasomal degradation. However, the biological function of DNAJA1 remains largely unknown. Here we show that DNAJA1 promotes tumor metastasis by accumulating unfolded mutp53. Levels of DNAJA1 in head and neck squamous cell carcinoma (HNSCC) tissues were higher than those in normal tissues. Knockdown of DNAJA1 in HNSCC cell lines carrying unfolded mutp53 significantly decreased the levels of mutp53, filopodia/lamellipodia formation, migratory potential, and active forms of CDC42/RAC1, which were not observed in HNSCC cells with DNA contact mutp53, wild-type p53, or p53 null. Such mutp53-dependent functions of DNAJA1 were supported by the observation that DNAJA1 selectively bound to unfolded mutp53. Moreover, DNAJA1 knockdown in HNSCC cells carrying unfolded mutp53 inhibited primary tumor growth and metastases to the lymph nodes and lungs. Our study suggests that DNAJA1 promotes HNSCC metastasis mainly in a manner dependent on mutp53 status, suggesting DNAJA1 as a potential therapeutic target for HNSCC harboring unfolded mutp53.


Subject(s)
Biomarkers, Tumor , HSP40 Heat-Shock Proteins/genetics , HSP40 Heat-Shock Proteins/metabolism , Mutant Proteins/metabolism , Neoplasms/genetics , Neoplasms/metabolism , Tumor Suppressor Protein p53/metabolism , Animals , Disease Models, Animal , Disease Progression , Disease Susceptibility , Gene Expression , Gene Knockdown Techniques , Heterografts , Humans , Mice , Mutant Proteins/genetics , Neoplasm Metastasis , Neoplasm Staging , Neoplasms/pathology , Oncogenes/genetics , Tumor Suppressor Protein p53/genetics , Unfolded Protein Response/genetics , cdc42 GTP-Binding Protein/metabolism , rac1 GTP-Binding Protein/metabolism
6.
Adv Healthc Mater ; 10(9): e2001988, 2021 05.
Article in English | MEDLINE | ID: mdl-33694289

ABSTRACT

Systems for "protein transduction," intracellular delivery of functional proteins, are needed to address deliverability challenges of protein therapeutics. However, in vivo protein transduction remains challenging because of instability in serum, extracellular protease digestion and rapid excretion from the bloodstream. Here, a magnetically guided in vivo protein transduction using magnetic nanogel chaperone (MC) composed of iron oxide nanoparticles and a polysaccharide nanogel, a protein carrier inspired by "catch and release" mechanisms of molecular chaperones is demonstrated. The MC system enables efficient delivery of anti-cancer proteins, saporin and RNaseA, into cultured tumor lines and inhibits cell proliferation, mainly via apoptosis. Magnetic in vivo protein transduction via intravenous whole body administration is demonstrated in a fibrosarcoma model. By in vivo optical imaging, MC accumulated in tumor tissues under magnetic field three times more than without irradiation. With subcutaneous injection, saporin is delivered by MC to the cytoplasm in magnetically targeted tissues. In an oral cancer model, MC-delivered magnetically targeted saporin decreased tumor volume without significant body weight changes and no regrowth of tumor at 3 months after complete regression. Protein transduction with MC shows promise for cancer therapeutics and, potentially, for regenerative medicine and other biomedical applications.


Subject(s)
Ferric Compounds , Magnetics , Molecular Chaperones , Nanogels
7.
Int J Clin Exp Pathol ; 13(8): 2211-2217, 2020.
Article in English | MEDLINE | ID: mdl-32922622

ABSTRACT

Secretory carcinoma (SC) of the salivary gland was identified in 2010, and it is characterized by a specific ETV6 gene arrangement. The most common primary site for SC is the parotid gland; however, SC around the Stensen's duct is rare. Here we describe a rare case of a SC around the Stensen's duct that was initially misdiagnosed as a salivary duct cyst. A 59-year-old woman presented with a mass in the region of the left parotid papilla. Magnetic resonance imaging (MRI) revealed a well-circumscribed lesion and enhancement with a rim and an inner wall-like part that appeared in the late phase. Based on the initial clinical and imaging findings, a salivary duct cyst of the parotid gland was diagnosed. However, the lesion was histopathologically diagnosed as a SC based on immunohistochemical findings. The tumor cells showed diffuse positive staining for AE1/AE3, vimentin, and mammaglobin and focal positive staining for S-100 protein, SOX-10, and DOG-1. Fluorescence in-situ hybridization revealed ETV6 gene rearrangement in the tumor. In cases of cystic lesions around the Stensen's duct, clinicians should bear in mind that the possibility that they could be minor salivary gland cancers, such as SC.

