In preclinical studies, the cynomolgus monkey (CM) model is frequently used to predict the pharmacokinetics of drugs in the human small intestine, because of its evolutionary closeness to humans. Intestinal organoids that mimic the intestinal tissue have attracted attention in regenerative medicine and drug development. In this study, we generated intestinal organoids from CM induced pluripotent stem (CMiPS) cells and analyzed their pharmacokinetic functions. CMiPS cells were induced into the hindgut; then, the cells were seeded on microfabricated culture vessel plates to form spheroids. The resulting floating spheroids were differentiated into intestinal organoids in a medium containing small-molecule compounds. The mRNA expression of intestinal markers and pharmacokinetic-related genes was markedly increased in the presence of small-molecule compounds. The organoids possessed a polarized epithelium and contained various cells constituting small intestinal tissues. The intestinal organoids formed functional tight junctions and expressed drug transporter proteins. In addition, in the organoids generated, cytochrome P450 3A8 (CYP3A8) activity was inhibited by the specific inhibitor ketoconazole and was induced by rifampicin. Therefore, in the present work, we successfully generated intestinal organoids, with pharmacokinetic functions, from CMiPS cells using small-molecule compounds.
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References
1.
KomoriM, KikuchiO, SakumaT, FunakiJ, KitadaM and KamatakiT. (1992). Molecular cloning of monkey liver cytochrome P-450 cDNAs: similarity of the primary sequences to human cytochromes P-450. Biochim Biophys Acta, 1171:141–146.
2.
ShimadaT, MimuraM, InoueK, NakamuraS, OdaH, OhmoriS and YamazakiH. (1997). Cytochrome P450-dependent drug oxidation activities in liver microsomes of various animal species including rats, guinea pigs, dogs, monkeys, and humans. Arch Toxicol, 71:401–408.
3.
UnoY, HosakaS, MatsunoK, NakamuraC, KitoG, KamatakiT and NagataR. (2007). Characterization of cynomolgus monkey cytochrome P450 (CYP) cDNAs: is CYP2C76 the only monkey-specific CYP gene responsible for species differences in drug metabolism?. Arch Biochem Biophys, 466:98–105.
4.
OgasawaraA, KumeT and KazamaE. (2007). Effect of oral ketoconazole on intestinal first-pass effect of midazolam and fexofenadine in cynomolgus monkeys. Drug Metab Dispos, 35:410–418.
5.
OgasawaraA, UtohM, NiiK, UedaA, YoshikawaT, KumeT and FukuzakiK. (2009). Effect of oral ketoconazole on oral and intravenous pharmacokinetics of simvastatin and its acid in cynomolgus monkeys. Drug Metab Dispos, 37:122–128.
6.
WardKW and SmithBR. (2004). A comprehensive quantitative and qualitative evaluation of extrapolation of intravenous pharmacokinetic parameters from rat, dog, and monkey to humans. II. Volume of distribution and mean residence time. Drug Metab Dispos, 32:612–619.
7.
NagillaR and WardKW. (2004). A comprehensive analysis of the role of correction factors in the allometric predictivity of clearance from rat, dog, and monkey to humans. J Pharm Sci, 93:2522–2534.
8.
ChoiJS, ChoiI and ChoiDH. (2013). Effects of nifedipine on the pharmacokinetics of repaglinide in rats: possible role of CYP3A4 and P-glycoprotein inhibition by nifedipine. Pharmacol Rep, 65:1422–1430.
9.
YoshidaK, MaedaK and SugiyamaY. (2013). Hepatic and intestinal drug transporters: prediction of pharmacokinetic effects caused by drug-drug interactions and genetic polymorphisms. Annu Rev Pharmacol Toxicol, 53:581–612.
10.
KostewiczES, AbrahamssonB, BrewsterM, BrouwersJ, ButlerJ, CarlertS, DickinsonPA, DressmanJ, HolmR, et al. (2014). In vitro models for the prediction of in vivo performance of oral dosage forms. Eur J Pharm Sci, 57:342–366.
11.
NishimuraM, KoedaA, MorikawaH, SatohT, NarimatsuS and NaitoS. (2009). Tissue-specific mRNA expression profiles of drug-metabolizing enzymes and transporters in the cynomolgus monkey. Drug Metab Pharmacokinet, 24:139–144.
12.
ZhangQY, DunbarD, OstrowskaA, ZeisloftS, YangJ and KaminskyLS. (1999). Characterization of human small intestinal cytochromes P-450. Drug Metab Dispos, 27:804–809.
13.
