299 results on '"Philippa T. K. Saunders"'
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2. The Influence of Sex Hormones in Liver Function and Disease
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Alvile Kasarinaite, Matthew Sinton, Philippa T. K. Saunders, and David C. Hay
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liver ,NAFLD ,sex hormones ,estrogen ,testosterone ,HRT ,Cytology ,QH573-671 - Abstract
The liver performs a multitude of bodily functions, whilst retaining the ability to regenerate damaged tissue. In this review, we discuss sex steroid biology, regulation of mammalian liver physiology and the development of new model systems to improve our understanding of liver biology in health and disease. A major risk factor for the development of liver disease is hepatic fibrosis. Key drivers of this process are metabolic dysfunction and pathologic activation of the immune system. Although non-alcoholic fatty liver disease (NAFLD) is largely regarded as benign, it does progress to non-alcoholic steatohepatitis in a subset of patients, increasing their risk of developing cirrhosis and hepatocellular carcinoma. NAFLD susceptibility varies across the population, with obesity and insulin resistance playing a strong role in the disease development. Additionally, sex and age have been identified as important risk factors. In addition to the regulation of liver biochemistry, sex hormones also regulate the immune system, with sexual dimorphism described for both innate and adaptive immune responses. Therefore, sex differences in liver metabolism, immunity and their interplay are important factors to consider when designing, studying and developing therapeutic strategies to treat human liver disease. The purpose of this review is to provide the reader with a general overview of sex steroid biology and their regulation of mammalian liver physiology.
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- 2023
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3. Improving the Diagnosis of Endometrial Hyperplasia Using Computerized Analysis and Immunohistochemical Biomarkers
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Peter A. Sanderson, Arantza Esnal-Zufiaurre, Mark J. Arends, C. Simon Herrington, Frances Collins, Alistair R. W. Williams, and Philippa T. K. Saunders
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endometrial hyperplasia ,intraepithelial neoplasia ,carcinoma ,HAND2 ,stromal to epithelial ratio ,Reproduction ,QH471-489 ,Medicine (General) ,R5-920 - Abstract
Endometrial hyperplasia (EH) is a precursor lesion to endometrial carcinoma (EC). Risks for EC include genetic, hormonal and metabolic factors most notably those associated with obesity: rates are rising and there is concern that cases in pre-menopausal women may remain undetected. Making an accurate distinction between benign and pre-malignant disease is both a challenge for the pathologist and important to the gynecologist who wants to deliver the most appropriate care to meet the needs of the patient. Premalignant change may be recognized by histological changes of endometrial hyperplasia (which may occur with or without atypia) and endometrial intraepithelial neoplasia (EIN). In this study we created a tissue resource of EH samples diagnosed between 2004 and 2009 (n = 125) and used this to address key questions: 1. Are the EIN/WHO2014 diagnostic criteria able to consistently identify premalignant endometrium? 2. Can computer aided image analysis inform identification of EIN? 3. Can we improve diagnosis by incorporating analysis of protein expression using immunohistochemistry. Our findings confirmed the inclusion of EIN in diagnostic criteria resulted in a better agreement between expert pathologists compared with the previous WHO94 criteria used for the original diagnosis of our sample set. A computer model based on assessment of stromal:epithelial ratio appeared most accurate in classification of areas of tissue without EIN. From an extensive panel of putative endometrial protein tissue biomarkers a score based on assessment of HAND2, PTEN, and PAX2 was able to identify four clusters one of which appeared to be more likely to be benign. In summary, our study has highlighted new opportunities to improve diagnosis of pre-malignant disease in endometrium and provide a platform for further research on this important topic.
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- 2022
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4. Mechanisms of Scarless Repair at Time of Menstruation: Insights From Mouse Models
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Phoebe M. Kirkwood, Isaac W. Shaw, and Philippa T. K. Saunders
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hypoxia ,endometrium ,mesenchyme to epithelial transition (MET) ,inflammation ,cytokine ,angiogenesis ,Reproduction ,QH471-489 ,Medicine (General) ,R5-920 - Abstract
The human endometrium is a remarkable tissue which may experience up to 400 cycles of hormone-driven proliferation, differentiation and breakdown during a woman's reproductive lifetime. During menstruation, when the luminal portion of tissue breaks down, it resembles a bloody wound with piecemeal shedding, exposure of underlying stroma and a strong inflammatory reaction. In the absence of pathology within a few days the integrity of the tissue is restored without formation of a scar and the endometrium is able to respond appropriately to subsequent endocrine signals in preparation for establishment of pregnancy if fertilization occurs. Understanding mechanisms regulating scarless repair of the endometrium is important both for design of therapies which can treat conditions where this is aberrant (heavy menstrual bleeding, fibroids, endometriosis, Asherman's syndrome) as well as to provide new information that might allow us to reduce fibrosis and scar formation in other tissues. Menstruation only occurs naturally in species that exhibit spontaneous stromal cell decidualization during the fertile cycle such as primates (including women) and the Spiny mouse. To take advantage of genetic models and detailed time course analysis, mouse models of endometrial shedding/repair involving hormonal manipulation, artificial induction of decidualization and hormone withdrawal have been developed and refined. These models are useful in modeling dynamic changes across the time course of repair and have recapitulated key features of endometrial repair in women including local hypoxia and immune cell recruitment. In this review we will consider the evidence that scarless repair of endometrial tissue involves changes in stromal cell function including mesenchyme to epithelial transition, epithelial cell proliferation and multiple populations of immune cells. Processes contributing to endometrial fibrosis (Asherman's syndrome) as well as scarless repair of other tissues including skin and oral mucosa are compared to that of menstrual repair.
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- 2022
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5. Identification of Altered Evoked and Non-Evoked Responses in a Heterologous Mouse Model of Endometriosis-Associated Pain
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Miguel A. Tejada, Ana I. Santos-Llamas, Lesley Escriva, Juan J. Tarin, Antonio Cano, Maria J. Fernández-Ramírez, Paulina Nunez-Badinez, Bianca De Leo, Philippa T. K. Saunders, Victor Vidal, Florent Barthas, Katy Vincent, Patrick J. Sweeney, Rowland R. Sillito, James Douglas Armstrong, Jens Nagel, and Raúl Gomez
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endometriosis ,heterologous model ,pain ,evoked and non-evoked response ,Biology (General) ,QH301-705.5 - Abstract
The aim of this study was to develop and refine a heterologous mouse model of endometriosis-associated pain in which non-evoked responses, more relevant to the patient experience, were evaluated. Immunodeficient female mice (N = 24) were each implanted with four endometriotic human lesions (N = 12) or control tissue fat (N = 12) on the abdominal wall using tissue glue. Evoked pain responses were measured biweekly using von Frey filaments. Non-evoked responses were recorded weekly for 8 weeks using a home cage analysis (HCA). Endpoints were distance traveled, social proximity, time spent in the center vs. outer areas of the cage, drinking, and climbing. Significant differences between groups for von Frey response, climbing, and drinking were detected on days 14, 21, and 35 post implanting surgery, respectively, and sustained for the duration of the experiment. In conclusion, a heterologous mouse model of endometriosis-associated evoked a non-evoked pain was developed to improve the relevance of preclinical models to patient experience as a platform for drug testing.
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- 2022
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6. A pilot randomised double blind controlled trial of the efficacy of purified fatty acids for the treatment of women with endometriosis-associated pain (PurFECT): study protocol
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Ibtisam M. Abokhrais, Philippa T. K. Saunders, Fiona C. Denison, Ann Doust, Linda Williams, and Andrew W. Horne
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Endometriosis ,Purified fatty acids ,Chronic pelvic pain ,Pilot trial ,Medicine (General) ,R5-920 - Abstract
Abstract Background Endometriosis affects 6–10% of women and is associated with debilitating pelvic pain. It costs the UK > £2.8 billion per year in loss of productivity. Endometriosis can be managed by surgical excision or medically by ovarian suppression. However, ~ 75% symptoms recur after surgery and available medical treatments have undesirable side effects and are contraceptive. Omega-3 purified fatty acids (PUFA) have been shown in animal models to reduce factors that are thought to lead to endometriosis-associated pain, have minimal side effects, and no effects on fertility. This paper presents a protocol for a two-arm, pilot parallel randomised controlled trial (RCT) which aims to inform the planning of a future multicentre trial to evaluate the efficacy of Omega-3 PUFA in the management of endometriosis-associated pain in women. Methods The study will recruit women with endometriosis over a 12-month period in the National Health Service (NHS) Lothian, UK, and randomise them to 8 weeks of treatment with Omega-3 PUFA or comparator (olive oil). The primary objective is to assess recruitment and retention rates. The secondary objectives are to determine the effectiveness/acceptability to participants of the proposed methods of recruitment/randomisation/treatments/questionnaires, to inform the sample size calculation and to refine the research methodology for a future large randomised controlled trial. Response to treatment will be monitored by pain scores and questionnaires assessing physical and emotional function compared at baseline and 8 weeks. Discussion We recognise that there may be potential difficulties in mounting a large randomised controlled trial for endometriosis to assess Omega-3 PUFA because they are a dietary supplement readily available over the counter and already used by women with endometriosis. We have therefore designed this pilot study to assess practical feasibility and following the ‘Initiative on Methods, Measurement, and Pain Assessment in Clinical Trials’ recommendations for the design of chronic pain trials. Trial registration ISRCTN44202346
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- 2018
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7. Hypoxia and hypoxia inducible factor-1α are required for normal endometrial repair during menstruation
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Jacqueline A. Maybin, Alison A. Murray, Philippa T. K. Saunders, Nikhil Hirani, Peter Carmeliet, and Hilary O. D. Critchley
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Science - Abstract
About a quarter of pre-menopausal women will suffer from heavy menstrual bleeding in their lives. Here, Maybin and colleagues show hypoxia and subsequent activation of HIF-1α during menses are required for normal endometrial repair, and identify pharmacological stabilisation of HIF-1α as a potential therapeutic strategy for this debilitating condition.
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- 2018
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8. Chlamydia trachomatis infection of human endometrial stromal cells induces defective decidualisation and chemokine release
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Sevi Giakoumelou, Nick Wheelhouse, Jeremy Brown, Jean Wade, Ioannis Simitsidellis, Douglas Gibson, Philippa T. K. Saunders, Patrick Horner, Gary Entrican, Sarah E. M. Howie, and Andrew W. Horne
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Medicine ,Science - Abstract
Abstract Miscarriage affects ~20% of pregnancies and maternal infections account for ~15% of early miscarriages. Chlamydia trachomatis (Ct) has been associated with miscarriage but the underlying mechanisms are unknown. Successful implantation requires endometrial stromal cell (ESC) decidualisation. Maintenance of pregnancy requires angiogenesis, establishment of the correct cellular milieu and trophoblast invasion, all of which involve the action of chemokines. Our objective was to determine whether Ct infection impacts upon ESC decidualisation and chemokine secretion. Human primary ESC were decidualised in-vitro, infected with Ct serovar E, and changes in expression of genes of interest were measured using RT-PCR, proteomic array and ELISA. We demonstrate for the first time that Ct can infect and proliferate in ESC. Expression of the decidualisation marker prolactin was decreased in Ct-infected ESC at both mRNA and protein levels. Ct infection altered the chemokine profile of decidualised ESC as shown by proteomic array. Chemokines CXCL12 and CXCL16, important for trophoblast invasion, were analysed further and expression was reduced in infected decidualised cells at mRNA and protein levels. Our data indicate that Ct infection of ESC impairs decidualisation and alters chemokine release. These findings at least partially explain how Ct infection could result in adverse pregnancy outcomes.
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- 2017
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9. Effects of Boswellia Serrata Roxb. and Curcuma longa L. in an In Vitro Intestinal Inflammation Model Using Immune Cells and Caco-2
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Paolo Governa, Maddalena Marchi, Veronica Cocetta, Bianca De Leo, Philippa T. K. Saunders, Daniela Catanzaro, Elisabetta Miraldi, Monica Montopoli, and Marco Biagi
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Boswellia serrata Roxb. ,Curcuma longa L. ,intestinal bowel diseases (IBD) ,Caco-2 ,PBMC ,HMC-1.1 ,mast cells ,cytokines ,trans epithelial electrical resistance (TEER) ,reactive oxygen species (ROS) ,Medicine ,Pharmacy and materia medica ,RS1-441 - Abstract
Inflammatory bowel diseases, which consist of chronic inflammatory conditions of the colon and the small intestine, are considered a global disease of our modern society. Recently, the interest toward the use of herbal therapies for the management of inflammatory bowel diseases has increased because of their effectiveness and favourable safety profile, compared to conventional drugs. Boswellia serrata Roxb. and Curcuma longa L. are amongst the most promising herbal drugs, however, their clinical use in inflammatory bowel diseases is limited and little is known on their mechanism of action. The aim of this work was to investigate the effects of two phytochemically characterized extracts of B. serrata and C. longa in an in vitro model of intestinal inflammation. Their impact on cytokine release and reactive oxygen species production, as well as the maintenance of the intestinal barrier function and on intestinal mucosa immune cells infiltration, has been evaluated. The extracts showed a good protective effect on the intestinal epithelium at 1 µg/mL, with TEER values increasing by approximately 1.5 fold, compared to LPS-stimulated cells. C. longa showed an anti-inflammatory mechanism of action, reducing IL-8, TNF-α and IL-6 production by approximately 30%, 25% and 40%, respectively, compared to the inflammatory stimuli. B. serrata action was linked to its antioxidant effect, with ROS production being reduced by 25%, compared to H2O2-stimulated Caco-2 cells. C. longa and B. serrata resulted to be promising agents for the management of inflammatory bowel diseases by modulating in vitro parameters which have been identified in the clinical conditions.
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- 2018
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10. A role for steroid 5 alpha-reductase 1 in vascular remodeling during endometrial decidualization
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Isaac W, Shaw, Phoebe M, Kirkwood, Diane, Rebourcet, Fiona L, Cousins, Rebecca J, Ainslie, Dawn E W, Livingstone, Lee B, Smith, Philippa T K, Saunders, and Douglas A, Gibson
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Vascular Endothelial Growth Factor A ,Mice ,Cholestenone 5 alpha-Reductase ,Endometrium ,3-Oxo-5-alpha-Steroid 4-Dehydrogenase ,Pregnancy ,Endocrinology, Diabetes and Metabolism ,Androgens ,Decidua ,Animals ,Female ,Dihydrotestosterone ,Vascular Remodeling - Abstract
Decidualization is the hormone-dependent process of endometrial remodeling that is essential for fertility and reproductive health. It is characterized by dynamic changes in the endometrial stromal compartment including differentiation of fibroblasts, immune cell trafficking and vascular remodeling. Deficits in decidualization are implicated in disorders of pregnancy such as implantation failure, intra-uterine growth restriction, and pre-eclampsia. Androgens are key regulators of decidualization that promote optimal differentiation of stromal fibroblasts and activation of downstream signaling pathways required for endometrial remodeling. We have shown that androgen biosynthesis, via 5α-reductase-dependent production of dihydrotestosterone, is required for optimal decidualization of human stromal fibroblasts in vitro, but whether this is required for decidualization in vivo has not been tested. In the current study we used steroid 5α-reductase type 1 (SRD5A1) deficient mice (Srd5a1-/- mice) and a validated model of induced decidualization to investigate the role of SRD5A1 and intracrine androgen signaling in endometrial decidualization. We measured decidualization response (weight/proportion), transcriptomic changes, and morphological and functional parameters of vascular development. These investigations revealed a striking effect of 5α-reductase deficiency on the decidualization response. Furthermore, vessel permeability and transcriptional regulation of angiogenesis signaling pathways, particularly those that involved vascular endothelial growth factor (VEGF), were disrupted in the absence of 5α-reductase. In Srd5a1-/- mice, injection of dihydrotestosterone co-incident with decidualization restored decidualization responses, vessel permeability, and expression of angiogenesis genes to wild type levels. Androgen availability declines with age which may contribute to age-related risk of pregnancy disorders. These findings show that intracrine androgen signaling is required for optimal decidualization in vivo and confirm a major role for androgens in the development of the vasculature during decidualization through regulation of the VEGF pathway. These findings highlight new opportunities for improving age-related deficits in fertility and pregnancy health by targeting androgen-dependent signaling in the endometrium.
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- 2022
11. Endometriosis among African women
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Ezekiel O Mecha, Joseph N Njagi, Roselydiah N Makunja, Charles O A Omwandho, Philippa T K Saunders, and Andrew W Horne
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General Medicine - Abstract
Endometriosis has long been wrongly perceived to be rare among women of African descent. The misconception about the prevalence of endometriosis among African women has significantly contributed to long diagnostic delays, limited access to diagnosis and care, and a scarcity of research on the condition among African women. In this commentary, we highlight the prevalence of endometriosis among African women, the state of endometriosis care in Africa, and the gaps in knowledge that need to be addressed. Based on the available data, the prevalence of endometriosis in Africa is likely higher than previously thought, with varying subtypes. There is a long diagnostic delay of endometriosis among African women. Additionally, endometriosis care in Africa from the general population and health practitioners is poor; this can be attributed to the high diagnostic cost, scarcity of trained specialists, as well as patients’ inability to express their symptoms due to societal taboos surrounding menstrual health. Public sensitization on endometriosis may help improve endometriosis diagnosis and care in Africa. Lay summary Endometriosis is a condition in which tissue like the uterine lining is found outside the uterus, causing women to experience pain especially before, during, or after menstruation. Although endometriosis affects an estimated 176 million women worldwide, it has been wrongly reported that endometriosis is a rare condition among African women, mainly due to lack of awareness among healthcare providers and historical bias. In the current commentary, we discuss the prevalence of endometriosis, the diagnostic delays, and the care of endometriosis among black African women living in the African continent. Much of the literature has demonstrated (falsely) that endometriosis is rare in Black women compared to White ethnicity. African women experience a long diagnostic delay and do not receive appropriate care. Public awareness of endometriosis may help improve diagnosis delay and endometriosis care in Africa.
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- 2022
12. Female Reproductive Systems: Hormone Dependence and Receptor Expression
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Kevin K W, Kuan and Philippa T K, Saunders
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Mice ,Receptors, Steroid ,Androgens ,Animals ,Humans ,Estrogens ,Female ,Genitalia, Female ,Progestins ,Ligands ,Progesterone - Abstract
The female reproductive system which consists of the ovaries, uterus (myometrium, endometrium), Fallopian tubes, cervix and vagina is exquisitely sensitive to the actions of steroid hormones. The ovaries play a key role in the synthesis of bioactive steroids (oestrogens, androgens, progestins) that act both within the tissue (intracrine/paracrine) as well as on other reproductive organs following release into the blood stream (endocrine action). Sex steroid receptors encoded by the oestrogen (ESR1, ESR2), progesterone (PR) and androgen (AR) receptor genes, which are members of the superfamily of ligand activated transcription factors are widely expressed within these tissues. These receptors play critical role(s) in regulation of cell proliferation, ovulation, endometrial receptivity, myometrial cell function and inflammatory cell infiltration. Our understanding of their importance has been informed by studies on human tissues and cells, which have employed immunohistochemistry as well as a wide range of molecular and genetic methods to identify which processes are dependent steroid ligand activation. The development of mice with targeted deletions of each of these receptors has provided complementary data that has extended our appreciation of cell-cell interactions in the fine tuning of reproductive tissue function. This large body of work has formed the basis of new and improved therapeutics to treat conditions such as infertility.
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- 2022
13. Preclinical models of endometriosis and interstitial cystitis/bladder pain syndrome: an Innovative Medicines Initiative-PainCare initiative to improve their value for translational research in pelvic pain
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Florent Barthas, Paulina Nunez-Badinez, Raúl Gómez, Francisco Cruz, Jens Nagel, Ana Charrua, Katy Vincent, Alexis Laux-Biehlmann, Stephen B. McMahon, Jane Meijlink, Lone Hummelshoj, J. Douglas Armstrong, Patrick J. Sweeney, Anja Hoffmann, Ioannis Simitsidellis, Laure Lo Re, Philippa T. K. Saunders, Thomas M. Zollner, Miguel Angel Tejada, Judy Birch, Rolf-Detlef Treede, and Bianca De Leo
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Oncology ,medicine.medical_specialty ,Bladder Pain Syndrome ,Cystitis, Interstitial ,Endometriosis ,MEDLINE ,Translational research ,Pelvic Pain ,Translational Research, Biomedical ,03 medical and health sciences ,Interstitial cystitis/bladder pain syndrome ,0302 clinical medicine ,Internal medicine ,In vivo ,medicine ,Humans ,Animal model ,Rodent ,030219 obstetrics & reproductive medicine ,business.industry ,Pelvic pain ,Chronic pain ,Reproducibility of Results ,Interstitial cystitis ,medicine.disease ,3. Good health ,Transplantation ,Anesthesiology and Pain Medicine ,Neurology ,Preclinical research ,ComputingMethodologies_DOCUMENTANDTEXTPROCESSING ,Female ,Neurology (clinical) ,Narrative Review ,medicine.symptom ,business ,030217 neurology & neurosurgery - Abstract
Supplemental Digital Content is Available in the Text., Endometriosis (ENDO) and interstitial cystitis/bladder pain syndrome (IC/BPS) are chronic pain conditions for which better treatments are urgently needed. Development of new therapies with proven clinical benefit has been slow. We have conducted a review of existing preclinical in vivo models for ENDO and IC/BPS in rodents, discussed to what extent they replicate the phenotype and pain experience of patients, as well as their relevance for translational research. In 1009 publications detailing ENDO models, 41% used autologous, 26% syngeneic, 18% xenograft, and 11% allogeneic tissue in transplantation models. Intraperitoneal injection of endometrial tissue was the subcategory with the highest construct validity score for translational research. From 1055 IC/BPS publications, most interventions were bladder centric (85%), followed by complex mechanisms (8%) and stress-induced models (7%). Within these categories, the most frequently used models were instillation of irritants (92%), autoimmune (43%), and water avoidance stress (39%), respectively. Notably, although pelvic pain is a hallmark of both conditions and a key endpoint for development of novel therapies, only a small proportion of the studies (models of ENDO: 0.5%-12% and models of IC/BPS: 20%-44%) examined endpoints associated with pain. Moreover, only 2% and 3% of publications using models of ENDO and IC/BPS investigated nonevoked pain endpoints. This analysis highlights the wide variety of models used, limiting reproducibility and translation of results. We recommend refining models so that they better reflect clinical reality, sharing protocols, and using standardized endpoints to improve reproducibility. We are addressing this in our project Innovative Medicines Initiative-PainCare/Translational Research in Pelvic Pain.
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- 2021
14. Pelvic pain correlates with peritoneal macrophage abundance not endometriosis
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Frances Collins, Bianca De Leo, Douglas A Gibson, Philippa T. K. Saunders, and Andrew W Horne
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endometriosis ,Pathology ,medicine.medical_specialty ,peritoneal ,lcsh:QH471-489 ,Endometriosis ,macrophage ,Pelvic Pain ,lcsh:Gynecology and obstetrics ,Abundance (ecology) ,medicine ,Macrophage ,lcsh:Reproduction ,Ascitic Fluid ,Humans ,pain ,lcsh:RG1-991 ,business.industry ,Pelvic pain ,Research ,General Medicine ,medicine.disease ,Chronic Disease ,Macrophages, Peritoneal ,Female ,medicine.symptom ,Peritoneum ,business - Abstract
Endometriosis is a chronic neuroinflammatory pain condition affecting ~180 million women worldwide. Surgical removal or hormonal suppression of endometriosis lesions only relieves pain symptoms in some women and symptomatic relapse following treatment is common. Identifying factors that contribute to pain is key to developing new therapies. We collected peritoneal fluid samples and clinical data from a cohort of women receiving diagnostic laparoscopy for suspected endometriosis (n = 52). Peritoneal fluid immune cells were analysed by flow cytometry and data compared with pain scores determined using the pain domain of the Endometriosis Health Profile Questionnaire (EHP-30) in order to investigate the association between peritoneal immune cells and pain symptoms. Pain scores were not different between women with or without endometriosis, nor did they differ according to disease stage; consistent with a poor association between disease presentation and pain symptoms. However, linear regression and correlation analysis demonstrated that peritoneal macrophage abundance correlated with the severity of pelvic pain. CD14high peritoneal macrophages negatively correlated with pain scores whereas CD14low peritoneal macrophages were positively correlated, independent of diagnostic outcome at laparoscopy. Stratification by pain subtype, rather than endometriosis diagnosis, resulted in the most robust correlation between pain and macrophage adundance. Pain score strongly correlated with CD14high (P = 0.007) and CD14low (P = 0.008) macrophages in patients with non-menstrual pain and also in patients who reported dysmennorhea (CD14high P = 0.021, CD14low P = 0.019) or dysparunia (CD14high P = 0.027, CD14low P = 0.031). These results provide new insight into the association between peritoneal macrophages and pelvic pain which may aid the identification of future therapeutic targets. Lay summary Endometriosis is a common condition where cells similar to those that line the womb are found elsewhere in the body. It is associated with inflammation and pain in the pelvis and affects ~180 million women worldwide. Current treatments are not effective for all patients and we, therefore, need to understand what causes pain in order to develop new treatments. We investigated the types of immune cells present within the pelvis of women undergoing investigation for suspected endometriosis. Disease diagnosis and stage (I–IV) was recorded along with pain score determined by questionnaire. We characterised the immune cells present and compared them to disease stage and pain score. We found that pelvic pain was linked to the abundance of immune cells but, surprisingly, not to disease stage. These findings suggest that immune cells are closely associated with pain severity in endometriosis and may be good targets for future endometriosis treatments.
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- 2021
15. Androgens, oestrogens and endometrium: a fine balance between perfection and pathology
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Philippa T. K. Saunders, Frances Collins, Douglas A Gibson, and Ioannis Simitsidellis
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medicine.medical_specialty ,Intracrine ,Stromal cell ,Endocrinology, Diabetes and Metabolism ,Endometriosis ,Endometrium ,Endocrinology ,Internal medicine ,medicine ,Humans ,Aromatase ,biology ,business.industry ,Endometrial cancer ,Estrogen Receptor alpha ,Estrogens ,medicine.disease ,Androgen receptor ,medicine.anatomical_structure ,Androgens ,Cancer research ,biology.protein ,Female ,business ,GPER - Abstract
The endometrium is a complex multicellular tissue that is exquisitely sensitive to the actions of sex steroids synthesised in the ovary (endocrine system). Recent studies have highlighted a previously under-appreciated role for local (intracrine) metabolism in fine-tuning tissue function in both health and disease. In this review we have focused on the impact of oestrogens and androgens on endometrial function summarising data from studies on normal endometrial physiology and disorders including infertility, endometriosis and cancer. We consider the evidence that expression of enzymes including aromatase, sulphatase and AKR1C3 by endometrial cells plays an important role in tissue function and malfunction and discuss results from studies using drugs targeting intracrine pathways to treat endometrial disorders. We summarise studies exploring the spatial and temporal expression of oestrogen receptors (ERalpha/ESR1, ERbeta/ESR2 and GPER) and their role in mediating the impact of endogenous and synthetic ligands on cross-talk between vascular, immune, epithelial and stromal cells. There is a single androgen receptor gene and androgens play a key role in stromal-epithelial cross-talk, scar-free healing of endometrium during menstruation and regulation of cell proliferation. The development of new receptor-selective drugs (SERMs, SARMs, SARDs) has reinvigorated interest in targeting receptor subtypes in treatment of disorders including endometriosis and endometrial cancer and some show promise as novel therapies. In summary, understanding the mechanisms regulated by sex steroids provides the platform for improved personalised treatment of endometrial disorders as well as novel insights into the impact of steroids on processes such as tissue repair and regeneration.
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- 2020
16. Endometriosis: etiology, pathobiology and therapeutic prospects
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Andrew W Horne and Philippa T. K. Saunders
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Infertility ,Adult ,medicine.medical_specialty ,Delayed Diagnosis ,Endometriosis ,Tissue Adhesions ,Biology ,Pelvic Pain ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Endometrium ,0302 clinical medicine ,medicine ,Humans ,Intensive care medicine ,030304 developmental biology ,Inflammation ,0303 health sciences ,Pelvic pain ,Uterus ,Chronic pain ,Middle Aged ,medicine.disease ,Hormones ,Clinical trial ,Natural history ,Surgical Procedures, Operative ,Etiology ,Biomarker (medicine) ,Female ,medicine.symptom ,030217 neurology & neurosurgery - Abstract
Endometriosis is a common condition associated with infertility that causes chronic pain in many, but not all, women. It is defined by the presence of endometrial-like tissue outside the uterus. Although the cause and natural history of the disorder remain uncertain, hormonal, neurological, and immunological factors are all implicated in the mechanisms contributing to development of symptoms. Because definitive diagnosis requires surgery, there is often a long diagnostic delay after onset of symptoms. Current interventions for endometriosis have limited efficacy and unacceptable side effects/risks and are associated with high rates of symptom recurrence. Here, we review recent advances in our understanding of the etiology of endometriosis, discuss current diagnostic and treatment strategies, highlight current clinical trials, and consider how recent results offer new avenues for the identification of endometriosis biomarkers and the development of effective non-surgical therapies that are fertility-sparing.
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- 2021
17. The transcription factor EGR2 is indispensable for tissue-specific imprinting of alveolar macrophages in health and tissue repair
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David H. Dockrell, Gareth-Rhys Jones, Nik Hirani, Ananda S. Mirchandani, Bernard Malissen, Frédéric Fercoq, Sandrine Henri, Calum C. Bain, Anna M. Hoy, Jack McCowan, Leo M. Carlin, Philippa T. K. Saunders, Stephen J. Jenkins, Richard Cunningham, Phoebe M. Kirkwood, Sarah R. Walmsley, Connar M. Mawer, Wouter T’Jonck, and Carsten G. Hansen
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Lung ,Transforming growth factor beta ,respiratory system ,Biology ,Lung injury ,Colony-stimulating factor ,Cell biology ,medicine.anatomical_structure ,Alveolar macrophage ,biology.protein ,medicine ,Transcription factor ,Homeostasis ,Function (biology) - Abstract
Alveolar macrophages are the most abundant macrophages in the healthy lung where they play key roles in homeostasis and immune surveillance against air-borne pathogens. Tissue-specific differentiation and survival of alveolar macrophages relies on niche-derived factors, such as colony stimulating factor 2 (CSF-2) and transforming growth factor beta (TGF-β). However, the nature of the downstream molecular pathways that regulate the identity and function of alveolar macrophages and their response to injury remains poorly understood. Here, we identify that the transcriptional factor EGR2 is an evolutionarily conserved feature of lung alveolar macrophages and show that cell-intrinsic EGR2 is indispensable for the tissue-specific identity of alveolar macrophages. Mechanistically, we show that EGR2 is driven by TGF-β and CSF-2 in a PPAR-γ-dependent manner to control alveolar macrophage differentiation. Functionally, EGR2 was dispensable for lipid handling, but crucial for the effective elimination of the respiratory pathogen Streptococcus pneumoniae. Finally, we show that EGR2 is required for repopulation of the alveolar niche following sterile, bleomycin-induced lung injury and demonstrate that EGR2-dependent, monocyte-derived alveolar macrophages are vital for effective tissue repair following injury. Collectively, we demonstrate that EGR2 is an indispensable component of the transcriptional network controlling the identity and function of alveolar macrophages in health and disease.One Sentence SummaryEGR2 controls alveolar macrophage function in health and disease
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- 2021
18. Repurposing dichloroacetate for the treatment of women with endometriosis
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Ioannis Simitsidellis, Andrew W Horne, Nicholas M. Morton, Philippa T. K. Saunders, Chloe Hogg, S. Furquan Ahmad, Roderick N. Carter, and Erin Greaves
- Subjects
endometriosis ,medicine.medical_specialty ,Medical Sciences ,Pyruvate dehydrogenase kinase ,repurposing ,Endometriosis ,Uterus ,Lesion ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,medicine ,Animals ,Humans ,Glycolysis ,dichloroacetate ,Cells, Cultured ,Cell Proliferation ,030304 developmental biology ,Endometrial Stromal Cell ,0303 health sciences ,Multidisciplinary ,Dichloroacetic Acid ,business.industry ,Peritoneal fluid ,Drug Repositioning ,Biological Sciences ,glycolysis ,Pyruvate dehydrogenase complex ,medicine.disease ,3. Good health ,body regions ,Disease Models, Animal ,medicine.anatomical_structure ,Endocrinology ,030220 oncology & carcinogenesis ,Female ,RG ,Peritoneum ,medicine.symptom ,Extracellular Space ,business - Abstract
Endometriosis is a chronic pain condition affecting ∼176 million women worldwide. It is defined by the presence of endometrium-like tissue (lesions) outside the uterus, most commonly on the pelvic peritoneum. There is no cure for endometriosis. All endometriosis drug approvals to date have been contraceptive, limiting their use in women of child-bearing age. We have shown that human peritoneal mesothelial cells (HPMCs) recovered from the pelvic peritoneum of women with endometriosis exhibit significantly higher glycolysis, lower mitochondrial respiration, decreased enzymatic activity of pyruvate dehydrogenase (PDH), and increased production of lactate compared to HPMCs from women without disease. Transforming growth factor-β1 (TGF-β1) is elevated in the peritoneal fluid from women with endometriosis, and exposure of HPMCs to TGF-β1 exacerbates this abnormal phenotype. Treatment of endometriosis HPMCs with the pyruvate dehydrogenase kinase (PDK) inhibitor/PDH activator dichloroacetate (DCA) normalizes HPMC metabolism, reduces lactate secretion, and abrogates endometrial stromal cell proliferation in a coculture model. Oral DCA reduced peritoneal fluid lactate concentrations and endometriosis lesion size in a mouse model. These findings provide the rationale for targeting metabolic processes as a noncontraceptive treatment for women with endometriosis either as a primary nonhormonal treatment or to prevent recurrence after surgery.
- Published
- 2019
19. Optimization of Endometrial Decidualization in the Menstruating Mouse Model for Preclinical Endometriosis Research
- Author
-
F O Dorien, Thomas D'Hooghe, Chloë Goossens, Amelie Fassbender, Philippa T. K. Saunders, Joris Vriens, Daniëlle Peterse, Katrien De Clercq, and M. Mercedes Binda
- Subjects
0301 basic medicine ,medicine.medical_specialty ,medicine.drug_class ,medicine.medical_treatment ,Endometriosis ,Reproductive medicine ,Stimulus (physiology) ,03 medical and health sciences ,0302 clinical medicine ,Laparotomy ,Decidua ,medicine ,Animals ,Laparoscopy ,Progesterone ,Gynecology ,030219 obstetrics & reproductive medicine ,medicine.diagnostic_test ,business.industry ,fungi ,food and beverages ,Obstetrics and Gynecology ,Decidualization ,Estrogens ,medicine.disease ,Menstruation ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,medicine.anatomical_structure ,Estrogen ,Vagina ,Female ,business ,Sesame Oil - Abstract
To induce endometrial decidualization in rodents, an intrauterine oil stimulus can be delivered via the nontraumatic vagina or via the traumatic laparotomy. However, there is considerable variation in amount of decidualization using these inducing methods. Therefore, we studied which oil delivery route could achieve the highest rate of endometrial decidualization along the full length of both uterine horns.To induce decidualization, ovariectomized C57Bl/6J mice were injected with estrogen (100 ng/day; 3 days). A progesterone pellet (5 mg) was implanted subcutaneously, followed by estrogen injections (5 ng/day; 3 days). Oil (20 µL/horn) was injected in the uterus via laparotomy, laparoscopy, or vagina. Four days later, the pellet was removed, followed by hysterectomy after 4 to 6 hours. Endometrial decidualization was evaluated macroscopically and microscopically using hematoxylin and eosin and desmin staining. Furthermore, uterine weight and hormone levels were measured.The proportion of animals with macroscopic bicornuate decidualization was higher after laparoscopic (83%) and laparotomic (89%) injection than after sham injection (11%). Furthermore, macroscopic bicornuate decidualization was significantly higher after laparotomic injection (89%) compared to the vaginal injection (38%). Uterine weight and endometrial surface area were significantly higher in both laparotomy and laparoscopy groups compared to the sham group, while the relative desmin-positive endometrial surface area was only significantly different between the laparotomy and the sham animals.Methods using laparoscopic and laparotomic intrauterine oil injection resulted in a higher amount of decidualized endometrium compared to sham injection, although further optimization is needed to reach full bicornuate decidualization.
