In the mammalian genome, the clustered protocadherin (cPCDH) locus provides a paradigm for stochastic gene expression with the potential to generate a unique cPCDH combination in every neuron. Here we report a chromatin-based mechanism that emerges during the transition from the naive to the primed states of cell pluripotency and reduces, by orders of magnitude, the combinatorial potential in the human cPCDH locus. This mechanism selectively increases the frequency of stochastic selection of a small subset of cPCDH genes after neuronal differentiation in monolayers, 10-month-old cortical organoids and engrafted cells in the spinal cords of rats. Signs of these frequent selections can be observed in the brain throughout fetal development and disappear after birth, except in conditions of delayed maturation such as Down’s syndrome. We therefore propose that a pattern of limited cPCDH-gene expression diversity is maintained while human neurons still retain fetal-like levels of maturation.
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The sequencing datasets generated in this study have been deposited in GEO under accession number GSE106872.
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We thank M. G. Rosenfeld and W. Dillmann for providing a nurturing research environment to perform this study, and the many colleagues who helped us to conduct or understand our research (in alphabetical order): A. Becskei, A. Gamliel, T. Haaf, A. Holder, K. Jepsen, E. Kothari, S. Linker, C. Marchetto, S. Marsala, J. Mertens, I. Narvaiza, F. Neri, D. Meluzzi, A. Muotri, P. Negraes, K. Ohgi, M. Parast, S. Sathe, D. Skowronska-Krawczyk, C. Trujillo, R. Tsunemoto, R. Van der Kant, G. Yeo; the team at the UCSD Human Embryonic Stem Cell Core Facility for reagents and technical assistance, and the ENCODE and BrainSpan Consortia for sharing data. Special thanks to H. Garcia Garcia in representation of the many deceased anonymous donors who altruistically donate their bodies for the advancement of science; these donors have made our work possible. Q.M. was supported by the postdoctoral fellowship from the American Cancer Society. C.A. was supported by the postdoctoral fellowship Sara Borrell. Study supported by the US Department of Defense (DoD) (AZ140064) to A.A.-Q., the Sanford Stem Cell Clinical Center (SANPORC) to M.M. and NIH/NIA (1RF1AG048083-01 and 5P50AG005131-34) to L.S.B.G. The Department of Medicine, School of Medicine (UCSD) supported I.G.-B.
M.M. is the scientific founder of Neurgain Technologies and has an equity interest in the company. In addition, M.M. serves as a consultant to Neurgain Technologies and receives compensation for these services. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies.
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Almenar-Queralt, A., Merkurjev, D., Kim, H.S. et al. Chromatin establishes an immature version of neuronal protocadherin selection during the naive-to-primed conversion of pluripotent stem cells. Nat Genet 51, 1691–1701 (2019). https://doi.org/10.1038/s41588-019-0526-4