3290587920230816
2589-00422392020Sep25iScienceiScienceSOX1 Is Required for the Specification of Rostral Hindbrain Neural Progenitor Cells from Human Embryonic Stem Cells.10147510147510147510.1016/j.isci.2020.101475Region-specific neural progenitor cells (NPCs) can be generated from human embryonic stem cells (hESCs) by modulating signaling pathways. However, how intrinsic transcriptional factors contribute to the neural regionalization is not well characterized. Here, we generate region-specific NPCs from hESCs and find that SOX1 is highly expressed in NPCs with the rostral hindbrain identity. Moreover, we find that OTX2 inhibits SOX1 expression, displaying exclusive expression between the two factors. Furthermore, SOX1 knockout (KO) leads to the upregulation of midbrain genes and downregulation of rostral hindbrain genes, indicating that SOX1 is required for specification of rostral hindbrain NPCs. Our SOX1 chromatin immunoprecipitation sequencing analysis reveals that SOX1 binds to the distal region of GBX2 to activate its expression. Overexpression of GBX2 largely abrogates SOX1-KO-induced aberrant gene expression. Taken together, this study uncovers previously unappreciated role of SOX1 in early neural regionalization and provides new information for the precise control of the OTX2/GBX2 interface.© 2020 The Author(s).LiuXinyuanXCAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai Institute of Nutrition and Health, CAS Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.FangZhuoqingZCAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai Institute of Nutrition and Health, CAS Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.WenJingJCAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai Institute of Nutrition and Health, CAS Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.TangFanFDepartment of Histoembryology, Genetics and Developmental Biology, Shanghai Key Laboratory of Reproductive Medicine, Shanghai JiaoTong University School of Medicine, 225 South Chongqing Road, Shanghai 200025, China.LiaoBingBDepartment of Histoembryology, Genetics and Developmental Biology, Shanghai Key Laboratory of Reproductive Medicine, Shanghai JiaoTong University School of Medicine, 225 South Chongqing Road, Shanghai 200025, China.JingNaiheNState Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China.School of Life Science and Technology, ShanghaiTech University, 100 Haike Road, Shanghai 201210, China.LaiDongmeiDInternational Peace Maternity and Child Health Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200030, China.JinYingYCAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai Institute of Nutrition and Health, CAS Center for Excellence in Molecular Cell Science, University of Chinese Academy of Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.Department of Histoembryology, Genetics and Developmental Biology, Shanghai Key Laboratory of Reproductive Medicine, Shanghai JiaoTong University School of Medicine, 225 South Chongqing Road, Shanghai 200025, China.Basic Clinical Research Center, Renji Hospital, Shanghai JiaoTong University School of Medicine, 160 Pujian Road, Shanghai 200127, China.School of Life Science and Technology, ShanghaiTech University, 100 Haike Road, Shanghai 201210, China.engJournal Article20200820
United StatesiScience1017240382589-0042Developmental NeuroscienceMolecular GeneticsThe authors declare that they have no conflicts of interest with the contents of this article.201912102020711202081720209106120209106020209920102020820epublish32905879PMC748643310.1016/j.isci.2020.101475S2589-0042(20)30667-2Acampora D., Mazan S., Lallemand Y., Avantaggiato V., Maury M., Simeone A., Brulet P. Forebrain and midbrain regions are deleted in Otx2-/- mutants due to a defective anterior neuroectoderm specification during gastrulation. Development. 1995;121:3279–3290.7588062Acampora D., Avantaggiato V., Tuorto F., Simeone A. Genetic control of brain morphogenesis through Otx gene dosage requirement. Development. 1997;124:3639–3650.9342056Ang S.L., Jin O., Rhinn M., Daigle N., Stevenson L., Rossant J. A targeted mouse Otx2 mutation leads to severe defects in gastrulation and formation of axial mesoderm and to deletion of rostral brain. Development. 