- Published
- 2018
20. Dichloroacetate as a possible treatment for endometriosis-associated pain: a single-arm open-label exploratory clinical trial (EPiC)
- Author
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M-C Jones, H. W. Leow, Linda Williams, Magda Koscielniak, Lucy Whitaker, Y. Bagger, Andrew W Horne, Ann Doust, Jane P Daniels, Philippa T. K. Saunders, and G. D. Ferguson
- Subjects
medicine.medical_specialty ,Endometriosis ,Medicine (miscellaneous) ,EPIC ,law.invention ,Gynaecology ,Study Protocol ,03 medical and health sciences ,0302 clinical medicine ,Randomized controlled trial ,Feasibility trial ,law ,Internal medicine ,medicine ,lcsh:R5-920 ,030219 obstetrics & reproductive medicine ,Medical treatment ,business.industry ,Pelvic pain ,Retention rate ,medicine.disease ,Clinical trial ,030220 oncology & carcinogenesis ,medicine.symptom ,Open label ,lcsh:Medicine (General) ,business ,Glycolysis ,Repurposing ,Chronic pelvic pain - Abstract
Background Endometriosis (where endometrial-like tissue is found outside the uterus) affects ~ 176 million women worldwide and can lead to debilitating pelvic pain. There is an unmet need for new medical treatment options for endometriosis. Pelvic peritoneal mesothelial cells of women with endometriosis exhibit detrimental metabolic reprogramming that creates an environment favouring the formation and survival of endometriosis lesions. We have generated powerful preclinical proof-of-concept data to show that it is possible to correct this metabolic phenotype using dichloroacetate (DCA), a non-hormonal compound previously used to treat rare metabolic disorders in children. We plan a single-arm, open-label, single site exploratory clinical trial to inform the design of a future randomised controlled trial (RCT) to determine the efficacy of DCA for the treatment of endometriosis-associated pain. Methods We will recruit 30 women with endometriosis-associated pain over a 6-month period. All participants will receive approximately 6.25 mg/kg oral DCA capsules twice daily for 6 weeks, with a dose increase to approximately 12.5 mg/kg twice daily for a further 6 weeks if their pain has not been adequately controlled on this dose regime and side-effects are acceptable. If pain is adequately controlled with minimal side-effects, the lower dose will be continued for a further 6 weeks. The primary objective is to determine whether it is possible to achieve acceptable recruitment and retention rates within the defined exclusion and inclusion criteria. Secondary objectives are to determine the acceptability of the trial to participants, including the proposed methods of recruitment, treatment, follow-up frequency and number of questionnaires. The recruitment rate will be determined by the proportion of patients recruited from the pool of eligible women. The retention rate will be determined by the proportion of participants who attended the final trial visit. Discussion This is a feasibility study to explore effectiveness and acceptability of the proposed field methodology (recruitment, retention, study processes and compliance with treatment). The results will be used to inform the design of a future RCT. Trial registration ClinicalTrials.gov, NCT04046081 Registered 6 August 2019
- Published
- 2021
21. What Have We Learned from Animal Models of Endometriosis and How Can We Use the Knowledge Gained to Improve Treatment of Patients?
- Author
-
Philippa T K, Saunders
- Subjects
Disease Models, Animal ,Endometriosis ,Animals ,Humans ,Female ,Ovarian Diseases ,Peritoneal Diseases ,Infertility, Female - Abstract
Endometriosis is a complex disorder with a high socio-economic impact. Development of effective novel drug therapies which can be given to women to relieve chronic pain symptoms without side effects such as hormone suppression is urgently required, but progress has been slow. Several different rodent models of 'endometriosis' have been developed, the majority of which mimic aspects of peritoneal disease (e.g. 'lesions' in peritoneal cavity either surgically or spontaneously attached to wall, mesentery, fat). Results obtained using these models have informed our understanding of aetiology including evidence for differential expression of regulatory factors in lesions and impacts on pain perception and fertility. Refinement of these models to ensure reproducibility, extension of models to replicate ovarian and deep disease, complementary in vitro approaches and robust experimental design are all needed to ensure preclinical drug testing results in positive findings in clinical trials and translation for patient benefit.
- Published
- 2020
22. Endometriosis-Associated Pain - Do Preclinical Rodent Models Provide a Good Platform for Translation?
- Author
-
Erin, Greaves, Matthew, Rosser, and Philippa T K, Saunders
- Subjects
Disease Models, Animal ,Mice ,Uterus ,Endometriosis ,Animals ,Humans ,Female ,Pelvic Pain ,Rats - Abstract
Pelvic pain is a common symptom of endometriosis. Our understanding of its etiology remains incomplete and medical management is limited by poor translation from preclinical models to clinical trials. In this review, we briefly consider the evidence, or lack thereof, that different subtypes of lesion, extra-uterine bleeding, and neuropathic pathways add to the complex and heterogeneous pain experience of women with the condition. We summarize the studies in rodent models of endometriosis that have used behavioral endpoints (evoked and non-evoked) to explore mechanisms of endometriosis-associated pain. Lesion innervation, activation of nerves by pronociceptive molecules released by immune cells, and a role for estrogen in modulating hyperalgesia are key endometriosis-associated pain mechanisms replicated in preclinical rodent models. The presence of ectopic (full thickness uterus or endometrial) tissue may be associated with changes in the spinal cord and brain, which appear to model changes reported in patients. While preclinical models using rats and mice have yielded insights that appear relevant to mechanisms responsible for the development of endometriosis-associated pain, they are limited in scope. Specifically, most studies are based on models that only resulted in the formation of superficial lesions and use induced (evoked) behavioral 'pain' tests. We suggest that translation for patient benefit will be improved by new approaches including models of ovarian and deep infiltrating disease and measurement of spontaneous pain behaviors. Future studies must also capitalize on new advances in the wider field of pain medicine to identify more effective treatments for endometriosis-associated pain.
- Published
- 2020
23. Single-cell RNA sequencing redefines the mesenchymal cell landscape of mouse endometrium
- Author
-
Philippa T. K. Saunders, Olympia Kelepouri, James R Smith, Arantza Esnal-Zufiaurre, Douglas A Gibson, Neil C. Henderson, Ross Dobie, John R. Wilson-Kanamori, and Phoebe M. Kirkwood
- Subjects
0301 basic medicine ,Stromal cell ,Mesenchyme ,Green Fluorescent Proteins ,CD34 ,Biology ,Biochemistry ,Receptor, Platelet-Derived Growth Factor beta ,03 medical and health sciences ,Endometrium ,Mice ,0302 clinical medicine ,Genetics ,medicine ,Animals ,Homeostasis ,Progenitor cell ,CXCL14 ,Molecular Biology ,Sequence Analysis, RNA ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,CD146 ,Female ,Pericyte ,Single-Cell Analysis ,Transcriptome ,030217 neurology & neurosurgery ,Biomarkers ,Biotechnology - Abstract
The endometrium is a dynamic tissue that exhibits remarkable resilience to repeated episodes of differentiation, breakdown, regeneration, and remodeling. Endometrial physiology relies on a complex interplay between the stromal and epithelial compartments with the former containing a mixture of fibroblasts, vascular, and immune cells. There is evidence for rare populations of putative mesenchymal progenitor cells located in the perivascular niche of human endometrium, but the existence of an equivalent cell population in mouse is unclear. We used the Pdgfrb-BAC-eGFP transgenic reporter mouse in combination with bulk and single-cell RNA sequencing to redefine the endometrial mesenchyme. In contrast to previous reports we show that CD146 is expressed in both PDGFRβ + perivascular cells and CD31 + endothelial cells. Bulk RNAseq revealed cells in the perivascular niche which express the high levels of Pdgfrb as well as genes previously identified in pericytes and/or vascular smooth muscle cells (Acta2, Myh11, Olfr78, Cspg4, Rgs4, Rgs5, Kcnj8, and Abcc9). scRNA-seq identified five subpopulations of cells including closely related pericytes/vascular smooth muscle cells and three subpopulations of fibroblasts. All three fibroblast populations were PDGFRα+/CD34 + but were distinct in their expression of Ngfr/Spon2/Angptl7 (F1), Cxcl14/Smoc2/Rgs2 (F2), and Clec3b/Col14a1/Mmp3 (F3), with potential functions in the regulation of immune responses, response to wounding, and organization of extracellular matrix, respectively. Immunohistochemistry was used to investigate the spatial distribution of these populations revealing F1/NGFR + cells in most abundance beside epithelial cells. We provide the first definitive analysis of mesenchymal cells in the adult mouse endometrium identifying five subpopulations providing a platform for comparisons between mesenchymal cells in endometrium and other adult tissues which are prone to fibrosis.
