NIHRDataInsights/HRCSData huggingface dataset:
Title: Lysosomes in the mammary gland : their role in mediating cell death
Funder: Medical Research Council

Abstract: Recently, we have demonstrated that cell death during involution of the mammary gland is not classical apoptosis but occurs rather by a lysosomal mediated and executioner caspase independent pathway of cell death (LM-PCD). We showed that Stat3 leads to a striking upregulation of the lysosomal proteases cathepsin B and cathepsin L while concomitantly downregulating the expression of an endogenous cathepsin inhibitor Spi2a. This is accompanied by destabilisation of the lysosomal membrane, efflux of cathepsins and subsequent cell death. Importantly, inhibition or deletion of executioner caspases has no effect, while inhibition of cathepsin B abolishes cell death. This is the first time that a mechanism for Stat3 mediated cell death has been elucidated, as well as the first time that LM-PCD has been demonstrated to drive a physiological pathway of cell death. We aim to discover the mechanism of lysosomal leakiness in vivo and how this is controlled. There are two questions that need to be addressed: 1) what is the mechanism by which lysosomes become leaky and 2) which pathways and molecules control LMP? The aim of this programme is to answer these questions. We will utilise two main approaches to uncover the mechanism of LMP: candidate gene analysis and sophisticated proteomics approaches. We have preliminary data on Stat3 targets upregulated during involution and will use the quantitative proteomics techniques, LOPIT and SILAC, to identify novel regulators of LMP both in vivo and in cell culture models of differentiated mammary epithelium. These candidates will be investigated further using fluorescence, time-lapse, and electron microscopy to confirm their lysosomal localisation and we will investigate their function using lentivirus-mediated gene knockdown in culture models and in primary cells transplanted into mammary fat pads. The implications of this work for understanding cell death mechanisms and developing new therapeutic approaches for cancer are considerable


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Poly(A)-binding proteins highlight the importance of regulated mRNA translation and stability in determining a functional materno-fetal interface
Funder: Medical Research Council

Abstract: The majority of mammalian mRNAs are now known to be regulated at the level of mRNA translation and/or stability, with dysregulation contributing to a wide range of diseases e.g. metabolic, neurological, and neoplastic disorders. However, the mechanisms of regulation are poorly understood. Poly(A)-binding protein (PABP) 1, the prototypical member of the PABP family, acts as a global determinant of mRNA translation and stability, regulates mRNA-specific translation and is required for miRNA-mediated repression and nonsense-mediated decay. However little is known about the biological roles of this central regulator of gene expression and other family members remain poorly characterised. As an important step toward determining the role of PABPs in human health, we are exploring their biological functions in vivo, focusing on the two widely expressed PABP proteins, PABP1 and PABP4. Intriguingly, PABP4 deficiency results in a specific defect in mammalian pregnancy, revealing a surprising role for maternally expressed PABP4 in fetal growth and viability during mid to late gestation. Thus, we will investigate the cellular and molecular roles of PABP4, define its mRNA targets and also determine the extent to which they overlap with PABP1 to test their functional relatedness. This will involve developmental, cellular, molecular and systems biology (in vivo; ex vivo; in vitro), patient sample analysis and several specialised techniques e.g. translational profiling using deep sequencing, proteomics, longitudinal analysis of pregnancy/placental function using ultrasound analysis/Doppler imaging. This will shed light on the role of misregulated translation/mRNA stability in intrauterine growth restriction, late miscarriage and stillbirth. Stillbirth affects almost 3 million couples annually and our work may provide a molecular diagnosis and be a first step towards pursuing potential therapeutic avenues, as our model relates not to fetal problems but to the biology of the mother.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Regulation of oocyte cohesin and chromosome segregation defects in the female germ line
Funder: Medical Research Council