8.
PLoS One ; 15(2): e0222689, 2020.
Article in English | MEDLINE | ID: mdl-32012175

ABSTRACT

Autoimmune regulator (AIRE) is a transcriptional regulator that is primarily expressed in medullary epithelial cells, where it induces tissue-specific antigen expression. Under pathological conditions, AIRE expression is induced in epidermal cells and promotes skin tumor development. This study aimed to clarify the role of AIRE in the pathogenesis of oral squamous cell carcinoma (OSCC). AIRE expression was evaluated in six OSCC cell lines and in OSCC tissue specimens. Expression of STAT1, ICAM1, CXCL10, CXCL11, and MMP9 was elevated in 293A cells stably expressing AIRE, and conversely, was decreased in AIRE-knockout HSC3 OSCC cells when compared to the respective controls. Upregulation of STAT1, and ICAM in OSCC cells was confirmed in tissue specimens by immunohistochemistry. We provide evidence that AIRE exerts transcriptional control in cooperation with ETS1. Expression of STAT1, ICAM1, CXCL10, CXCL11, and MMP9 was increased in 293A cells upon Ets1 transfection, and coexpression of AIRE further increased the expression of these proteins. AIRE coprecipitated with ETS1 in a modified immunoprecipitation assay using formaldehyde crosslinking. Chromatin immunoprecipitation and quantitative PCR analysis revealed that promoter fragments of STAT1, ICAM1, CXCL10, and MMP9 were enriched in the AIRE precipitates. These results indicate that AIRE is induced in OSCC and supports cancer-related gene expression in cooperation with ETS1. This is a novel function of AIRE in extrathymic tissues under the pathological condition.


Subject(s)
Carcinoma, Squamous Cell/genetics , Mouth Neoplasms/genetics , Transcription Factors/genetics , Transcriptional Activation , Autoimmunity , Cell Line, Tumor , Chromatin Immunoprecipitation , Gene Expression , HEK293 Cells , Humans , Promoter Regions, Genetic , Proto-Oncogene Protein c-ets-1/genetics , AIRE Protein
9.
J Oral Pathol Med ; 49(3): 235-242, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31762177

ABSTRACT

BACKGROUND: This study aimed to elucidate the correlation between gene amplification, protein expression of receptor tyrosine kinase, and prognosis of patients with oral squamous cell carcinoma (OSCC) using immunohistochemistry (IHC) and next-generation sequencing data. METHODS: We evaluated data pertaining to 208 patients with OSCC using IHC for epidermal growth factor receptor (EGFR) and mesenchymal-epithelial transition factor (MET). RESULTS: High expressions of EGFR and MET were detected in 60 and 41 patients, respectively. We evaluated the association of clinicopathological variables with high expressions of EGFR and/or MET. Distant metastasis was found in 9 of 41 patients (22.0%) and 6 of 15 patients (40.0%) with high expression of MET and high co-expressions of EGFR and MET, respectively; statistically significant differences were detected in both high expression of MET (P = .003) and high co-expressions of EGFR and MET (P = 3.41 × 10-5 ). The cumulative 5-year survival rate of patients with high and low expressions of EGFR or MET was approximately 65% and 85%, respectively. Conversely, among cases with high expressions of EGFR or MET, there was no additional decrease in the survival rate of patients harboring TP53 mutations. Moreover, the survival rate of patients with high co-expression of both EGFR and MET was very poor (22.0%) (P < 1.0 × 10-9 ). CONCLUSION: Our findings suggest that evaluation of protein expressions of EGFR and MET may facilitate prognostic assessment of patients with OSCC; in addition, patients with OSCC should be screened for enrollment in clinical trials of combination therapy with EGFR and MET inhibitors.


