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Title: Novel allelic mutations in murine Serca2 induce differential development of squamous cell tumors

Abstract

Dominant mutations in the Serca2 gene, which encodes sarco(endo)plasmic reticulum calcium-ATPase, predispose mice to gastrointestinal epithelial carcinoma [1–4] and humans to Darier disease (DD) [14–17]. In this study, we generated mice harboring N-ethyl-N-nitrosourea (ENU)-induced allelic mutations in Serca2: three missense mutations and one nonsense mutation. Mice harboring these Serca2 mutations developed tumors that were categorized as either early onset squamous cell tumors (SCT), with development similar to null-type knockout mice [2,4] (aggressive form; M682, M814), or late onset tumors (mild form; M1049, M1162). Molecular analysis showed no aberration in Serca2 mRNA or protein expression levels in normal esophageal cells of any of the four mutant heterozygotes. There was no loss of heterozygosity at the Serca2 locus in the squamous cell carcinomas in any of the four lines. The effect of each mutation on Ca{sup 2+}-ATPase activity was predicted using atomic-structure models and accumulated mutated protein studies, suggesting that putative complete loss of Serca2 enzymatic activity may lead to early tumor onset, whereas mutations in which Serca2 retains residual enzymatic activity result in late onset. We propose that impaired Serca2 gene product activity has a long-term effect on squamous cell carcinogenesis from onset to the final carcinoma stage through an as-yetmore » unrecognized but common regulatory pathway. -- Highlights: •Novel mutations in murine Serca2 caused early onset or late onset of tumorigenesis. •They also caused higher or lower incidence of Darier Disease phenotype. •3D structure model suggested the former mutations led to severer defect on ATPase. •Driver gene mutations via long-range effect on Ca2+ distributions are suggested.« less

Authors:
; ; ; ; ;  [1];  [2];  [3];  [4];  [2];  [5];  [3];  [6];  [1];  [7]
  1. Team for Advanced Development and Evaluation of Human Disease Models, Riken BioResource Center (BRC), Tsukuba, Ibaraki (Japan)
  2. Technology and Development Team for Mouse Phenotype Analysis, Riken BRC, Tsukuba, Ibaraki (Japan)
  3. Mutagenesis and Genomics Team, Riken BRC, Tsukuba, Ibaraki (Japan)
  4. Department of Molecular Pathology, Tohoku University Graduate School of Medicine, Sendai, Miyagi (Japan)
  5. Department of Biochemistry, Asahikawa Medical University, Asahikawa, Hokkaido (Japan)
  6. Mammalian Genetics Laboratory, National Institute of Genetics, Mishima, Shizuoka (Japan)
  7. (Japan)
Publication Date:
OSTI Identifier:
22598784
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 476; Journal Issue: 4; Other Information: Copyright (c) 2016 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CALCIUM IONS; CARCINOGENESIS; CARCINOMAS; DOMINANT MUTATIONS; ENDOPLASMIC RETICULUM; ESOPHAGUS; GENE MUTATIONS; GENES; KNOCK-OUT REACTIONS; MESSENGER-RNA; MICE; MUTAGENESIS; MUTANTS; PHENOTYPE; PROTEINS

Citation Formats

Toki, Hideaki, Minowa, Osamu, Inoue, Maki, Motegi, Hiromi, Karashima, Yuko, Ikeda, Ami, Kaneda, Hideki, Sakuraba, Yoshiyuki, Saiki, Yuriko, Wakana, Shigeharu, Suzuki, Hiroshi, Gondo, Yoichi, Shiroishi, Toshihiko, Noda, Tetsuo, E-mail: tnoda@jfcr.or.jp, and Department of Cell Biology, Cancer Institute, The Japanese Foundation for Cancer Research, Tokyo. Novel allelic mutations in murine Serca2 induce differential development of squamous cell tumors. United States: N. p., 2016. Web. doi:10.1016/J.BBRC.2016.04.136.