Abstract: Based on recent findings from our lab (Lister et al, 2010 Curr. Biol., 20(17)) and others (Chiang et al, 2010, Curr. Biol., 20(17)), we propose that loss of chromosome-associated cohesin during prophase and prometaphase of meiosis I is a key molecular link between female ageing and chromosome missegregation. In this project we will (1) Determine whether chromosomal cohesin is lost during all stages of oogenesis by comparing primordial, growing and prometaphase oocytes from Rec8-Myc females of different ages. Chromosome-associated Rec8-Myc will be measured by immunofluorescence and, in the case of primordial oocytes, by ChIP. (2) Test the hypothesis that cohesin loss is due to a low level of separase activity during prophase/prometaphase. In support of this, we find that separase is expressed during all stages of oogenesis. We will test the functional significance of this using oocytes in which separase is specifically deleted from primordial stage oocytes. (3) Determine whether age-related cohesin loss and disruption of bivalent structure occurs in human oocytes. We anticipate that 800-1000 oocytes will be donated per year. (4) Test the hypothesis that depletion of chromosome cohesin during prophase occurs independently of germ cell depletion. We propose to test this using a mouse model in which depletion of the germ cell pool is uncoupled from female ageing. In the case of human oocytes, we will determine whether the level of cohesin is correlated with circulating AMH, which is indicative of the size of the germ cell pool. The work outlined here will greatly advance our understanding of basic biological mechanisms of female reproductive senescence. It will also yield clinically significant insights that will lead to improved family planning and fertility advice to women, and will shed light on the possibility of developing strategies to prevent or reduce the effects of female ageing on reproductive fitness.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Is Vitamin D deficiency a mechanistic driver of Acute Lung Injury (ALI)?
Funder: Medical Research Council

Abstract: Acute Lung Injury (ALI) is the final common pathway of immunological response to a variety of direct and indirect pulmonary insults. Vitamin D has been shown to have profound effects on human immunity as a result of its actions on both the innate and adaptive immune systems By acting as an immune system modulator, preventing excessive expression of inflammatory cytokines and increasing the 'oxidative burst' potential of macrophages and therefore enhancing bacterial killing. Transthoracic oesophagectomy involves an extended period of one lung ventilation and has been suggested as a model with which to study the development of lung injury as there is a high postoperative incidence (28%) of ALI Using archived clinical samples we have measured 25-OH vitamin D levels in 50 patients with ALI and 50 patients undergoing oesophagectomy. All ALI cases and 48/50 oesophagectomy cases had deficient levels of vitamin D. Very low 25-OH vitamin D levels in the oesophagectomy cohort were associated with elevated plasma levels of IL-6, sRAGE (a type I epithelial cell marker) and HMGB-1 in the immediate post-operative period. Sequential plasma and bronchoalveolar lavage fluid (BALF) will be obtained from patients undergoing one lung ventilation for oesophagectomy and ALI as well as murine models that are genetically and dietarily vitamin D deficient to investigate the effects of vitamin D deficiency and its replacement on acute lung injury. This translational work aims to confirm if vitamin D deficiency is of mechanistic importance in the development of ALI by determining the influence of vitamin D levles upon primary alveolar epithelial cell dysfunction, antibacterial peptide release and phagocytosis/efferocytosis. Combining this with murine studies and complimenting the MRC DCS study we intend the results from this study to form the scientific rationale for a randomised controlled trial of vitamin D replacement in patients with ALI.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: The role of DHEA and androgens in the regulation of ovarian follicular development
Funder: Medical Research Council

