2. Academic Validation
  3. Schlafen 12 Interaction with SerpinB12 and Deubiquitylases Drives Human Enterocyte Differentiation

Schlafen 12 Interaction with SerpinB12 and Deubiquitylases Drives Human Enterocyte Differentiation

  • Cell Physiol Biochem. 2018;48(3):1274-1290. doi: 10.1159/000492019.
Marc D Basson 1,  Qinggang Wang 1,  Lakshmi S Chaturvedi 1 2,  Shyam More 1,  Emilie E Vomhof-DeKrey 1,  Sarmad Al-Marsoummi 1,  Kelian Sun 1,  Leslie A Kuhn 3,  Pavlo Kovalenko 1 4,  Matti Kiupel 5
Affiliations

Affiliations

  • 1 Departments of Surgery, Pathology, and Biomedical Sciences, University of North Dakota School of Medicine and the Health Sciences, Cambridge, Massachusetts, USA.
  • 2 Currently at Departments of Pharmaceutical Sciences and Biomedical Sciences-College of Pharmacy, Departments of Basic Sciences and Surgery-College of Medicine, California Northstate University, Cambridge, Massachusetts, USA.
  • 3 Department of Biochemistry and Molecular Biology, Colleges of National Science, Human Medicine, Osteopathic Medicine and Engineering, Michigan State University, Cambridge, Massachusetts, USA.
  • 4 Currently at Sarepta Therapeutics, Cambridge, Massachusetts, USA.
  • 5 Department of Pathobiology and Diagnostic Investigation, College of Veterinary Medicine, Michigan State University, Lansing, Michigan, USA.
PMID: 30045019 DOI: 10.1159/000492019
Abstract

Background/aims: Human enterocytic differentiation is altered during development, fasting, adaptation, and bariatric surgery, but its intracellular control remains unclear. We hypothesized that Schlafen 12 (SLFN12) regulates enterocyte differentiation.

Methods: We used laser capture dissection of epithelium, qRT-PCR, and immunohistochemistry to evaluate SLFN12 expression in biopsies of control and fasting human duodenal mucosa, and viral overexpression and siRNA to trace the SLFN12 pathway in human Caco-2 and HIEC6 intestinal epithelial cells.

Results: Fasting human duodenal mucosa expressed less SLFN12 mRNA and protein, accompanied by decreases in enterocytic markers like sucrase-isomaltase. SLFN12 overexpression increased Caco-2 sucrase-isomaltase promoter activity, mRNA, and protein independently of proliferation, and activated the SLFN12 putative promoter. SLFN12 coprecipitated Serpin B12 (SERPB12). An inactivating SLFN12 point mutation prevented both SERPB12 binding and sucrase-isomaltase induction. SERPB12 overexpression also induced sucrase-isomaltase, while reducing SERPB12 prevented the SLFN12 effect on sucrase-isomaltase. Sucrase-isomaltase induction by both SLFN12 and SERPB12 was attenuated by reducing UCHL5 or USP14, and blocked by reducing both. SERPB12 stimulated USP14 but not UCHL5 activity. SERPB12 coprecipitated USP14 but not UCHL5. Moreover, SLFN12 increased protein levels of the sucrase-isomaltase-promoter-binding transcription factor cdx2 without altering Cdx2 mRNA. This was prevented by reducing UCHL5 and USP14. We further validated this pathway in vitro and in vivo. SLFN12 or SERPB12 overexpression induced sucrase-isomaltase in human non-malignant HIEC-6 enterocytes.

Conclusions: SLFN12 regulates human enterocytic differentiation by a pathway involving SERPB12, the deubiquitylases, and Cdx2. This pathway may be targeted to manipulate human enterocytic differentiation in mucosal atrophy, short gut or obesity.

Keywords

Differentiation; Epithelium; Intestine; Schlafen 12; Serpin B12; Signaling; UCHL5; USP14.

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