Intestinal Failure and Aberrant Lipid Metabolism in Patients With DGAT1 Deficiency.
Intestinal Failure and Aberrant Lipid Metabolism in Patients With DGAT1 Deficiency.
复制标题
DOI：
10.1053/j.gastro.2018.03.040
复制
发表时间：
2018-07
期刊：
影响因子：
29.4
通讯作者：
Boztug K
中科院分区：
文献类型：
--
作者：
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;Lichtenbelt KD;Massink MPG;Duran KJ;Verheij JBGM;Lugtenberg D;Nikkels PGJ;Brouwer HGF;Verkade HJ;Scheenstra R;Spee B;Nieuwenhuis EES;Coffer PJ;Janecke AR;van Haaften G;Houwen RHJ;Müller T;Middendorp S;Boztug K
先天性腹泻是一种罕见的遗传性肠道疾病，其特征是顽固性，有时危及生命，腹泻和营养吸收不良;有些与二酰基甘油酰基转移酶1（DGAT 1）的突变有关，DGAT 1催化从二酰基甘油和酰基辅酶A形成三酰基甘油。我们研究了DGAT 1缺乏导致肠衰竭的机制，使用患者来源的类器官。我们收集了来自6个不相关家系的10名患者的血液样本，这些患者表现为早发性严重腹泻和/或呕吐、低白蛋白血症和/或（致死性）蛋白丢失性肠病伴肠衰竭;我们对8名患者的DNA进行了下一代测序分析。从来自3名患者和3名健康个体（对照）的十二指肠活检产生类器官。用表达全长或突变形式的DGAT 1或全长DGAT 2的载体转染或转导Caco-2细胞和患者来源的真皮成纤维细胞。我们在对照肠类器官中进行了DGAT 1的CRISPR/Cas9引导的破坏。通过免疫印迹、免疫荧光、流式细胞术、色谱法、定量实时聚合酶链反应以及半胱天冬酶3和7的活性分析细胞和类器官。在10例患者中，我们确定了5个DGAT 1双等位基因功能丧失突变。在患者来源的成纤维细胞和类器官中，突变降低了DGAT 1蛋白的表达并改变了三酰甘油代谢，导致油酸添加后脂滴形成减少。患者来源的成纤维细胞中全长DGAT 2的表达恢复了脂滴的形成。来自DGAT 1突变患者的类器官比对照类器官更容易受到脂质诱导的细胞死亡的影响。我们确定了一个大型的先天性胃肠疾病患者队列，这些患者的DGAT 1突变降低了其产物的表达;这些患者的真皮成纤维细胞和肠类器官改变了脂质代谢，容易发生脂质诱导的细胞死亡。全长野生型DGAT 1或DGAT 2的表达恢复了这些细胞的正常脂质代谢。这些发现表明DGAT 1在肠上皮的脂肪代谢和脂毒性中的重要性。无脂饮食可能作为DGAT 1表达降低患者的一线治疗。识别与先天性尿道疾病相关的遗传变异对于正确诊断和选择治疗策略是重要的。
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. 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. 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. 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.
登录
查看更多内容
DOI：
10.1056/nejmoa1615887
发表时间：
2017-07-06
期刊：
The New England journal of medicine
影响因子：
--
作者：
Ozen A;Comrie WA;Ardy RC;Domínguez Conde C;Dalgic B;Beser ÖF;Morawski AR;Karakoc-Aydiner E;Tutar E;Baris S;Ozcay F;Serwas NK;Zhang Y;Matthews HF;Pittaluga S;Folio LR;Unlusoy Aksu A;McElwee JJ;Krolo A;Kiykim A;Baris Z;Gulsan M;Ogulur I;Snapper SB;Houwen RHJ;Leavis HL;Ertem D;Kain R;Sari S;Erkan T;Su HC;Boztug K;Lenardo MJ
通讯作者：
Lenardo MJ
影响因子：
4.5
作者：
Meyers CD;Tremblay K;Amer A;Chen J;Jiang L;Gaudet D
通讯作者：
Gaudet D
影响因子：
30.8
作者：
Kircher, Martin;Witten, Daniela M.;Jain, Preti;O'Roak, Brian J.;Cooper, Gregory M.;Shendure, Jay
通讯作者：
Shendure, Jay
影响因子：
10.1
作者：
Salzer, Elisabeth;Daschkey, Svenja;Seidel, Markus G.
通讯作者：
Seidel, Markus G.
DOI：
10.1038/nrgastro.2015.44
发表时间：
2015-05
期刊：
Nature reviews. Gastroenterology & hepatology
影响因子：
--
作者：
Canani RB;Castaldo G;Bacchetta R;Martín MG;Goulet O
通讯作者：
Goulet O