Subject(s)
Carcinoma, Squamous Cell/pathology , Mouth Neoplasms/pathology , Proto-Oncogene Proteins c-met/genetics , Carcinoma, Squamous Cell/diagnosis , Carcinoma, Squamous Cell/genetics , ErbB Receptors/genetics , Female , High-Throughput Nucleotide Sequencing , Humans , Immunohistochemistry , Male , Middle Aged , Mouth Neoplasms/diagnosis , Mouth Neoplasms/genetics , Prognosis , Survival Rate
10.
Exp Cell Res ; 372(2): 129-140, 2018 11 15.
Article in English | MEDLINE | ID: mdl-30266659

ABSTRACT

Notch signaling functions in diverse developmental and homeostatic processes, including stem cell self-renewal and cell fate determination. Notch1-inactivating mutations are frequently detected in skin and oro-esophageal cancers, suggesting a role for Notch1 as a tumor suppressor. Here, we clarify the contribution of Notch1 deficiency to oro-esophageal tumorigenesis using a physiological experimental model. Tongue and esophageal tumors induced in mice by 4-nitroquinoline-1-oxide (4-NQO) showed pathophysiological similarities to human tumors, including decreased Notch1 expression in the basal cells. We created mutant mice (N1cKO), in which the Notch1 gene was disrupted specifically in the squamous epithelium. The epithelium formed normally in N1cKO mice, and although multiple skin tumors were detected at 65 weeks, no tumors developed in the tongue and esophagus. However, 4-NQO-induced tumorigenesis assays revealed that tumor onset occurred earlier in N1cKO mice than in wild-type littermates, and the tumors arose preferentially from the Notch1-negative epithelium, indicating the tumor susceptibility of Notch1-deficient epithelium. Notch1 regulates the expression of TERT, and age-related telomere erosion was more rapid in Notch1-deficient basal cells. Our results indicated that although Notch1 deficiency had little effect on squamous epithelium formation, it predisposed the affected epithelium to tumor development, at least in part through accelerated telomere erosion.


Subject(s)
Carcinogenesis/genetics , Esophageal Neoplasms/genetics , Receptor, Notch1/genetics , Tongue Neoplasms/genetics , 4-Nitroquinoline-1-oxide/toxicity , Animals , Disease Models, Animal , Epithelium/metabolism , Epithelium/pathology , Esophageal Neoplasms/chemically induced , Esophageal Neoplasms/pathology , Esophagus/metabolism , Esophagus/pathology , Gene Expression Regulation, Neoplastic/genetics , Humans , Mice , Mice, Knockout , Telomerase/genetics , Telomere/genetics , Tongue Neoplasms/chemically induced , Tongue Neoplasms/pathology
11.
Pathol Int ; 68(2): 63-90, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29431262

ABSTRACT

Genome research using appropriately collected pathological tissue samples is expected to yield breakthroughs in the development of biomarkers and identification of therapeutic targets for diseases such as cancers. In this connection, the Japanese Society of Pathology (JSP) has developed "The JSP Guidelines on the Handling of Pathological Tissue Samples for Genomic Research" based on an abundance of data from empirical analyses of tissue samples collected and stored under various conditions. Tissue samples should be collected from appropriate sites within surgically resected specimens, without disturbing the features on which pathological diagnosis is based, while avoiding bleeding or necrotic foci. They should be collected as soon as possible after resection: at the latest within about 3 h of storage at 4°C. Preferably, snap-frozen samples should be stored in liquid nitrogen (about -180°C) until use. When intending to use genomic DNA extracted from formalin-fixed paraffin-embedded tissue, 10% neutral buffered formalin should be used. Insufficient fixation and overfixation must both be avoided. We hope that pathologists, clinicians, clinical laboratory technicians and biobank operators will come to master the handling of pathological tissue samples based on the standard operating procedures in these Guidelines to yield results that will assist in the realization of genomic medicine.