1996;122:243–252.8565836Aubert J., Stavridis M.P., Tweedie S., O'Reilly M., Vierlinger K., Li M., Ghazal P., Pratt T., Mason J.O., Roy D. Screening for mammalian neural genes via fluorescence-activated cell sorter purification of neural precursors from Sox1-gfp knock-in mice. Proc. Natl. Acad. Sci. U S A. 2003;100:11836–11841.PMC30409512923295Bally-Cuif L., Cholley B., Wassef M. Involvement of Wnt-1 in the formation of the mes/metencephalic boundary. Mech. Dev. 1995;53:23–34.8555108Bally-Cuif L., Gulisano M., Broccoli V., Boncinelli E. c-otx2 is expressed in two different phases of gastrulation and is sensitive to retinoic acid treatment in chick embryo. Mech. Dev. 1995;49:49–63.7748789Bouillet P., Chazaud C., Oulad-Abdelghani M., Dolle P., Chambon P. Sequence and expression pattern of the Stra7 (Gbx-2) homeobox-containing gene induced by retinoic acid in P19 embryonal carcinoma cells. Dev. Dyn. 1995;204:372–382.8601031Broccoli V., Boncinelli E., Wurst W. The caudal limit of Otx2 expression positions the isthmic organizer. Nature. 1999;401:164–168.10490025Chambers S.M., Fasano C.A., Papapetrou E.P., Tomishima M., Sadelain M., Studer L. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat. Biotechnol. 2009;27:275–280.PMC275672319252484Fang Z., Liu X., Wen J., Tang F., Zhou Y., Jing N., Jin Y. SOX21 ensures rostral forebrain identity by suppression of WNT8B during neural regionalization of human embryonic stem cells. Stem Cell Reports. 2019;13:1038–1052.PMC691584331761677Fasano C.A., Chambers S.M., Lee G., Tomishima M.J., Studer L. Efficient derivation of functional floor plate tissue from human embryonic stem cells. Cell Stem Cell. 2010;6:336–347.PMC433680020362538Hanks M., Wurst W., Anson-Cartwright L., Auerbach A.B., Joyner A.L. Rescue of the en-1 mutant phenotype by replacement of en-1 with en-2. Science. 1995;269:679–682.7624797Hunt P., Gulisano M., Cook M., Sham M.H., Faiella A., Wilkinson D., Boncinelli E., Krumlauf R. A distinct Hox code for the branchial region of the vertebrate head. Nature. 1991;353:861–864.1682814Imaizumi K., Sone T., Ibata K., Fujimori K., Yuzaki M., Akamatsu W., Okano H. Controlling the regional identity of hPSC-derived neurons to uncover neuronal subtype specificity of neurological disease phenotypes. Stem Cell Reports. 2015;5:1010–1022.PMC468212326549851Joyner A.L., Liu A., Millet S. Otx2, Gbx2 and Fgf8 interact to position and maintain a mid-hindbrain organizer. Curr. Opin. Cell Biol. 2000;12:736–741.11063941Katahira T., Sato T., Sugiyama S., Okafuji T., Araki I., Funahashi J., Nakamura H. Interaction between Otx2 and Gbx2 defines the organizing center for the optic tectum. Mech. Dev. 2000;91:43–52.10704829Kiecker C., Niehrs C. A morphogen gradient of Wnt/beta-catenin signalling regulates anteroposterior neural patterning in Xenopus. Development. 2001;128:4189–4201.11684656Kiecker C., Niehrs C. The role of prechordal mesendoderm in neural patterning. Curr. Opin. Neurobiol. 2001;11:27–33.11179869Kirkeby A., Grealish S., Wolf D.A., Nelander J., Wood J., Lundblad M., Lindvall O., Parmar M. Generation of regionally specified neural progenitors and functional neurons from human embryonic stem cells under defined conditions. Cell Rep. 2012;1:703–714.22813745Kondoh H., Kamachi Y. SOX-partner code for cell specification: regulatory target selection and underlying molecular mechanisms. Int. J. Biochem. Cell Biol. 2010;42:391–399.19747562Kriks S., Shim J.W., Piao J., Ganat Y.M., Wakeman D.R., Xie Z., Carrillo-Reid L., Auyeung G., Antonacci C., Buch A. Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson's disease. Nature. 2011;480:547–551.PMC324579622056989Li C., Yang Y., Lu X., Sun Y., Gu J., Feng Y., Jin Y. Efficient derivation of Chinese human embryonic stem cell lines from frozen embryos. In Vitro Cell. Dev. Biol. Anim. 2010;46:186–191.20186511Luu B., Ellisor D., Zervas M. The lineage contribution and role of Gbx2 in spinal cord development. PLoS One. 2011;6:e20940.PMC311686021698205Maden M. Retinoic acid in the development, regeneration and maintenance of the nervous system. Nat. Rev. Neurosci. 2007;8:755–765.17882253Malas S., Duthie S.M., Mohri F., Lovell-Badge R., Episkopou V. Cloning and mapping of the human SOX1: a highly conserved gene expressed in the developing brain. Mamm. Genome. 1997;8:866–868.9337405Malas S., Postlethwaite M., Ekonomou A., Whalley B., Nishiguchi S., Wood H., Meldrum B., Constanti A., Episkopou V. Sox1-deficient mice suffer from epilepsy associated with abnormal ventral forebrain development and olfactory cortex hyperexcitability. Neuroscience. 