@article{6534fe956a9d457dabd3c92a920ca7c9,
title = "hPSC-derived sacral neural crest enables rescue in a severe model of Hirschsprung's disease",
abstract = "The enteric nervous system (ENS) is derived from both the vagal and sacral component of the neural crest (NC). Here, we present the derivation of sacral ENS precursors from human PSCs via timed exposure to FGF, WNT, and GDF11, which enables posterior patterning and transition from posterior trunk to sacral NC identity, respectively. Using a SOX2::H2B-tdTomato/T::H2B-GFP dual reporter hPSC line, we demonstrate that both trunk and sacral NC emerge from a double-positive neuro-mesodermal progenitor (NMP). Vagal and sacral NC precursors yield distinct neuronal subtypes and migratory behaviors in vitro and in vivo. Remarkably, xenografting of both vagal and sacral NC lineages is required to rescue a mouse model of total aganglionosis, suggesting opportunities in the treatment of severe forms of Hirschsprung's disease.",
keywords = "GDF11, Hirschsprung's disease, axial patterning, axial progenitors, cell therapy, directed differentiation, enteric nervous system, nervous system disorder, neural crest development, pluripotent stem cells, regenerative medicine, sacral neural crest",
author = "Yujie Fan and James Hackland and Arianna Baggiolini and Hung, {Lin Y.} and Huiyong Zhao and Paul Zumbo and Polina Oberst and Minotti, {Andrew P.} and Emiliano Hergenreder and Sarah Najjar and Zixing Huang and Cruz, {Nelly M.} and Aaron Zhong and Mega Sidharta and Ting Zhou and {de Stanchina}, Elisa and Doron Betel and White, {Richard M.} and Michael Gershon and Margolis, {Kara Gross} and Lorenz Studer",
note = "Funding Information: We acknowledge use of the Integrated Genomics Operation Core, funded by the NCI Cancer Center Support Grant (CCSG, P30 CA08748), Cycle for Survival, and the Marie-Jos{\'e}e and Henry R. Kravis Center for Molecular Oncology, and assistance from the Bioinformatics Core also funded in part by P30 CA008748. We are grateful to the SKI Stem Cell Research Facility for creating the GFP and mCherry expressing and the SOX2::Tomato/T::GFP dual reporter hESC lines; Angela Pepe-Caprio for assistance with animal protocols and laboratory management; K. Lertpiriyapong, C. Aguinaga, K. Chan, and the colony management group at MSKCC for support of mouse studies; molecular cytology core at MSKCC for preparation paraffin slides and H&E staining; E. Chan for the script of automatic measurement of migration distance and counting of colocalization. Figures contain images created with BioRender.com. Y.F. was supported by Frank Lappin Horsfall, Jr. Fellowship; A.B. by the Alan and Sandra Gerry Metastasis and Tumor Ecosystems Center (GMTEC) Scholars Fellowship. A.P.M. by NIA/NIH F31AG067709 fellowship and S.N by NIDDK/NIH (F32DK132810) fellowship. This work was supported by NINDS/NIH (R01NS099270) and a sponsored research agreement from BlueRock Therapeutics (SK2018-1304) to L.S. by NINDS/NIH (RO1NS015547) to M.D.G. and by NIDDK/NIH (RO1DK126644) to K.M. Y.F. and L.S. developed the experiments and interpreted results. Y.F. performed most experiments and collected and analyzed data. J.H. performed GDF11-related experiments and the analysis of RNA-seq data and assisted in manuscript preparation. A.B. contributed to sequencing experiments and melanocyte-related experiments and assisted in manuscript preparation. L.Y.H. performed the gut motility experiments, whole-mount dissections and immunostaining experiments with assistance of S.N. and Z.H. K.G.M. and M.G. helped with experimental design, results, and discussion. A.P.M. and P.O. performed synapse quantification analysis. H.Z. performed the mouse transplantation studies. P.Z. and D.B. performed computational analyses. E.H. performed and analyzed the MEA experiment. N.M.C. performed melanocyte-related experiments. A.Z. M.S. and T.Z. generated the reporter lines used in this study. Y.F. and L.S. wrote the manuscript. L.S. acquired funding for the project. All authors read and edited the manuscript. L.S. is a scientific co-founder and consultant and has received sponsored research support for work related to this study from Bluerock Therapeutics. L.S. and Y.F. are inventors of a patent application filed by Memorial Sloan Kettering Cancer Center on the methods described in this study. We support inclusive, diverse, and equitable conduct of research. Funding Information: We acknowledge use of the Integrated Genomics Operation Core, funded by the NCI Cancer Center Support Grant (CCSG, P30 CA08748), Cycle for Survival, and the Marie-Jos{\'e}e and Henry R. Kravis Center for Molecular Oncology, and assistance from the Bioinformatics Core also funded in part by P30 CA008748. We are grateful to the SKI Stem Cell Research Facility for creating the GFP and mCherry expressing and the SOX2::Tomato/T::GFP dual reporter hESC lines; Angela Pepe-Caprio for assistance with animal protocols and laboratory management; K. Lertpiriyapong, C. Aguinaga, K. Chan, and the colony management group at MSKCC for support of mouse studies; molecular cytology core at MSKCC for preparation paraffin slides and H&E staining; E. Chan for the script of automatic measurement of migration distance and counting of colocalization. Figures contain images created with BioRender.com. Y.F. was supported by Frank Lappin Horsfall, Jr. Fellowship; A.B. by the Alan and Sandra Gerry Metastasis and Tumor Ecosystems Center (GMTEC) Scholars Fellowship. A.P.M. by NIA/NIH F31AG067709 fellowship and S.N by NIDDK/NIH (F32DK132810) fellowship. This work was supported by NINDS/NIH (R01NS099270) and a sponsored research agreement from BlueRock Therapeutics (SK2018-1304) to L.S., by NINDS/NIH (RO1NS015547) to M.D.G., and by NIDDK/NIH (RO1DK126644) to K.M. Publisher Copyright: {\textcopyright} 2023 The Authors",
year = "2023",
month = mar,
day = "2",
doi = "10.1016/j.stem.2023.02.003",
language = "English (US)",
volume = "30",
pages = "264--282.e9",
journal = "Cell Stem Cell",
issn = "1934-5909",
publisher = "Cell Press",
number = "3",
}