Toki, Hideaki, Minowa, Osamu, Inoue, Maki, Motegi, Hiromi, Karashima, Yuko, Ikeda, Ami, Kaneda, Hideki, Sakuraba, Yoshiyuki, Saiki, Yuriko, Wakana, Shigeharu, Suzuki, Hiroshi, Gondo, Yoichi, Shiroishi, Toshihiko, Noda, Tetsuo, E-mail: tnoda@jfcr.or.jp, & Department of Cell Biology, Cancer Institute, The Japanese Foundation for Cancer Research, Tokyo. Novel allelic mutations in murine Serca2 induce differential development of squamous cell tumors. United States. doi:10.1016/J.BBRC.2016.04.136.
Toki, Hideaki, Minowa, Osamu, Inoue, Maki, Motegi, Hiromi, Karashima, Yuko, Ikeda, Ami, Kaneda, Hideki, Sakuraba, Yoshiyuki, Saiki, Yuriko, Wakana, Shigeharu, Suzuki, Hiroshi, Gondo, Yoichi, Shiroishi, Toshihiko, Noda, Tetsuo, E-mail: tnoda@jfcr.or.jp, and Department of Cell Biology, Cancer Institute, The Japanese Foundation for Cancer Research, Tokyo. 2016. "Novel allelic mutations in murine Serca2 induce differential development of squamous cell tumors". United States. doi:10.1016/J.BBRC.2016.04.136.
@article{osti_22598784,
title = {Novel allelic mutations in murine Serca2 induce differential development of squamous cell tumors},
author = {Toki, Hideaki and Minowa, Osamu and Inoue, Maki and Motegi, Hiromi and Karashima, Yuko and Ikeda, Ami and Kaneda, Hideki and Sakuraba, Yoshiyuki and Saiki, Yuriko and Wakana, Shigeharu and Suzuki, Hiroshi and Gondo, Yoichi and Shiroishi, Toshihiko and Noda, Tetsuo, E-mail: tnoda@jfcr.or.jp and Department of Cell Biology, Cancer Institute, The Japanese Foundation for Cancer Research, Tokyo},
abstractNote = {Dominant mutations in the Serca2 gene, which encodes sarco(endo)plasmic reticulum calcium-ATPase, predispose mice to gastrointestinal epithelial carcinoma [1–4] and humans to Darier disease (DD) [14–17]. In this study, we generated mice harboring N-ethyl-N-nitrosourea (ENU)-induced allelic mutations in Serca2: three missense mutations and one nonsense mutation. Mice harboring these Serca2 mutations developed tumors that were categorized as either early onset squamous cell tumors (SCT), with development similar to null-type knockout mice [2,4] (aggressive form; M682, M814), or late onset tumors (mild form; M1049, M1162). Molecular analysis showed no aberration in Serca2 mRNA or protein expression levels in normal esophageal cells of any of the four mutant heterozygotes. There was no loss of heterozygosity at the Serca2 locus in the squamous cell carcinomas in any of the four lines. The effect of each mutation on Ca{sup 2+}-ATPase activity was predicted using atomic-structure models and accumulated mutated protein studies, suggesting that putative complete loss of Serca2 enzymatic activity may lead to early tumor onset, whereas mutations in which Serca2 retains residual enzymatic activity result in late onset. We propose that impaired Serca2 gene product activity has a long-term effect on squamous cell carcinogenesis from onset to the final carcinoma stage through an as-yet unrecognized but common regulatory pathway. -- Highlights: •Novel mutations in murine Serca2 caused early onset or late onset of tumorigenesis. •They also caused higher or lower incidence of Darier Disease phenotype. •3D structure model suggested the former mutations led to severer defect on ATPase. •Driver gene mutations via long-range effect on Ca2+ distributions are suggested.},
doi = {10.1016/J.BBRC.2016.04.136},
journal = {Biochemical and Biophysical Research Communications},
number = 4,
volume = 476,
place = {United States},
year = 2016,
month = 8
}