Abstract: AIMS: To study the role of DHEA and androgens in follicular development OBJECTIVES: Follicular androgen levels appear to be crucial for follicular development, oocyte release and fertility. I hypothesize that increased androgen levels promote early stage folliculogenesis, but result in follicular arrest at the antral stage. I plan to test this hypothesis, which may help to elucidate mechanisms underlying impaired follicular development in prevalent conditions such as polycystic ovary syndrome (PCOS) and low ovarian reserve. METHODOLOGY: In vitro studies I will employ a unique murine in vitro follicular maturation model (collaboration Professor Teresa Woodruff, Chicago). This model allows detailed examination of all stages of folliculogenesis after alginate encapsulation of immature follicles harvested from prepubertal mice. I will expose the follicles to the universal androgen precursor DHEA and to the most potent androgen, dihydrotestosterone, with additional manipulation by inhibitors that block androgen synthesis or action. I will assess follicular development and oocyte quality by microscopy. I will analyse mRNA expression of genes involved in follicular development and steroidogenesis employing microfluidic card realtime PCR. Finally, I will assess steroidogenic capacity of the follicles by mass spectrometry based steroid assays in follicular fluid and cell culture supernatant samples. Ex vivo studies I will have access to human biomaterial including follicular fluid and cumulus granulosa cells from women with PCOS, normal and low ovarian reserve and also to clinical data on androgen status and fertility outcome. This will allow me to test results from my mouse based studies in the human situation. SCIENTIFIC /MEDICAL OPPORTUNITIES: PCOS and low ovarian reserve are the leading causes of anovulatory infertility and enhancing our understanding of underlying mechanisms may identify novel diagnostic markers or even targets for therapeutic interventions.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Assessing the Risk of Spontaneous Premature Birth by Electrical Impedance Spectroscopy of the Cervix.
Funder: Medical Research Council

Abstract: The timing of birth is crucial to reproduction. 8-13% of babies are born prematurely at annual costs to the NHS of £1bn more than for term babies. Preterm birth(PTB) accounts for > 2/3 of deaths of babies, and contributes huge costs to hospital services during the first 5 years of life. A quarter of babies born <28 weeks suffer handicaps, with socioeconomic burdens on society, parents often having to give up work to care for their child. Prevention of PTB is limited by the lack of accurate prediction of risk. Electrical impedance spectroscopy(EIS) is a technique which may enable prediction of PTB risk. It involves the use of sensing electrodes to assess tissue impedance following the injection of a small electrical current. In 40 women at risk of PTB we recently showed that cervical impedance(CI) between 20 and 28 weeks gestation predicts PTB before 34 wks [sensitivity 100%, specificity 90%, PPV 63, NPV 100, LR+ 9.7, LR- 0.0,P < 0.0001] and 37 weeks [sens 85%, spec 85%, PPV 60%, NPV 96, LR+ 5.8, LR- 0.17,P < 0.0001]. We aim to: a)improve the accuracy of the device by standardising the contact pressure on the cervix, improving the device firmware and online spectral quality checks(Phase 1); b)determine the accuracy of CI-based prediction of PTB in 250 high-risk women studied at 20-22 and 26-28 wks, and 250 women with no risk factors, comparing CI to cervix length and the fibronectin test used in some clinical settings, and assessing whether determining vaginal microbiota using global array technology would prove a screening tool for PTB secondary to CI (phase 2); c)assess the cost-effectiveness & acceptability of CI, and design a study incorporating CI into current PTB screening pathways (Phase 3). A patient group held an inaugural meeting recently, proffered advice on this application and will inform the entire research project. Prompt identification of PTB risk by CI will enable prevention, better hospital bed utilisation, and care in appropriate centres.


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Community intervention to improve growth among children under 2 in rural India
Funder: Medical Research Council

Abstract: Study objectives To determine: 1. The impact and cost-effectiveness of a community intervention with participatory women's groups and home visits on linear growth among children 0-18 months in rural India 2. Whether the intervention leads to improvements in feeding, infection control and caring practices for children 0-18 months 3. Whether the intervention leads to increases in food intake and dietary diversity for women during pregnancy and lactation 4. The operational factors that affect the intervention's delivery and impact 5. The scalability, operational requirements, cost and impact of the intervention beyond the trial area Design: Cluster-randomised controlled trial with 120 clusters (villages and hamlets of c.1000 population), 60 per arm. Participants: Birth cohort of 2520 children and their mothers followed up at seven time points: third trimester, 0, 3, 6, 9, 12 and 18 months Interventions Intervention arm only: a community-based agent will carry out two activities: (a) monthly home visits for growth monitoring and promotion for improved feeding, infection control and caregiving with all children 0-2 years and their mothers; (b) a monthly participatory women's group meeting to catalyse individual and community action on maternal and child health and nutrition. 120 villages will be randomised to two trial arms Intervention & control arms: a capacity building intervention with Village and Health Sanitation and Nutrition Committees Primary outcome: length-for-age Z scores at 18 months. Selected secondary outcomes: weight-for-height and weight-for-age z scores at birth, 3, 6, 9, 12 and 18 months; growth velocity; dietary diversity and food intake for pregnant and lactating women; feeding, caring and infection control practices for infants/children aged 0-18 months. A process evaluation will address objective 4. An economic evaluation will address objective 1 and a sub-study on scalability will address objective 5.


