A role for Lin28 in primordial germ-cell development and germ-cell malignancy

Abstract

The rarity and inaccessibility of the earliest primordial germ cells (PGCs) in the mouse embryo thwart efforts to investigate molecular mechanisms of germ-cell specification. stella (also called Dppa3) marks the rare founder population of the germ lineage1,2. Here we differentiate mouse embryonic stem cells carrying a stella transgenic reporter into putative PGCs in vitro. The Stella+ cells possess a transcriptional profile similar to embryo-derived PGCs, and like their counterparts in vivo, lose imprints in a time-dependent manner. Using inhibitory RNAs to screen candidate genes for effects on the development of Stella+ cells in vitro, we discovered that Lin28, a negative regulator of let-7 microRNA processing3,4,5,6, is essential for proper PGC development. Furthermore, we show that Blimp1 (also called Prdm1), a let-7 target and a master regulator of PGC specification7,8,9, can rescue the effect of Lin28 deficiency during PGC development, thereby establishing a mechanism of action for Lin28 during PGC specification. Overexpression of Lin28 promotes formation of Stella+ cells in vitro and PGCs in chimaeric embryos, and is associated with human germ-cell tumours. The differentiation of putative PGCs from embryonic stem cells in vitro recapitulates the early stages of gamete development in vivo, and provides an accessible system for discovering novel genes involved in germ-cell development and malignancy.

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Figure 1: ES cell differentiation into putative PGCs in vitro is marked by stella expression.
Figure 2: Lin28 regulates PGC development.
Figure 3: Lin28 and Blimp1 regulate PGC development in vivo.
Figure 4: LIN28 expression in human germ-cell tumours.

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Gene Expression Omnibus

Data deposits

The microarray data have been deposited in the Gene Expression Omnibus (GEO) and given the series accession number GSE7948.

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Acknowledgements

We thank M. W. Lensch for comments on this manuscript; D. K. Gifford, G. Gerber, C. Reeder and J. Baughman for comments and input regarding microarray analysis; G. Losyev for flow cytometry expertise; and S. Winkler of the British Consulate for providing support for collaboration between the Daley and Surani laboratories. This study was supported by grants from the NIH, the NIH Director’s Pioneer Award of the NIH Roadmap for Medical Research, and by support from the germ cell program of the Harvard Stem Cell Institute. G.Q.D. is a recipient of the Burroughs Wellcome Fund Clinical Scientist Award in Translational Research.

Author Contributions J.A.W., project planning, experimental work, manuscript preparation; S.R.V., A.Y., A.T., K.C., I.-H.P., J.E.S., H.Z., A.P.-A., A.L.F., experimental work; M.A.S., contributed reagents and critical feedback; G.Q.D., project planning, data analysis and manuscript preparation.

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Correspondence to George Q. Daley.

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West, J., Viswanathan, S., Yabuuchi, A. et al. A role for Lin28 in primordial germ-cell development and germ-cell malignancy. Nature 460, 909–913 (2009). https://doi.org/10.1038/nature08210

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