SOX2 is essential for in vivo reprogramming of seminoma-like TCam-2 cells to an embryonal carcinoma-like fate.
Type II germ cell cancers (GCC) are divided into seminomas, which are highly similar to primordial germ cells and embryonal carcinomas (EC), often described as malignant counterparts to embryonic stem cells.Previously, we demonstrated that the development of GCCs is a highly plastic process and strongly influenced by the microenvironment. While orthotopic transplantation into the testis promotes seminomatous growth of the seminoma-like cell line TCam-2, ectopic xenotransplantation into the flank initiates reprogramming into an EC-like fate.During this reprogramming, BMP signaling is inhibited, leading to induction of NODAL signaling, upregulation of pluripotency factors and downregulation of seminoma markers, like SOX17. The pluripotency factor and EC-marker SOX2 is strongly induced.Here, we adressed the molecular role of SOX2 in this reprogramming. Using CRISPR/Cas9-mediated genome-editing, we established SOX2-deficient TCam-2 cells. Xenografting of SOX2-deficient cells into the flank of nude mice resulted in maintenance of a seminoma-like fate, indicated by the histology and expression of OCT3/4, SOX17, TFAP2C, PRDM1 and PRAME. In SOX2-deficient cells, BMP signaling is inhibited, but NODAL signaling is not activated. Thus, SOX2 appears to be downstream of BMP signaling but upstream of NODAL activation. So, SOX2 is an essential factor in acquiring an EC-like cell fate from seminomas.A small population of differentiated cells was identified resembling a mixed non-seminoma. Analyses of these cells revealed downregulation of the pluripotency and seminoma markers OCT3/4, SOX17, PRDM1 and TFAP2C. In contrast, the pioneer factor FOXA2 and its target genes were upregulated, suggesting that FOXA2 might play an important role in induction of non-seminomatous differentiation.
Institute of Pathology, Department of Developmental Pathology, University of Bonn Medical School, Bonn, Germany.
Daniel
D
Nettersheim
Institute of Pathology, Department of Developmental Pathology, University of Bonn Medical School, Bonn, Germany.
Alena
A
Heimsoeth
Institute of Pathology, Department of Developmental Pathology, University of Bonn Medical School, Bonn, Germany.
Sina
S
Jostes
Institute of Pathology, Department of Developmental Pathology, University of Bonn Medical School, Bonn, Germany.
Simon
S
Schneider
Institute of Pathology, Department of Developmental Pathology, University of Bonn Medical School, Bonn, Germany.
Martin
M
Fellermeyer
Institute of Human Genetics, University of Bonn Medical School, Bonn, Germany.
Andrea
A
Hofmann
Institute of Pathology, Department of Developmental Pathology, University of Bonn Medical School, Bonn, Germany.
Hubert
H
Schorle
D000818:Animals; D018236:Carcinoma, Embryonal / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002454:Cell Differentiation / Q000502:physiology; D045744:Cell Line, Tumor; D065150:Cellular Reprogramming / Q000502:physiology; D055786:Gene Knockout Techniques; D064593:Heterografts; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D055457:Nodal Protein / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D018239:Seminoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D014162:Transfection
C526007:NODAL protein, human; D055457:Nodal Protein; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors
SOX2/SOX17; TCam-2; embryonal carcinoma; reprogramming; seminoma