Investigating the human myometrial acetylome and identifying potential new targets for treating preterm labour

早产在英国每年发生50万例，仍然是英国新生儿死亡和发病的主要原因。存活下来的早产儿，其长期身体或精神残疾的风险较高。世卫组织估计，在全球范围内，每年有多达1500万例早产，约有100万例新生儿死亡。因此，开发更好的治疗早产的方法是一个主要的研究目标，它有助于解决这种疾病对健康和社会经济的重大影响。早产发生率的一个主要因素是子宫收缩的过早激活，即使在没有其他临床风险关联的情况下，如感染，但仍然没有对母亲和/或新生儿无风险因素的广泛有效的治疗方法。目前，与其他器官系统相比，我们对足月或早产子宫激活控制机制的理解较弱，这反映在我们无法在不危及新生儿结局的情况下开出限制子宫过早收缩的药物。本项目旨在解决这些问题。最近高通量蛋白质组学的技术和实验进展使我们意识到可能调节细胞功能的蛋白质修饰范围不断扩大。我们实验室发表的证据表明，肌丝相关蛋白乙酰化是调节人类子宫肌收缩性的重要机制。通过抑制赖氨酸去乙酰化酶（kdac）来增加蛋白质乙酰化的药物不仅在体外降低人子宫肌收缩力，而且在早产动物模型中延缓子宫激活，同时提高新生儿存活率。在目前的研究中，我们将采用先进的蛋白质组学技术来鉴定这些过程中乙酰化的靶蛋白。利用跨学科的方法，包括科学和临床专业知识，以及人类离体活检与体内早产动物模型的实验，我们将在KDAC抑制或赖氨酸乙酰化酶（KAT）激活的治疗下，提供肌层蛋白乙酰化变化的新鉴定，包括单个乙酰化位点的鉴定。同时，我们将确定KDAC抑制或KAT激活延缓实验性早产和改善新生儿存活率和预后的能力。由于这些是开发任何新的抗早产药物的明确愿望，我们的数据不仅将告诉我们涉及调节足月和早产的新分子机制，而且还将告诉我们有希望的蛋白质靶点，用于开发未来的早产治疗方法。除了对妊娠和妊娠并发症感兴趣的直接科学和临床研究从业人员之外，这项工作还将提供一个公开可用的数据集，其中包含对人类和豚鼠细胞蛋白质乙酰化的广泛评估，这将使比较生物学家和蛋白质组学生物信息学家感兴趣。

Preterm birth occurs in >50,000 cases per annum in the UK and is still the major cause of neonatal mortality and morbidity in the UK. Preterm babies that survive do so with an elevated risk of long-term physical or mental handicap. Globally, the WHO estimate that up to 15 million births per annum occur prematurely with approximately 1 million neonatal deaths. The development of better therapeutics in managing preterm birth, therefore, is a prime research aim that serves to tackle the considerable health and socio-economic impacts of this disorder. A major contributor to the incidence of preterm birth is premature activation of uterine contractions, even in the absence of other clinical risk associations such as infection, yet there still is no broadly efficient therapeutic treatment free of risk factors for the mother and/or neonate. At present, our understanding of the mechanisms that control activation of the uterus at term or preterm is weak compared to other organ systems and this is reflected in our inability to prescribe drugs that limit premature uterine contractility without jeopardising neonatal outcome. This project seeks to address these issues.Recent technical and experimental advances in high throughput proteomics have made us aware of an expanding range of protein modifications that may regulate cell function. Published evidence from our laboratory has indicated that myofilament-associated protein acetylation is an important mechanism for regulating human myometrial contractility. Drugs which act to increase protein acetylation by inhibiting lysine deacetylases (KDACs) not only reduce human myometrial contractility in vitro but also delay uterine activation in an animal model of preterm labour whilst improving neonatal survival. In the present study we will employ advanced proteomic techniques to identify proteins that are targets for acetylation in these processes. Utilising a cross-disciplinary approach, encompassing scentific and clinical expertise, and experimentation on human ex vivo biopsies in alliance with in vivo animal model of preterm labour, we will provide novel identification of changes in myometrial protein acetylation, including the identification of individual acetylation sites, with treatment by KDAC inhibition or lysine acetylase (KAT) activation. In concert, we will establish the ability of KDAC inhibition or KAT activation to delay experimentally-induced preterm labour and improve neonatal survival and outcome. As these are the defining aspirations of developing any new tocolytic drugs, our data will not only inform us of novel molecular mechanisms involved in regulating term and preterm labour, but will also inform us of promising protein targets for development of future therapeutics for preterm labour. Beyond the immediate scientific and clinical research practitioners interested in pregnancy and pregnancy complications, this work will also provide a publicly-available dataset with a broad scale assessment of human and guinea pig cell protein acetylation which will be of interest to comparative biologists and proteomic bioinformaticians.