Pediatric Gastroenterology, Hepatology & Nutrition

The Korean Society of Pediatric Gastroenterology, Hepatology and Nutrition (대한소아소화기영양학회)
권호 표지
DOI QR코드

DOI QR Code

Establishment and Characterization of Patient-Derived Intestinal Organoids from Pediatric Crohn's Disease Patients

  • Sunghyun An (Department of Pediatrics, Hallym University Industry Academic Cooperation Foundation) ;
  • Homin Huh (Department of Pediatrics, Seoul National University College of Medicine) ;
  • Jae Sung Ko (Department of Pediatrics, Seoul National University College of Medicine) ;
  • Jin Soo Moon (Department of Pediatrics, Seoul National University College of Medicine) ;
  • Ky Young Cho (Department of Pediatrics, Hallym University Kangnam Sacred Heart Hospital)
  • Received : 2024.09.22
  • Accepted : 2024.10.07
  • Published : 2024.11.15
Download PDF
⟨ Previous Next ⟩

Abstract

Purpose: This study aimed to establish and characterize patient-derived intestinal organoids (PDOs) from children with Crohn's disease (CD). Methods: To generate PDOs, endoscopic biopsy specimens were obtained from non-inflamed duodenal bulbs of normal controls and CD patients. To verify the presence of PDOs, histological staining and quantitative reverse transcription polymerase chain reaction (RT-qPCR) analyses were performed. Results: PDOs were successfully established in normal controls (n=2) and CD patients (n=2). Hematoxylin and eosin staining of formalin-fixed, paraffin-embedded PDO sections revealed crypt and villus structures, whereas immunofluorescence staining with EpCAM and DAPI confirmed the epithelial-specific architecture of the PDOs. RT-qPCR results revealed a significant increase in Lgr5, Si, and Chga gene expression and a decrease in Olfm4 and Muc2 expression in CD patients compared to normal controls, suggesting altered stem cell activity and mucosal barrier function (p<0.05). Conclusion: We successfully established and characterized PDOs in children with CD, providing a valuable tool for understanding the pathophysiology of the disease and evaluating potential therapeutic approaches. The differential gene expression of PDOs in CD patients might be caused by the complex interplay between epithelial adaptation and inflammation in the intestinal epithelium.

Keywords

Acknowledgement

This research was supported and funded by SNUH Lee Kun-hee Child Cancer & Rare Disease Project, Republic of Korea (22C-017-0200), Hallym University Medical Center Research Fund, and the Korean Society of Pediatric Gastroenterology, Hepatology and Nutrition Research Fund. The funding bodies had no role in the study design, collection, analysis, or interpretation of the data or writing of the manuscript.