Subject(s)
DNA/analysis , Genomics , Guidelines as Topic , Neoplasms/pathology , Tissue Fixation/standards , Formaldehyde , Humans , Japan , Research/standards , Tissue Fixation/methods
12.
PLoS One ; 12(6): e0180224, 2017.
Article in English | MEDLINE | ID: mdl-28658279

ABSTRACT

Calcifying cystic odontogenic tumors (CCOTs) are benign cystic tumors that form abnormally keratinized ghost cells. Mutations in CTNNB1, which encodes beta-catenin, have been implicated in the development of these tumors, but a causal relationship has not been definitively established. Thus, mutational hot spots in 50 cancer genes were examined by targeted next-generation sequencing in 11 samples of CCOT. Mutations in CTNNB1, but not in other genes, were observed in 10 of 11 cases. These mutations constitutively activate beta-catenin signaling by abolishing the phosphorylation sites Asp32, Ser33, or Ser37, and are similar to those reported in pilomatrixoma and adamantinomatous craniopharyngioma. In contrast, BRAF or NRAS mutations were observed in 12 and two control samples of ameloblastoma, respectively. In HEK293 cells, overexpression of mutated CTNNB1 also upregulated hair keratin, a marker of ghost cells. Furthermore, ghost cells were present in two cases of ameloblastoma with BRAF and CTNNB1 mutations, indicating that ghost cells form due to mutations in CTNNB1. The data suggest that mutations in CTNNB1 are the major driver mutations of CCOT, and that CCOT is the genetic analog of pilomatrixoma and adamantinomatous craniopharyngioma in odontogenic tissue.


Subject(s)
Jaw Neoplasms/genetics , Odontogenic Cyst, Calcifying/genetics , Adolescent , Adult , Aged , Ameloblastoma/genetics , Blotting, Western , Child , DNA, Neoplasm/genetics , Female , HEK293 Cells , High-Throughput Nucleotide Sequencing , Humans , Jaw Neoplasms/pathology , Male , Middle Aged , Mutation, Missense/genetics , Odontogenic Cyst, Calcifying/pathology , Polymerase Chain Reaction , Proto-Oncogene Proteins B-raf/genetics , Young Adult , beta Catenin/genetics
13.
J Oral Pathol Med ; 46(3): 223-231, 2017 Mar.
Article in English | MEDLINE | ID: mdl-27529842

ABSTRACT

BACKGROUND: LAMC2 plays an important role in cancer invasion. The aim of this study was to (i) compare the immunoexpression of LAMC2 in different stages of oral squamous cell carcinoma (OSCC), early and advanced, and (ii) to evaluate LAMC2 as a marker of malignant transformation in leukoplakia. MATERIALS AND METHODS: The expression of LAMC2 was examined by immunohistochemistry in 50 surgical specimens of advanced OSCC assembled as tissue microarrays, and by cDNA microarray in 43 surgical specimens of advanced OSCC. LAMC2 expression was further examined in 39 surgical specimens of early OSCC and in 93 incisional biopsy specimens of leukoplakia of the tongue, which exhibited epithelial dysplasia. The relationship of LAMC2 expression score with clinico-pathological characteristics was analyzed. RESULTS: LAMC2 was remarkably upregulated in OSCC at the cancer-stroma interface. The grade of LAMC2 expression was significantly associated with the pattern and depth of invasion of OSCC. Foci of LAMC2-positive cells were observed in some cases of leukoplakia. The number and size of LAMC2-positive foci were significantly associated with the grade of dysplasia. The presence of LAMC2-positive foci was a significant predictive factor for the malignant progression of leukoplakia. LAMC2-positive leukoplakia had an approximately 11-fold increased risk of malignancy compared with LAMC2-negative leukoplakia. CONCLUSIONS: The results of this study highlight the value of LAMC2 as a marker of cancer invasion. LAMC2-positive foci in leukoplakia suggest an imminent risk of cancer. LAMC2 immunostaining is expected to contribute to a more precise assessment of the malignancy of leukoplakia.


Subject(s)
Biomarkers, Tumor/metabolism , Carcinoma, Squamous Cell/metabolism , Laminin/metabolism , Leukoplakia, Oral/metabolism , Aged , Biomarkers, Tumor/genetics , Biopsy , Carcinoma, Squamous Cell/pathology , Cell Transformation, Neoplastic/pathology , Disease Progression , Female , Humans , Immunohistochemistry , Laminin/genetics , Leukoplakia, Oral/pathology , Male , Middle Aged , Mouth Mucosa/pathology , Oligonucleotide Array Sequence Analysis , Proportional Hazards Models , RNA/metabolism , Tongue Neoplasms/metabolism , Tongue Neoplasms/pathology
14.
Cancer Sci ; 108(2): 256-266, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27889930

