CAG repeat instability in embryonic stem cells and derivative spermatogenic cells of transgenic Huntington’s disease monkey

Abstract

Purpose

The expansion of CAG (glutamine; Q) trinucleotide repeats (TNRs) predominantly occurs through male lineage in Huntington’s disease (HD). As a result, offspring will have larger CAG repeats compared to their fathers, which causes an earlier onset of the disease called genetic anticipation. This study aims to develop a novel in vitro model to replicate CAG repeat instability in early spermatogenesis and demonstrate the biological process of genetic anticipation by using the HD stem cell model for the first time.

Methods

HD rhesus monkey embryonic stem cells (rESCs) were cultured in vitro for an extended period. Male rESCs were used to derive spermatogenic cells in vitro with a 10-day differentiation. The assessment of CAG repeat instability was performed by GeneScan and curve fit analysis.

Results

Spermatogenic cells derived from rESCs exhibit progressive expansion of CAG repeats with high daily expansion rates compared to the extended culture of rESCs. The expansion of CAG repeats is cell type–specific and size-dependent.

Conclusions

Here, we report a novel stem cell model that replicates genome instability and CAG repeat expansion in in vitro derived HD monkey spermatogenic cells. The in vitro spermatogenic cell model opens a new opportunity for studying TNR instability and the underlying mechanism of genetic anticipation, not only in HD but also in other TNR diseases.

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Data availability

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Code availability

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Acknowledgements

All materials and experiments were performed at the Yerkes National Primate Research Center (YNPRC). We would like to thank YNPRC staff, Jinjing Yang, Siran Tian, Dr. Kanchana Punyawai, and other Dr. Chan’s Lab members.

Funding

YNPRC is supported by the Office of Research and Infrastructure Program (ORIP)/OD P51OD11132. The Transgenic Huntington’s Disease Monkey Resource “THDMR” and this study were supported in part by grants awarded by the ORIP/NIH (OD010930) and NINDS/NIH (NS101701) to AWSC. This study was also supported in part by OD020182 and Georgia Partners in Regenerative Medicine Seed Grant to AWSC and CAE. We also received the support by the Emory University Research Council and Arthur and Sarah Merrill Foundation to AWSC and IKC. Sujittra Khampang and Rangsun Parnpai are supported by SUT-PhD scholarship, Suranaree University of Technology, Nakhorachasrima, Thailand.

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Anthony W. S. Chan, In Ki Cho, and Sujittra Khampang conceptualized the study, designed experimentations, and wrote the manuscript. Sujittra Khampang and In Ki Cho performed experiments, data collection, and data analysis. Anthony W. S. Chan and Charles A. Easley IV supervised the spermatogenic cell differentiation, characterization, and experimental procedures. Wiriya Mahikul supervised the data analysis and statistical interpretation. Rangsun Parnpai supervised and reviewed the manuscript. All authors reviewed and approved the manuscript.

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Correspondence to In Ki Cho or Anthony W.S. Chan.

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This work was prepared while Anthony W.S. Chan was employed at Yerkes National Primate Research Center and Emory University. The opinions expressed in this article are the author's own and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government.

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Khampang, S., Parnpai, R., Mahikul, W. et al. CAG repeat instability in embryonic stem cells and derivative spermatogenic cells of transgenic Huntington’s disease monkey. J Assist Reprod Genet (2021). https://doi.org/10.1007/s10815-021-02106-3

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Keywords

  • Huntington’s disease
  • Spermatogenic cells
  • CAG repeat instability
  • HD rhesus monkey embryonic stem cells (rESCs)
  • Genetic anticipation