NIHRDataInsights/HRCSData huggingface dataset:
Title: Ca2+ signalling in regulation and therapeutic manipulation of human sperm behaviour
Funder: Medical Research Council

Abstract: To ascend the female tract, penetrate the layers that surround the oocyte and fertilise it a sperm must employ diverse motility patterns, switching between behaviours in response to cues. [Ca2+]i signaling, the central regulator of sperm motility, mediates these behavioural choices. Impaired Ca2+ signaling renders the sperm infertile. This study addresses regulation of human sperm motility and how it may be pharmacologically enhanced in subfertile men. Activation of CatSper channels and release of Ca2+ from a store at the sperm neck are both implicated in motility regulation. We have shown that, while both raise [Ca2+]i, they elicit functionally different behaviours in human sperm. CatSper activation causes subtle changes in flagellar beat that enhance penetration into viscous medium whereas mobilisation of stored Ca2+ induces robust hyperactivation. Since CatSper is activated by diverse stimuli (pHi, hormones, prostaglandins, Em) whereas the Ca2+ store is sensitive primarily to Ca2+-induced-Ca2+-release, we propose that (a) CatSper functions as a multimodal Ca2+-signaling node that secondarily recruits release of the stored Ca2+ and (b) availability of Ca2+ store release (and the behaviours it regulates) is regulated by capacitation and by stimuli encountered in the female tract such as NO. Using advanced imaging, Ca2+ uncaging, patch clamp and sperm tracking to assess signaling and behavioural regulation in human sperm we will (i) characterise generation/regulation of Ca2+ signals mediated by CatSper, stored Ca2+ and putative non-CatSper conductances in response to endogenous stimuli (ii) elucidate the diversity of sperm behaviours, their functional significance and their 'selection' by Ca2+ signals induced by stimuli encountered in the female tract and (iii) characterise incidence and types of Ca2+-signaling lesions in men with motility-based subfertility and identify agents for targeted manipulation of Ca2+ signaling components to reverse/bypass these lesions.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Role of growth factors of the TGFbeta superfamily in aberrant follicle development in polycystic ovary syndrome
Funder: Medical Research Council

Abstract: Polycystic ovary syndrome (PCOS) is the commonest endocrine disorder in women and is the major cause of anovulatory infertility. PCOS is characterized by hypersecretion of ovarian androgens but women with PCOS also carry an increased risk of metabolic dysfunction and have 2-4 fold increased risk of developing type 2 diabetes mellitus (T2DM). Anovulation in PCOS is characterized by arrest of antral follicles, and appears to a function of the abnormal endocrine environment, but there is increasing evidence for aberrations in preantral follicle development which may predispose to later, antral follicle dysfunction. Our recent studies using human ovarian tissue point to a key role for growth factors of the TGFbeta (TGFb) superfamily in development of preantral follicles. Our overall objective is to test the hypothesis that aberrant expression of TGFb ligands and their receptors plays a key role in abnormal preantral follicle development and function in PCOS. We suggest that the findings in ovarian tissue will provide insight into the putative role of aberrant TGFb signalling in metabolic dysfunction (and in predisposition to T2DM), a proposal that can be explored in parallel studies of adipose tissue. The specific aims are to study (1) the stage related pattern of gene expression of TGFb ligands, their receptors and endogenous antagonists in preantral follicles from ovaries of women with PCOS compared with that in normal ovaries (using next generation sequencing and qPCR); (2) where and when proteins of TGFb ligands, receptors, antagonists, and Smads are expressed in polycystic compared with normal ovaries; (3) the effects of TGFb ligands and antagonists on early follicle development in cultured slices of human normal and PCOS ovarian cortex; (4) the effect of androgen treatment on expression of TGFb ligands, receptors, antagonists and Smads in mouse ovaries and (5) Do other key growth factors (IGFs, EGFs) in the ovary affect TGFb signalling pathways?