  • A monosaccharide precursor of Escherichia coli lipid A, designated lipid X, which is a diacylglucosamine 1-phosphate with ..beta..-hydroxymyristoyl groups at positions 2 and 3, was shown to have the ability to induce the production of tumor necrosis factor (TNF)-like tumor-cytotoxic factor by a murine macrophage-like cell line, J774.1. This cytotoxic factor was released from J774.1 cells grown in the presence of lipid X and related compounds, and it was assayed as to its lytic activity against (/sup 3/H)thymidine-labeled L929 cells. Dose-response studies revealed that lipid X induced the production of smaller amounts of the tumor-cytotoxic factor than LPS at lowmore » concentrations, but it induced that of considerable amounts at and over 1 ..mu..g/ml. Elimination of 1-phosphate or 3-O-..beta..-hydroxymyristoyl group from lipid X completely prevented the induction of producing this factor by the macrophages. Therefore, it is suggested that both 1-phosphate and 3-O-..beta..-hydroxymyristoyl groups are essential for the biologic activity of lipid X, as to the induction of the tumor-cytotoxic factor production in the macrophages.« less
  • The purpose of this study is to test the hypothesis that gold nanoparticle (AuNP, nanogold)-enhanced radiation therapy (nanogold radiation therapy, NRT) is efficacious when treating the radiation resistant and highly aggressive mouse head and neck squamous cell carcinoma model, SCCVII, and to identify parameters influencing the efficacy of NRT. Subcutaneous (sc) SCCVII leg tumors in mice were irradiated with x-rays at the Brookhaven National Laboratory (BNL) National Synchrotron Light Source (NSLS) with and without prior intravenous (iv) administration of AuNPs. Variables studied included radiation dose, beam energy, temporal fractionation and hyperthermia. AuNP-mediated NRT was shown to be effective for themore » sc SCCVII model. AuNPs were more effective at 42 Gy than at 30 Gy (both at 68 keV median beam energy) compared to controls without gold. Similarly, at 157 keV median beam energy, 50.6 Gy NRT was more effective than 44 Gy NRT. At the same radiation dose (42 Gy), 68 keV was more effective than 157 keV. Hyperthermia and radiation therapy (RT) were synergistic and AuNPs enhanced this synergy, thereby further reducing TCD50 s (tumor control dose 50%) and increasing long-term survivals. It is concluded that gold nanoparticles enhance the radiation therapy of a radioresistant mouse squamous cell carcinoma. The data show that radiation dose, energy and hyperthermia influence efficacy and better define the potential utility of gold nanoparticles for cancer x-ray therapy.« less
  • Histone deacetylase (HDAC) inhibitors are potent anticancer agents and show efficacy against various human neoplasms. Vorinostat is a potent HDAC inhibitor and has shown potential to inhibit growth of human xenograft tumors. However, its effect on the growth of skin neoplasm remains undefined. In this study, we show that vorinostat (2 μM) reduced expression of HDAC1, 2, 3, and 7 in epidermoid carcinoma A431 cells. Consistently, it increased acetylation of histone H3 and p53. Vorinostat (100 mg/kg body weight, IP) treatment reduced human xenograft tumor growth in highly immunosuppressed nu/nu mice. Histologically, the vorinostat-treated tumor showed features of well-differentiation withmore » large necrotic areas. Based on proliferating cell nuclear antigen (PCNA) staining and expression of cyclins D1, D2, E, and A, vorinostat seems to impair proliferation by down-regulating the expression of these proteins. However, it also induced apoptosis. The mechanism by which vorinostat blocks proliferation and makes tumor cells prone to apoptosis, involved inhibition of mTOR signaling which was accompanied by reduction in cell survival AKT and extracellular-signal regulated kinase (ERK) signaling pathways. Our data provide a novel mechanism-based therapeutic intervention for cutaneous squamous cell carcinoma (SCC). Vorinostat may be utilized to cure skin neoplasms in organ transplant recipient (OTR). These patients have high morbidity and surgical removal of these lesions which frequently develop in these patients, is difficult. -- Highlights: ► Vorinostat reduces SCC growth in a xenograft murine model. ► Vorinostat dampens proliferation and induces apoptosis in tumor cells. ► Diminution in mTOR, Akt and ERK signaling underlies inhibition in proliferation. ► Vorinostat by inhibiting HDACs inhibits epithelial–mesenchymal transition.« less
