Introduction: Ulcerative colitis (UC) is an inflammatory bowel disease characterized by repeated remissions and relapses. Immunosuppressive drugs have facilitated the induction and maintenance of remission in many patients with UC. However, immunosuppressive drugs cannot directly repair impaired intestinal mucosa and are insufficient for preventing relapse. Therefore, new treatment approaches to repair the damaged epithelium in UC have been attempted through the transplantation of intestinal organoids, which can be differentiated into mucosa by embedding in Matrigel, generated from patient-derived intestinal stem cells. The method, however, poses the challenge of yielding sufficient cells for UC therapy, and patient-derived cells might already have acquired pathological changes. In contrast, human induced pluripotent stem (iPS) cells generated from healthy individuals are infinitely proliferated and can be differentiated into target cells. Recently developed human iPS cell-derived intestinal organoids (HIOs) aim to generate organoids that closely resemble the adult intestine. However, no study till date has reported HIOs injected into in vivo inflammatory models, and it remains unclear whether HIOs with cells that closely resemble the adult intestine or with intestinal stem cells retain the better ability to repair tissue in colitis.
Methods: We generated two types of HIOs via suspension culture with and without small-molecule compounds: HIOs that include predominantly more intestinal stem cells [HIO (A)] and those that include predominantly more intestinal epithelial and secretory cells [HIO (B)]. We examined whether the generated HIOs engrafted in vivo and compared their ability to accelerate recovery of the damaged tissue.
Results: Findings showed that the HIOs expressed intestinal-specific markers such as caudal-type homeobox 2 (CDX2) and villin, and HIOs engrafted under the kidney capsules of mice. We then injected HIOs into colitis-model mice and found that the weight and clinical score of the mice injected with HIO (A) recovered earlier than that of the mice in the sham group. Further, the production of mucus and the expression of cell proliferation markers and tight junction proteins in the colon tissues of the HIO (A) group were restored to levels similar to those observed in healthy mice. However, neither HIO (A) nor HIO (B) could be engrafted into the colon.
Conclusions: Effective cell therapy should directly repair tissue by engraftment at the site of injury. However, the difference in organoid property impacting the rate of tissue repair in transplantation without engraftment observed in the current study should be considered a critical consideration in the development of regenerative medicine using iPS-derived organoids.
Keywords: 5-aza, 5-aza-2′-deoxycytidine; A-83-01, 3-(6-methyl-2-pyridinyl)-N-phenyl-4-(4-quinolinyl)-1H-pyrazole-1-carbothioamide; CDX2, caudal-type homeobox 2; CHGA, chromogranin A; Cell therapy; DAPI, 4′,6-diamidino-2-phenylindole; DAPT, N-[(3,5-difluorophenyl)acetyl]-L-alanyl-2-phenyl-1,1-dime-thylethyl ester-glycine; DSS, dextran sodium sulfate; FBS, fetal bovine serum; HIO, human induced pluripotent stem cell-derived intestinal organoid; HLA, human leukocyte antigen; HPRT, hypoxanthine phosphoribosyltransferase; Human induced pluripotent stem cell; Inflammatory bowel disease; Intestinal organoid; LGR5, leucine-rich repeat-containing G-protein-coupled receptor 5; MUC2, mucin 2; NSG, NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ; OLFM4, olfactomedin 4; PBS, phosphate-buffered saline; PD98059, 2-(2-amino-3-methoxyphenyl)4-H-1-benzopyran-4-one; SCID-Beige, CB17.Cg-PrkdcscidLystbg-J/CrlCrlj; Suspension culture; UC, ulcerative colitis; Ulcerative colitis; VIL1, villin 1; Y-27632, (+)-(R)-trans-4-(1-amino-ethyl)-N-(4-pyridyl) cyclohexanecarboxamide dihydrochloride; iPS, induced pluripotent stem; qPCR, quantitative polymerase chain reaction; α-SMA, α-smooth muscle actin.
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