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Learning from health systems strengthening in maternal and newborn health (MNH) in China to inform accelerated progress for saving lives in Africa
Funder: Medical Research Council

Abstract: The aim of this project is to examine China's progress in reducing maternal and neonatal mortality to draw lessons applicable to the MNH context in sub-Saharan Africa. The focus will be on understanding the health systems inputs and outputs that have enabled progress towards comprehensive obstetric care in health facilities, particularly for the rural poor and those living in remote areas, and the financing mechanisms that facilitated reimbursement of hospital delivery charges. We will contribute knowledge in three areas: 1. We will assess implementation strength, an emerging approach to programme evaluation which aims to characterise the intensity of programme effort required to achieving programme goals. This will be done through a quantitative analysis of the relationship between health system inputs/outputs, contextual factors and maternal and neonatal mortality across all 3255 counties in China between 1996 and 2013, using routinely available data. Multilevel statistical models will identify thresholds for MNH health systems indicators, incorporating time varying covariates where possible. 2. A case study in eight counties in Western China will provide an in depth understanding of the relationship between MNH inputs that are more difficult to capture through routine data, including the mix and levels of the midwifery workforce, referrals, levels and allocation of health care financing, and selected service coverage and outcome indicators. Eight counties will be selected purposively to reflect various levels of maternal and neonatal mortality, matching counties as far as possible within provinces on contextual factors. Data will be collected at province, county and facility levels and methods will include document review, in-depth interviews with key stakeholders and facility and patient surveys. 3. We will adapt lessons from China's experience in MNH to low income countries, by analysing the transferability of policy lessons from China to Tanzania


Research activity codes: 8.1 ORGANISATION AND DELIVERY OF SERVICES
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Determinants of effectiveness of a novel community health workers programme in improving maternal and child health in Nigeria
Funder: Medical Research Council

Abstract: The AIM of this project is to inform strengthening and scaling up of community health worker (CHW) programmes. This will be done by investigating two implementations (with and without conditional cash transfers) of a Nigerian CHW programme, to understand what factors, under what conditions, promote equitable access to quality services, and improve maternal and child health outcomes. We will: 1. Understand of the context and the process of implementation of the interventions; 2. Identify, assess and compare the intervention outputs and outcomes; 3. Develop a model of complex relations between the actors, context, implementation process, outputs and outcomes of the interventions; 4. Develop transferable best practices for scalability and generalizability of the interventions. This five-year study will be implemented in two States in Nigeria - Niger State in the North and Anambra State in the South, identified in consultation with the Federal MOH and SUREP national programme officer. The project is designed as a multi-disciplinary and mixed methods study, using qualitative and quantitative methods. Realist evaluation will be an overall platform for the study, which will use economic evaluation, social sciences and statistical analysis. Qualitative data will be collected using in-depth interviews with facility managers, community health workers, PHC staff; facility exit interviews and focus group discussions with service users and their families. Quantitative data will include existing data from HMIS and SURE-P programme and structured facility exit survey with pregnant women. Analysis of quantitative and qualitative datasets will be integrated, to allow in-depth exploration of emerging issues and continuous triangulation of findings. Project results will be disseminated widely through development of policy briefs, presentations at management meetings, newsletters and press-releases, to ensure their uptake in policy and practice in Nigeria and wider.


Research activity codes: 8.1 ORGANISATION AND DELIVERY OF SERVICES
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Aberrant first trimester placental endothelial cell biology in fetal growth restriction
Funder: Medical Research Council

