Investigating the role of PIP4K2B, nuclear phosphoinositides and TAF3 in transcription and genome organisation during myogenic differentiation

研究 PIP4K2B、核磷酸肌醇和 TAF3 在成肌分化过程中转录和基因组组织中的作用

基本信息

批准号：
BB/P003508/1
负责人：
Nullin Divecha
金额：
$ 72.72万
依托单位：
University of Southampton
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2016
资助国家：
英国
起止时间：
2016 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FP003508%2F1
关键词：
Investigating role PIP4K2B nuclear phosphoinositides

项目摘要

Cells are constantly being instructed (programmed) to modulate their behaviour because of changes to environment of the body. In particular, continual repair of muscle tissue is essential to our daily lives but unfortunately is a process which deteriorates as we age. Deterioration in muscle function not only decreases our movement and function but also makes us more susceptible to metabolic type diseases such as diabetes and cardiovascular disease. Satellite cells are specialised cells that are present in small numbers within muscle tissue. When muscles are damaged either by exercise or during disease, the satellite cells become active and produce more muscle cells, a process called differentiation that helps to repair muscle tissue. The activation of satellite cells is complex and depends on signals generated within the environment of the damaged muscle. As organisms age the ability of satellite cells to respond to these signals and generate more muscle cells decreases and in part is responsible for age-induced deterioration in muscle function. Using a model of muscle cell generation we have found that by controlling an enzyme called PIP4K2B we can increase the ability of a cell to differentiate to produce muscle cells. PIP4K2B controls the levels of naturally occurring molecules called phosphoinositides that are present in the cells. It is well established knowledge that phosphoinositides are present in the plasma membrane of cells where they control many different cellular functions. However, over the years there has been growing evidence that phosphoinositides are also present in the nucleus, the cells' control centre, where their levels change in response to different environments. In fact in response to signals generated during muscle cell differentiation the levels of phosphoinositides in the nucleus go up. We now know that nuclear phosphoinositides interact with and change the function of special proteins in the nucleus that are involved in generating instructions that control the behaviour of the cells. One such protein is TAF3, which is part of different protein complexes that control the programming of cells. Interaction of TAF3 with phosphoinositides reprograms the cell to increase muscle cell differentiation. In this proposal we will use a novel state of the art technology called CRISPR CAS to change the DNA of cells so that we can investigate how PIP4K2B and nuclear phosphoinositides control TAF3 and its various complexes to increase muscle differentiation. We believe that specialised TAF3 complexes direct specific instructions to the cell in response to the interaction of TAF3 with phosphoinositides and we can investigate this using a technique called CHiP-Seq. We also think that nuclear phosphoinositides together with TAF3 act as a platform that helps to organise which instructions are given and how these are coordinated to increase muscle differentiation. We will investigate this using a novel technique called promoter capture HiC which will allow us to understand the three dimensional aspect of DNA in the nucleus that is used to generate these instructions. PIP4K2B is a very druggable protein as is the site of interaction between phosphoinositides and TAF3 and we hope that eventually we might be able to use our knowledge to develop drugs to harness this control process within a cell's nucleus and help satellite cells differentiate more effectively, thus aiding the process of muscle tissue repair.

由于身体环境的变化，细胞不断被指示（编程）来调节其行为。特别是，肌肉组织的持续修复对于我们的日常生活至关重要，但不幸的是，这个过程会随着年龄的增长而恶化。肌肉功能恶化不仅会降低我们的运动和功能，还使我们更容易患代谢型疾病，例如糖尿病和心血管疾病。卫星细胞是肌肉组织中少量存在的特殊细胞。当肌肉因运动或疾病而受损时，卫星细胞会变得活跃并产生更多的肌肉细胞，这一过程称为分化，有助于修复肌肉组织。卫星细胞的激活很复杂，取决于受损肌肉环境中产生的信号。随着生物体年龄的增长，卫星细胞对这些信号做出反应并产生更多肌肉细胞的能力下降，这在一定程度上是年龄引起的肌肉功能退化的原因。使用肌肉细胞生成模型，我们发现通过控制一种名为 PIP4K2B 的酶，我们可以提高细胞分化产生肌肉细胞的能力。 PIP4K2B 控制细胞中存在的称为磷酸肌醇的天然分子的水平。众所周知，磷酸肌醇存在于细胞质膜中，它们控制许多不同的细胞功能。然而，多年来，越来越多的证据表明磷酸肌醇也存在于细胞核（细胞的控制中心）中，其水平会根据不同的环境而变化。事实上，响应肌细胞分化过程中产生的信号，细胞核中磷酸肌醇的水平会上升。我们现在知道，核磷酸肌醇与细胞核中特殊蛋白质相互作用并改变其功能，这些蛋白质参与生成控制细胞行为的指令。 TAF3 就是这样的一种蛋白质，它是控制细胞编程的不同蛋白质复合物的一部分。 TAF3 与磷酸肌醇的相互作用可重新编程细胞以增加肌肉细胞分化。在本提案中，我们将使用一种称为 CRISPR CAS 的新颖的最先进技术来改变细胞的 DNA，以便我们可以研究 PIP4K2B 和核磷酸肌醇如何控制 TAF3 及其各种复合物以增加肌肉分化。我们相信，专门的 TAF3 复合物会针对 TAF3 与磷酸肌醇的相互作用向细胞发出特定指令，我们可以使用称为 CHiP-Seq 的技术对此进行研究。我们还认为核磷酸肌醇与 TAF3 一起充当一个平台，帮助组织发出哪些指令以及如何协调这些指令以增加肌肉分化。我们将使用一种称为启动子捕获 HiC 的新技术来研究这一问题，该技术将使我们能够了解细胞核中用于生成这些指令的 DNA 的三维方面。 PIP4K2B 是一种非常适合药物化的蛋白质，也是磷酸肌醇和 TAF3 之间相互作用的位点，我们希望最终能够利用我们的知识来开发药物来利用细胞核内的这种控制过程，并帮助卫星细胞更有效地分化，从而帮助肌肉组织修复过程。