Anopthalmia Spectrum Disorders

无视症谱系疾病

基本信息

批准号：
7771496
负责人：
ANNE M. SLAVOTINEK
金额：
$ 23.18万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-01 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7771496
关键词：
Affect Animal Model Animals Anophthalmos Choroid Chromosome Deletion Chromosomes Coloboma Congenital Abnormality Counseling Defect Development Disease Epithelium Eye Eye Development Failure Family Fishes Gene Expression Genes Genetic Genetic Counseling Goals Grant Healthcare Histone H3 Human Immunohistochemistry In Situ Hybridization Individual Injection of therapeutic agent Knock-out Knockout Mice Knowledge Label Larva Lead Life Maintenance Medical Messenger RNA Microphthalmos Modeling Morphology Mus Optics Partner in relationship Pathogenesis Pathway interactions Patients Pattern Peripheral Phenotype Research Retina Retinal Retinal Cone Staining method Stains System Testing Thioredoxin Tissue-Specific Gene Expression Tissues United States Visual impairment Zebrafish cell type disorder prevention eye formation gene function improved interest lens loss of function male mortality mouse model mutant neuroepithelium novel public health relevance research study social therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Anophthalmia, or absence of the eye, is found in 1 in 5,000 to 10,000 individuals and is a devastating birth defect because of the resulting visual impairment. Anopththalmia is closely related to microphthalmia (small eyes) and coloboma (failure of the choroid fissure or the optic fissure to close). We have studied a male patient with severe microphthalmia who had a novel chromosome deletion at chromosome 18q22.1. We have implicated one of the deleted genes at 18q22.1, TXNDC10, in eye development. In situ hybridization showed that Txndc10 was expressed in the retinal neuroepithelium and lens epithelium in the developing murine eye. We re-sequenced TXNDC10 in 66 patients with anophthalmia or microphthalmia, and found c.260G>A, predicting a missense substitution, p.R39Q, in an unrelated patient with microphthalmia and retinal coloboma. We used antisense morpholinos targeted against the zebrafish Txndc10 orthologue, Zgc110025, to show that the morphant larvae had microphthalmia and coloboma. In-situ hybridization with Pax2 showed increased choroid fissure staining in morphants compared to controls, suggesting that Pax2 dysregulation was involved in the coloboma formation. The morphant retinas had increased anti-histone H3 staining in the peripheral retina and we observed fewer late-differentiating cell types in the morphant retinas. Co-injection of human wild type TXNDC10 mRNA with a morpholino rescued the morphant phenotype, whereas co-injection of human TXNDC10/(p.R39Q) mutant mRNA with morpholino did not. Our results suggest that haploinsufficiency for TXNDC10 perturbs eye development. We propose to continue our research on TXNDC10 in two directions. We hypothesize that Txndc10 may be part of a known pathway for ventral retinal development, and would like to use zebrafish to determine whether Zgc110025 interacts with other known genes that are involved in choroid fissure closure and the formation of the ventral retina, such as Vax1, Vax2 and Shh (Syu). We will use in-situ hybridization, dual injections of antisense morpholinos and mRNA rescue experiments to accomplish this aim. Our purpose is to try to uncover the mechanism whereby Txndc10 affects eye development. Secondly, we propose to make a mouse model of loss of Txndc10 function to examine the eye phenotype in a genetic null model rather than in an animal model with only partial loss of gene function. We hypothesize that loss of Txndc10 function will produce mice with microphthalmia and coloboma, and we will use the resultant knock-out mice to examine eye morphology in a Txndc10 null animal and to perform expression arrays to evaluate for differences in gene expression between Txndc10 null mice and wildtype litter mates. Our ultimate goal is to improve the existing knowledge about the genes that are required for ventral retina formation and choroid fissure closure so that human patients with anophthalmia and related eye defects can be offered improved genetic counseling and testing. PUBLIC HEALTH RELEVANCE: Birth defects affect an estimated 120,000 (1 in 33) babies born in the United States each year, and are the leading case of mortality in the first year of life. Anophthalmia (absent eye), microphthalmia (small eye) and coloboma (failure of the choroid fissure or the optic fissure to close) can result in a significant burden for affected individuals and their families because of the medical and social effects of reduced vision. This application seeks to improve the existing knowledge pertaining to the genetic causes of these eye defects so that patient counseling, management and treatment can be improved.

描述（由申请人提供）：在5,000至10,000个个人中发现1个或缺乏眼睛的人，由于视觉障碍而是毁灭性的先天缺陷。 Anopthalmia与微感染（小眼睛）和coloboma（脉络膜裂缝的失败或关闭的视觉裂缝）密切相关。我们研究了一名男性患有严重的微观心脏病患者，该患者在18q22.1染色体时具有新型的染色体缺失。我们已经在眼睛发育中牵涉到18Q22.1 TXNDC10的一个已删除基因。原位杂交表明，TXNDC10在发育中的鼠眼中的视网膜神经上皮和晶状体上皮中表达。我们在66例心血症或微观心脏病患者中重新测试了TXNDC10，并发现了一个无关的微粒细胞和视网膜成骨的患者，发现了C.260G> a，预测错义取代，P.R39Q。我们使用了针对斑马鱼TxnDC10直系同源物ZGC110025的反义形态学，以表明形态的幼虫具有微观粒细胞和古罗巴马。与对照组相比，与PAX2与PAX2的原位杂交显示形态的脉络裂染色增加，这表明PAX2失调涉及coloboma的形成。形态视网膜在周围视网膜中增加了抗固件H3染色，我们观察到形态视网膜中较少的晚分化细胞类型。将人类野生型TXNDC10 mRNA与形态的注射拯救了形态表型，而人类TXNDC10/（p.r39q）突变体mRNA与morpholino共同注射没有。我们的结果表明，TXNDC10的单倍不使眼睛发育。我们建议在两个方向上继续对TXNDC10的研究。我们假设TXNDC10可能是腹侧视网膜发育的已知途径的一部分，并希望使用斑马鱼来确定ZGC110025是否与其他已知基因相互作用，这些基因与脉络膜裂口闭合和腹侧视网膜的形成相互作用，例如Vax1，Vax1，vax2，vax2和shh（Syu）。我们将使用原位杂交，反义形态学的双重注射和mRNA救援实验来实现这一目标。我们的目的是尝试发现TXNDC10影响眼睛发育的机制。其次，我们建议使TXNDC10功能丧失的小鼠模型在遗传无效模型中检查眼表型，而不是在仅部分丧失基因功能的动物模型中。我们假设TXNDC10功能的丧失将产生带有微观心脏的小鼠和可骨瘤的小鼠，我们将使用所得的敲除小鼠检查TXNDC10无效动物中的眼睛形态，并进行表达阵列，以评估TXNDC10 Null小鼠和Will WirdTypepepepepepepepepepe littype-litter litter sates null null小鼠之间的基因表达差异。我们的最终目标是提高有关腹视网膜形成和脉络膜裂缝所需的基因的现有知识，以便可以改善患有障碍性的人类患者和相关眼缺陷的患者。公共卫生相关性：每年在美国出生的估计有120,000（三分之一）的婴儿，这是生命第一年死亡的主要案例。由于视力减少视力的医疗和社会影响，会导致受影响的个人及其家人造成沉重的负担，因此，由于视力降低而导致受影响的个体及其家人的重大负担。该应用程序旨在改善与这些眼睛缺陷的遗传原因有关的现有知识，从而可以改善患者咨询，管理和治疗。