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

• 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

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，细胞不断地被指示（编程）来调节它们的行为。特别是，肌肉组织的持续修复对我们的日常生活至关重要，但不幸的是，这是一个随着年龄增长而恶化的过程。肌肉功能的恶化不仅会降低我们的运动和功能，还会使我们更容易患上代谢型疾病，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化细胞。当肌肉因运动或疾病而受损时，卫星细胞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。卫星细胞的激活是复杂的，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，部分原因是年龄引起的肌肉功能退化。使用肌肉细胞生成的模型，我们发现通过控制一种称为PIP 4K 2B的酶，我们可以增加细胞分化产生肌肉细胞的能力。PIP 4K 2B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，在那里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中，细胞的控制中心，它们的水平会随着不同的环境而变化。事实上，响应于肌肉细胞分化期间产生的信号，细胞核中磷酸肌醇的水平上升。我们现在知道，核磷酸肌醇与细胞核中的特殊蛋白质相互作用并改变其功能，这些蛋白质参与产生控制细胞行为的指令。其中一种蛋白质是TAF 3，它是控制细胞编程的不同蛋白质复合物的一部分。TAF 3与磷酸肌醇的相互作用重新编程细胞以增加肌细胞分化。在这项提案中，我们将使用一种名为CRISPR CAS的最新技术来改变细胞的DNA，这样我们就可以研究PIP 4K 2B和核磷酸肌醇如何控制TAF 3及其各种复合物来增加肌肉分化。我们相信，专门的TAF 3复合物会响应TAF 3与磷酸肌醇的相互作用，向细胞发出特定的指令，我们可以使用一种名为CHiP-Seq的技术来研究这一点。我们还认为，核磷酸肌醇与TAF 3一起作为一个平台，有助于组织给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点，这将使我们能够了解用于产生这些指令的细胞核中DNA的三维方面。PIP 4K 2B是一种非常可药用的蛋白质，也是磷酸肌醇和TAF 3之间相互作用的位点，我们希望最终能够利用我们的知识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。

项目成果

Mechanisms of SARS-CoV-2 Inactivation Using UVC Laser Radiation.

• DOI: 10.1021/acsphotonics.3c00828
• 发表时间: 2024-01-17
• 期刊: ACS PHOTONICS
• 影响因子: 7
• 作者: Devitt, George;Johnson, Peter B.;Hanrahan, Niall;Lane, Simon I. R.;Vidale, Magdalena C.;Sheth, Bhavwanti;Allen, Joel D.;Humbert, Maria V.;Spalluto, Cosma M.;Herve, Rodolphe C.;Staples, Karl;West, Jonathan J.;Forster, Robert;Divecha, Nullin;McCormick, Christopher J.;Crispin, Max;Hempler, Nils;Malcolm, Graeme P. A.;Mahajan, Sumeet
• 通讯作者: Mahajan, Sumeet

Phosphatidylinositol-5-Phosphate 4-Kinases Regulate Cellular Lipid Metabolism By Facilitating Autophagy.

• DOI: 10.1016/j.molcel.2018.03.037
• 发表时间: 2018-05-03
• 期刊: Molecular cell
• 影响因子: 16
• 作者: Lundquist MR;Goncalves MD;Loughran RM;Possik E;Vijayaraghavan T;Yang A;Pauli C;Ravi A;Verma A;Yang Z;Johnson JL;Wong JCY;Ma Y;Hwang KS;Weinkove D;Divecha N;Asara JM;Elemento O;Rubin MA;Kimmelman AC;Pause A;Cantley LC;Emerling BM
• 通讯作者: Emerling BM

Exploring the controversial role of PI3K signalling in CD4+ regulatory T (T-Reg) cells.

• DOI: 10.1016/j.jbior.2020.100722
• 发表时间: 2020-04
• 期刊: Advances in biological regulation
• 作者: A. Poli;R. Fiume;S. Mongiorgi;A. Zaurito;B. Sheth;M. C. Vidalle;Shidqiyyah Abdul Hamid;Scott T. Kimber;F. Campagnoli;Stefano Ratti;Isabella Rusciano;I. Faenza;L. Manzoli;N. Divecha

PIP4K2B is mechanoresponsive and controls heterochromatin-driven nuclear softening through UHRF1.

PIP4K2B是机械响应性的，可以通过UHRF1控制异染色质驱动的核软化。

• DOI: 10.1038/s41467-023-37064-0
• 发表时间: 2023-03-14
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Poli, Alessandro;Pennacchio, Fabrizio A.;Ghisleni, Andrea;di Gennaro, Mariagrazia;Lecacheur, Margaux;Nastaly, Paulina;Crestani, Michele;Pramotton, Francesca M.;Iannelli, Fabio;Beznusenko, Galina;Mironov, Alexander A.;Panzetta, Valeria;Fusco, Sabato;Sheth, Bhavwanti;Poulikakos, Dimos;Ferrari, Aldo;Gauthier, Nils;Netti, Paolo A.;Divecha, Nullin;Maiuri, Paolo
• 通讯作者: Maiuri, Paolo

PIP4K2B: Coupling GTP Sensing to PtdIns5P Levels to Regulate Tumorigenesis.

PIP4K2B：将 GTP 传感与 PtdIns5P 水平耦合以调节肿瘤发生。

• DOI: 10.1016/j.tibs.2016.04.003
• 发表时间: 2016
• 期刊: Trends in biochemical sciences
• 影响因子: 13.8
• 作者: Fiume R