DMRT1-mediated reprogramming drives development of cancer resembling human germ cell tumors with features of totipotency

In vivo reprogramming provokes a wide range of cell fate conversion. Here, we discover that in vivo induction of higher levels of OSKM in mouse somatic cells leads to increased expression of primordial germ cell (PGC)-related genes and provokes genome-wide erasure of genomic imprinting, which takes place exclusively in PGCs. Moreover, the in vivo OSKM reprogramming results in development of cancer that resembles human germ cell tumors. Like a subgroup of germ cell tumors, propagated tumor cells can differentiate into trophoblasts. Moreover, these tumor cells give rise to induced pluripotent stem cells (iPSCs) with expanded differentiation potential into trophoblasts. Remarkably, the tumor-derived iPSCs are able to contribute to non-neoplastic somatic cells in adult mice. Mechanistically, DMRT1, which is expressed in PGCs, drives the reprogramming and propagation of the tumor cells in vivo. Furthermore, the DMRT1-related epigenetic landscape is associated with trophoblast competence of the reprogrammed cells and provides a therapeutic target for germ cell tumors. These results reveal an unappreciated route for somatic cell reprogramming and underscore the impact of reprogramming in development of germ cell tumors.


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For ATAC-seq analysis, adaptor sequences in reads were trimmed using Trim Galore (version 0.3.7.) Then trimmed reads were aligned to the mouse reference genome (mm10) build using bowtie (version 0.12.7) with default parameters. Duplicated reads were removed using samtools (version 0.1.18).
All data of WGBS, Methyl-seq, RNA-seq and ATAC-seq used in this study were deposited in the Gene Expression Omnibus ( No statistical methods were used to predetermine sample size. Sample size was chosen based on standards in the field. Sample size is described in methods, figures or figure legends. qRT-PCR experiments were performed in biological triplicate. ATAC-qPCR experiments were performed in biological duplicate. We have had enough number of sequencing reads for WGBS, Methyl-seq, RNA-seq and ATAC-seq.
Since we used chimeric mice to evaluate pathological data, we excluded mice with no chimeric contribution. We also excluded a few mice which unexpectedly died during Dox treatment. We didn't excluded any other data from the analysis to avoid the arbitrary selection.
We were able to confirm the reproducibility of our results. See experimental materials and methods section.
In animal experiments, chimeric mice used were allocated to arrange coat color chimerism to avoid the effect of chimerism. For quantification of histological features, HE-stained sections and immunostained sections were randomly photographed at 100× magnification. Four or five images from each sample were processed with ImageJ software (NIH) to evaluate the region of interest.
We were not blinded during data collection and analysis. We did not considered blinding required in these type of experiments.

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April 2020 Note that full information on the approval of the study protocol must also be provided in the manuscript. Validation of dilution was determined in accordance with manufacturer's directions. In some primary antibodies, we optimized the dilution to fit our staining protocols.
We authenticated that each cell lines have correct genotypes using PCR, immunostaining and western blot.
All samples used were tested for mycoplasma contamination. We confirmed that our cell lines are negative to mycoplasma contamination.
No commonly misidentified lines were used in this study.
We performed in vivo reprogramming experiments using 4-8 week old chimeric mice. In pathological investigations, we used both male and female mice. Pseudopregnant ICR and male/female ICR (8-10 week old) mice were obtained from Japan SLC to make chimeric mice. C57BL/6 mice were also obtained from Japan SLC. BALB/c-nu/nu mice were obtained from Japan CLEA.