Detection and validation of novel mutations in MERTK in a simplex case of retinal degeneration using WGS and hiPSC-RPEs model.
Detection and validation of novel mutations in MERTK in a simplex case of retinal degeneration using WGS and hiPSC-RPEs model.
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使用WGS和hiPSC-RPE模型检测和验证单纯性视网膜变性病例中MERTK的新突变。
DOI：
10.1002/humu.24146
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发表时间：
2021-03
期刊：
影响因子：
3.9
通讯作者：
Ayyagari R
中科院分区：
文献类型：
--
作者：
Biswas P;Borooah S;Matsui H;Voronchikhina M;Zhou J;Zawaydeh Q;Raghavendra PB;Ferreyra H;Riazuddin SA;Wahlin K;Frazer KA;Ayyagari R
遗传性视网膜变性(IRD)是一组具有广泛临床表型异质性的遗传异质性疾病。分析了一个欧洲血统的核心家庭，其中一个人患有进行性视网膜变性。对先证者及其未受影响的同胞进行全基因组测序，发现视网膜色素上皮Mer酪氨酸激酶原癌基因c.1297_1451del；p.K433_G484fsTer3存在一个新的内含子8供体剪接位点变异(c.1296+1G>；A)和一个新的包含外显子9(chr2：112751488-112752218)的731碱基对缺失。先证者携带两种变异体，处于杂合子状态，这与家系中的疾病分离。这些MERTK变异分别导致外显子8的剪接缺陷和外显子9的缺失。为了评估这些新变异体的影响，从先证者和亲代外周血单个核细胞(PBMC)重新编程人诱导多能干细胞系(HiPSC)，并将其分化为hiPSC-RPE。先证者的HiPSC-RPE分析显示，与亲本的HiPSC-RPE相比，MERTK转录本和蛋白的缺失以及吞噬功能异常。总之，WGS发现MERTK中新的复合杂合变异是先证者IRD的根本原因。此外，使用HiPSC-RPE模型的分析建立了新的MERTK突变的功能影响，并揭示了这些突变导致的潜在病理机制。
Inherited Retinal degenerations (IRD) are a group of genetically heterogeneous conditions with broad clinical phenotypic heterogeneity. A nuclear family of European ancestry with one individual affected by progressive retinal degeneration was analyzed. Whole genome sequencing of the proband and her unaffected sibling identified a novel intron 8 donor splice site variant (c.1296+1G>A) and a novel 731 base pair deletion encompassing exon 9 (Chr2:112751488–112752218) resulting in c.1297_1451del; p.K433_G484fsTer3 in the Mer tyrosine kinase protooncogene (MERTK) expressed in retinal pigment epithelium (RPE). The proband carried both variants in the heterozygous state which segregated with disease in the pedigree. These MERTK variants are predicted to result in the defective splicing of exon 8 and loss of exon 9 respectively. To evaluate the impact of these novel variants, human induced pluripotent stem cell lines (hiPSC) were reprogrammed from the proband and parental peripheral blood mononuclear cells (PBMCs) and differentiated to hiPSC-RPE. Analysis of the proband’s hiPSC-RPE revealed the absence of MERTK transcript and protein as well as abnormal phagocytosis when compared with the parental hiPSC-RPE. In summary, WGS identified novel compound heterozygous variants in MERTK as the underlying cause of IRD in the proband. Further, analysis using an hiPSC-RPE model established a functional impact of novel MERTK mutations and revealed the potential mechanism underlying pathology due to these mutations.
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影响因子：
12.6
作者：
Gamal, W.;Borooah, S.;Smith, S.;Underwood, I.;Srsen, V.;Chandran, S.;Bagnaninchi, P. O.;Dhillon, B.
通讯作者：
Dhillon, B.
影响因子：
3.9
作者：
Audo, Isabelle;Mohand-Said, Saddek;Zeitz, Christina
通讯作者：
Zeitz, Christina
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：
1.2
作者：
Cingolani, Pablo;Platts, Adrian;Ruden, Douglas M.
通讯作者：
Ruden, Douglas M.
影响因子：
5.3
作者：
Chekuri, Anil;Guru, Aditya A.;Ayyagari, Radha
通讯作者：
Ayyagari, Radha