Abstract: Poor placental vascular development at term is seen in pregnancies complicated by fetal growth restriction (FGR). FGR affects 5-8% of human pregnancies worldwide and is a major cause of neonatal morbidity and mortality. Although the pathological changes responsible are believed to occur early in pregnancy, studies directed towards the cellular and molecular mechanisms have been hampered by the lack of appropriate cellular models and the ability to determine, at a clinically relevant time, which pregnancies are at risk of developing FGR. Studies have established a correlation between high first trimester uterine artery Doppler resistance indices (hRI) and birth weight due to placental insufficiency. We will use Doppler ultrasound in the first trimester as a proxy measure of placental function. Using this unique approach and methods developed here to isolate first trimester placental endothelial cells (PEC), we have found that PEC isolated from pregnancies with hRI are more sensitive to apoptotic factors than cells from pregnancies with a normal RI (nRI). It is our hypothesis that poor placental capillary network development is the result of changes to PEC migration/motility and/or sensitivity to apoptotic stimulation and we will seek to establish the mechanism of these differences at a cellular and molecular level. We will investigate whether these differences are the result of changes in gene and/or protein expression in PEC. As placental angiogenesis and PEC survival may also be dependent on factors secreted by surrounding placental stromal cells (PSC) we will determine whether they support an anti-angiogenic, pro-apoptotic phenotype in hRI pregnancies by characterizing differences in gene expression and protein secretion profiles. In vitro findings will be supported by immunohistochemical studies performed on first trimester placental tissue sections.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Sphingosine-1-phosphate and vitamin D as modifiable essential mediators of human placental development
Funder: Medical Research Council

Abstract: The development of a functioning placental unit is dependent upon extravillous trophoblast (EVT) invasion and remodelling of maternal blood vessels; failure of this process leads to the pregnancy complications pre-eclampsia (PE) and fetal growth restriction (FGR) and a significant risk of immediate mortality and long term morbidity for both mother and child. The molecular mechanisms responsible for coordinating trophoblast and vascular cell interactions are unknown. However, evidence from other systems suggests a key role for the bioactive lipid, sphingosine-1-phosphate (S1P). Our preliminary data show that S1P inhibits EVT migration via activation of S1P receptor 2 (S1PR2), suggesting that aberrant control of signalling through S1PR2 could contribute to unfavourable trophoblast behaviour; indeed the levels of a known S1PR2 regulator - vitamin D - are perturbed in PE/FGR. Excitingly, we have also found that vitamin D attenuates the S1P/S1PR2 pathway in an EVT cell line. Here we will to define the role of S1P signalling in directing EVT invasion and vessel transformation and test the potential of novel and repurposed therapeutic interventions as strategies for enhancing vascular remodelling. Specifically we will use human first trimester placenta and rodent models in ex vivo and in vivo assays of normal and abnormal placentation to test our hypotheses that: (1) S1P orchestrates EVT invasion and spiral artery transformation in normal placental development; (2) Low vitamin D levels favour inappropriate S1P signalling contributing to the shallow placentation and vascular abnormalities associated with PE/FGR; (3) Correcting S1P signalling by direct (e.g. S1PR agonist/antagonists) or indirect (e.g. vitamin D supplementation) intervention strategies will improve placental development and outcomes in PE/FGR.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Delineating the roles of NSun proteins at the onset of mouse embryogenesis
Funder: Medical Research Council

Abstract: Our objective is to understand how NSuns regulate the mammalian gamete-to-embryo transition at fertilisation. This phase governs balanced chromosome segregation, transgenerational inheritance and the establishment of totipotency. NSun function will be dissected in living mouse eggs using a unique integrated molecular, cellular and embryological approach established by us since returning to the UK. We will reduce protein levels in oocytes using morpholino- or Cas9-based strategies, or elevate them with cRNA constructs encoding NSun protein fusions to fluorescent tags (eg mCherry, Venus). This approach relies on micromanipulation of mouse oocytes, but because conventional sperm injection typically kills mouse oocytes, we use piezo-actuated microinjection (piezo), which is not widely mastered outside of Asia but is routine in our laboratory. We have generated transgenic lines expressing fluorescent markers in gametes, including H3-mCherry and Venus-tubulin. When combined with piezo, these trangenic tools will allow us to monitor protein dynamics after NSun levels have been altered, thereby mapping each NSun to molecular behaviours and developmental phenotypes, which we can also determine. We have developed a microbead technology to dissect NSun interactions in vivo at the onset of development, which will be extended in a collaboration with Shobbir Hussain; Dr. Hussain has devised a powerful method to identify at single-base resolution NSun RNA substrates. We will start to evaluate the biomedical applications implied by the biology of NSuns. We will determine whether NSuns are dysregulated in infertility and cancer patients following knowledge transfer to our collaborators. These approaches that will reveal the biomedical function and relevance of NSuns.


