NF-κB Inducing Kinase Attenuates Colorectal Cancer by Regulating Noncanonical NF-κB Mediated Colonic Epithelial Cell Regeneration

Holly A Morrison; Kristin Eden; Brie Trusiano; Daniel E Rothschild; Yufeng Qin; Paul A Wade; Audrey J Rowe; Christina Mounzer; Morgan C Stephens; Katherine M Hanson; Stephan L Brown; Eda K Holl; Irving C Allen

doi:10.1016/j.jcmgh.2024.05.004

NF-κB Inducing Kinase Attenuates Colorectal Cancer by Regulating Noncanonical NF-κB Mediated Colonic Epithelial Cell Regeneration

Cell Mol Gastroenterol Hepatol. 2024 May 13:S2352-345X(24)00110-3. doi: 10.1016/j.jcmgh.2024.05.004. Online ahead of print.

Authors

Affiliations

¹ Virginia Tech, Virginia Maryland College of Veterinary Medicine, Department of Biomedical Science and Pathobiology, Blacksburg VA 24061.
² Virginia Tech, Virginia Maryland College of Veterinary Medicine, Department of Biomedical Science and Pathobiology, Blacksburg VA 24061; Virginia Tech, Department of Basic Science Education, Virginia Tech Carilion School of Medicine, Roanoke VA 24016.
³ National Institute of Environmental Health Sciences, Research Triangle Park NC 27709.
⁴ Via College of Osteopathic Medicine, Department of Cell Biology and Physiology, Spartanburg SC 29303.
⁵ Duke University, Department of Surgery, Durham NC 27708.
⁶ Virginia Tech, Virginia Maryland College of Veterinary Medicine, Department of Biomedical Science and Pathobiology, Blacksburg VA 24061; Virginia Tech, Department of Basic Science Education, Virginia Tech Carilion School of Medicine, Roanoke VA 24016; Graduate Program in Translational Biology, Medicine and Health, Virginia Polytechnic Institute and State University, Roanoke, VA, 24016. Electronic address: icallen@vt.edu.

PMID: 38750899
DOI: 10.1016/j.jcmgh.2024.05.004

Abstract

Background & aims: Dysregulated colonic epithelial cell (CEC) proliferation is a critical feature in the development of colorectal cancer. We show that NF-κB-inducing kinase (NIK) attenuates colorectal cancer through coordinating CEC regeneration/differentiation via noncanonical NF-κB signaling that is unique from canonical NF-κB signaling.

Methods: Initial studies evaluated crypt morphology/functionality, organoid generation, transcriptome profiles, and the microbiome. Inflammation and inflammation-induced tumorigenesis was initiated in whole-body NIK knockout mice (Nik^-/-)and conditional-knockout mice following administration of azoxymethane and dextran sulfate sodium (AOM/DSS).

Results: Human transcriptomic data revealed dysregulated noncanonical NF-κB signaling. In vitro studies evaluating Nik^-/- crypts and organoids derived from mature, nondividing CECs and colonic stem cells (CSCs) exhibited increased accumulation and stunted growth, respectively. Transcriptomic analysis of Nik^-/- cells revealed gene expression signatures associated with altered differentiation-regeneration. When assessed in vivo, Nik^-/- mice exhibited more severe colitis with DSS administration and an altered microbiome characterized by increased colitogenic microbiota. In the inflammation-induced tumorigenesis model, we observed both increased tumor burdens and inflammation in mice where NIK is knocked out in CECs (Nik^ΔCEC). Interestingly, this was not recapitulated when NIK was conditionally knocked out in myeloid cells (Nik^ΔMYE). Surprisingly, conditional knockout of the canonical pathway in myeloid cells (RelA^ΔMYE) revealed decreased tumor burden and inflammation and no significant changes when conditionally knocked out in CECs (RelA^ΔCEC).

Conclusions: Dysregulated noncanonical NF-κB signaling is associated with the development of colorectal cancer in a tissue dependent manner and defines a critical role for NIK in regulating gastrointestinal inflammation and regeneration associated with colorectal cancer.

Keywords: colitis-associated tumorigenesis; differentiation; epithelial cells; organoids.