Aberrant differentiation of Tsc2-deficient teratomas associated with activation of the mTORC1-TFE3 pathway

Kawano,Haruna; Ito,Yoshitaka; Kanai,Fumio; Nakamura,Eri; Tada,Norihiro; Takai,Setsuo; Horie,Shigeo; Kobayashi,Toshiyuki; Hino,Okio

doi:10.3892/or.2015.4254

Aberrant differentiation of Tsc2-deficient teratomas associated with activation of the mTORC1-TFE3 pathway

Authors:
- Haruna Kawano
- Yoshitaka Ito
- Fumio Kanai
- Eri Nakamura
- Norihiro Tada
- Setsuo Takai
- Shigeo Horie
- Toshiyuki Kobayashi
- Okio Hino
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Affiliations: Department of Urology, Juntendo University Graduate School of Medicine, Tokyo 113-8431, Japan, Department of Molecular Pathogenesis, Juntendo University Graduate School of Medicine, Tokyo 113-8431, Japan, Laboratory of Genome Research, Research Institute for Diseases of Old Age, Juntendo University Graduate School of Medicine, Tokyo 113-8431, Japan, Department of Clinical Radiology, Faculty of Health Sciences, Hiroshima International University, Hiroshima 724-0695, Japan
Published online on: September 8, 2015 https://doi.org/10.3892/or.2015.4254
Pages: 2251-2258
Copyright: © Kawano et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The model animal of renal cell carcinoma (RCC), the Eker rat, has a germline mutation in the tuberous sclerosis 2 (Tsc2) gene. Heterozygous mutants develop RCCs by second hit in the wild-type Tsc2 allele, whereas homozygous mutants are embryonic lethal. In the present study, a new cell differentiation model was developed to study the mechanism of Tsc2 mutation-associated pathogenesis by generating Tsc2-deficient embryonic stem cells (ESCs) from Eker rats. Tsc2+/+, Tsc2+/- and Tsc2-/- ESCs were all capable of generating three germ layers: mesoderm, ectoderm, and endoderm. Interestingly, epithelial tumor-like abnormal ductal structures were reproducibly observed in Tsc2-/- teratomas from different ESC lines. Immunohistochemical analysis revealed that mammalian target of rapamycin complex 1 (mTORC1) signaling was activated in abnormal ducts of Tsc2-/- teratomas, on the basis of positive staining for p-S6 and p-4EBP1. In these abnormal ducts, expression levels of epithelial markers (i.e., megalin and cubilin) and the cytoplasmic localization of E-cadherin and β-catenin were similar to those in Eker rat RCCs. Moreover, a transcription factor regulated by mTORC1, named TFE3, was located in the nuclei of abnormal ducts and Eker rat RCCs. As a negative regulator of ESC differentiation, TFE3 may result in tissue-specific differentiation defects related to tumorigenesis in Eker rats and Tsc2-/- teratomas. The present study suggests that ESCs derived from Eker rats constitute a novel experimental tool with which to analyze differentiation defects and cell-type specific pathogenesis associated with Tsc2 deficiency.

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