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: Understanding the role of the Glycine Cleavage System in Neural Tube Defects
Funder: Medical Research Council

Abstract: This project aims to understand how abnormal folate one-carbon metabolism (FOCM) causes neural tube defects (NTDs). We focus on a mitochondrial-specific component of FOCM, the glycine cleavage system (GCS), which has been implicated in human NTDs. Mice lacking expression of the GCS component glycine decarboxylase (Gldc) exhibit abnormal FOCM, elevated glycine levels and NTDs. Main goals are: 1. To understand the metabolic basis of NTDs in Gldc-deficient embryos, we will adopt complementary approaches: (i) metabolic labelling to track one-carbon units; (ii) mass spectrometry-based quantification of folate profiles in differing subcellular compartments and cell types; (iii) gene expression/pathway analysis. 2. Genetic approaches, involving conditional knockout and rescue of Gldc expression, will be used to determine whether there is tissue heterogeneity in the requirement for GCS activity in FOCM and to test whether Gldc function in the neuroepithelium is necessary and sufficient for neural tube closure. 3. Outputs of FOCM include provision of one-carbon units for nucleotide biosynthesis and/or methylation reactions. We will determine which of these outputs are compromised in Gldc-deficient embryos and whether they mechanistically contribute to NTDs. Downstream effects on DNA copy number/integrity and DNA/protein methylation will be evaluated. The relative importance of diminished methylation and nucleotide biosynthesis in Gldc-mediated NTDs will be tested by supplementation strategies and genetic approaches that channel one-carbon units towards either output. 4. Mitochondrial FOCM contributes to production of NADPH, required for redox balance. We will ask whether Gldc-deficiency results in redox imbalance and oxidative stress using mass spectrometry based read-outs of reactive oxygen species, as well as imaging and reporter assays in embryos. The functional contribution of oxidative stress to NTDs will be assessed using cellular markers and rescue experiments.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Childhood Maltreatment: Emotional Consequences and Potential Intervention
Funder: Medical Research Council

Abstract: Childhood maltreatment is a significant global problem. Most estimates of prevalence are from surveys in high-income countries with less known about the scope of childhood maltreatment in lower income countries. The first objective of our research is to investigate the frequency of different forms and severity of child abuse and neglect and co-occurring psychopathology in India and Nepal. We will target child and adolescent labourers, as these represent one of the most disadvantaged sectors of society, with high rates of maltreatment. The second objective of our research is to extend our understanding of the cognitive phenotype of childhood maltreatment. Prior work (mostly from high-income countries) finds that child maltreatment - in particular physical abuse - are associated with greater attention capture by, and difficulties directing attention away from threatening cues. Physically-abused youth also show distorted appraisal styles, selecting more threatening or hostile interpretations of ambiguous information. Many of these studies do not consider the confounding effects of mood, anxiety or trauma symptoms. We will investigate whether in Indian and Nepalese youth, physical abuse is also associated with differences in attention and appraisal biases relative to their non-physically abused peers after controlling for co-occurring psychopathology. These data lay the groundwork for considering novel strategies for targeting distorted information-processing in physically abused youth. Cognitive bias modification (CBM) training has been used successfully to reduce attention and appraisal differences among anxious and aggressive children and young people, with changes in behaviour. In adults, training has resulted in brain-based changes in regions involved in emotion. Our last objective is to extend CBM investigations to physically-abused youth in the UK, India and Nepal and investigate the neural substrates of training changes.


Research activity codes: 6.6 PSYCHOLOGICAL AND BEHAVIOURAL
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Epigenetic mechanisms linking maternal pre-conceptional micronutrient supplementation with offspring health in India and The Gambia
Funder: Medical Research Council

Abstract: Animal and in vitro studies show that the nutrient milieu of the embryo affects the establishment of epigenetic signatures regulating fetal development, providing a plausible mechanism to explain the developmental origins of health and disease (DOHaD). We have preliminary evidence of such effects in humans and now propose to create an India/Gambia/UK partnership to extend this work by studying children of mothers randomised to receive pre- and peri-conceptional micronutrient supplements or placebo. We will analyse DNA methylation in 5-8 year old children whose mothers took part in the Mumbai Maternal Nutrition Project in India (MMNP; ISRCTN 62811278; n=1100 children) and the Peri-conceptional Multiple Micronutrient Supplementation Trial in rural Gambia (PMMST; ISRCTN 13687662; n=350) to identify differences between intervention and control groups. In an initial discovery phase we will measure genome-wide methylation using the Illumina Human Infinium 450K bead chip employing screens for differentially methylated positions (DMPs), regions (DMRs), and variably methylated regions (VMRs) with validation of top hits by bisulfite pyrosequencing. Candidate loci not present on the 450K array will also be assayed by pyrosequencing. We hope to confirm that maternal micronutrient supplementation reduces disrupted methylation of key genes. We will further correlate differences in methylation with health-related outcomes in the children (body composition, cardiometabolic risk markers and cognitive ability). The project will build capacity for next-generation epigenetic research in India and sub-Saharan Africa. It offers a highly efficient means to explore fundamental questions of human developmental biology. If we show that improving a mother's peri-conceptional diet by micronutrient supplementation minimises epigenetic disruption and improves metabolic health in the child, with potential lifelong benefit, our findings will offer a scalable pathway to impact in mothers worldwide.


Research activity codes: 3.3 NUTRITION AND CHEMOPREVENTION
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Ethnic specific risk stratification in early pregnancy for identifying mothers at risk of gestational diabetes mellitus in India and Kenya.
Funder: Medical Research Council

Abstract: Hyperglycaemia in Pregnancy or Gestational Diabetes Mellitus (GDM) is one of the most common obstetric medical conditions which when undetected can cause significant adverse outcomes for the mother and the offspring. Diagnosis is typically made between 24-28 weeks of pregnancy using oral glucose tolerance test (OGTT). Therefore, some damage might have already happened prior to detection. Although universal screening is recommended by many guidelines, this is not uniformly followed across the world, partly because of doubts about cost-effectiveness. Only selective screening is followed based on presence of at least one of the high risk factors (age, BMI, previous history, etc). This strategy can miss up to 50% of GDM. In addition, no data exists in India and Kenya. In low and middle-income countries (LMICs), where majority live in rural settings, the major limitations are difficulty in conducting OGTT, which requires prompt access to laboratory facilities. Combining the clinical and easily analysable biochemical markers (composite risk score) could improve the prediction and if proven, could help to prevent the onset of GDM. Fasting glucose levels (at non-diabetes levels) in early pregnancy could predict future GDM. HbA1c in early pregnancy can be a better marker as it can be done point-of-care and does not require patients to be in a fasting state. The overall objective of the proposed project is to develop a composite risk score to predict GDM in early pregnancy using a combination of easily identifiable risk factors such as age, BMI, family history of Type 2 Diabetes along with HbA1c in Indians and Kenyans. The project will recruit pregnant women in early pregnancy from South India (n=3400) and Western Kenya (n=4000). Contribution of individual risk factors as well as the composite risk score on the risk of developing GDM will be assessed. Detailed health economic analyses will enable policy makers to make informed decision based on local data.


Research activity codes: 4.4 POPULATION SCREENING
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH