H2020Индивидуална стипендия2017–2019

NEURO_NMD · Functional impact of alternative splicing coupled to nonsense-mediated decay in developing neurons

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2017-05-01 → 2019-12-04
Финансиране от ЕС: 183 455 €
Участници: 1
Схема: MSCA-IF-EF-ST

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Накратко на български

Механизмите за регулиране на гените при развитието на неврони се изследват чрез процеси като алтернативен сплайсинг и разграждане на РНК, например при протеините на актиновия скелет. Това помага да се разбере как стволови клетки се превръщат в специализирани нервни клетки.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

Functional impact of alternative splicing coupled to nonsense-mediated decay in developing neurons

Embryonic stem cells (ESCs) give rise to all cell types in multicellular eukaryotes. Since cells in the same organism typically share virtually invariant genomic sequences, distinct ESC differentiation outcomes require different sets of genes to be turned on and off in a highly regulated manner. A growing body of evidence points to the importance of post-transcriptional mechanisms in this process. This is particularly evident during development of mammalian neurons, where changes in RNA splicing and stability appear to control gene expression on a truly global scale.. Many important aspects of post-transcriptional regulation in general and a crosstalk between alternative splicing (AS) and RNA destabilization mechanism known as nonsense-mediated decay (NMD) in particular remain poorly understood. This project aimed to uncover post-transcriptional mechanisms orchestrating neuronal differentiation, with two main technical objectives: 1) Understanding how AS-NMD regulation of newly identified Ptbp1 targets contributes to neuronal development and function. 2) Understanding how NMD target repertoire changes in developing neurons using unbiased time-resolved approaches. In order to accomplish the objectives of the action, we combined advanced genetic, biochemical, high-throughput and bioinformatics techniques and obtained two main results. 1) We showed that a subset of genes encoding regulators and components of the actin cytoskeleton are co-ordinately downregulated during neuronal differentiation through Ptbp1-dependent AS-NMD. 2) We discovered a novel and unexpected role for Ptbp1 in facilitating co-transcriptional intron removal in a large group of genes expressed in mouse ESCs. The RNA binding protein Ptbp1 coordinates a large number of key alternative splicing events during early neuronal development. Deregulation of many RNA-based processes has been linked to neurodevelopmental and neurodegenerative diseases. Defects in the NMD machinery itself are known to lead to mental retardation and severe psychiatric conditions including autism, attention deficit hyperactivity disorder and schizophrenia. This project unveiled new important molecular regulation processes in stem cell state maintenance and neuronal development and thus improved our understanding of molecular mechanisms underlying the onset of these serious conditions. Moreover, we produced novel data and tools that should become important resources for the wide scientific community.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

Differentiation of precursor cells into mature neurons relies on transcriptome-wide changes in gene expression that have to be coordinated in a precise spatiotemporal fashion. Alternative pre-mRNA splicing coupled to nonsense-mediated decay (AS-NMD) is a widespread post-transcriptional mechanism known to orchestrate gene expression dynamics in developmental contexts. Earlier studies identified several neural targets of this pathway; however, in most cases, the extent to which AS-NMD contributes to the overall gene expression dynamics and biological significance of this regulation is poorly understood. Moreover, whether AS-NMD target repertoire undergoes considerable changes in developing brain and how this might fit to the global regulation network underlying neuronal differentiation remains unclear. I will address these questions using two separate approaches. First, I will investigate novel AS-NMD targets encoding actin cytoskeleton factors and controlled by an important regulator of neuronal alternative splicing, Ptbp1. I will elucidate the extent of AS-NMD regulation in these genes by modulating the inclusion of the NMD-promoting exons with corresponding antisense oligonucleotides. in mouse embryonic stem cells undergoing neuronal differentiation, neural stem cells and primary neurons. Second, I will systematically analyse how NMD contributes to different stages of neuronal development by acutely inhibiting this pathway in a time-resolved manner using genetic means. I will then identify gene expression effects and functional consequences of NMD inactivation using transcriptome sequencing (RNA-Seq) and appropriate cell biological methods. All in all, this work will provide critical quantitative insights into AS-NMD functions and uncover novel mechanisms allowing neurons to attain their unique morphological and functional properties.

Оригинален текст от CORDIS (на английски).

Участници

KING'S COLLEGE LONDON · LondonКоординаторОбединеното кралство