Villegas-Ruiz Vanessa, Hendlmeier Ferdinand, Buentello-Volante Beatriz, Rodríguez-Loaiza José L, Miranda-Duarte Antonio, Zenteno Juan C
Department of Genetics-Research Unit, Institute of Ophthalmology "Conde de Valenciana", Mexico City, Mexico.
Medizinische Fakultät Mannheim, Universität Heidelberg, Mannheim, Germany.
Exp Eye Res. 2017 Feb;155:99-106. doi: 10.1016/j.exer.2017.01.004. Epub 2017 Jan 27.
Diabetic retinopathy (DR) affects approximately one third of all diabetic subjects and is the leading cause of blindness in young to middle-aged adults in the developed world. While early diagnosis is crucial for preventing DR-associated visual loss, the identification of accessible biomarkers that could lead to presymptomatic recognition of the disease is of great clinical importance. The aim of this work was to investigate the possible involvement of alternative splicing events in DR development by performing a genome-wide transcriptional profiling comparing blood-derived RNA from DR subjects and from diabetic-non DR controls. A total of 95 RNA samples, 67 from patients with bilateral DR and 28 from diabetic patients without DR after a period of at least 10 years with type 2 DM, were compared in a genome-wide transcriptome analysis using the GeneChip Human Gene 2.0 ST Array which contains probe sets covering all exons of ∼33,500 coding transcripts of annotated genes. Microarray data analysis followed by RT-PCR and cDNA sequencing identified important differential splicing events in TUBD1 (Tubulin, Delta-1) isoforms between DR and DM samples. Specifically, the co-expression of particular TUBD1 isoforms was significantly associated with NPDR risk (p = 0.039 by Pearson's chi-squared test; OR (CI 95%): 8.1 (1.0-72.7)). Analysis of TUBD1 signal pathways and regulating networks using a MetaCore platform showed that HIF-1, a molecule playing an important role in the pathogenesis of DR, is a direct regulator of TUBD1 expression. In conjunction, our data suggest that TUBD1 mRNA isoform expression profile in peripheral blood could be an accessible biomarker for predicting the risk for diabetic retinopathy development.
糖尿病视网膜病变（DR）影响着约三分之一的糖尿病患者，是发达国家中青年成年人失明的主要原因。虽然早期诊断对于预防与DR相关的视力丧失至关重要，但识别可用于疾病症状前识别的易获取生物标志物具有重大临床意义。这项工作的目的是通过对DR患者和糖尿病非DR对照的血液来源RNA进行全基因组转录谱分析，研究可变剪接事件在DR发展中的可能作用。在一项全基因组转录组分析中，使用包含覆盖约33,500个注释基因编码转录本所有外显子的探针集的基因芯片人类基因2.0 ST阵列，比较了总共95个RNA样本，其中67个来自双侧DR患者，28个来自患有2型糖尿病至少10年的无DR糖尿病患者。微阵列数据分析随后进行RT-PCR和cDNA测序，确定了DR和糖尿病样本之间TUBD1（微管蛋白，δ-1）异构体中重要的差异剪接事件。具体而言，特定TUBD1异构体的共表达与非增殖性糖尿病视网膜病变（NPDR）风险显著相关（Pearson卡方检验p = 0.039；OR（95%CI）：8.1（1.0 - 72.7））。使用MetaCore平台对TUBD1信号通路和调控网络进行分析表明，在DR发病机制中起重要作用的分子HIF-1是TUBD1表达的直接调节因子。综合来看，我们的数据表明外周血中TUBD1 mRNA异构体表达谱可能是预测糖尿病视网膜病变发展风险的易获取生物标志物。