ABSTRACT

This study aimed to clarify the genomic factors associated with the diagnosis and prognosis of oral squamous cell carcinoma via next-generation sequencing. We evaluated data from 220 cases of oral squamous cell carcinoma. Genomic DNA was eluted using formalin-fixed, paraffin-embedded samples, and targeted resequencing of 50 cancer-related genes was performed. In total, 311 somatic mutations were detected in 220 patients, consisting of 68 synonymous mutations and 243 non-synonymous mutations. Genes carrying mutations included TP53, CDKN2A, and PIK3CA in 79 (35.9%), 35 (15.9%), and 19 patients (8.6%), respectively. Copy number analysis detected amplification of PIK3CA and AKT1 in 38 (17.3%) and 11 patients (5.0%), respectively. Amplification of receptor tyrosine kinases was found in 37 patients (16.8%). Distant metastasis was noted in nine of 37 patients (24%) with receptor tyrosine kinase amplification, accounting for 43% of the 21 cases of distant metastasis. The cumulative 5-year survival rate was 64.6% in the receptor tyrosine kinase amplification group vs 85.2% in the no receptor tyrosine kinase amplification group. Moreover, we identified significantly poorer prognosis in the TP53 mutation/receptor tyrosine kinase amplification group, for which the cumulative 5-year survival rate was 41.6%. In conclusion, the results of this study demonstrated that receptor tyrosine kinase amplification is a prognostic factor for distant metastasis of oral squamous cell carcinoma, indicating the necessity of using next-generation sequencing in clinical sequencing.


Subject(s)
Carcinoma, Squamous Cell/secondary , Gene Amplification , Genes, p16 , Genes, p53 , Mouth Neoplasms/pathology , Mutation , Phosphatidylinositol 3-Kinases/genetics , Adult , Aged , Carcinoma, Squamous Cell/mortality , Class I Phosphatidylinositol 3-Kinases , DNA Mutational Analysis , Female , Humans , Male , Middle Aged , Mouth Neoplasms/mortality , Prognosis , Proportional Hazards Models , Receptor Protein-Tyrosine Kinases/genetics , Survival Rate , Time Factors , Young Adult
15.
Hum Genome Var ; 3: 16003, 2016.
Article in English | MEDLINE | ID: mdl-27081569

ABSTRACT

Stickler syndrome (STL) is an autosomal, dominantly inherited, clinically variable and genetically heterogeneous connective tissue disorder characterized by ocular, auditory, orofacial and skeletal abnormalities. We conducted targeted resequencing using a next-generation sequencer for molecular diagnosis of a 2-year-old girl who was clinically suspected of having STL with Pierre Robin sequence. We detected a novel heterozygous missense mutation, NM_001854.3:n.4838G>A [NM_001854.3 (COL11A1_v001):c.4520G>A], in COL11A1, resulting in a Gly to Asp substitution at position 1507 [NM_001854.3(COL11A1_i001)] within one of the collagen-like domains of the triple helical region. The same mutation was detected in her 4-year-old brother with cleft palate and high-frequency sensorineural hearing loss.

16.
Monoclon Antib Immunodiagn Immunother ; 35(2): 109-16, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26937552

ABSTRACT

Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein, which possesses a platelet aggregation-stimulating (PLAG) domain in its N-terminus. Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for the binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. Although many anti-PDPN monoclonal antibodies (mAbs) have been established, almost all mAbs bind to PLAG domains. We recently established CasMab technology to produce mAbs against membranous proteins. Using CasMab technology, we produced a novel anti-PDPN mAb, LpMab-17, which binds to non-PLAG domains. LpMab-17 clearly detected endogenous PDPN of cancer cells and normal cells in Western-blot, flow cytometry, and immunohistochemistry. LpMab-17 recognized glycan-deficient PDPN in flow cytometry, indicating that the interaction between LpMab-17 and PDPN is independent of its glycosylation. The minimum epitope of LpMab-17 was identified as Gly77-Asp82 of PDPN using enzyme-linked immunosorbent assay. Of interest, LpMab-17 did not bind to monkey PDPN, whereas the homology is 94% between human PDPN and monkey PDPN, indicating that the epitope of LpMab-17 is unique compared with the other anti-PDPN mAbs. The combination of different epitope-possessing mAbs could be advantageous for the PDPN-targeting diagnosis or therapy.


Subject(s)
Antibodies, Monoclonal/immunology , Antibody Specificity/immunology , Membrane Glycoproteins/immunology , Protein Domains/immunology , Animals , Enzyme-Linked Immunosorbent Assay , Epitope Mapping , Epitopes/immunology , Flow Cytometry , HEK293 Cells , Haplorhini/immunology , Humans , Membrane Glycoproteins/isolation & purification , Mice
17.
J Oral Pathol Med ; 45(10): 730-739, 2016 Nov.
Article in English | MEDLINE | ID: mdl-26850833

ABSTRACT

BACKGROUND: THBS1 (thrombospondin-1) is the extracellular matrix (ECM) protein that affects diverse cellular activities. It constitutes the tumor stroma, but the role of THBS1 in oral squamous cell carcinoma (OSCC) development is unclear. The aim of this study was to clarify the relevance of THBS1 in the pathogenesis of OSCC. MATERIALS AND METHODS: The expression of THBS1 was examined in 44 OSCC by immunohistochemical analysis and in 43 OSCC by cDNA microarray analysis. Cell culture experiments were conducted using human OSCC cell lines HSC3 and HO1N1 and mouse fibroblast ST2 cells to examine the effect of TGFB1 on THBS1 expression, and the effect of THBS1 on cellular behaviors. RESULTS: THBS1 was specifically induced in the tumor microenvironment of OSCC. THBS1 appeared to be produced mainly by the stromal cells, but also by OSCC cells. TGFB1 stimulated THBS1 expression in ST2, primary fibroblasts, and the OSCC cells. THBS1 promoted migration and invasion of HSC3 and HO1N1 in transwell migration assays. THBS1 stimulated the expression of MMP3 (matrix metalloprotease 3), MMP9, MMP11, and MMP13 in ST2 cells and MMP3, MMP11, and MMP13 in HO1N1 cells. The RGD peptide suppressed the THBS1-stimulated migration and upregulation of MMP11 and MMP13. CONCLUSIONS: THBS1 is a tumor-specific ECM protein that is induced by TGFB1 and promotes migration of cancer cells and stimulates the expression of MMPs partly through the integrin signaling, thereby favoring OSCC invasion.


Subject(s)
Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/pathology , Head and Neck Neoplasms/metabolism , Head and Neck Neoplasms/pathology , Mouth Neoplasms/metabolism , Mouth Neoplasms/pathology , Thrombospondin 1/biosynthesis , Transforming Growth Factor beta1/pharmacology , Animals , Carcinoma, Squamous Cell/enzymology , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , DNA, Complementary/metabolism , Head and Neck Neoplasms/enzymology , Humans , Immunohistochemistry , Metalloendopeptidases/biosynthesis , Mice , Mouth Neoplasms/enzymology , Squamous Cell Carcinoma of Head and Neck , Stromal Cells/enzymology , Stromal Cells/metabolism , Stromal Cells/pathology , Up-Regulation
18.
PLoS One ; 10(11): e0140480, 2015.
Article in English | MEDLINE | ID: mdl-26544948

ABSTRACT

Gorlin syndrome (GS) is an autosomal dominant disorder that predisposes affected individuals to developmental defects and tumorigenesis, and caused mainly by heterozygous germline PTCH1 mutations. Despite exhaustive analysis, PTCH1 mutations are often unidentifiable in some patients; the failure to detect mutations is presumably because of mutations occurred in other causative genes or outside of analyzed regions of PTCH1, or copy number alterations (CNAs). In this study, we subjected a cohort of GS-affected individuals from six unrelated families to next-generation sequencing (NGS) analysis for the combined screening of causative alterations in Hedgehog signaling pathway-related genes. Specific single nucleotide variations (SNVs) of PTCH1 causing inferred amino acid changes were identified in four families (seven affected individuals), whereas CNAs within or around PTCH1 were found in two families in whom possible causative SNVs were not detected. Through a targeted resequencing of all coding exons, as well as simultaneous evaluation of copy number status using the alignment map files obtained via NGS, we found that GS phenotypes could be explained by PTCH1 mutations or deletions in all affected patients. Because it is advisable to evaluate CNAs of candidate causative genes in point mutation-negative cases, NGS methodology appears to be useful for improving molecular diagnosis through the simultaneous detection of both SNVs and CNAs in the targeted genes/regions.


Subject(s)
Basal Cell Nevus Syndrome/genetics , Biomarkers/analysis , DNA Copy Number Variations/genetics , High-Throughput Nucleotide Sequencing/methods , Polymorphism, Single Nucleotide/genetics , Adolescent , Adult , Aged , Child , Cohort Studies , Female , Humans , Male , Pedigree
19.
Oral Oncol ; 51(11): 1020-1025, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26410021

ABSTRACT

UNLABELLED: The standard treatments for oral leukoplakia range from careful observation to complete resection. No surgical intervention is chosen for several supposable reasons. Surgical treatment and no surgical treatment for oral leukoplakia have no defined basis for comparisons, and few studies have reported on the long-term outcomes of oral leukoplakia without surgery. OBJECTIVES: This study aimed to identify the important factors using a long-term wait-and-see policy in patients with oral leukoplakia. MATERIALS AND METHODS: In total, 237 lesions from 218 patients selected for non-surgical therapy between 2001 and 2010 were analyzed. On the basis of long-term follow-up data, lesions were classified as unchanged, reduced, disappeared, expanded, and malignantly transformed. RESULTS: In total, 135 (57.0%) lesions remained unchanged, 30 (12.7%) lesions were characterized by a reduction in size or clinical severity, and 44 (18.6%) lesions had disappeared. Another 17 (7.2%) lesions resulted in spread or clinical deterioration, and 11 (4.6%) lesions developed oral squamous cell carcinoma. CONCLUSIONS: We demonstrated a cumulative malignant transformation rate of 11.6% in 10years without resection. The lesions that were nonhomogeneous, and higher degree of epithelial dysplasia, located on the tongue were likely to progress into cancer. In addition, 32.5% of lesions without surgical treatment were reduced or disappeared. There is a possibility that removal of considerable irritation for a long time contributes to the treatment of this disease. The development of appropriate treatments for oral leukoplakia is required, which will enable successful differentiation between surgical and observation cases.


Subject(s)
Disease Progression , Leukoplakia, Oral/pathology , Watchful Waiting , Carcinoma, Squamous Cell/pathology , Female , Follow-Up Studies , Humans , Male , Mouth Neoplasms/pathology , Retrospective Studies , Risk Factors , Treatment Outcome
20.
Asian Pac J Cancer Prev ; 15(10): 4135-41, 2014.
Article in English | MEDLINE | ID: mdl-24935359

ABSTRACT

BACKGROUND: Human papillomaviruses (HPV) may play an important role as one of the possible etiologies of oral squamous cell carcinoma (OSCC). The present study aimed to investigate the association between HPV and OSCC in young Japanese patients by examining the presence of HPV DNA and surrogate markers in OSCC tissues. MATERIALS AND METHODS: Forty young patients with OSCC whose surgical specimens were available were analyzed and compared with 40 patients randomly recruited from a pool of patients aged >40 years. HPV DNA was detected using the polymerase chain reaction-based AMPLICOR(®) HPV test, and surrogate markers of HPV infection were analyzed using immunohistochemical techniques to detect p16(INK4a) and p53. RESULTS: Only two (5%) young patients and one (2.5%) older patient were positive for HPV DNA. p16(INK4a) overexpression was identified in six (15%) young patients. p53 staining levels were not high in tissues of most young patients (27 patients, 67.5%). HPV DNA status did not significantly correlate with p16(INK4a) expression levels. Profiles of increased levels of p16(INK4a) expression with diminished levels of p53 staining were not associated with the presence of HPV DNA. The combined p53 with p16(INK4a) profiles were significantly correlated with alcohol consumption in younger patients (p=0.006). CONCLUSIONS: RESULTS of the present study indicate that HPV is less likely to cause OSCC in young Japanese patients, and the p16(INK4a) expression level is not an appropriate surrogate marker for HPV infection in OSCC.


Subject(s)
Carcinoma, Squamous Cell/virology , Cyclin-Dependent Kinase Inhibitor p16/biosynthesis , DNA, Viral/blood , Mouth Neoplasms/virology , Papillomavirus Infections/etiology , Adult , Age Factors , Alcohol Drinking , Biomarkers, Tumor/blood , Cyclin-Dependent Kinase Inhibitor p16/genetics , Female , Human Papillomavirus DNA Tests , Humans , Japan , Male , Papillomaviridae
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