title: Iterative use of nuclear receptor Nr5a2 regulates multiple stages of liver and pancreas development.
abstract: The stepwise progression of common endoderm progenitors into differentiated liver and pancreas organs is regulated by a dynamic array of signals that are not well understood. The nuclear receptor subfamily 5, group A, member 2 gene nr5a2, also known as Liver receptor homolog-1 (Lrh-1) is expressed in several tissues including the developing liver and pancreas. Here, we interrogate the role of Nr5a2 at multiple developmental stages using genetic and chemical approaches and uncover novel pleiotropic requirements during zebrafish liver and pancreas development. Zygotic loss of nr5a2 in a targeted genetic null mutant disrupted the development of the exocrine pancreas and liver, while leaving the endocrine pancreas intact. Loss of nr5a2 abrogated exocrine pancreas markers such as trypsin, while pancreas progenitors marked by ptf1a or pdx1 remained unaffected, suggesting a role for Nr5a2 in regulating pancreatic acinar cell differentiation. In the developing liver, Nr5a2 regulates hepatic progenitor outgrowth and differentiation, as nr5a2 mutants exhibited reduced hepatoblast markers hnf4α and prox1 as well as differentiated hepatocyte marker fabp10a. Through the first in vivo use of Nr5a2 chemical antagonist Cpd3, the iterative requirement for Nr5a2 for exocrine pancreas and liver differentiation was temporally elucidated: chemical inhibition of Nr5a2 function during hepatopancreas progenitor specification was sufficient to disrupt exocrine pancreas formation and enhance the size of the embryonic liver, suggesting that Nr5a2 regulates hepatic vs. pancreatic progenitor fate choice. Chemical inhibition of Nr5a2 at a later time during pancreas and liver differentiation was sufficient to block the formation of mature acinar cells and hepatocytes. These findings define critical iterative and pleiotropic roles for Nr5a2 at distinct stages of pancreas and liver organogenesis, and provide novel perspectives for interpreting the role of Nr5a2 in disease.
authors.affiliation: Gastroenterology Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Dana-Farber Cancer Institute, Boston, MA 02115, USA.
authors.forename: Sahar
authors.initials: S
authors.lastname: Nissim
authors.affiliation: Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Olivia
authors.initials: O
authors.lastname: Weeks
authors.affiliation: Departments of Molecular Genetics and Biological Chemistry and Pharmacology, Ohio State University, Columbus, OH 43210, USA.
authors.forename: Jared C
authors.initials: JC
authors.lastname: Talbot
authors.affiliation: Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: John W
authors.initials: JW
authors.lastname: Hedgepeth
authors.affiliation: Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Julia
authors.initials: J
authors.lastname: Wucherpfennig
authors.affiliation: Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Schatzman-Bone
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Ian
authors.initials: I
authors.lastname: Swinburne
authors.affiliation: Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Cortes
authors.affiliation: Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Kristen
authors.initials: K
authors.lastname: Alexa
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Sean
authors.initials: S
authors.lastname: Megason
authors.affiliation: Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Trista E
authors.initials: TE
authors.lastname: North
authors.affiliation: Departments of Molecular Genetics and Biological Chemistry and Pharmacology, Ohio State University, Columbus, OH 43210, USA.
authors.forename: Sharon L
authors.initials: SL
authors.lastname: Amacher
authors.affiliation: Gastroenterology Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Dana-Farber Cancer Institute, Boston, MA 02115, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: wgoessling@partners.org.
authors.forename: Wolfram
authors.initials: W
authors.lastname: Goessling
mesh_terms: D061354:Acinar Cells / Q000166:cytology*; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D004707:Endoderm / Q000166:cytology; D050556:Fatty Acid-Binding Proteins / Q000378:metabolism; D055785:Gene Knockdown Techniques; D051557:Hepatocyte Nuclear Factor 4 / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology*; D043143:Hepatopancreas / Q000196:embryology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000196:embryology*; D060172:Morpholinos / Q000235:genetics; D046790:Pancreas, Exocrine / Q000196:embryology*; D018160:Receptors, Cytoplasmic and Nuclear / Q000037:antagonists & inhibitors / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014357:Trypsin / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics; D029961:Zebrafish Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D000098551:Prospero-Related Homeobox 1 Protein
chemical_list: D050556:Fatty Acid-Binding Proteins; D051557:Hepatocyte Nuclear Factor 4; D018398:Homeodomain Proteins; D060172:Morpholinos; C515056:NR5A2 protein, zebrafish; D018160:Receptors, Cytoplasmic and Nuclear; D015534:Trans-Activators; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D029961:Zebrafish Proteins; C513422:fatty acid-binding protein 10, zebrafish; C084010:pancreatic and duodenal homeobox 1 protein; D000098551:Prospero-Related Homeobox 1 Protein; C062602:transcription factor PTF1; D014357:Trypsin
keywords: Endoderm development; Exocrine pancreas; Hepatopancreas progenitors; Liver; Nuclear receptor