Scattered Crypt Intestinal Epithelial Cell Apoptosis Induces Necrotizing Enterocolitis Via Intricate Mechanisms

Authors

Saravanan Subramanian, Pediatric Mucosal Inflammation and Regeneration Research Program, Center for Pediatric Translational Research and Education, Department of Pediatrics, College of Medicine, University of Illinois at Chicago, Chicago, Illinois.
Heng-Fu Bu, Pediatric Mucosal Inflammation and Regeneration Research Program, Center for Pediatric Translational Research and Education, Department of Pediatrics, College of Medicine, University of Illinois at Chicago, Chicago, Illinois.
Pauline M. Chou, Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois.
Xiao Wang, Pediatric Mucosal Inflammation and Regeneration Research Program, Center for Pediatric Translational Research and Education, Department of Pediatrics, College of Medicine, University of Illinois at Chicago, Chicago, Illinois.
Hua Geng, Pediatric Mucosal Inflammation and Regeneration Research Program, Center for Pediatric Translational Research and Education, Department of Pediatrics, College of Medicine, University of Illinois at Chicago, Chicago, Illinois.
Suhail Akhtar, A.T. Still UniversityFollow
Chao Du, Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, Illinois.
Stephanie C. Tan, Department of Medical Education, Stritch School of Medicine, Loyola University Chicago, Maywood, Illinois.
Justin Eze Ideozu, Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, Illinois.
Aasrita Tulluri, Pediatric Mucosal Inflammation and Regeneration Research Program, Center for Pediatric Translational Research and Education, Department of Pediatrics, College of Medicine, University of Illinois at Chicago, Chicago, Illinois.
Yuxiang Sun, Department of Nutrition, Texas A&M University, College Station, Texas.
Wen-Xen Ding, Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, Kansas.
Isabelle G. De Plaen, Division of Neonatology, Department of Pediatrics, Feinberg School of Medicine, Northwestern University, Chicago, Illinois.
Xiao-Di Tan, Pediatric Mucosal Inflammation and Regeneration Research Program, Center for Pediatric Translational Research and Education, Department of Pediatrics, College of Medicine, University of Illinois at Chicago, Chicago, Illinois; Department of Research & Development, Jesse Brown Veterans Affairs Medical Center, Chicago, Illinois.

Document Type

Article

Abstract

Abstract

Background & aims: Necrotizing enterocolitis (NEC) is a life-threatening disease affecting mostly the ileum of preemies. Intestinal epithelial cell (IEC) apoptosis contributes to NEC pathogenesis. However, how scattered crypt IEC apoptosis leads to NEC with excessive villus epithelial necrosis remains unclear.

Methods: A novel triple-transgenic mouse model, namely, 3xTg-iAPcIEC (inducible apoptosis phenotype in crypt-IEC), was developed to induce IEC-specific overexpression of Fasl transgene using doxycycline (Dox)-inducible tetO-rtTA system and villin-cre technology. The 3-days-old neonatal 3xTg-iAPcIEC mice and their littermate controls were subcutaneously (s.c.) challenged with a single dose of Dox. Intestinal tissues were processed at different time points to examine scattered crypt IEC apoptosis-mediated NEC development. Gene knockout technology, antibody-mediated cell depletion, and antibiotic-facilitated Gram-positive bacteria depletion were used to study mechanisms.

Results: Treatment of 3xTg-iAPcIEC mouse pups with Dox induces scattered crypt IEC apoptosis followed by crypt inflammation and excessive villous necrosis resembling NEC. This progression correlated with elevated Ifng, Rip3, CD8+ T cells, and Gram-positive bacteria in the ileum. Mechanistically, IFN-γ and RIP3-activated signals mediate the effect of scattered crypt IEC apoptosis on the induction of intestinal crypt inflammation and villous necrosis. Meanwhile, pathophysiological events of CD8+ T cell infiltration and dysbiosis with Gram-positive bacteria primarily contribute to excessive villous inflammation and necrosis. Notably, blocking any of these events protects against NEC development in 3xTg-iAPcIEC mouse pups, underlining their central roles in NEC pathogenesis.

Conclusions: Scattered crypt IEC apoptosis induces NEC in mouse pups via IFN-γ, RIP3, CD8+ T cells, and Gram-positive bacteria-mediated comprehensive pathophysiological events. Our findings may advance knowledge in the prevention and treatment of NEC.

DOI

10.1016/j.jcmgh.2024.05.012.

Publication Date

5-2023

Comments

Subramanian, S., Bu, H. F., Chou, P. M., Wang, X., Geng, H., Akhtar, S., Du, C., Tan, S. C., Ideozu, J. E., Tulluri, A., Sun, Y., Ding, W. X., De Plaen, I. G., & Tan, X. D. (2024). Scattered Crypt Intestinal Epithelial Cell Apoptosis Induces Necrotizing Enterocolitis Via Intricate Mechanisms. Cellular and molecular gastroenterology and hepatology, 18(3), 101364.

https://doi.org/10.1016/j.jcmgh.2024.05.012

No Information on institutional repositories; open access

Recommended Citation

Subramanian, S., Bu, H. F., Chou, P. M., Wang, X., Geng, H., Akhtar, S., Du, C., Tan, S. C., Ideozu, J. E., Tulluri, A., Sun, Y., Ding, W. X., De Plaen, I. G., & Tan, X. D. (2024). Scattered Crypt Intestinal Epithelial Cell Apoptosis Induces Necrotizing Enterocolitis Via Intricate Mechanisms. Cellular and molecular gastroenterology and hepatology, 18(3), 101364.