NIHRDataInsights/HRCSData huggingface dataset:
Title: Implications of the Vaginal Microbiome in Preterm Premature Rupture of Membranes (PPROM)
Funder: Medical Research Council

Abstract: Preterm Premature Rupture of Membranes (PPROM) precedes preterm birth (PTB) in 30-40% of cases and is a cause of neonatal mortality and morbidity. A relationship exists between ascending vaginal infection, PPROM and preterm labour (PTL). The vaginal microbiota of pregnancy is dominated by Lactobacillus spp., which inhibit the growth of pathogens through lactic acid and bacteriocin secretion. Bacterial exposure to the neonate occurs during delivery therefore the composition of vaginal microbiota is an important factor in neonatal colonisation. The hypothesis of this study is that a dysbiosis of the vaginal microbiome is a determinate of PPROM and influences the risk of subsequent infection, PTL and neonatal colonisation. The objectives of the study are to: i) Characterise the "normal" vaginal microbiome throughout pregnancy using 454 pyrosequencing methods and determine how advancing gestational age affects the composition of the vaginal microbiome. ii) Assess how the vaginal microbiome differs in pregnancies complicated by PPROM and examine how vaginal microbiota may influence time to delivery (latency), risk of ascending infection, neonatal outcome and determine how current clinical treatment (erythromycin) impacts upon the vaginal microbiome. iii) Understand how vaginal microbiota may be functionally implicit in PPROM by testing for their presence in gestational tissues following delivery and identifying which inflammatory pathways are activated in these tissues. iv) Use MS and NMR-based metabolic profiling to explore the inter-relationship between the vaginal microbiome and vaginal metabolome and assess how host-microbe interaction is reflected in the urinary metabolic profile. v) Identify urinary metabolites with prognostic and diagnostic potential for PPROM. By applying a systems biology approach to PPROM, this proposal will improve understanding of the vaginal microbiome in PPROM should lead to improved clinical management and treatment strategies.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Investigating the human myometrial acetylome and identifying potential new targets for treating preterm labour
Funder: Medical Research Council

Abstract: Data from our group indicate the importance of myofilament-associated protein acetylation in regulating human myometrial contractility. A multi-faceted strategy will be followed to determine the extent of myometrial acetylated proteins utilising human biopsies from pregnant women at term and tissues obtained from a clinically relevant animal model of human pregnancy/preterm labour. Our aim is to identify myometrial cell proteins that change their acetylation status by pharmacological manipulation. These may thus serve as potential novel targets for the future development of agents to treat preterm labour. The Stable Isotope Labelling of Cells in Culture (SILAC) of primary myometrial cells treated to increase protein acetylation with the lysine deacetylase (KDAC) inhibitor TSA or lysine acetyltransferase (KAT) agonist CTB, will be established. Protein lysates from these cultures will be trypsin digested and acetylated peptides immunoprecipitated using lysine acetylated-specific antibodies and subsequently fractionated using an Agilent 3100 OffGel fractionater and analysed using LCMSMS on a Dionex U3000 nano-HPLC system coupled to an Orbitrap LTQ XL Discovery mass spectrometer. Peptides will be distinguished by their unique mass/charge ratios that will enable (i) identification of any newly acetylated peptide sequences and (ii) assessment of the relative changes in abundance of acetylated species. The peptides from SILAC will than serve as internal standards for MS analysis of the protein acetylation changes in the native human tissues at rest or treated with TSA/CTB. Our preliminary data also suggest a potential tocolytic benefit of TSA treatment in the Guinea pig model of preterm labour. Similar proteomic procedures will, therefore, be performed in myometrial tissues from animals undergoing preterm labour both before and after treatment with TSA/CTB. The neonatal outcome will be subsequently established by following pups for 3 months post-natally.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: MOTIF: MObile Technology for Improved Family Planning
Funder: Medical Research Council

Abstract: MOTIF: MObile Technology for Improved Family Planning services is a research project that includes a multisite individual RCT that will reliably establish the impact of a mobile phone-based intervention on use of post-abortion family planning (PAFP) in Cambodia. Unsafe abortion accounts for one in eight maternal deaths in developing countries. Clients attending for abortion often have an unmet need for family planning. In Cambodia, over 50% of clients do not use a reliable method of PAFP, and around 25% will have a repeat abortion within five years. Mobile phones allow personalised support to be delivered inexpensively wherever the person is located. MOTIF is the first trial to evaluate a mobile phone-based family planning intervention in a low-income setting. Key achievements to date include: (1) Conducting formative research to understand why women attend for abortion and reasons for not using PAFP (2) Developing mobile phone-based intervention to support PAFP in Cambodia (3) Commencing a RCT with 500 participants to evaluate the intervention Key objectives of the proposed research are to: (1) Conduct in-depth analysis of 4 month trial follow up data which will have been already collected (2) Conduct 12 and 24-month follow up of trial participants already recruited in order to (a) evaluate the long-terms health outcomes of the MOTIF intervention to support PAFP and (b) conduct prospective analysis to calculate rates and risk factors of/for repeat abortion, pregnancy (3) Conduct additional qualitative and cost-effectiveness analysis of the intervention (4) Conduct a Cochrane review entitled 'Mobile phone-based interventions for improving adherence and acceptability of contraception' (5) Conduct analysis of contraceptive discontinuation rates from the Cambodia Demographic and Health Survey 2010 (6) Provide the applicant with rigorous research training at LSHTM in order to achieve the above objectives and become a world leader in the field


Research activity codes: 6.6 PSYCHOLOGICAL AND BEHAVIOURAL;6.8 COMPLEMENTARY
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Long term health outcomes for women and their children after assisted reproductive technologies: a data linkage study for England
Funder: Medical Research Council

Abstract: The main aim of this project is to assess the long-term health outcomes of mothers and children after the use of assisted reproductive technologies (ART) in England. A linked dataset combining information from the Clinical Practice Research Datalink and the Human Fertilisation and Embryology Authority (HFEA) register will be created. About 280,000 babies, born 1991-2009, and their mothers, who are registered at GP practices in England, will be included. Duration of follow-up varies from a few months to over 20 years, depending on date of delivery. The main 'exposure' is ART, which comprises IVF, ICSI, and ovulation induction; other techniques (e.g. blastocyst transfer) will be examined, where possible. Outcome data include all consultations, diagnoses, prescriptions and referrals recorded by GPs, supplemented by data on hospital episodes and mental health treatment. Key outcomes for children include growth, illness and infection, atopic and allergic conditions, behaviour and developmental outcomes; and for mothers, physical and mental health. Multivariable regression models will be used to estimate adjusted rate ratios and odds ratios as appropriate, comparing outcomes in the ART and non-ART groups. An economic analysis will compare differences in NHS expenditure on the long-term provision of care for ART and non-ART pairs, and model future costs. Concern among researchers that low consent rates since 2009 adveresly affect the representativeness of the HFEA dataset, means that the newest data are under-utilised. Methodological work to explore the impact of missingness, and to develop methods to reduce the potential bias, will be conducted. Qualitative research, exploring patients' decisions to consent (or not) to studies of ART will also be completed, through face-to-face interviews and thematic analysis. This study will contribute to the evidence for the safety and costs of ART in England, and provide tools for future work using these rich data sources.


Research activity codes: 7.3 MANAGEMENT AND DECISION MAKING
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Dual functions of WDR11 in the pathogenesis of hypogonadotrophic hypogonadism
Funder: Medical Research Council

Abstract: Reproductive failure occurs in 10-15% of all couples and nearly 30% of them have unexplained infertility. Defects in the hypothalamic-pituitary-gonadal axis are likely to underlie a significant proportion of the aetiologies in these patients, but a critical barrier to proper diagnosis and treatment is the lack of understanding of the specific mechanisms of impairment. WDR11 gene was identified by positional cloning from normosmic hypogonadotrophic hypogonadism (nHH) and Kallmann syndrome (KS) patients. The mechanism of how WDR11 regulates reproductive function is not known, except that WDR11 mutations interfere with its binding to EMX1. Based on the findings of human mutations, we hypothesise that it will affect GnRH neuron migration and/or regulate GnRH secretion or action. Recently, several genes have been shown to affect not only hypothalamic function, but also directly affect gonadal function. In fact, our preliminary data suggests that WDR11 be one such protein that affects the migration of primordial germ cells (PGCs), resulting in primary gonadal defects. We hypothesise that WDR11 is important for normal hypothalamic function and PGC migration. To test this hypothesis, we will determine the phenotype of the Wdr11 KO mouse and confirm the effects of WDR11 deficiency on the development and establishment of GnRH neurons and PGCs. We will also investigate the role of WDR11 on the migratory behaviour of PGCs using Stella-GFP transgenic mouse that allows purification and live monitoring of PGCs. We will also determine the molecular mechanisms of WDR11 and EMX1 in human and mouse cell lines and test the potential involvement of Hedgehog signal pathway. These studies will reveal novel insights into the function of WDR11 in the regulation of GnRH neuronal migration and establishment of PGCs in the gonads, disruptions of which underlie human reproductive disorders.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: MICA: Identification of age-related and age-independent changes to meiotic chromosome structure and their association with aneuploidy in human oocytes
Funder: Medical Research Council

Abstract: Changes to chromosome structure during the prolonged meiotic arrest of human oocytes have been proposed to underlie age-related aneuploidy of human foetuses. Age-related aneuploidy poses major risks to reproductive health. The goal of this project is to characterise chromosomal changes directly in human oocytes from women of different ages. We focus on several important themes based upon studies in model organisms, aiming to increase knowledge of the mechanisms responsible for oocyte aneuploidy directly in human. In possession of this knowledge, future development of clinical pre-gestational tests for aneuploidy risk will become a possibility. In this work, we shall test whether meiotic chromosome structure deteriorates with advanced maternal age in human oocytes, such as precocious separation of homologous chromosomes or sister chromatids. Such structural changes would strongly predispose chromosomes to missegregate at meiosis I or II, resulting in chromosomally imbalanced oocytes. We will also study the localization pattern of several candidate protein complexes to examine whether their localisation or relative abundance correlates with such aberrant chromosome structures. These candidate complexes include cohesin, condensin, synaptonemal complex and DNA repair factors; all implicated as major factors in chromosome structure and segregation from studies from model organisms. To achieve our objectives, we have assembled a world-class cross-disciplinary team of discovery scientists, statisticians, embryologists, as well as clinicians. We propose a logical and realistic plan of work, governed by high ethical standards and rigorous methodology to address important scientific and clinical questions.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Life course epidemiology of women's reproductive health and its relation to chronic disease
Funder: Medical Research Council

Abstract: Reproductive health is important to a woman's overall health and well-being and is increasingly recognised as a sentinel for chronic disease in later life. However important knowledge gaps in the life course epidemiology of female reproductive health and in the understanding of the nature of its relation to chronic disease remain. Here I will study multiple exposures from foetal life to adulthood in relation to reproductive health and potential as measured by reproductive hormones in early adulthood, a critical time for family planning. I will use information on multiple reproductive indicators such as menstrual cycle characteristics, parity, pregnancy complications, gynaecological disorders and their treatment, to better characterize women's reproductive health. I will study relations between these reproductive indicators (separately and in combination), reproductive hormones and major chronic disease outcomes (including breast cancer, cardiovascular disease, diabetes, osteoporotic fractures, depression, dementia, and lung disease) and disease markers to study causality and identify mechanistic pathways. Finally, I will examine whether readily available information on indicators of female reproductive health can improve risk stratification in women. I will use data from multiple complimentary studies and a variety of analytical approaches to identify causal pathways, as well as novel statistical methods to formally compare different life course models of the relationship between early life exposures, reproductive health and later disease outcomes. This work has the potential to inform preventive strategies, to identify treatment targets for female reproductive health as well as related chronic diseases, and to improve risk stratification in women at increased risk of diabetes, cardiovascular disease, osteoporosis and dementia. This is important because most are preventable or have a better prognosis if treated earlier and all are costly to health services.


Research activity codes: 2.5 RESEARCH DESIGN AND METHODOLOGIES (AETIOLOGY)
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Complement as a novel regulator of Wnt signalling during craniofacial development
Funder: Medical Research Council

Abstract: The importance of Wnt signalling in embryonic development has been well documented. In adult life, Wnt signalling continues to be critically important for processes such as governing cell fate decisions and maintaining the stem cell niche. During these processes, as in neural crest induction, levels of Wnt activity must be precisely controlled in time and space. Yet many of the molecular details underlying this regulation remain poorly understood. Our preliminary data suggest that the complement cascade, normally associated with the immune response, may be an important regulator of Wnt signalling during neural crest induction and may, therefore, play an important role in craniofacial development. As the complement cascade contains more than 35 proteins we will focus our attention on the terminal effector of the pathway, complement component C9. In this project we will propose the novel hypothesis that C9 controls Wnt signalling during neural crest and craniofacial development. The aim of this project is to identify the molecular mechanism by which C9 controls Wnt signalling in general, and how this is important for neural crest and craniofacial development. The major contributions to be expected from this project are: i) The discovery of a novel regulator of Wnt activity, a signalling pathway essential for normal development and critical in disease. ii) The characterization of a completely new and unexpected role for C9 in neural crest and craniofacial formation. iii) The description of the mechanism by which C9 regulates Wnt signalling.j iv) The identification of novel C9 partners involved in neural crest formation that could also be partners of C9 in the immune system. Our findings could have important implications for human health, as it may suggest the avoidance of complement inhibitors to treat immune diseases during pregnancy


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Spectroscopic Probes of Energy Regulation & Metabolism (SPERM): Using High Resolution MR spectroscopy to identify biomarkers of male fertility
Funder: Medical Research Council

Abstract: This project aims to use the techniques of high-resolution Nuclear Magnetic Resonance (NMR) spectroscopy to examine human sperm energy metabolism. The aim is to shed new light on an age-old debate of how sperm are appropriately fuelled and whether dysfunctions of sperm metabolism are contributing to the male infertility related to poor sperm motility (asthenozoospermia). We propose using the following techniques: (i) High Resolution Magic Angle Spinning (HR-MAS) 1H NMR spectroscopy at 9.4T of sperm pellet in a zirconium rotor and spun at a frequency of 3-6 KHz at an angle thetam = 54.74 degrees to the magnetic field direction; (ii) High Resolution NMR spectroscopy at 9.4T of sperm incubated with 13C labelled substrates including 13C1-pyruvic acid, 13C2-pyruvic acid, 13C1-lactate, 13C5-glutamine, 13C1-6,d7-glucose, 13C2-fructose, and 13C1-butyric acid (fatty acid analogue); and (iii) Dissolution Dynamic Nuclear Polarisation enhanced 13C NMR spectroscopy using a HyperSense DNP polariser linked to a 7T Bruker MRI/S system to examine relatively short-term metabolic changes in sperm. For each technique, sperm will be obtained from healthy male volunteers (aged 18 to 65) and their motility assessed by computer to quantify the proportion and speed of motile sperm (and define asthenozoospermia in a robust way). Sperm will then be isolated from semen using a two-step 45/95% (v/v) Percoll gradient suspended in phosphate buffered saline and then removing any residual leukocytes by subjecting sperm found both in the pellet and at the 45/95 interface to magnetic cell sorting with CD45 Dynabeads. For each of our objectives, we will scan sperm from at least 6 different males to compare both between and within male differences in metabolite concentrations, metabolism of labelled substrates and real time metabolic kinetics. Metabolites will be identified from spectra using the dedicated NMR processing packages (Bruker Topspin) and custom software (Matlab).


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Studies on the dose-dependent functions of TGFbeta-Smad2/3 signalling
Funder: Medical Research Council

Abstract: During embryonic development TGFbeta signalling is essential for gastrulation and patterning, including specification of body axes. In the adult it plays roles in tissue homeostasis by promoting differentiation and counteracting proliferation, it also exhibits tumour-suppressing functions in several tissues. Deregulation of signalling is implicated in various diseases, including vascular disorders, fibrosis and cancer. TGFbeta regulates epithelial to mesenchymal transition (EMT), a migratory cellular behaviour required during gastrulation and organogenesis, while within tumours EMT promotes metastasis. The TGFbeta ligand Nodal is essential for gastrulation and anterior-posterior (A-P) patterning of the vertebrate embryo. A-P axis specification is the best-characterised dose-dependent function of this pathway, with high levels required for anterior and lower for more posterior structures. However, it is not known how this is established. Several regulatory factors have been identified suggesting that TGFb/Nodal signalling levels are tightly regulated. In mice, loss of function mutations in single intracellular negative regulators of the pathway, such as SnoN, Ski, Smad7 and Smad6, failed to impair A-P patterning raising concerns about their normal function in vivo. On the contrary, loss of Arkadia/RNF111, which has been shown to degrade all the above negative factors to enhance intracellular Nodal signalling, causes severe anterior truncation of the A-P embryonic axis. This implies that in its absence there is hyperactivity of negative regulators that reduces signalling. We propose to examine this using mouse genetics and A-P patterning in the embryo, along with the differentiation of embryonic stem cells towards anterior endoderm-foregut cell fates in culture. We expect our results to shed light on the dose-dependent functions of Nodal by revealing the importance of intracellular regulation in establishing the different responses and the underlying mechanisms.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Identifying sperm-egg receptor pairs essential for mammalian fertilization to select new targets for fertility treatment and contraception.
Funder: Medical Research Council

Abstract: Fertilization involves the cellular recognition and subsequent fusion of haploid gametes to create a diploid zygote. We know little about the molecular events involved in mammalian fertilization, including the identity of the sperm-egg receptor pairs involved. Largely, this is due to technical challenges due to the rarity of eggs and because the interactions between extracellular receptors are weak requiring specialised techniques to identify them. We have developed methods based on creating highly avid binding probes to detect weak extracellular interactions and have applied them to the problem of sperm-egg recognition and identified the first sperm-egg receptor pair that is essential for mammalian fertilization (Bianchi et al. Nature 2014 v508 p483). There is good evidence to suggest that there are additional important sperm-egg recognition molecules and this grant application outlines how we intend to apply the same approach to identify additional sperm-egg receptor pairs that could be useful for developing novel contraceptives and fertility treatments. We will use a library of ~40 highly avid recombinant proteins that represents the secretome and cell surface receptor repertoire of the mouse spermatozoon and systematically test them for interactions with the oolemma and zona pellucida. For those sperm probes that exhibit positive staining with the egg, we will identify the molecular nature of the egg receptor using expression cloning (an approach we have successfully used in the past) and other receptor matching techniques that we have developed in the laboratory. The function of any identified interactions will be determined using transgenic mice.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;5.2 CELLULAR AND GENE THERAPIES
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH