title: Deterministic HOX patterning in human pluripotent stem cell-derived neuroectoderm.
abstract: Colinear HOX expression during hindbrain and spinal cord development diversifies and assigns regional neural phenotypes to discrete rhombomeric and vertebral domains. Despite the precision of HOX patterning in vivo, in vitro approaches for differentiating human pluripotent stem cells (hPSCs) to posterior neural fates coarsely pattern HOX expression thereby generating cultures broadly specified to hindbrain or spinal cord regions. Here, we demonstrate that successive activation of fibroblast growth factor, Wnt/β-catenin, and growth differentiation factor signaling during hPSC differentiation generates stable, homogenous SOX2(+)/Brachyury(+) neuromesoderm that exhibits progressive, full colinear HOX activation over 7 days. Switching to retinoic acid treatment at any point during this process halts colinear HOX activation and transitions the neuromesoderm into SOX2(+)/PAX6(+) neuroectoderm with predictable, discrete HOX gene/protein profiles that can be further differentiated into region-specific cells, e.g., motor neurons. This fully defined approach significantly expands capabilities to derive regional neural phenotypes from diverse hindbrain and spinal cord domains.
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53706, USA; Wisconsin Institute for Discovery, University of Wisconsin, Madison, WI 53706, USA.
authors.forename: Ethan S
authors.initials: ES
authors.lastname: Lippmann
authors.affiliation: Genome Center of Wisconsin, University of Wisconsin, Madison, WI 53706, USA; Department of Biomolecular Chemistry, University of Wisconsin, Madison, WI 53706, USA.
authors.forename: Clay E
authors.initials: CE
authors.lastname: Williams
authors.affiliation: Department of Neuroscience, University of Wisconsin, Madison, WI 53706, USA; Howard Hughes Medical Institute, University of Wisconsin, Madison, WI 53706, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Ruhl
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53706, USA; Wisconsin Institute for Discovery, University of Wisconsin, Madison, WI 53706, USA.
authors.forename: Maria C
authors.initials: MC
authors.lastname: Estevez-Silva
authors.affiliation: Department of Neuroscience, University of Wisconsin, Madison, WI 53706, USA; Howard Hughes Medical Institute, University of Wisconsin, Madison, WI 53706, USA.
authors.forename: Edwin R
authors.initials: ER
authors.lastname: Chapman
authors.affiliation: Genome Center of Wisconsin, University of Wisconsin, Madison, WI 53706, USA; Department of Biomolecular Chemistry, University of Wisconsin, Madison, WI 53706, USA; Department of Chemistry, University of Wisconsin, Madison, WI 53706, USA.
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Coon
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53706, USA; Wisconsin Institute for Discovery, University of Wisconsin, Madison, WI 53706, USA. Electronic address: rashton2@wisc.edu.
authors.forename: Randolph S
authors.initials: RS
authors.lastname: Ashton
mesh_terms: D005346:Fibroblast Growth Factors / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D054258:Neural Plate / Q000378:metabolism*; D009928:Organ Specificity / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D015533:Transcriptional Activation; D059467:Transcriptome; D014212:Tretinoin / Q000494:pharmacology; D051153:Wnt Proteins / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism
chemical_list: D018398:Homeodomain Proteins; D051153:Wnt Proteins; D051176:beta Catenin; D014212:Tretinoin; D005346:Fibroblast Growth Factors
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