  • Purpose: To evaluate the efficacy of low-dose pulsed radiation therapy (PRT) in 2 head and neck squamous cell carcinoma (HNSCC) xenografts and to investigate the mechanism of action of PRT compared with standard radiation therapy (SRT). Methods and Materials: Subcutaneous radiosensitive UT-SCC-14 and radioresistant UT-SCC-15 xenografts were established in athymic NIH III HO female mice. Tumors were irradiated with 2 Gy/day by continuous standard delivery (SRT: 2 Gy) or discontinuous low-dose pulsed delivery (PRT: 0.2 Gy × 10 with 3-min pulse interval) to total doses of 20 Gy (UT14) or 40 Gy (UT15) using a clinical 5-day on/2-day off schedule. Treatment response was assessed by changes inmore » tumor volume, {sup 18}F-fluorodeoxyglucose (FDG) (tumor metabolism), and {sup 18}F-fluoromisonidazole (FMISO) (hypoxia) positron emission tomography (PET) imaging before, at midpoint, and after treatment. Tumor hypoxia using pimonidazole staining and vascular density (CD34 staining) were assessed by quantitative histopathology. Results: UT15 and UT14 tumors responded similarly in terms of growth delay to either SRT or PRT. When compared with UT14 tumors, UT15 tumors demonstrated significantly lower uptake of FDG at all time points after irradiation. UT14 tumors demonstrated higher levels of tumor hypoxia after SRT when compared with PRT as measured by {sup 18}F-FMISO PET. By contrast, no differences were seen in {sup 18}F-FMISO PET imaging between SRT and PRT for UT15 tumors. Histologic analysis of pimonidazole staining mimicked the {sup 18}F-FMISO PET imaging data, showing an increase in hypoxia in SRT-treated UT14 tumors but not PRT-treated tumors. Conclusions: Differences in {sup 18}F-FMISO uptake for UT14 tumors after radiation therapy between PRT and SRT were measurable despite the similar tumor growth delay responses. In UT15 tumors, both SRT and PRT were equally effective at reducing tumor hypoxia to a significant level as measured by {sup 18}F-FMISO and pimonidazole.« less
  • Purpose: To assess the efficacy of 3-week schedules of low-dose pulsed radiation treatment (PRT) and standard radiation therapy (SRT), with concurrent cisplatin (CDDP) in a head and neck squamous cell carcinoma xenograft model. Methods and Materials: Subcutaneous UT-SCC-14 tumors were established in athymic NIH III HO female mice. A total of 30 Gy was administered as 2 Gy/d, 5 d/wk for 3 weeks, either by PRT (10 × 0.2 Gy/d, with a 3-minute break between each 0.2-Gy dose) or SRT (2 Gy/d, uninterrupted delivery) in combination with concurrent 2 mg/kg CDDP 3 times per week in the final 2 weeks of radiation therapy. Treatment-induced growth delays were defined from twice-weeklymore » tumor volume measurements. Tumor hypoxia was assessed by {sup 18}F-fluoromisonidazole positron emission tomography imaging, and calculated maximum standardized uptake values compared with tumor histology. Tumor vessel density and hypoxia were measured by quantitative immunohistochemistry. Normal tissues effects were evaluated in gut and skin. Results: Untreated tumors grew to 1000 mm{sup 3} in 25.4 days (±1.2), compared with delays of 62.3 days (±3.5) for SRT + CDDP and 80.2 days (±5.0) for PRT + CDDP. Time to reach 2× pretreatment volume ranged from 8.2 days (±1.8) for untreated tumors to 67.1 days (±4.7) after PRT + CDDP. Significant differences in tumor growth delay were observed for SRT versus SRT + CDDP (P=.04), PRT versus PRT + CDDP (P=.035), and SRT + CDDP versus PRT + CDDP (P=.033), and for survival between PRT versus PRT + CDDP (P=.017) and SRT + CDDP versus PRT + CDDP (P=.008). Differences in tumor hypoxia were evident by {sup 18}F-fluoromisonidazole positron emission tomography imaging between SRT and PRT (P=.025), although not with concurrent CDDP. Tumor vessel density differed between SRT + CDDP and PRT + CDDP (P=.011). No differences in normal tissue parameters were seen. Conclusions: Concurrent CDDP was more effective in combination PRT than SRT at restricting tumor growth. Significant differences in tumor vascular density were evident between PRT and SRT, suggesting a preservation of vascular network with PRT.« less