Research activity codes: 3.1 PRIMARY PREVENTION INTERVENTIONS TO MODIFY BEHAVIOURS OR PROMOTE WELL-BEING
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Effectiveness of antibiotic prophylaxis during surgical evacuation of the uterus for miscarriage management in low income countries. (AIMS Trial)
Funder: Medical Research Council

Abstract: Objective: To evaluate the effects of prophylactic antibiotics (Doxycycline 400mg and metronidazole 400mg, given orally, as a single dose prior to surgery) in women undergoing surgical management of miscarriage in low income countries, with the primary outcome of pelvic infection. Short title: The AIMS (Antibiotics in Miscarriage Surgery) Trial Study design: Randomised, double-blind, placebo-controlled, multicentre, multi-national trial. Unit of randomisation is at the individual patient level. Study duration: 36 months. Research sites: a) Malawi: Queen Elizabeth Central Hospital, Blantyre. b) Uganda: Mbale Regional Hospital. c) Tanzania: St Francis Hospital (Ifakara) and Bagamoyo District Hospital. Trial sponsorship: University of Birmingham Trial management: Jointly hosted by University of Birmingham Clinical Trials Unit (BCTU) and College of Medicine, Blantyre, Vaccine & Clinical Trials Unit (VCTU). Population: Women undergoing surgical evacuation of the uterus for miscarriage management are eligible to take part in the study. Intervention: Doxycycline 400mg oral and Metronidazole 400mg oral, as single dose, taken 2 hours before surgery. Control: Identical placebo Randomisation: Computerised third-party randomisation with allocation concealment. Primary outcome: Pelvic infection within 14 days of surgery, defined as two or more of a) purulent vaginal discharge, b) pyrexia >38.0, c) uterine tenderness, and d) a white cell count >15x109 /l. Sample size: To randomise 2400 women (Will give 90% power at p=0.05 to detect a 4% reduction in absolute risk of pelvic infection from a baseline risk of 10% (a relative risk of 0.6, based on our systematic review of antibiotic prophylaxis in induced abortion surgery)). Dissemination: By publication of findings and dissemination through FIGO, WHO, RCOG, National societies, dissemination meetings in all countries and the study website. We will also update the Cochrane review


Research activity codes: 6.1 PHARMACEUTICALS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Use of nanoparticles to deliver growth signals to the placenta
Funder: Medical Research Council

Abstract: Failure of placental growth causes impaired fetal growth which has effects on health in childhood and later life. We now seek funding to develop a novel approach to therapy in utero: the use of maternally administered nanoparticles to deliver a growth stimulus to the placenta. The placenta is an attractive target for nanoparticle-mediated therapies as it filters material in maternal circulation to prevent toxins from accessing fetal tissues. Enhanced placental growth would rescue defective fetal growth and particulate material would be confined to the placenta and discarded at birth. Using an explant model of the early human placenta in which its barrier properties are maintained, we have shown that the growth factors (GF) IGF and TGF can stimulate placental cell proliferation. We also have data to suggest this stimulus can be delivered on a quantum dot (QD) but targeting GF-QD to the placenta and overcoming the adverse effects caused by some nanoparticles will be a pre-requisite for therapeutic exploitation. Here we plan to 1. (a) monitor the effect the effect of GF-QD on human placental trophoblast proliferation and differentiation in order to (b) generate formulations that retain GF activity once labelled with our recently identified placental "homing" peptides. 2. (a) track GF-QD (+/- homing peptides) through the placenta and identify key signalling mediators of altered trophoblast function; (b) investigate whether growth signals generated by trophoblast exposure to QD-GF are transduced to other cellular compartments within placenta. 3. (a) assess the potential for GF-QD to elicit damaging effects (DNA strand breaks and chromosomal abnormalities) and (b) explore ways of offsetting detrimental outcomes. At the end of the project we will have developed an efficacious and safe strategy for manipulating placental growth and / or function that will form the basis for therapeutic intervention in pregnancy disorders characterised by placental abnormalities.


