1. Intestinal failure and aberrant lipid metabolism in patients with DGAT1 deficiency
- Author
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van Rijn, Jorik M, Ardy, Rico Chandra, Kuloğlu, Zarife, Härter, Bettina, van Haaften-Visser, Désirée Y, van der Doef, Hubert P J, van Hoesel, Marliek, Kansu, Aydan, van Vugt, Anke H M, Ng, Marini, Kokke, Freddy T M, Krolo, Ana, Başaran, Meryem Keçeli, Kaya, Neslihan Gurcan, Ünlüsoy Aksu, Aysel, Dalgıç, Buket, Ozcay, Figen, Baris, Zeren, Kain, Renate, Stigter, Edwin C A, Lichtenbelt, Klaske D, Massink, Maarten P G, Duran, Karen J, Verheij, Joke B G M, Lugtenberg, Dorien, Nikkels, Peter G J, Brouwer, Henricus G F, Verkade, Henkjan J, Scheenstra, Rene, Spee, Bart, Nieuwenhuis, Edward E S, Coffer, Paul J, Janecke, Andreas R, van Haaften, Gijs, Houwen, Roderick H J H, Müller, Thomas, Middendorp, Sabine, Boztug, Kaan, van Rijn, Jorik M, Ardy, Rico Chandra, Kuloğlu, Zarife, Härter, Bettina, van Haaften-Visser, Désirée Y, van der Doef, Hubert P J, van Hoesel, Marliek, Kansu, Aydan, van Vugt, Anke H M, Ng, Marini, Kokke, Freddy T M, Krolo, Ana, Başaran, Meryem Keçeli, Kaya, Neslihan Gurcan, Ünlüsoy Aksu, Aysel, Dalgıç, Buket, Ozcay, Figen, Baris, Zeren, Kain, Renate, Stigter, Edwin C A, Lichtenbelt, Klaske D, Massink, Maarten P G, Duran, Karen J, Verheij, Joke B G M, Lugtenberg, Dorien, Nikkels, Peter G J, Brouwer, Henricus G F, Verkade, Henkjan J, Scheenstra, Rene, Spee, Bart, Nieuwenhuis, Edward E S, Coffer, Paul J, Janecke, Andreas R, van Haaften, Gijs, Houwen, Roderick H J H, Müller, Thomas, Middendorp, Sabine, and Boztug, Kaan
- Abstract
BACKGROUND & AIMS: Congenital diarrheal disorders are rare inherited intestinal disorders characterized by intractable, sometimes life-threatening, diarrhea and nutrient malabsorption; some have been associated with mutations in diacylglycerol-acyltransferase 1 (DGAT1), which catalyzes formation of triacylglycerol from diacylglycerol and acyl-CoA. We investigated the mechanisms by which DGAT1 deficiency contributes to intestinal failure using patient-derived organoids.METHODS: We collected blood samples from 10 patients, from 6 unrelated pedigrees, who presented with early-onset severe diarrhea and/or vomiting, hypoalbuminemia, and/or (fatal) protein-losing enteropathy with intestinal failure; we performed next-generation sequence analysis of DNA from 8 patients. Organoids were generated from duodenal biopsies from 3 patients and 3 healthy individuals (controls). Caco-2 cells and patient-derived dermal fibroblasts were transfected or transduced with vectors that express full-length or mutant forms of DGAT1 or full-length DGAT2. We performed CRISPR/Cas9-guided disruption of DGAT1 in control intestinal organoids. Cells and organoids were analyzed by immunoblot, immunofluorescence, flow cytometry, chromatography, quantitative real-time PCR, and for activities of caspases 3 and 7.RESULTS: In the 10 patients, we identified 5 bi-allelic loss-of-function mutations in DGAT1. In patient-derived fibroblasts and organoids, the mutations reduced expression of DGAT1 protein and altered triacylglycerol metabolism, resulting in decreased lipid droplet formation after oleic acid addition. Expression of full-length DGAT2 in patient-derived fibroblasts restored formation of lipid droplets. Organoids derived from patients with DGAT1 mutations were more susceptible to lipid-induced cell death than control organoids.CONCLUSIONS: We identified a large cohort of patients with congenital diarrheal disorders with mutations in DGAT1 that reduced expression of its
- Published
- 2018