Luhmann Ulrich F O, Carvalho Livia S, Holthaus Sophia-Martha Kleine, Cowing Jill A, Greenaway Simon, Chu Colin J, Herrmann Philipp, Smith Alexander J, Munro Peter M G, Potter Paul, Bainbridge James W B, Ali Robin R
Department of Genetics and
Department of Genetics and.
Hum Mol Genet. 2015 Jan 1;24(1):128-41. doi: 10.1093/hmg/ddu424. Epub 2014 Aug 21.
Understanding phenotype-genotype correlations in retinal degeneration is a major challenge. Mutations in CRB1 lead to a spectrum of autosomal recessive retinal dystrophies with variable phenotypes suggesting the influence of modifying factors. To establish the contribution of the genetic background to phenotypic variability associated with the Crb1(rd8/rd8) mutation, we compared the retinal pathology of Crb1(rd8/rd8)/J inbred mice with that of two Crb1(rd8/rd8) lines backcrossed with C57BL/6JOlaHsd mice. Topical endoscopic fundal imaging and scanning laser ophthalmoscopy fundus images of all three Crb1(rd8/rd8) lines showed a significant increase in the number of inferior retinal lesions that was strikingly variable between the lines. Optical coherence tomography, semithin, ultrastructural morphology and assessment of inflammatory and vascular marker by immunohistochemistry and quantitative reverse transcriptase-polymerase chain reaction revealed that the lesions were associated with photoreceptor death, Müller and microglia activation and telangiectasia-like vascular remodelling-features that were stable in the inbred, variable in the second, but virtually absent in the third Crb1(rd8/rd8) line, even at 12 months of age. This suggests that the Crb1(rd8/rd8) mutation is necessary, but not sufficient for the development of these degenerative features. By whole-genome SNP analysis of the genotype-phenotype correlation, a candidate region on chromosome 15 was identified. This may carry one or more genetic modifiers for the manifestation of the retinal pathology associated with mutations in Crb1. This study also provides insight into the nature of the retinal vascular lesions that likely represent a clinical correlate for the formation of retinal telangiectasia or Coats-like vasculopathy in patients with CRB1 mutations that are thought to depend on such genetic modifiers.
了解视网膜变性中的表型-基因型相关性是一项重大挑战。CRB1基因突变会导致一系列常染色体隐性视网膜营养不良,其表型各异,这表明存在修饰因子的影响。为了确定遗传背景对与Crb1(rd8/rd8)突变相关的表型变异性的作用,我们比较了Crb1(rd8/rd8)/J近交系小鼠与两个与C57BL/6JOlaHsd小鼠回交的Crb1(rd8/rd8)品系的视网膜病理学。所有三个Crb1(rd8/rd8)品系的局部内镜眼底成像和扫描激光眼底镜检查图像显示,视网膜下方病变数量显著增加,且品系间差异显著。光学相干断层扫描、半薄切片、超微结构形态学以及通过免疫组织化学和定量逆转录聚合酶链反应对炎症和血管标记物的评估显示,这些病变与光感受器死亡、Müller细胞和小胶质细胞活化以及毛细血管扩张样血管重塑有关,这些特征在近交系中稳定,在第二代中可变,但在第三个Crb1(rd8/rd8)品系中即使在12个月大时也几乎不存在。这表明Crb1(rd8/rd8)突变对于这些退行性特征的发展是必要的,但并不充分。通过对基因型-表型相关性进行全基因组SNP分析,在15号染色体上确定了一个候选区域。该区域可能携带一个或多个与Crb1突变相关的视网膜病理学表现的遗传修饰因子。这项研究还深入了解了视网膜血管病变的本质,这些病变可能与CRB1突变患者视网膜毛细血管扩张或Coats样血管病变的形成存在临床关联,而这些病变被认为依赖于此类遗传修饰因子。