2003;119:421–432.12770556Mandegar M.A., Huebsch N., Frolov E.B., Shin E., Truong A., Olvera M.P., Chan A.H., Miyaoka Y., Holmes K., Spencer C.I. CRISPR interference efficiently induces specific and reversible gene silencing in human iPSCs. Cell Stem Cell. 2016;18:541–543.PMC483069726971820Maroof A.M., Keros S., Tyson J.A., Ying S.W., Ganat Y.M., Merkle F.T., Liu B., Goulburn A., Stanley E.G., Elefanty A.G. Directed differentiation and functional maturation of cortical interneurons from human embryonic stem cells. Cell Stem Cell. 2013;12:559–572.PMC368152323642365Matsuo I., Kuratani S., Kimura C., Takeda N., Aizawa S. Mouse Otx2 functions in the formation and patterning of rostral head. Genes Dev. 1995;9:2646–2658.7590242Millet S., Bloch-Gallego E., Simeone A., Alvarado-Mallart R.M. The caudal limit of Otx2 gene expression as a marker of the midbrain/hindbrain boundary: a study using in situ hybridisation and chick/quail homotopic grafts. Development. 1996;122:3785–3797.9012500Millet S., Campbell K., Epstein D.J., Losos K., Harris E., Joyner A.L. A role for Gbx2 in repression of Otx2 and positioning the mid/hindbrain organizer. Nature. 1999;401:161–164.10490024Nakayama Y., Kikuta H., Kanai M., Yoshikawa K., Kawamura A., Kobayashi K., Wang Z., Khan A., Kawakami K., Yamasu K. Gbx2 functions as a transcriptional repressor to regulate the specification and morphogenesis of the mid-hindbrain junction in a dosage- and stage-dependent manner. Mech. Dev. 2013;130:532–552.23933069Nishiguchi S., Wood H., Kondoh H., Lovell-Badge R., Episkopou V. Sox1 directly regulates the gamma-crystallin genes and is essential for lens development in mice. Genes Dev. 1998;12:776–781.PMC3166329512512Pevny L.H., Sockanathan S., Placzek M., Lovell-Badge R. A role for SOX1 in neural determination. Development. 1998;125:1967–1978.9550729Prince V., Lumsden A. Hoxa-2 expression in normal and transposed rhombomeres: independent regulation in the neural tube and neural crest. Development. 1994;120:911–923.7600967Rada-Iglesias A., Bajpai R., Swigut T., Brugmann S.A., Flynn R.A., Wysocka J. A unique chromatin signature uncovers early developmental enhancers in humans. Nature. 2011;470:279–283.PMC444567421160473Schneider-Maunoury S., Topilko P., Seitandou T., Levi G., Cohen-Tannoudji M., Pournin S., Babinet C., Charnay P. Disruption of Krox-20 results in alteration of rhombomeres 3 and 5 in the developing hindbrain. Cell. 1993;75:1199–1214.7903221Shamim H., Mason I. Expression of Gbx-2 during early development of the chick embryo. Mech. Dev. 1998;76:157–159.9767156Simeone A. Otx1 and Otx2 in the development and evolution of the mammalian brain. EMBO J. 1998;17:6790–6798.PMC11710269843484Simeone A., Acampora D., Gulisano M., Stornaiuolo A., Boncinelli E. Nested expression domains of four homeobox genes in developing rostral brain. Nature. 1992;358:687–690.1353865Thomson J.A., Itskovitz-Eldor J., Shapiro S.S., Waknitz M.A., Swiergiel J.J., Marshall V.S., Jones J.M. Embryonic stem cell lines derived from human blastocysts. Science. 1998;282:1145–1147.9804556Tsankov A.M., Gu H., Akopian V., Ziller M.J., Donaghey J., Amit I., Gnirke A., Meissner A. Transcription factor binding dynamics during human ES cell differentiation. Nature. 2015;518:344–349.PMC449933125693565Uchikawa M., Yoshida M., Iwafuchi-Doi M., Matsuda K., Ishida Y., Takemoto T., Kondoh H. B1 and B2 Sox gene expression during neural plate development in chicken and mouse embryos: universal versus species-dependent features. Dev. Growth Differ. 2011;53:761–771.21762129Wassarman K.M., Lewandoski M., Campbell K., Joyner A.L., Rubenstein J.L., Martinez S., Martin G.R. Specification of the anterior hindbrain and establishment of a normal mid/hindbrain organizer is dependent on Gbx2 gene function. Development. 1997;124:2923–2934.9247335Waters S.T., Lewandoski M. A threshold requirement for Gbx2 levels in hindbrain development. Development. 2006;133:1991–2000.16651541Wilkinson D.G., Bhatt S., Chavrier P., Bravo R., Charnay P. Segment-specific expression of a zinc-finger gene in the developing nervous system of the mouse. Nature. 1989;337:461–464.2915691Wood H.B., Episkopou V. Comparative expression of the mouse Sox1, Sox2 and Sox3 genes from pre-gastrulation to early somite stages. Mech. Dev. 1999;86:197–201.10446282Xuan S., Baptista C.A., Balas G., Tao W., Soares V.C., Lai E. Winged helix transcription factor BF-1 is essential for the development of the cerebral hemispheres. Neuron. 1995;14:1141–1152.7605629