Although mutations in hundreds (>200) of genes have been causally linked to neural tube defects (NTDs) in mouse, in humans the elucidation of genetic factors associated to NTD has remained elusive. In addition, both environmental and epigenetic components are also believed to contribute to the disease pathogenesis. Recently, circulating microRNAs (miRNAs) have been analyzed in samples from a small cohort of pregnant women and their use as non-invasive markers for NTDs diagnosis was suggested. This study aims at investigating the role of miRNAs in neural tube development applying both a candidate-based and a systematic unbiased approach on human samples. From a database of 534 consecutive fetal autopsies performed after legal termination of pregnancy during the second trimester, we selected singleton pregnancies with fetal neural tube defects (NTD) in Caucasian women. Defects were prenatally detected by ultrasound. All cases underwent a complete autopsy performed by feto-pathologists, according to standard protocols, including X-rays, photographs, macroscopical and histological examination of all organs. Fetuses with abnormal karyotype or other associated malformations were excluded. Sixteen cases were eligible. Seven cases were lost due to unavailability of specimens, therefore study population included 9 fetuses (7 myelomeningocele, 2 anencephaly). Neural tube was analyzed from cervical to sacral vertebral segments using serial paraffin sections. Spine defects were located on occipital, thoracic, or lumbosacral vertebrae. On one side, using a combination of bioinformatics tools, such as the MirWalk target predictor, the Co-expression Meta-analysis of miRNA Targets (CoMeTa), and the Database for Annotation, Visualization and Integrated Discovery (DAVID), we have identified 4 candidate miRNAs whose predicted targets are significantly enriched for functional pathways related to neurulation. Data were then integrated to build a model of a miRNA-regulated gene network possibly involved in NTDs. miRNAs levels will be analyzed on NTD samples and matched controls. On the other, a systematic analysis of all miRNAs expressed in NTDs is being performed using a Small RNA sequencing approach, which implements “next-generation” technologies (ie. Miseq platform, Illumina) for unbiased miRNA discovery and profiling. The results will be validated by TaqMan MicroRNA assays targeting the single specific miRNA. Finally, we will use the zebrafish model system to functionally validate miRNAs related to neural tube development.
|Titolo:||Role of miRNAs in neural tube development|
|Data di pubblicazione:||2013|
|Settore Scientifico Disciplinare:||Settore BIO/13 - Biologia Applicata|
|Citazione:||Role of miRNAs in neural tube development / G. Soldà, L. Avagliano, G. Fazio, A. Pistocchi, A. Gallina, G. Cazzaniga, G. Bulfamante, V. Massa. ((Intervento presentato al convegno International Conference of Neural Tube Defects tenutosi a Austin, Texas (USA) nel 2013.|
|Appare nelle tipologie:||14 - Intervento a convegno non pubblicato|