■为了调查遗传原因,临床特征,和婴儿角膜肌纤维瘤病的潜在治疗目标。
■具有遗传和功能体外分析的案例系列。
■研究了来自2个无关家庭的4名具有角膜肌纤维瘤病临床体征的个体。
■在各指标患者中对血小板衍生生长因子受体β基因(PDGFRB)和notch同源蛋白3基因(NOTCH3)进行基于外显子组测序。通过Sanger测序测试了每个家族的一个临床上受影响的成员在各自的索引中检测到的致病变体。对切除的角膜组织进行免疫组织化学染色。使用酪氨酸激酶抑制剂通过转染的猪主动脉内皮细胞上的荧光素酶报告基因测定在体外进行个体PDGFRB变体的功能分析。通过Western印迹分析突变的PDGFRB的蛋白表达分析。
■测序数据,免疫组织化学染色,PDGFRB变体的功能分析,和蛋白质表达分析。
■我们确定了2本小说,来自2个无关的角膜肌纤维瘤病家族的4例PDGFRB中的杂合功能获得变异。免疫组织化学显示α-平滑肌肌动蛋白和β-连环蛋白阳性,Ki-67的低增殖率(<5%),Desmin的边际积极性,Caldesmon和CD34阴性染色。在所有患者中,角膜手术后疾病复发。当在培养细胞中转染时,PDGFRB变体在不存在其配体的情况下赋予受体组成活性,并且对酪氨酸激酶抑制剂伊马替尼敏感.根据美国医学遗传学和基因组学学院的分类标准,这些变体都可以被分类为可能的致病性。
■我们描述了4例由常染色体显性传递的新型PDGFRB变异引起的角膜肌纤维瘤病。伊马替尼体外敏感性提示了靶向治疗预防未来复发的前景。
■专有或商业披露可在本文末尾的脚注和披露中找到。
UNASSIGNED: To investigate the genetic cause, clinical characteristics, and potential therapeutic targets of infantile corneal myofibromatosis.
UNASSIGNED: Case series with genetic and functional in vitro analyses.
UNASSIGNED: Four individuals from 2 unrelated families with clinical signs of corneal myofibromatosis were investigated.
UNASSIGNED: Exome-based panel sequencing for platelet-derived growth factor receptor beta gene (PDGFRB) and notch homolog protein 3 gene (NOTCH3) was performed in the respective index patients. One clinically affected member of each family was tested for the pathogenic variant detected in the respective index by Sanger sequencing. Immunohistochemical staining on excised corneal tissue was conducted. Functional analysis of the individual PDGFRB variants was performed in vitro by luciferase reporter assays on transfected porcine aortic endothelial cells using tyrosine kinase inhibitors. Protein expression analysis of mutated PDGFRB was analyzed by Western blot.
UNASSIGNED: Sequencing data, immunohistochemical stainings, functional analysis of PDGFRB variants, and protein expression analysis.
UNASSIGNED: We identified 2 novel, heterozygous gain-of-function variants in PDGFRB in 4 individuals from 2 unrelated families with corneal myofibromatosis. Immunohistochemistry demonstrated positivity for alpha-smooth muscle actin and β-catenin, a low proliferation rate in Ki-67 (< 5%), marginal positivity for Desmin, and negative staining for Caldesmon and CD34. In all patients, recurrence of disease occurred after corneal surgery. When transfected in cultured cells, the PDGFRB variants conferred a constitutive activity to the receptor in the absence of its ligand and were sensitive to the tyrosine kinase inhibitor imatinib. The variants can both be classified as likely pathogenic regarding the American College of Medical Genetics and Genomics classification criteria.
UNASSIGNED: We describe 4 cases of corneal myofibromatosis caused by novel PDGFRB variants with autosomal dominant transmission. Imatinib sensitivity in vitro suggests perspectives for targeted therapy preventing recurrences in the future.
UNASSIGNED: Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.