An investigation into the functional role of the RhoGAP protein SH3BP1/ARHGAP43 in Autosomal Dominant Polycystic Kidney Disease

RhoGAP 蛋白 SH3BP1/ARHGAP43 在常染色体显性多囊肾病中的功能作用研究

• 批准号: MR/W015579/1
• 负责人: Joshua Griffiths
• 金额: $ 34.43万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FW015579%2F1
• 关键词: investigation; into; functional; role; RhoGAP

Autosomal Dominant Polycystic Kidney Disease (ADPKD) is a potentially life changing and life-threatening disease that affects up to 1:1000 people. The disease causes fluid filled cysts to grow in the kidneys where they continue to grow to huge sizes through a patient's life. These cysts slowly reduce the function of both kidneys until patients reach end stage kidney failure, most commonly by the ages of 50 to 60. When patients reach end stage kidney failure, they will need dialysis or kidney transplant to survive. ADPKD is caused by a problem in a patient's genetic code which leads to defects in the proteins Polycystin 1 (PC1) or Polycystin 2 (PC2). Parents with one of these genetic problems have a 50:50 chance of passing this down to their children and there is currently no cure. There is only one treatment that can delay developing end stage kidney failure, but it has significant side effects and only slows the disease a small amount. Even though we have known what damaged proteins cause ADPKD for a number of years, how damage to these proteins causes cysts to grow remains a mystery. By looking at the cells lining these these cysts we can see they grow and divide faster but also move slower and are less adherent to each other than normal cells. However, how PC1 or PC2 damage leads to these changes in cells is not currently know.This project will aim to understand what the functions of PC1 and PC2 do when not defective and how defects in these proteins lead to the known changes in cell behaviour and therefore cyst growth. We have newly discovered that PC1 interacts with another protein, SH3BP1. This protein has been shown to be important for cell movement and cell to cell adhesion. Further, it is also important in the uptake into the cells of certain receptors that stimulate cell growth after being activated. This process has only recently been discovered and its potential role in ADPKD has not yet been explored. SH3BP1 provides an exciting potential explanation for why defective PC1 leads to the changes in cells we see under the microscope and therefore how cysts develop.I will use lab-based techniques to establish what parts of PC1 and SH3BP1 interact specifically and if this interaction changes how either protein works. I will also be able to screen for any other proteins that may also be involved in this interaction using mass spectrometry. Using high powered microscopy, I will look at how specific growth receptors are taken up by cells in real time and evaluate if this is different in cells with damaged PC1 or SH3BP1. Finally, to confirm the role of the interaction of PC1 and SH3BP1 in cyst growth, I will see if increasing or decreasing SH3BP1 in cells with damaged PC1 will reduce the growth of cysts in 3D. By completing this project, we will have a better understanding of how cysts grow in ADPKD and this may lead to better treatments for the disease which are desperately needed.

常染色体显性多囊性肾脏疾病（ADPKD）是一种潜在的改变生命和威胁生命的疾病，可影响1：1000人。该疾病会导致填充液体囊肿在肾脏中生长，并在患者的生活中继续生长到巨大的尺寸。这些囊肿逐渐降低了两个肾脏的功能，直到患者达到末期肾衰竭，最常见于50至60岁。当患者达到末期肾衰竭时，他们需要透析或肾脏移植才能生存。 ADPKD是由患者的遗传密码中的问题引起的，该问题导致蛋白质Polycystin 1（PC1）或Polycystin 2（PC2）中的缺陷。有一个遗传问题之一的父母有50:50的机会将其传给孩子，目前无法治愈。只有一种治疗方法可以延迟发展末期肾脏衰竭，但是它具有重大的副作用，只会减慢疾病的量。即使我们知道蛋白质受损的蛋白质会导致ADPKD多年，但这些蛋白质的损害如何导致囊肿生长仍然是一个谜。通过查看这些囊肿的细胞，我们可以看到它们生长和分裂更快，但移动速度也较慢，并且比正常细胞更不依赖彼此。但是，当前不知道PC1或PC2损伤如何导致细胞中的这些变化。此项目的目的是了解PC1和PC2的功能在不存在的情况下的功能以及这些蛋白质中的缺陷如何导致细胞行为的已知变化以及囊肿的生长。我们新发现，PC1与另一种蛋白质SH3BP1相互作用。该蛋白已被证明对细胞运动和细胞对细胞粘附至关重要。此外，在激活后刺激细胞生长的某些受体的细胞中，它也很重要。直到最近才发现此过程，并且尚未探索其在ADPKD中的潜在作用。 SH3BP1提供了一个令人兴奋的潜在解释，说明为什么有缺陷的PC1导致我们在显微镜下看到的细胞变化，因此囊肿如何开发。我将使用基于实验室的技术来确定PC1和SH3BP1的哪些部分特定相互作用，以及这种相互作用是否会改变任何一种蛋白质的作用。我还将能够筛选使用质谱法参与这种相互作用的任何其他蛋白质。使用高功率显微镜，我将研究细胞如何实时吸收特定的生长受体，并评估这在受损的PC1或SH3BP1的细胞中是否有所不同。最后，为了确认PC1和SH3BP1在囊肿生长中的相互作用的作用，我将看到PC1受损细胞中SH3BP1的增加或减少会减少3D中囊肿的生长。通过完成该项目，我们将更好地了解ADPKD中囊肿的生长方式，这可能会为迫切需要的疾病提供更好的治疗方法。