title: Intestinal organoids: a model of intestinal fibrosis for evaluating anti-fibrotic drugs.
abstract: OBJECTIVE
  Intestinal fibrosis is a critical complication of Crohn's disease (CD). Current in vitro models of intestinal fibrosis cannot model the complex intestinal architecture, while in vivo rodent models do not fully recapitulate human disease and have limited utility for large-scale screening. Here, we exploit recent advances in stem cell derived human intestinal organoids (HIOs) as a new human model of fibrosis in CD.
  
  
  METHODS
  Human pluripotent stem cells were differentiated into HIOs. We identified myofibroblasts, the key effector cells of fibrosis, by immunofluorescence staining for alpha-smooth muscle actin (αSMA), vimentin, and desmin. We examined the fibrogenic response of HIOs by treatment with transforming growth factor beta (TGFβ) in the presence or absence of the anti-fibrotic drug spironolactone. Fibrotic response was assayed by expression of fibrogenic genes (COL1A1 (collagen, type I, alpha 1), ACTA2 (alpha smooth muscle actin), FN1 (fibronectin 1), MYLK (myosin light chain kinase), and MKL1 (megakaryoblastic leukemia (translocation) 1)) and proteins (αSMA).
  
  
  RESULTS
  Immunofluorescent staining of organoids identified a population of myofibroblasts within the HIO mesenchyme. TGFβ stimulation of HIOs produced a dose-dependent pro-fibrotic response. Spironolactone treatment blocked the fibrogenic response of HIOs to TGFβ.
  
  
  CONCLUSIONS
  HIOs contain myofibroblasts and respond to a pro-fibrotic stimulus in a manner that is consistent with isolated human myofibroblasts. HIOs are a promising model system that might bridge the gap between current in vitro and in vivo models of intestinal fibrosis in IBD.
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Eva S
authors.initials: ES
authors.lastname: Rodansky
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Johnson
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Sha
authors.initials: S
authors.lastname: Huang
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA; Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Jason R
authors.initials: JR
authors.lastname: Spence
authors.affiliation: Division of Gastroenterology, Department of Internal Medicine, University of Michigan, Ann Arbor, MI, USA. Electronic address: phiggins@med.umich.edu.
authors.forename: Peter D R
authors.initials: PD
authors.lastname: Higgins
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D000090602:Collagen Type I, alpha 1 Chain; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D005355:Fibrosis / Q000188:drug therapy; D006801:Humans; D007422:Intestines / Q000187:drug effects* / Q000473:pathology; D058628:Myofibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000235:genetics / Q000378:metabolism; D009940:Organoids / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects; D013148:Spironolactone / Q000494:pharmacology* / Q000627:therapeutic use; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000494:pharmacology
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D002135:Calcium-Binding Proteins; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; C067625:FN1 protein, human; D005353:Fibronectins; C431575:MRTFA protein, human; D015534:Trans-Activators; D016212:Transforming Growth Factor beta; D013148:Spironolactone; C499378:MYLK protein, human; D009219:Myosin-Light-Chain Kinase
keywords: Drug screening; Fibrosis model; Myofibroblasts; Organoid; TGFβ
references.citation: Med Clin North Am. 2003 Mar;87(2):441-57
references.pmid: 12693733
references.citation: Int J Biochem Cell Biol. 2012 Jan;44(1):158-69
references.pmid: 22041029
references.citation: Inflamm Bowel Dis. 2014 Jan;20(1):154-65
references.pmid: 24280883
references.citation: J Heart Lung Transplant. 1994 Jul-Aug;13(4):S107-12
references.pmid: 7947865
references.citation: Inflamm Bowel Dis. 2012 Jul;18(7):1315-24
references.pmid: 22081497
references.citation: Gastroenterology. 2008 Mar;134(3):768-80
references.pmid: 18325390
references.citation: Nature. 2011 Feb 3;470(7332):105-9
references.pmid: 21151107
references.citation: Ultrasound Med Biol. 2008 Jun;34(6):902-12
references.pmid: 18294759
references.citation: Am J Physiol. 1999 Aug;277(2 Pt 1):C183-201
references.pmid: 10444394
references.citation: Cell Res. 2008 Oct;18(10):997-1006
references.pmid: 18766170
references.citation: Cell Rep. 2014 Mar 27;6(6):1046-1058
references.pmid: 24613355
references.citation: Gastroenterol Clin North Am. 1999 Jun;28(2):255-81, vii
references.pmid: 10372268
references.citation: Infect Immun. 2015 Jan;83(1):138-45
references.pmid: 25312952
references.citation: Nat Protoc. 2011 Nov 10;6(12):1920-8
references.pmid: 22082986
references.citation: Adv Drug Deliv Rev. 2014 Apr;69-70:1-18
references.pmid: 24613390
references.citation: Perit Dial Int. 2007 Jul-Aug;27(4):424-31
references.pmid: 17602151
references.citation: N Engl J Med. 1999 Sep 2;341(10):709-17
references.pmid: 10471456
references.citation: Am J Gastroenterol. 2003 Dec;98(12):2712-8
references.pmid: 14687822
references.citation: J Cell Biol. 2010 Aug 23;190(4):693-706
references.pmid: 20733059
references.citation: Ren Fail. 2006;28(6):509-14
references.pmid: 16928621
references.citation: Adv Drug Deliv Rev. 2014 Apr;69-70:19-28
references.pmid: 24582599
references.citation: Am J Physiol Renal Physiol. 2008 May;294(5):F1084-93
references.pmid: 18353870
references.citation: PLoS One. 2013;8(1):e54543
references.pmid: 23382912
references.citation: Gastroenterol Hepatol (N Y). 2011 Oct;7(10):661-71
references.pmid: 22298959
references.citation: Exp Mol Pathol. 2012 Aug;93(1):91-8
references.pmid: 22542712
references.citation: J Biomech. 2010 Jan 5;43(1):146-55
references.pmid: 19800625
references.citation: Am J Kidney Dis. 2010 Jan;55(1 Suppl 1):S1-420, A6-7
references.pmid: 20082919
references.citation: J Immunol. 2004 Mar 1;172(5):2731-8
references.pmid: 14978070
references.citation: Inflamm Bowel Dis. 2013 Apr;19(5):891-903
references.pmid: 23502354
references.citation: J Med Econ. 2012;15(5):829-35
references.pmid: 22455577
references.citation: Inflamm Bowel Dis. 2012 Mar;18(3):460-71
references.pmid: 21761511
references.citation: Int Rev Immunol. 2000;19(1):51-62
references.pmid: 10723677
references.citation: PLoS One. 2011;6(5):e19929
references.pmid: 21637769
references.citation: Gut. 2007 Jan;56(1):130-9
references.pmid: 17172588