Research activity codes: 2.2 FACTORS RELATING TO PHYSICAL ENVIRONMENT
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Genetic and transcriptional characterization of a region on chromosome 7p15.2 that shows pleiotropy between endometriosis and fat distribution
Funder: Medical Research Council

Abstract: We have identified a pleiotropic signal in an inter-genic region of ~ 900 Kb on chromosome 7p15.2, from genome-wide association (GWA) studies of moderate-severe endometriosis and differential fat distribution (waist-hip ratio, adjusted for body mass index; WHR-adjBMI). Genome-wide expression analysis in subcutaneous adipose tissue identified a local transcript that was significantly associated with the WHR-adjBMI GWA SNP. It is unclear whether: 1) this transcript is differentially expressed in relevant tissues from women with endometriosis; 2) there are other associated known or novel transcripts in the region; and 3) which genetic variants in the region are involved in their regulation. We are collecting endometriotic tissue, endometrium, sc abdominal fat, and whole blood from women undergoing a laparoscopy for: a) symptoms suggestive of endometriosis; and b) sterilisation. We propose to assess differential expression of novel and known transcripts in the 1Mb region of interest in tissues from 100 stage B endometriosis cases and 200 controls, and investigate association with endometriosis and WHR-adjBMI. Genetic variants genotyped and imputed across the region will be tested for association with transcripts (eQTL analyses); eQTL variants significantly associated will be tested for association with phenotypes in 10,936 endometriosis cases and controls. The information to be gained from the proposed study is crucial for the understanding of the GWA findings and their translation into clinically meaningful results, which include the development of non-invasive diagnostic methods (biomarkers) for endometriosis and the development of novel treatments.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH
Title: Physiological studies of sperm from normal and sub fertile men
Funder: Medical Research Council

Abstract: Ion channels in spermatozoa are essential for fertilization since they allow hormones (e.g. progesterone, PGE1) that are found in the female reproductive tract to modulate functional motility. The Cation Channel of Sperm (CatSper), the best characterised such channel, is thought to allow such hormones to evoke Ca influx from the external fluid and the resultant rise in intracellular free Ca seems to trigger motility changes essential for fertilization. In mice, CatSper gene deletion thus abolishes the fertilizing capacity of sperm whilst naturally occurring mutations in CatSper gene family members are associated with infertility in humans. The identification of CatSper has therefore established a new paradigm in reproductive biology and work from this laboratory has shown that progesterone induces abnormally small Ca2+-signals in spermatozoa from men with clinically identified fertility defects. The idea that CatSper is of central importance to the control of intracellular Ca therefore raises the possibility that reduced fertility of these patients might reflect CatSper dysfunction. If this is true then it may be possible to increase the fertilizing capacity of such spermatozoa using drugs that activate CatSper. CatSper blockade, on the other hand, could provide a new contraceptive strategy. Since it is important to understand the physiological role of the ion channels present in spermatozoa and we therefore propose to undertake physiological studies of spermatozoa from sub fertile men and healthy volunteers that will directly characterise the channels expressed by normal sperm and compare their properties with the equivalent channels in men with clinically identified sub fertility. We shall also use fluorimetric techniques to monitor hormone-induced changes in intracellular Ca. It is hoped that the new information provided by these studies will facilitate the development of new ways to treat male factor infertility.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: REPRODUCTIVE HEALTH AND CHILDBIRTH