HilgendorfC, AhlinG, SeithelA, ArturssonP, UngellAL and KarlssonJ. (2007). Expression of thirty-six drug transporter genes in human intestine, liver, kidney, and organotypic cell lines. Drug Metab Dispos, 35:1333–1340.
14.
BaldrickP. (2017). Pharmacokinetic and toxicology comparator testing of biosimilar drugs—assessing need. Regul Toxicol Pharmacol, 86:386–391.
15.
TakahashiM, WashioT, SuzukiN, IgetaK and YamashitaS. (2009). The species differences of intestinal drug absorption and first-pass metabolism between cynomolgus monkeys and humans. J Pharm Sci, 98:4343–4353.
16.
NishimuraT, AmanoN, KuboY, OnoM, KatoY, FujitaH, KimuraY and TsujiA. (2007). Asymmetric intestinal first-pass metabolism causes minimal oral bioavailability of midazolam in cynomolgus monkey. Drug Metab Dispos, 35:1275–1284.
17.
SatoT, VriesRG, SnippertHJ, van de WeteringM, BarkerN, StangeDE, van EsJH, AboA, KujalaP, PetersPJ and CleversH. (2009). Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature, 459:262–265.
18.
SpenceJR, MayhewCN, RankinSA, KuharMF, VallanceJE, TolleK, HoskinsEE, KalinichenkoVV, WellsSI, et al. (2011). Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro. Nature, 470:105–109.
19.
MizutaniT, NakamuraT, MorikawaR, FukudaM, MochizukiW, YamauchiY, NozakiK, YuiS, NemotoY, et al. (2012). Real-time analysis of P-glycoprotein-mediated drug transport across primary intestinal epithelium three-dimensionally cultured in vitro. Biochem Biophys Res Commun, 419:238–243.
20.
OkabayashiK and AsashimaM. (2003). Tissue generation from amphibian animal caps. Curr Opin Genet Dev, 13:502–507.
21.
Juuti-UusitaloK, KlunderLJ, SjollemaKA, MackovicovaK, OhgakiR, HoekstraD, DekkerJ and van IjzendoornSC. (2011). Differential effects of TNF (TNFSF2) and IFN-γ on intestinal epithelial cell morphogenesis and barrier function in three-dimensional culture. PLoS One, 6:e22967.
NakatsujiN and SuemoriH. (2002). Embryonic stem cell lines of nonhuman primates. ScientificWorldJournal, 2:1762–1773.
24.
SatoH, IdirisA, MiwaT and KumagaiH. (2016). Microfabric vessels for embryoid body formation and rapid differentiation of pluripotent stem cells. Sci Rep, 6:31063.
25.
KimS, DinchukJE, AnthonyMN, OrcuttT, ZoecklerME, SauerMB, MosureKW, VuppugallaR, GraceJEJr., et al. (2010). Evaluation of cynomolgus monkey pregnane X receptor, primary hepatocyte, and in vivo pharmacokinetic changes in predicting human CYP3A4 induction. Drug Metab Dispos, 38:16–24.
26.
BollerK, ArpinM, PringaultE, MangeatP and ReggioH. (1988). Differential distribution of villin and villin MRNA in mouse intestinal epithelial cells. Differentiation, 39:51–57.
27.
GuN, AdachiT, MatsunagaT, TsujimotoG, IshiharaA, YasudaK and TsudaK. (2007). HNF-1α participates in glucose regulation of sucrase-isomaltase gene expression in epithelial intestinal cells. Biochem Biophys Res Commun, 353:617–622.
28.
ReisCA, DavidL, CorreaP, CarneiroF, de BolósC, GarciaE, MandelU, ClausenH and Sobrinho-SimõesM. (1999). Intestinal metaplasia of human stomach displays distinct patterns of mucin (MUC1, MUC2, MUC5AC, and MUC6) expression. Cancer Res, 59:1003–1007.
29.
CetinY, Müller-KöppelL, AunisD, BaderMF and GrubeD. (1989). Chromogranin A (CgA) in the gastro-entero-pancreatic (GEP) endocrine system. II. CgA in mammalian entero-endocrine cells. Histochemistry, 92:265–275.
30.
WardM, FergusonA and EastwoodMA. (1979). Jejunal lysozyme activity and the Paneth cell in coeliac disease. Gut, 20:55–58.
31.
van der FlierLG, HaegebarthA, StangeDE, van de WeteringM and CleversH. (2009). OLFM4 is a robust marker for stem cells in human intestine and marks a subset of colorectal cancer cells. Gastroenterology, 137:15–17.
32.
SchuijersJ, van der FlierLG, van EsJ and CleversH. (2014). Robust cre-mediated recombination in small intestinal stem cells utilizing the olfm4 locus. Stem Cell Reports, 3:234–241.
33.
BerndorffD, GessnerR, KreftB, SchnoyN, Lajous-PetterAM, LochN, ReutterW, HortschM and TauberR. (1994). Liver-intestine cadherin: molecular cloning and characterization of a novel Ca(2+)-dependent cell adhesion molecule expressed in liver and intestine. J Cell Biol, 125:1353–1369.
34.
KosinskiC, StangeDE, XuC, ChanAS, HoC, YuenST, MifflinRC, PowellDW, CleversH, LeungSY and ChenX. (2010). Indian hedgehog regulates intestinal stem cell fate through epithelial-mesenchymal interactions during development. Gastroenterology, 139:893–903.
ThummelKE, BrimerC, YasudaK, ThottasseryJ, SennT, LinY, IshizukaH, KharaschE, SchuetzJ and SchuetzE. (2001). Transcriptional control of intestinal cytochrome P-4503A by 1α,25-dihydroxy vitamin D3. Mol Pharmacol, 60:1399–1406.
37.
DedhiaPH, Bertaux-SkeirikN, ZavrosY and SpenceJR. (2016). Organoid models of human gastrointestinal development and disease. Gastroenterology, 150:1098–1112.
38.
ZachosNC, KovbasnjukO, Foulke-AbelJ, InJ, BluttSE, de JongeHR, EstesMK and DonowitzM. (2016). Human enteroids/colonoids and intestinal organoids functionally recapitulate normal intestinal physiology and pathophysiology. J Biol Chem, 291:3759–3766.
39.
FinkbeinerSR and SpenceJR. (2013). A gutsy task: generating intestinal tissue from human pluripotent stem cells. Dig Dis Sci, 58:1176–1184.
40.
McCrackenKW, HowellJC, WellsJM and SpenceJR. (2011). Generating human intestinal tissue from pluripotent stem cells in vitro. Nat Protoc, 6:1920–1928.
41.
HwangYS, ChungBG, OrtmannD, HattoriN, MoellerHC and KhademhosseiniA. (2009). Microwell-mediated control of embryoid body size regulates embryonic stem cell fate via differential expression of WNT5a and WNT11. Proc Natl Acad Sci U S A, 106:16978–16983.
42.
IwaoT, KodamaN, KondoY, KabeyaT, NakamuraK, HorikawaT, NiwaT, KuroseK and MatsunagaT. (2015). Generation of enterocyte-like cells with pharmacokinetic functions from human induced pluripotent stem cells using small-molecule compounds. Drug Metab Dispos, 43:603–610.
43.
OgakiS, ShirakiN, KumeK and KumeS. (2013). Wnt and Notch signals guide embryonic stem cell differentiation into the intestinal lineages. Stem Cells, 31:1086–1096.
44.
LaharN, LeiNY, WangJ, JabajiZ, TungSC, JoshiV, LewisM, StelznerM, MartínMG and DunnJC. (2011). Intestinal subepithelial myofibroblasts support in vitro and in vivo growth of human small intestinal epithelium. PLoS One, 6:e26898.
45.
LeiNY, JabajiZ, WangJ, JoshiVS, BrinkleyGJ, KhalilH, WangF, JaroszewiczA, PellegriniM, et al. (2014). Intestinal subepithelial myofibroblasts support the growth of intestinal epithelial stem cells. PLoS One, 9:e84651.
46.
International Transporter Consortium, GiacominiKM, HuangSM, TweedieDJ, BenetLZ, BrouwerKL, ChuX, DahlinA, EversR, FischerV, et al. (2010). Membrane transporters in drug development. Nat Rev Drug Discov, 9:215–236.
47.
NakanishiY, MatsushitaA, MatsunoK, IwasakiK, UtohM, NakamuraC and UnoY. (2010). Regional distribution of cytochrome p450 mRNA expression in the liver and small intestine of cynomolgus monkeys. Drug Metab Pharmacokinet, 25:290–297.
48.
UnoY, UeharaS and YamazakiH. (2016). Utility of non-human primates in drug development: comparison of non-human primate and human drug-metabolizing cytochrome P450 enzymes. Biochem Pharmacol, 121:1–7.
49.
OkamotoR and WatanabeM. (2016). Role of epithelial cells in the pathogenesis and treatment of inflammatory bowel disease. J Gastroenterol, 51:11–21.
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