Mutations in RAD21 disrupt regulation of APOB in patients with chronic intestinal pseudo-obstruction.
Mutations in RAD21 disrupt regulation of APOB in patients with chronic intestinal pseudo-obstruction.
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DOI：
10.1053/j.gastro.2014.12.034
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发表时间：
2015-04
期刊：
影响因子：
29.4
通讯作者：
De Giorgio R
中科院分区：
文献类型：
--
作者：
Bonora E;Bianco F;Cordeddu L;Bamshad M;Francescatto L;Dowless D;Stanghellini V;Cogliandro RF;Lindberg G;Mungan Z;Cefle K;Ozcelik T;Palanduz S;Ozturk S;Gedikbasi A;Gori A;Pippucci T;Graziano C;Volta U;Caio G;Barbara G;D'Amato M;Seri M;Katsanis N;Romeo G;De Giorgio R
慢性假性肠梗阻（锡波）的特征是严重的肠动力障碍，类似于机械性亚闭塞，但没有肠梗阻的证据。我们寻找与锡波相关的遗传变异，以增加我们对其发病机制的理解并确定潜在的生物标志物。我们对家族性锡波综合征患者的基因组DNA进行了全外显子组测序。从患者和对照组（无锡波的个体）中收集血液和淋巴母细胞;通过定量逆转录PCR、免疫印迹和迁移率变化测定分析mRNA和蛋白质水平。将cDNA转染到HEK 293细胞中。使用剪接阻断吗啉代（rad 21 a MO）在斑马鱼胚胎中抑制rad 21的表达。收集并分析肠组织。我们在一个锡波家系中发现了一个RAD 21纯合突变（p.622，编码Ala>Thr）。与对照组相比，来自锡波患者的细胞中RAD 21靶点RUNX 1的表达减少。在斑马鱼中，抑制rad 21 a降低runx 1的表达;这种表型通过注射人RAD 21 mRNA而得到纠正，但不与突变的p.622等位基因的mRNA一起。rad 21 a MO斑马鱼的肠转运延迟，肠神经元数量大大减少，与锡波患者相似。这种缺陷在抑制ret和rad 21表达的斑马鱼中更大，表明它们在调节肠道神经发生中的相互作用。APOB的启动子区结合RAD 21但不结合RAD 21 p.622 Ala>Thr;与对照载体相比，HEK 293细胞中野生型RAD 21的表达抑制APOB的表达。APOB的肠道特异性同种型（APOB 48）在携带RAD 21突变的锡波患者血清中过表达。APOB 48也在血清和肠道活检中的散发性锡波中过表达。一些锡波患者携带RAD 21突变，破坏其产物调节RUNX 1和APOB等基因的能力。斑马鱼中rad 21的表达减少，以及这些靶基因的失调，破坏了肠道运输和肠道神经元的发育。
Chronic intestinal pseudo-obstruction (CIPO) is characterized by severe intestinal dysmotility that mimicks a mechanical sub-occlusion with no evidence of gut obstruction. We searched for genetic variants associated with CIPO to increase our understanding of its pathogenesis and indentify potential biomarkers. We performed whole-exome sequencing of genomic DNA from patients with familial CIPO syndrome. Blood and lymphoblastoid cells were collected from patients and controls (individuals without CIPO); levels of mRNA and proteins were analyzed by quantitative reverse transcription PCR, immunoblot, and mobility shift assays. cDNAs were transfected into HEK293 cells. Expression of rad21 was suppressed in zebrafish embryos using a splice-blocking morpholino (rad21a MO). Gut tissues were collected and analyzed. We identified a homozygous mutation (p.622, encodes Ala>Thr) in RAD21 in patients from a consanguineous family with CIPO. Expression of RUNX1, a target of RAD21, was reduced in cells from patients with CIPO compared with controls. In zebrafish, suppression of rad21a reduced expression of runx1; this phenotype was corrected by injection of human RAD21 mRNA, but not with the mRNA from the mutated p.622 allele. rad21a MO zebrafish had delayed intestinal transit and greatly reduced numbers of enteric neurons, similar to patients with CIPO. This defect was greater in zebrafish with suppressed expression of ret and rad21, indicating their interaction in regulation of gut neurogenesis. The promoter region of APOB bound RAD21 but not RAD21 p.622 Ala>Thr; expression of wild-type RAD21 in HEK293 cells repressed expression of APOB, compared with control vector. The gut-specific isoform of APOB (APOB48) is overexpressed in sera from patients with CIPO who carry the RAD21 mutation. APOB48 is also overexpressed in sporadic CIPO in sera and gut biopsies. Some patients with CIPO carry mutations in RAD21 that disrupt the ability of its product to regulate genes such as RUNX1 and APOB. Reduced expression of rad21 in zebrafish, and dysregulation of these target genes, disrupts intestinal transit and development of enteric neurons.
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DOI：
10.1016/j.bbagrm.2013.11.007
发表时间：
2014-01-01
影响因子：
4.7
作者：
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作者：
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影响因子：
3.5
作者：
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DOI：
10.1152/ajpgi.00189.2007
发表时间：
2007-09-01
影响因子：
4.5
作者：
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Black, Dennis D.