Enhancement of pancreatic beta cell generation in vitro and in vivo by post-translational regulation of Neurogenin3

通过 Neurogenin3 的翻译后调节增强体外和体内胰腺 β 细胞的生成

• 批准号: MR/K018329/1
• 负责人: Anna Philpott
• 金额: $ 48.05万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FK018329%2F1
• 关键词: Enhancement; pancreatic; beta; cell; generation

Many illnesses of middle and old age result from a progressive loss of tissue function including Alzheimers, Parkinsons Disease, heart failure and notably diabetes. These chronic conditions generally require long-term medication, but recent spectacular advances mostly grounded in basic biology have raised the possibility that defective cells could be directly replaced. An increase in the mass of beta islets cells, the type of cells that are missing or malfunctioning in diabetes, has been proposed to ameliorate type 1 diabetes, and is also likely to be an effective treatment for many sufferers of Type 2 diabetes. Indeed, successful islet transplantation has been shown to result in relief from a requirement for insulin therapy and relief from diabetes-related symptoms. So called stem cells, cells that can both proliferate indefinitely and can be induced to form more specialised cell types in a culture dish, could act as a source for beta cell replacement. However, a major obstacle remains in that stem cells must be induced to adopt beta cell fate and maintain that fate indefinitely when transplanted back into the patient, retaining functionality and avoiding the risk of cancerous tumour formation. It is widely acknowledged that for this to occur, stem cells must be taken down the developmental pathway that beta cells usually take when undergoing formation in the developing embryo. Based on these developmental pathways, protocols have been published where human embryonic stem cells can be induced to form relatively immature beta cells, while their in vivo implantation back into animals results in further maturation and beta cell functionality. However, a caveat to this approach is that cancer formation was also occasionally observed, indicating the need for tranplantation of beta cells that are differentiated to a more mature state to minimise this risk. This is currently a road-block to generating more mature beta cells in vitro that we may have uncovered a way of moving through.The protein Neurogenin 3 (Ngn3) is needed for formation of all beta cells during development and many experiments indicate that manipulating its activity could enhance beta cell formation from stem cells. However, normal Ngn3 only causes rather immature beta cells to be generated and these cannot respond to blood glucose levels in the same way as normal beta cells. We see that changing the chemical modifications of Neurogenin protein by preventing the addition of phosphates can significantly enhance the ability of Neurogenins to drive cell maturation and we hypothesise that this may make a modified form of Ngn3 much better at generating mature beta cells than the normal protein. This project is designed to test this hypothesis and, if correct, to understand the mechanism of this enhanced cellular maturation. To help us move this project towards clinical application, we are working with Prof Douglas Melton at the Harvard Stem Cell institute who is focussed on generating beta cells of sufficient quality for human trails.

许多中年和老年疾病是由于组织功能的进行性丧失而导致的，包括老年痴呆症，帕金森氏病，心力衰竭和尤其是糖尿病。这些慢性疾病通常需要长期药物，但是最近的壮观进展主要基于基本生物学，这增加了可以直接替换有缺陷的细胞的可能性。已经提出，β小岛细胞的质量增加，糖尿病中缺失或故障的细胞类型可以改善1型糖尿病，并且对于许多2型糖尿病患者而言也可能是一种有效的治疗方法。实际上，已证明成功的胰岛移植可缓解胰岛素治疗的要求和缓解与糖尿病相关的症状的缓解。所谓的干细胞，可以无限期增殖并可以在培养皿中形成更专业的细胞类型的细胞，可以作为β细胞替代的来源。但是，必须诱使干细胞采用β细胞命运，并在移植回患者时无限期地保持命运，保留功能并避免形成癌性肿瘤的风险时，必须无限期地保持命运。人们普遍认为，要发生这种情况，必须将干细胞删除β细胞在发育中的胚胎中进行形成时通常采取的发育途径。基于这些发育途径，已经发表了方案，其中可以诱导人类胚胎干细胞形成相对不成熟的β细胞，而它们的体内植入将其植入动物，从而导致进一步的成熟和β细胞功能。但是，这种方法的一种警告是，偶尔还观察到癌症形成，表明需要将β细胞的静态化与更成熟的状态区分开以最大程度地减少这种风险。目前，这是在体外生成更成熟的β细胞的一个路障，我们可能已经发现了一种移动方法。蛋白质神经蛋白3（NGN3）在发育过程中需要形成所有β细胞的蛋白质神经蛋白蛋白3（NGN3），许多实验表明操纵其活性可以增强干细胞的β细胞形成。但是，正常的NGN3仅导致相当不成熟的β细胞产生，并且这些β细胞无法与正常β细胞相同的方式对血糖水平响应。我们看到，通过防止磷酸盐的添加来改变神经蛋白蛋白的化学修饰可以显着增强神经蛋白驱动细胞成熟的能力，我们假设这可能使NGN3的修饰形式比正常蛋白更好地产生成熟的β细胞。该项目旨在检验这一假设，如果正确，则旨在了解这种增强的细胞成熟的机制。为了帮助我们将该项目转向临床应用，我们正在与哈佛干细胞研究所的道格拉斯·梅尔顿教授合作，他专注于为人类步道生成足够质量的β细胞。

项目成果

Subcellular localisation modulates ubiquitylation and degradation of Ascl1.

• DOI: 10.1038/s41598-018-23056-4
• 发表时间: 2018-03-15
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Gillotin S;Davies JD;Philpott A
• 通讯作者: Philpott A

Multi-site Neurogenin3 Phosphorylation Controls Pancreatic Endocrine Differentiation.

• DOI: 10.1016/j.devcel.2017.04.004
• 发表时间: 2017-05-08
• 期刊: Developmental cell
• 影响因子: 11.8
• 作者: Azzarelli R;Hurley C;Sznurkowska MK;Rulands S;Hardwick L;Gamper I;Ali F;McCracken L;Hindley C;McDuff F;Nestorowa S;Kemp R;Jones K;Göttgens B;Huch M;Evan G;Simons BD;Winton D;Philpott A
• 通讯作者: Philpott A

N-terminal phosphorylation of xHes1 controls inhibition of primary neurogenesis in Xenopus.

xHes1 的 N 末端磷酸化控制爪蟾初级神经发生的抑制。

• DOI: 10.17863/cam.35808
• 发表时间: 2019
• 作者: Hardwick L

The developmental origin of brain tumours: a cellular and molecular framework.

• DOI: 10.1242/dev.162693
• 发表时间: 2018-05-14
• 期刊: Development (Cambridge, England)
• 作者: Azzarelli R;Simons BD;Philpott A
• 通讯作者: Philpott A

Neurogenin3 phosphorylation controls reprogramming efficiency of pancreatic ductal organoids into endocrine cells.

• DOI: 10.1038/s41598-018-33838-5
• 发表时间: 2018-10-18
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Azzarelli R;Rulands S;Nestorowa S;Davies J;Campinoti S;Gillotin S;Bonfanti P;Göttgens B;Huch M;Simons B;Philpott A
• 通讯作者: Philpott A