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Genetic and functional studies of the microphthalmia with linear skin defects (MLS) syndrome

伴有线性皮肤缺损 (MLS) 综合征的小眼症的遗传和功能研究

基本信息

批准号：
195162804
负责人：
Professorin Dr. Kerstin Kutsche
金额：
--
依托单位：
Institut für Humangenetik
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2011
资助国家：
德国
起止时间：
2010-12-31 至 2020-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/195162804?language=en
关键词：
Genetic functional studies microphthalmia linear

项目摘要

MLS (microphthalmia with linear skin defects) syndrome is a rare, X-chromosomal neurocutaneous developmental disorder that usually manifests in females and is associated with male lethality in utero. Besides linear skin defects on face and neck, microphthalmia and sclerocornea other clinical features, such as congenital diaphragmatic hernia (CGH), can be present in affected individuals. The known disease genes HCCS, COX7B and NDUFB11, in which heterozygous mutations have been identified, code for proteins of the mitochondrial respiratory chain. Cells deficient of HCCS, COX7B or NDUFB11 show defects in assembly and activity of the respiratory chain enzymes as well as enhanced cell death. Downregulation of hccs in Medaka causes increased cell death via an apoptosome-independent caspase-9 activation which is triggered by reactive oxygen species (ROS).In the second funding period, we will focus on work packages to elucidate further the genetic basis and pathomechanism underlying MLS syndrome. By sequencing and copy number analysis of the HCCS gene in patients with CGH, we will answer the question if HCCS mutations cause an isolated form of CGH. To test if detected HCCS variants affect respiratory chain activity, we will use a yeast complementation assay. By whole-exome sequencing (WES) in six patients affected by MLS syndrome, we aim to identify novel disease genes. We will determine pathogenicity of a single sequence variant or characterize consequences of different variants in the same gene by performing functional studies. By WES in a male patient with typical features of MLS syndrome, we identified the p.(A217V) variant in the X-chromosomal gene HDAC6 which encodes the histone deacetylase 6. We will use patient and control fibroblasts to biochemically analyse histone deacetylase 6 activity and protein-protein interactions. The effect of the HDAC6 variant on cell death and respiratory chain activity will be studied using annexin V staining, measurement of ROS levels, analysis of the mitochondrial membrane potential, cytochrome c release from mitochondria, and measurement of caspase activity. Similar experiments will be performed with stable, NDUFB11-deficient HeLa cells. We will generate zebrafish models for cox7b and ndufb11 by morpholino-induced knockdown and by using the CRISPR-Cas9 technology to detect and characterize developmental defects in the fishes. Zebrafish mutants will also be used to study the effect of the genetic defect on cell death, cell proliferation and oxidative phosphorylation.

MLS（小眼伴线状皮肤缺损）综合征是一种罕见的X染色体神经皮肤发育障碍，通常在女性中出现，与子宫内男性死亡有关。除了面部和颈部的线性皮肤缺损，小眼症和巩膜其他临床特征，如先天性腹股沟疝（CGH），可能存在于受影响的个体中。已知的疾病基因HCCS、COX 7 B和NDUFB 11编码线粒体呼吸链的蛋白质，其中已经鉴定出杂合突变。缺乏HCCS、COX 7 B或NDUFB 11的细胞显示呼吸链酶的组装和活性缺陷以及增强的细胞死亡。在青鳉中hccs的下调通过活性氧（ROS）触发的不依赖于线粒体的caspase-9激活导致细胞死亡增加。在第二个资助期，我们将专注于工作包，以进一步阐明MLS综合征的遗传基础和病理机制。通过对CGH患者HCCS基因的测序和拷贝数分析，我们将回答HCCS突变是否会导致孤立形式的CGH的问题。为了测试检测到的HCCS变体是否影响呼吸链活性，我们将使用酵母互补试验。通过对6例MLS综合征患者进行全外显子组测序（WES），我们的目的是确定新的疾病基因。我们将通过进行功能研究来确定单个序列变体的致病性或表征同一基因中不同变体的后果。通过对1例具有典型MLS综合征特征的男性患者的WES检查，我们确定了P。（A217 V）X染色体基因HDAC 6中的变体，其编码组蛋白脱乙酰基酶6。我们将使用患者和对照组成纤维细胞进行组蛋白去乙酰化酶6活性和蛋白质-蛋白质相互作用的生化分析。HDAC 6变体对细胞死亡和呼吸链活性的影响将使用膜联蛋白V染色、ROS水平的测量、线粒体膜电位的分析、细胞色素c从线粒体的释放和半胱天冬酶活性的测量来研究。将用稳定的NDUFB 11缺陷型HeLa细胞进行类似的实验。我们将通过吗啉代诱导的敲低产生cox 7 b和ndufb 11的斑马鱼模型，并使用CRISPR-Cas9技术检测和表征鱼类的发育缺陷。斑马鱼突变体也将用于研究遗传缺陷对细胞死亡，细胞增殖和氧化磷酸化的影响。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Coinheritance of biallelic SLURP1 and SLC39A4 mutations cause a severe genodermatosis with skin peeling and hair loss all over the body

双等位基因 SLURP1 和 SLC39A4 突变的共同遗传导致严重的遗传性皮肤病，伴有全身皮肤脱皮和脱发

DOI：
10.1111/bjd.16912
发表时间：
2018
期刊：
British Journal of Dermatology
影响因子：
10.3
作者：
Harms FL;Nampoothiri S;Kortüm F;Thomas J;Panicker VV;Alawi M;Altmüller J;Yesodharan D;Kutsche K
通讯作者：
Kutsche K

Fragmented Elastic Fibers in Focal Dermal Hypoplasia (Goltz-Gorlin Syndrome) Without Focal Dermal Hypoplasia: Report of a Male Case and Review of the Literature.

无局灶性真皮发育不全的局灶性真皮发育不全（Goltz-Gorlin 综合征）中的弹性纤维断裂：一例男性病例报告及文献综述