当前位置： SCI文献检索 > GASTROENTEROLOGY期刊下所有文献 > Intestinal Failure and Aberrant Lipid Metabolism in Patients With DGAT1 Deficiency.

Intestinal Failure and Aberrant Lipid Metabolism in Patients With DGAT1 Deficiency.

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 sequencing 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 polymerase chain reaction, and for the activity 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 product; dermal fibroblasts and intestinal organoids derived from these patients had altered lipid metabolism and were susceptible to lipid-induced cell death. Expression of full-length wildtype DGAT1 or DGAT2 restored normal lipid metabolism in these cells. These findings indicate the importance of DGAT1 in fat metabolism and lipotoxicity in the intestinal epithelium. A fat-free diet might serve as the first line of therapy for patients with reduced DGAT1 expression. It is important to identify genetic variants associated with congenital diarrheal disorders for proper diagnosis and selection of treatment strategies.

journal_name

Gastroenterology

journal_title

Gastroenterology

authors

van Rijn JM,Ardy RC,Kuloğlu Z,Härter B,van Haaften-Visser DY,van der Doef HPJ,van Hoesel M,Kansu A,van Vugt AHM,Thian M,Kokke FTM,Krolo A,Başaran MK,Kaya NG,Aksu AÜ,Dalgıç B,Ozcay F,Baris Z,Kain R,Stigter ECA,Lich

doi

10.1053/j.gastro.2018.03.040

subject

Has Abstract

pub_date

2018-07-01 00:00:00

pages

130-143.e15

issue

eissn

0016-5085

issn

1528-0012

pii

S0016-5085(18)30347-0

journal_volume

155

pub_type

杂志文章

在线工具

Intestinal Failure and Aberrant Lipid Metabolism in Patients With DGAT1 Deficiency.