- Published
- 2020
24. Single cell RNA sequencing redefines the mesenchymal cell landscape of mouse endometrium
- Author
-
James R Smith, Philippa T. K. Saunders, Ross Dobie, Douglas A Gibson, Phoebe M. Kirkwood, Arantza Esnal-Zufiaurre, John R. Wilson-Kanamori, Neil C. Henderson, and Olympia Kelepouri
- Subjects
Extracellular matrix ,medicine.anatomical_structure ,Stromal cell ,Smooth muscle cell migration ,Mesenchyme ,Mesenchymal stem cell ,medicine ,CD146 ,Progenitor cell ,Biology ,Fibroblast ,Cell biology - Abstract
The endometrium is a dynamic tissue that exhibits remarkable resilience to repeated episodes of differentiation, breakdown, regeneration and remodelling. Endometrial physiology relies on a complex interplay between the stromal and epithelial compartments with the former containing a mixture of fibroblasts, vascular and immune cells. There is evidence for rare populations of putative mesenchymal progenitor cells located in the perivascular niche of human endometrium, but the existence of an equivalent cell population in mouse is unclear.In the current study we used the Pdgfrb-BAC-eGFP transgenic reporter mouse in combination with bulk and single cell RNA sequencing (scRNAseq) to redefine the endometrial mesenchyme. Contrary to previous reports we show that CD146 is expressed in both PDGFRβ+ perivascular cells as well as CD31+ endothelial cells. Bulk RNAseq revealed cells in the perivascular niche which express high levels of Pdgfrb as well as genes previously identified in pericytes and/or vascular smooth muscle cells (Acta2, Myh11, Olfr78, Cspg4, Rgs4, Rgs5, Kcnj8, Abcc9). scRNAseq identified five subpopulations of cells including closely related pericytes/vascular smooth muscle cells and three subpopulations of fibroblasts. All three fibroblast populations were PDGFRα+/CD34+ but were distinct in their expression of Spon2/Angptl7 (fibroblast 1), Smoc2/Rgs2 (fibroblast 2) and Clec3b/Col14a1/Mmp3 (fibroblast 3), with potential functions in regulation of immune responses, response to wounding and organisation of extracellular matrix respectively.In conclusion, these data are the first to provide a single cell atlas of the mesenchymal cell landscape in mouse endometrium. By identifying novel markers for subpopulations of mesenchymal cells we can use mouse models investigate their contribution to endometrial function, compare with other tissues and apply these findings to further our understanding of human endometrium.HighlightsGFP expression in the mouse endometrium, under the control of the Pdgfrb promoter, is restricted to two cell populations based on the intensity of GFP with GFPbright cells close to the vasculatureSingle cell RNAseq identified five subpopulations of GFP+ mesenchymal cells: pericytes, vascular smooth muscle cells (vSMC) and three closely related but distinct populations of fibroblastsBioinformatics revealed that pericytes and vSMC share functions associated with the circulatory system, actin-filament process and cell adhesion, and an apparent role for pericytes in smooth muscle cell migration and response to interferonsComparisons between the fibroblast subpopulations suggest distinct roles in regulation of immune response, response to wound healing and collagen organisation.Graphical Abstract
- Published
- 2020
25. Peritoneal macrophage phenotype correlates with pain scores in women with suspected endometriosis
- Author
-
Philippa T. K. Saunders, Douglas A Gibson, Andrew W Horne, Bianca De Leo, and Frances Collins
- Subjects
Infertility ,medicine.medical_specialty ,business.industry ,Pelvic pain ,Peritoneal fluid ,Endometriosis ,Disease ,medicine.disease ,Gastroenterology ,Pathophysiology ,Menstrual cycle phase ,Internal medicine ,medicine ,medicine.symptom ,Stage (cooking) ,business - Abstract
ObjectiveTo characterise peritoneal macrophage populations in women with suspected endometriosis and assess if they are correlated with severity of pelvic pain symptoms.DesignFlow cytometry analysis of peritoneal fluid samples and clinical data.SettingUniversity Research Institute.PatientsClinical questionnaires, surgical data and peritoneal fluid were collected with informed consent from women undergoing diagnostic laparoscopy for suspected endometriosis (n=54).Intervention(s)NoneMain Outcome Measure(s)Severity of pelvic pain symptoms was assessed by the EHP-30 questionnaire. Immune cells recovered from peritoneal fluid were analysed by flow cytometry.ResultsPain scores (pain domain of EHP30) did not differ according to endometriosis diagnosis, stage of endometriosis or whether or not women were receiving hormone treatment. Analysis of immune cells in peritoneal fluid revealed two populations of peritoneal macrophages: CD14highand CD14lowwhich were not altered by menstrual cycle phase or hormone treatment. CD14highperitoneal macrophages were increased in women with endometriosis compared to those without but were not altered by coincident reproductive health issues such as infertility or heavy menstrual bleeding. Peritoneal macrophage phenotype correlated with pelvic pain symptoms in women with suspected endometriosis. Notably, CD14highperitoneal macrophages negatively correlated with pain scores whereas CD14lowperitoneal macrophages were positively correlated. This association was independent of endometriosis diagnosis.ConclusionPeritoneal macrophage phenotypes correlate with pelvic pain symptoms in women with suspected endometriosis and are altered by presence of disease. These results provide new insight into the association between endometriosis pathophysiology and pelvic pain symptoms.
- Published
- 2020
26. Androgen action on renal calcium and phosphate handling: Effects of bisphosphonate treatment and low calcium diet
- Author
-
Brigitte Decallonne, Dieter Schollaert, Philippa T. K. Saunders, Dirk Vanderschueren, Rougin Khalil, Geert Carmeliet, Ioannis Simitsidellis, Na Ri Kim, Ludo Deboel, Ferran Jardi, and Frank Claessens
- Subjects
Male ,0301 basic medicine ,medicine.medical_specialty ,medicine.drug_class ,chemistry.chemical_element ,030209 endocrinology & metabolism ,Urinalysis ,Calcium ,Kidney ,urologic and male genital diseases ,Biochemistry ,Bone and Bones ,Bone resorption ,Phosphates ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Bone Density ,Internal medicine ,Calcium flux ,medicine ,Animals ,Hypercalciuria ,Bone Resorption ,Molecular Biology ,Diphosphonates ,Chemistry ,medicine.disease ,Androgen ,Diet ,Calcium, Dietary ,Mice, Inbred C57BL ,Androgen receptor ,030104 developmental biology ,Dihydrotestosterone ,Androgens ,Secondary hyperparathyroidism ,Orchiectomy ,medicine.drug - Abstract
Renal calcium and phosphate handling is an important contributor to mineral homeostasis and bone health and the androgen receptor (AR) is highly expressed in the kidney. We investigated the short term effects of androgen deprivation on renal calcium and phosphate reabsorption, independent of their effects on bone. Two weeks following orchidectomy (ORX) of adult mice, bone loss occurred along with hypercalciuria, which was similarly prevented by testosterone and dihydrotestosterone supplementation. Treatment with bisphosphonates prior to ORX also inhibited hypercalciuria, indicating that the calcium flux originated from the bone. Renal calcium and phosphate transporter expression was increased post-ORX, independent of bisphosphonates. Furthermore, an- drogen deprivation appeared to stimulate local synthesis of 1,25(OH)2D3. When bisphosphonate-treated mice were fed a low calcium diet, bone resorption was no longer blocked and secondary hyperparathyroidism de- veloped, which was more pronounced in ORX mice than sham-operated mice. In conclusion, this study shows that androgen deprivation increased renal calcium and phosphate transporter expression, independent of bone, and underlines the importance of adequate intestinal calcium supply in circumstances of androgen deprivation and bisphosphonate treatment.
- Published
- 2020
27. Targeting colony stimulating factor-1 receptor signalling to treat ectopic pregnancy
- Author
-
W. Colin Duncan, Philippa T. K. Saunders, Magda Koscielniak, Frances Collins, Andrew W Horne, Robyn E. Beaty, S. Furquan Ahmad, and Lisa Campbell
- Subjects
0301 basic medicine ,Endocrine reproductive disorders ,Cost effectiveness ,Reproductive disorders ,lcsh:Medicine ,Receptor, Macrophage Colony-Stimulating Factor ,Article ,Cell Line ,Colony stimulating factor 1 receptor ,03 medical and health sciences ,0302 clinical medicine ,Syncytiotrophoblast ,Cell Movement ,Pregnancy ,medicine ,Humans ,Cell migration ,Molecular Targeted Therapy ,lcsh:Science ,Receptor ,030219 obstetrics & reproductive medicine ,Multidisciplinary ,Cytotrophoblast ,Cell Death ,business.industry ,Macrophage Colony-Stimulating Factor ,lcsh:R ,Trophoblast ,Placentation ,Endocrine system and metabolic diseases ,3. Good health ,Pregnancy, Ectopic ,Trophoblasts ,030104 developmental biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Cancer research ,lcsh:Q ,Methotrexate ,Female ,business ,medicine.drug ,Signal Transduction - Abstract
1–2% of pregnancies are ectopic, the majority implanting in the Fallopian tube. A single, systemic dose of methotrexate, a DNA-synthesis (S phase) inhibitor, has been used since 1991 for outpatient treatment of women with stable EP. However, methotrexate has limited clinical and cost effectiveness, restricting its use to 25–30% of these women. There is an unmet need for better medical treatment for EP. Colony stimulating factor-1 (CSF-1) promotes placentation and creates a pro-inflammatory environment that is fundamental for the maintenance of a normal pregnancy. We hypothesised that CSF-1 is also involved in the placentation and maintenance of an EP. Herein, we demonstrate the immunolocalisation of the CSF-1 receptor (CSF-1R) as well as its ligand (CSF-1) in immortalised first trimester trophoblast cells. We show that a specific CSF-1R kinase inhibitor, GW2580, abolishes CSF-1 induced trophoblast cell proliferation and migration and can be cytotoxic. We then demonstrate the expression of CSF-1R and CSF-1 in the cytotrophoblast and syncytiotrophoblast within ectopic implantation sites from women with EP. Our data suggests that CSF-1 is involved in the survival and proliferation of trophoblast cells in EP. This suggests that pharmacological disruption of CSF-1/CSF-1R signaling axis could be the basis of a new therapeutic for EP.
- Published
- 2020
28. Research priorities for endometriosis differ among patients, clinicians, and researchers
- Author
-
Paula C. Brady, Andrew W Horne, Ann M. Thomas, Stacey A. Missmer, Philippa T. K. Saunders, and Leslie V. Farland
- Subjects
Infertility ,Adult ,Male ,medicine.medical_specialty ,Health Personnel ,Emotions ,Endometriosis ,MEDLINE ,Health personnel ,Cost of Illness ,Risk Factors ,Stakeholder Participation ,Surveys and Questionnaires ,medicine ,Cost of illness ,Humans ,Pain Management ,Family ,Reino unido ,business.industry ,Research ,Obstetrics and Gynecology ,Pain management ,Middle Aged ,medicine.disease ,Research Personnel ,United Kingdom ,Gynecology ,Family medicine ,Female ,business ,Infertility, Female - Abstract
Objective:Endometriosis—the implantation of endometrial-like tissue outside of the uterus—burdens an estimated 6-10% of women worldwide and is associated with chronic pelvic pain and infertility. Currently, the standard method of diagnosis is surgery, which creates barriers to care and delays in diagnosis. As a result, many questions fundamental to our understanding of endometriosis remain unanswered. The objective of this work is to identify which research questions are most meaningful to those personally affected by the disease, as compared to those treating and studying it.
- Published
- 2020
29. Profiling the expression and function of oestrogen receptor isoform ER46 in human endometrial tissues and uterine natural killer cells
- Author
-
Cristina Bajo-Santos, Arantza Esnal-Zufiaurre, Hilary O. D. Critchley, Frances Collins, Philippa T. K. Saunders, and Douglas A Gibson
- Subjects
Stromal cell ,splice variant ,Immunocytochemistry ,Population ,Estrogen receptor ,Biology ,Endometrium ,03 medical and health sciences ,0302 clinical medicine ,Pregnancy ,medicine ,Decidua ,Humans ,Protein Isoforms ,education ,Receptor ,030304 developmental biology ,ERα ,0303 health sciences ,education.field_of_study ,Reproductive Biology ,Rehabilitation ,Uterus ,Obstetrics and Gynecology ,uNK ,uterine natural killer cell ,Cell biology ,Killer Cells, Natural ,medicine.anatomical_structure ,Reproductive Medicine ,Receptors, Estrogen ,030220 oncology & carcinogenesis ,embryonic structures ,oestrogen receptor isoform 46 ,Immunohistochemistry ,Original Article ,Female ,oestrogen receptor alpha - Abstract
STUDY QUESTION Does the oestrogen receptor isoform, ER46, contribute to regulation of endometrial function? SUMMARY ANSWER ER46 is expressed in endometrial tissues, is the predominant ER isoform in first trimester decidua and is localised to the cell membrane of uterine natural killer (uNK) cells where activation of ER46 increases cell motility. WHAT IS KNOWN ALREADY Oestrogens acting via their cognate receptors are essential regulators of endometrial function and play key roles in establishment of pregnancy. ER46 is a 46-kDa truncated isoform of full length ERα (ER66, encoded by ESR1) that contains both ligand- and DNA-binding domains. Expression of ER46 in the human endometrium has not been investigated previously. ER46 is located at the cell membrane of peripheral blood leukocytes and mediates rapid responses to oestrogens. uNK cells are a phenotypically distinct (CD56brightCD16−) population of tissue-resident immune cells that regulate vascular remodelling within the endometrium and decidua. We have shown that oestrogens stimulate rapid increases in uNK cell motility. Previous characterisation of uNK cells suggests they are ER66-negative, but expression of ER46 has not been characterised. We hypothesise that uNK cells express ER46 and that rapid responses to oestrogens are mediated via this receptor. STUDY DESIGN, SIZE, DURATION This laboratory-based study used primary human endometrial (n = 24) and decidual tissue biopsies (n = 30) as well as uNK cells which were freshly isolated from first trimester human decidua (n = 18). PARTICIPANTS/MATERIALS, SETTING, METHODS Primary human endometrial and first trimester decidual tissue biopsies were collected using methods approved by the local institutional ethics committee (LREC/05/51104/12 and LREC/10/51402/59). The expression of ERs (ER66, ER46 and ERβ) was assessed by quantitative PCR, western blot and immunohistochemistry. uNK cells were isolated from first-trimester human decidua by magnetic bead sorting. Cell motility of uNK cells was measured by live cell imaging: cells were treated with 17β-oestradiol conjugated to bovine serum albumin (E2-BSA, 10 nM equivalent), the ERβ-selective agonist 2,3-bis(4-hydroxyphenyl)-propionitrile (DPN; 10 nM) or dimethylsulphoxide vehicle control. MAIN RESULTS AND THE ROLE OF CHANCE ER46 was detected in proliferative and secretory phase tissues by western blot and was the predominant ER isoform in first-trimester decidua samples. Immunohistochemistry revealed that ER46 was co-localised with ER66 in cell nuclei during the proliferative phase but detected in both the cytoplasm and cell membrane of stromal cells in the secretory phase and in decidua. Triple immunofluorescence staining of decidua tissues identified expression of ER46 in the cell membrane of CD56-positive uNK cells which were otherwise ER66-negative. Profiling of isolated uNK cells confirmed expression of ER46 by quantitative PCR and western blot and localised ER46 protein to the cell membrane by immunocytochemistry. Functional analysis of isolated uNK cells using live cell imaging demonstrated that activation of ER46 with E2-BSA significantly increased uNK cell motility. LARGE SCALE DATA N/A. LIMITATIONS, REASONS FOR CAUTION Expression pattern in endometrial tissue was only determined using samples from proliferative and secretory phases. Assessment of first trimester decidua samples was from a range of gestational ages, which may have precluded insights into gestation-specific changes in these tissues. Our results are based on in vitro responses of primary human cells and we cannot be certain that similar mechanisms occur in situ. WIDER IMPLICATIONS OF THE FINDINGS E2 is an essential regulator of reproductive competence. This study provides the first evidence for expression of ER46 in the human endometrium and decidua of early pregnancy. We describe a mechanism for regulating the function of human uNK cells via expression of ER46 and demonstrate that selective targeting with E2-BSA regulates uNK cell motility. These novel findings identify a role for ER46 in the human endometrium and provide unique insight into the importance of membrane-initiated signalling in modulating the impact of E2 on uNK cell function in women. Given the importance of uNK cells to regulating vascular remodelling in early pregnancy and the potential for selective targeting of ER46, this may be an attractive future therapeutic target in the treatment of reproductive disorders. STUDY FUNDING/COMPETING INTEREST(S) These studies were supported by Medical Research Council (MRC) Programme Grants G1100356/1 and MR/N024524/1 to PTKS. H.O.D.C. was supported by MRC grant G1002033. The authors declare no competing interests related to the published work.
- Published
- 2020
30. Endometriosis-Associated Pain – Do Preclinical Rodent Models Provide a Good Platform for Translation?
- Author
-
Erin Greaves, Philippa T. K. Saunders, and Matthew Rosser
- Subjects
0303 health sciences ,business.industry ,Pain medicine ,Pelvic pain ,Endometriosis ,Disease ,medicine.disease ,Bioinformatics ,Clinical trial ,Lesion ,03 medical and health sciences ,Nociception ,Hyperalgesia ,Medicine ,medicine.symptom ,business ,030304 developmental biology - Abstract
Pelvic pain is a common symptom of endometriosis. Our understanding of its etiology remains incomplete and medical management is limited by poor translation from preclinical models to clinical trials. In this review, we briefly consider the evidence, or lack thereof, that different subtypes of lesion, extra-uterine bleeding, and neuropathic pathways add to the complex and heterogeneous pain experience of women with the condition. We summarize the studies in rodent models of endometriosis that have used behavioral endpoints (evoked and non-evoked) to explore mechanisms of endometriosis-associated pain. Lesion innervation, activation of nerves by pronociceptive molecules released by immune cells, and a role for estrogen in modulating hyperalgesia are key endometriosis-associated pain mechanisms replicated in preclinical rodent models. The presence of ectopic (full thickness uterus or endometrial) tissue may be associated with changes in the spinal cord and brain, which appear to model changes reported in patients. While preclinical models using rats and mice have yielded insights that appear relevant to mechanisms responsible for the development of endometriosis-associated pain, they are limited in scope. Specifically, most studies are based on models that only resulted in the formation of superficial lesions and use induced (evoked) behavioral 'pain' tests. We suggest that translation for patient benefit will be improved by new approaches including models of ovarian and deep infiltrating disease and measurement of spontaneous pain behaviors. Future studies must also capitalize on new advances in the wider field of pain medicine to identify more effective treatments for endometriosis-associated pain.
- Published
- 2020
31. What Have We Learned from Animal Models of Endometriosis and How Can We Use the Knowledge Gained to Improve Treatment of Patients?
- Author
-
Philippa T. K. Saunders
- Subjects
Infertility ,0303 health sciences ,business.industry ,Endometriosis ,Chronic pain ,Disease ,medicine.disease ,Precision medicine ,Bioinformatics ,Menstruation ,Clinical trial ,03 medical and health sciences ,medicine ,Etiology ,business ,030304 developmental biology - Abstract
Endometriosis is a complex disorder with a high socio-economic impact. Development of effective novel drug therapies which can be given to women to relieve chronic pain symptoms without side effects such as hormone suppression is urgently required, but progress has been slow. Several different rodent models of 'endometriosis' have been developed, the majority of which mimic aspects of peritoneal disease (e.g. 'lesions' in peritoneal cavity either surgically or spontaneously attached to wall, mesentery, fat). Results obtained using these models have informed our understanding of aetiology including evidence for differential expression of regulatory factors in lesions and impacts on pain perception and fertility. Refinement of these models to ensure reproducibility, extension of models to replicate ovarian and deep disease, complementary in vitro approaches and robust experimental design are all needed to ensure preclinical drug testing results in positive findings in clinical trials and translation for patient benefit.
- Published
- 2020
32. A two-arm parallel double-blind randomised controlled pilot trial of the efficacy of Omega-3 polyunsaturated fatty acids for the treatment of women with endometriosis-associated pain (PurFECT1)
- Author
-
Andrew W Horne, Philippa T. K. Saunders, Lucy Whitaker, Ibtisam M. Abokhrais, Ann Doust, Linda Williams, and Fiona C. Denison
- Subjects
Questionnaires ,Endometriosis ,Pathology and Laboratory Medicine ,law.invention ,0302 clinical medicine ,Randomized controlled trial ,Quality of life ,law ,Plant Products ,Medicine and Health Sciences ,Medicine ,030212 general & internal medicine ,Young adult ,030219 obstetrics & reproductive medicine ,Multidisciplinary ,Pharmaceutics ,Eukaryota ,Olives ,Drugs ,Agriculture ,Contraceptives ,Plants ,3. Good health ,Research Design ,Engineering and Technology ,medicine.symptom ,Research Article ,Biotechnology ,medicine.medical_specialty ,Drug Research and Development ,Clinical Research Design ,Science ,Bioengineering ,Research and Analysis Methods ,Vegetable Oils ,Fruits ,03 medical and health sciences ,Pharmacotherapy ,Signs and Symptoms ,Drug Therapy ,Diagnostic Medicine ,Clinical Trials ,Adverse effect ,Pharmacology ,Survey Research ,business.industry ,Pelvic pain ,Organisms ,Biology and Life Sciences ,medicine.disease ,Agronomy ,Randomized Controlled Trials ,Sample size determination ,Physical therapy ,Lesions ,Medical Devices and Equipment ,Adverse Events ,Clinical Medicine ,business ,Crop Science - Abstract
BackgroundEndometriosis is defined by the presence of endometrial-like tissue (lesions) outside the uterus, commonly on the pelvic peritoneum. It affects 6-10% of women and is associated with debilitating pelvic pain. Current management options are often unsatisfactory. Omega-3 polyunsaturated fatty acids (O-PUFA) have the potential to reduce the painful symptoms associated with endometriosis, reduce lesion size, preserve the patient's ability to conceive, and have minimal side effects. We performed a two-arm, parallel double-blinded randomised controlled trial to inform the planning of a future multicentre randomised controlled trial to evaluate the efficacy of O-PUFA for endometriosis-associated pain.ObjectivesThe primary objectives of the trial were to assess recruitment and retention rates. The secondary objectives were to determine the acceptability to women of the proposed methods of recruitment, randomisation, treatments and questionnaires, to estimate the variability in the proposed primary endpoints to inform the sample size calculation and to refine the research methodology for the future definitive trial.MethodsWe recruited women with endometriosis from June 2016 to June 2017 and randomised them to eight weeks of treatment with O-PUFA or olive oil. Pain scores and quality of life questionnaires were collected at baseline and eight weeks. We calculated the proportion of eligible women randomised, and of randomised participants who were followed up to eight weeks. Acceptability questionnaires were used to evaluate women's experiences of the trial.ResultsThe proportion of eligible participants who were randomised was 45.2% (33/73) and 81.8% (27/33) completed the study. The majority of participants described their overall trial experience favourably and there were no adverse events in either group.ConclusionOur pilot trial supports the feasibility of a future larger trial to definitively evaluate the efficacy of O-PUFA for endometriosis-associated pain.Trial registrationThe trial was registered on the ISRCTN registry (registration number ISRCTN44202346).
- Published
- 2020
33. The ERβ5 splice variant increases oestrogen responsiveness of ERαpos Ishikawa cells
- Author
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Carol Fitzgerald, Frances Collins, Douglas A Gibson, Nozomi Itani, Philippa T. K. Saunders, and Arantza Esnal-Zufiaurre
- Subjects
0301 basic medicine ,Cancer Research ,Endocrinology, Diabetes and Metabolism ,Estrogen receptor ,carcinoma ,Adenocarcinoma ,Response Elements ,Endometrium ,Malignant transformation ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,estrogen ,Tumor Cells, Cultured ,medicine ,Estrogen Receptor beta ,Humans ,Receptor ,Chemistry ,Research ,Endometrial cancer ,Estrogens ,Transfection ,medicine.disease ,Endometrial Neoplasms ,3. Good health ,Gene Expression Regulation, Neoplastic ,Alternative Splicing ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Female ,Estrogen receptor alpha ,Immunostaining ,estrogen receptor - Abstract
Endometrial cancer is a common gynaeological malignancy: life time exposure to oestrogen is a key risk factor. Oestrogen action is mediated by receptors encoded by ESR1 (ERα) and ESR2 (ERβ): ERα plays a key role in regulating endometrial cell proliferation. A truncated splice variant isoform (ERβ5) encoded by ESR2 is highly expressed in cancers. This study explored whether ERβ5 alters oestrogen responsiveness of endometrial epithelial cells. Immunhistochemistry profiling of human endometrial cancer tissue biopsies identified epithelial cells co-expressing ERβ5 and ERα in stage I endometrial adenocarcinomas and post menopausal endometrium. Induced co-expression of ERβ5 in ERαpos endometrial cancer cells (Ishikawa) significantly increased ligand-dependent activation of an ERE-luciferase reporter stimulated by either E2 or the ERα-selective agonist 1,3,5-(4-hydroxyphenyl)-4-propyl-1H-pyrazole (PPT) compared to untransfected cells. Fluorescence recovery after photobleaching (FRAP) analysis of tagged yellow fluorescent protein (YFP)-ERβ5 transfected into Ishikawa cells revealed that incubation with E2 induced a transient reduction in intra-nuclear mobility characterised by punctate protein redistribution which phenocopied the behaviour of ERα following ligand activation with E2. In ERαneg MDA-MD-231 breast cancer cells, there was no E2-dependent change in mobility of YFP-ERβ5 and no activation of the ERE reporter in cells expressing ERβ5. In conclusion, we demonstrate that ERβ5 can act as heterodimeric partner to ERα in Ishikawa cells and increases their sensitivity to E2. We speculate that expression of ERβ5 in endometrial epithelial cells may increase the risk of malignant transformation and suggest that immunostaining for ERβ5 should be included in diagnostic assessment of women with early grade cancers.
- Published
- 2019
34. Profiling the expression and function of the truncated oestrogen receptor isoform ER46 in human endometrium
- Author
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Hilary O. D. Critchley, Cristina Bajo-Santos, Philippa T. K. Saunders, Arantza Esnal-Zufiaurre, Douglas A Gibson, and Frances Collins
- Subjects
Gene isoform ,Oestrogen receptor ,Biology ,Human endometrium ,Cell biology - Published
- 2019
35. The transcription factor EGR2 is indispensable for tissue-specific imprinting of alveolar macrophages in health and tissue repair #
- Author
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Ananda S. Mirchandani, Frédéric Fercoq, Connar M. Mawer, Wouter T’Jonck, Carsten G. Hansen, Jack McCowan, Sandrine Henri, Philippa T. K. Saunders, Richard Cunningham, Gareth-Rhys Jones, Lizi M. Hegarty, Leo M. Carlin, Stephen J. Jenkins, Bernard Malissen, Nik Hirani, Phoebe M. