Spontaneous Posterior Segment Vascular Disease Phenotype of a Mouse Model, rnv3, Is Dependent on the Crb1rd8 Allele.
Spontaneous Posterior Segment Vascular Disease Phenotype of a Mouse Model, rnv3, Is Dependent on the Crb1rd8 Allele.
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DOI：
10.1167/iovs.18-25046
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发表时间：
2018-10-01
影响因子：
4.4
通讯作者：
Nishina PM
中科院分区：
文献类型：
--
作者：
Chang B;FitzMaurice B;Wang J;Low BE;Wiles MV;Nishina PM
确定自发性视网膜血管化鼠模型 rnv3（视网膜血管化 3，又名 JR5558）中病变发展的分子基础。通过临床评估、视网膜电图 (ERG) 和光学显微镜分析的纵向研究检查了 rnv3 的疾病进展。通过 DNA 合并和全基因组连锁分析确定隐性 rnv3 突变的染色体位置。通过比较 rnv3 突变体和野生型 (WT) 对照的整个外显子组序列，发现了致病突变。为了确认致病突变，利用转录激活因子样效应核酸酶（TALEN）介导的寡核苷酸定向修复（ODR）来纠正突变等位基因。通过活体小鼠的眼底成像和光学相干断层扫描来评估表型校正。 rnv3 表现出早发性、多灶性色素脱失的视网膜病变，从 18 日龄开始通过眼底检查即可观察到。视网膜病变与 25 日龄左右的荧光素渗漏有关，约 4 周龄时渗漏达到峰值。 ERG 反应随着 rnv3 突变体年龄的增长而恶化，同时伴随着组织学可观察到的渐进性光感受器破坏和损失。遗传分析将 rnv3 定位于小鼠染色体 (Chr) 1。通过对 rnv3/rnv3 突变体的整个外显子组捕获文库进行高通量测序以及随后的序列分析，鉴定出 Crb1 [crumbs 家族成员 1] 基因中的单碱基缺失 (del)，此前曾报道该基因会导致视网膜变性 8。 TALEN介导的ODR挽救了后段血管化表型；杂合子 Crb1rd8+em1Boc/Crb1rd8 和纯合子 Crb1rd8+em1Boc/Crb1rd8+em1Boc 小鼠表现出正常的视网膜表型。此外，通过 TALEN 诱导的异常非同源末端连接产生的 Crb1 的六种新破坏表现出不同水平的血管化，表明等位基因效应。 rnv3模型和Crb1的六种新破坏模型都是可靠的新型小鼠模型，用于研究与后段血管化相关的早期和晚期事件，也可用于测试治疗人类眼血管疾病的药理学靶点的效果。对这些模型的进一步研究可能有助于更好地了解不同 Crb1 等位基因如何导致异常血管生成。
To determine the molecular basis of lesion development in a murine model of spontaneous retinal vascularization, rnv3 (retinal vascularization 3, aka JR5558). Disease progression of rnv3 was examined in longitudinal studies by clinical evaluation, electroretinography (ERG) and light microscopy analyses. The chromosomal position for the recessive rnv3 mutation was determined by DNA pooling and genome-wide linkage analysis. The causative mutation was discovered by comparison of whole exome sequences of rnv3 mutant and wild-type (WT) controls. In order to confirm the causative mutation, transcription activator-like effector nuclease (TALEN)-mediated oligonucleotide directed repair (ODR) was utilized to correct the mutant allele. Phenotypic correction was assessed by fundus imaging and optical coherence tomography of live mice. rnv3 exhibits early-onset, multifocal depigmented retinal lesions observable by fundus examination starting at 18 days of age. The retinal lesions are associated with fluorescein leakage around 25 days of age, with peak leakage at about 4 weeks of age. ERG responses deteriorate as rnv3 mutants age, concomitant with progressive photoreceptor disruption and loss that is observable by histology. Genetic analysis localized rnv3 to mouse chromosome (Chr) 1. By high throughput sequencing of a whole exome capture library of a rnv3/rnv3 mutant and subsequent sequence analysis, a single base deletion (del) in the Crb1 [crumbs family member 1] gene, which was previously reported to cause retinal degeneration 8, was identified. The TALEN-mediated ODR rescued the posterior segment vascularization phenotype; heterozygous Crb1rd8+em1Boc/Crb1rd8 and homozygous Crb1rd8+em1Boc/Crb1rd8+em1Boc mice showed a normal retinal phenotype. Additionally, six novel disruptions of Crb1 that were generated through aberrant non-homologous end joining induced by TALEN exhibited variable levels of vascularization, suggesting allelic effects. The rnv3 model and the models of six novel disruptions of Crb1 are all reliable, novel mouse models for the study of both early and late events associated with posterior segment vascularization and can also be used to test the effects of pharmacological targets for treating human ocular vascular disorders. Further study of these models may provide a greater understanding about how different Crb1 alleles result in aberrant angiogenesis.
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影响因子：
13.7
作者：
Hartnett, M. Elizabeth
通讯作者：
Hartnett, M. Elizabeth
影响因子：
82.9
作者：
Joyal JS;Sun Y;Gantner ML;Shao Z;Evans LP;Saba N;Fredrick T;Burnim S;Kim JS;Patel G;Juan AM;Hurst CG;Hatton CJ;Cui Z;Pierce KA;Bherer P;Aguilar E;Powner MB;Vevis K;Boisvert M;Fu Z;Levy E;Fruttiger M;Packard A;Rezende FA;Maranda B;Sapieha P;Chen J;Friedlander M;Clish CB;Smith LE
通讯作者：
Smith LE
纯合CRB1RD8/RD8小鼠中视网膜病理的严重程度取决于其他遗传因素。
影响因子：
3.5
作者：
Luhmann UF;Carvalho LS;Holthaus SM;Cowing JA;Greenaway S;Chu CJ;Herrmann P;Smith AJ;Munro PM;Potter P;Bainbridge JW;Ali RR
通讯作者：
Ali RR
影响因子：
12.3
作者：
Fairfield H;Gilbert GJ;Barter M;Corrigan RR;Curtain M;Ding Y;D'Ascenzo M;Gerhardt DJ;He C;Huang W;Richmond T;Rowe L;Probst FJ;Bergstrom DE;Murray SA;Bult C;Richardson J;Kile BT;Gut I;Hager J;Sigurdsson S;Mauceli E;Di Palma F;Lindblad-Toh K;Cunningham ML;Cox TC;Justice MJ;Spector MS;Lowe SW;Albert T;Donahue LR;Jeddeloh J;Shendure J;Reinholdt LG
通讯作者：
Reinholdt LG
影响因子：
4.1
作者：
Henderson, Robert H.;Mackay, Donna S.;Moore, Anthony T.
通讯作者：
Moore, Anthony T.