References

  1. Long D. Crohn's disease and ulcerative colitis: from pathophysiology to novel therapeutic approaches. Biomedicines 2024;12:689.
  2. Orfanoudaki E, Foteinogiannopoulou K, Theodoraki E, Koutroubakis IE. Recent advances in the optimization of anti-TNF treatment in patients with inflammatory bowel disease. J Clin Med 2023;12:2452.
  3. Al Meslamani AZ. Understanding patient variability to biologic treatments in inflammatory bowel disease. Expert Opin Biol Ther 2024;24:317-9.
  4. Wlazlo M, Meglicka M, Wiernicka A, Osiecki M, Kierkus J. Dual biologic therapy in moderate to severe pediatric inflammatory bowel disease: a retrospective study. Children (Basel) 2022;10:11.
  5. Scheurlen KM, Parks MA, Macleod A, Galandiuk S. Unmet challenges in patients with Crohn's disease. J Clin Med 2023;12:5595.
  6. Clinton JW, Cross RK. Personalized treatment for Crohn's disease: current approaches and future directions. Clin Exp Gastroenterol 2023;16:249-76.
  7. Urzi O, Gasparro R, Costanzo E, De Luca A, Giavaresi G, Fontana S, et al. Three-dimensional cell cultures: the bridge between in vitro and in vivo models. Int J Mol Sci 2023;24:12046.
  8. Kiani AK, Pheby D, Henehan G, Brown R, Sieving P, Sykora P, et al.; International Bioethics Study Group. Ethical considerations regarding animal experimentation. J Prev Med Hyg 2022;63(2 Suppl 3):E255-66.
  9. Rudolph JE, Zhong Y, Duggal P, Mehta SH, Lau B. Defining representativeness of study samples in medical and population health research. BMJ Med 2023;2:e000399.
  10. Ollivier A, Mahe MM, Guasch G. Modeling gastrointestinal diseases using organoids to understand healing and regenerative processes. Cells 2021;10:1331.
  11. Kim J, Koo BK, Knoblich JA. Human organoids: model systems for human biology and medicine. Nat Rev Mol Cell Biol 2020;21:571-84.
  12. Tian CM, Yang MF, Xu HM, Zhu MZ, Yue NN, Zhang Y, et al. Stem cell-derived intestinal organoids: a novel modality for IBD. Cell Death Discov 2023;9:255.
  13. Niklinska-Schirtz BJ, Venkateswaran S, Anbazhagan M, Kolachala VL, Prince J, Dodd A, et al. Ileal derived organoids from Crohn's disease patients show unique transcriptomic and secretomic signatures. Cell Mol Gastroenterol Hepatol 2021;12:1267-80.
  14. Laudadio I, Carissimi C, Scafa N, Bastianelli A, Fulci V, Renzini A, et al. Characterization of patient-derived intestinal organoids for modelling fibrosis in Inflammatory bowel disease. Inflamm Res 2024;73:1359-70.
  15. Gasparetto M, Guariso G. Crohn's disease and growth deficiency in children and adolescents. World J Gastroenterol 2014;20:13219-33.
  16. van Rheenen PF, Aloi M, Assa A, Bronsky J, Escher JC, Fagerberg UL, et al. The medical management of paediatric Crohn's disease: an ECCO-ESPGHAN guideline update. J Crohns Colitis 2020:jjaa161.
  17. Senger S, Ingano L, Freire R, Anselmo A, Zhu W, Sadreyev R, et al. Human fetal-derived enterospheres provide insights on intestinal development and a novel model to study necrotizing enterocolitis (NEC). Cell Mol Gastroenterol Hepatol 2018;5:549-68.
  18. Ouchi T, Morimura S, Dow LE, Miyoshi H, Udey MC. EpCAM (CD326) regulates intestinal epithelial integrity and stem cells via rho-associated kinase. Cells 2021;10:256.
  19. Basak O, Beumer J, Wiebrands K, Seno H, van Oudenaarden A, Clevers H. Induced quiescence of Lgr5+ stem cells in intestinal organoids enables differentiation of hormone-producing enteroendocrine cells. Cell Stem Cell 2017;20:177-90.e4.
  20. Sakahara M, Okamoto T, Srivastava U, Natsume Y, Yamanaka H, Suzuki Y, et al. Paneth-like cells produced from OLFM4+ stem cells support OLFM4+ stem cell growth in advanced colorectal cancer. Commun Biol 2024;7:27.
  21. Tsai YH, Wu A, Wu JH, Capeling MM, Holloway EM, Huang S, et al. Acquisition of NOTCH dependence is a hallmark of human intestinal stem cell maturation. Stem Cell Reports 2022;17:1138-53.
  22. Santos AJM, Lo YH, Mah AT, Kuo CJ. The intestinal stem cell niche: homeostasis and adaptations. Trends Cell Biol 2018;28:1062-78.
  23. Tucci SA, Boyland EJ, Halford JC. The role of lipid and carbohydrate digestive enzyme inhibitors in the management of obesity: a review of current and emerging therapeutic agents. Diabetes Metab Syndr Obes 2010;3:125-43.
  24. Rath E, Haller D. Intestinal epithelial cell metabolism at the interface of microbial dysbiosis and tissue injury. Mucosal Immunol 2022;15:595-604.
  25. Dorofeyev AE, Vasilenko IV, Rassokhina OA, Kondratiuk RB. Mucosal barrier in ulcerative colitis and Crohn's disease. Gastroenterol Res Pract 2013;2013:431231.
  26. Grondin JA, Kwon YH, Far PM, Haq S, Khan WI. Mucins in intestinal mucosal defense and inflammation: learning from clinical and experimental studies. Front Immunol 2020;11:2054.
  27. Eissa N, Hussein H, Tshikudi DM, Hendy GN, Bernstein CN, Ghia JE. Interdependence between chromogranin-A, alternatively activated macrophages, tight junction proteins and the epithelial functions. A human and in-vivo/in-vitro descriptive study. Int J Mol Sci 2020;21:7976.
  28. Jin HY, Lim JS, Lee Y, Choi Y, Oh SH, Kim KM, et al. Growth, puberty, and bone health in children and adolescents with inflammatory bowel disease. BMC Pediatr 2021;21:35.
  29. Benchimol EI, Fortinsky KJ, Gozdyra P, Van den Heuvel M, Van Limbergen J, Griffiths AM. Epidemiology of pediatric inflammatory bowel disease: a systematic review of international trends. Inflamm Bowel Dis 2011;17:423-39.
  30. Lahad A, Weiss B. Current therapy of pediatric Crohn's disease. World J Gastrointest Pathophysiol 2015;6:33-42.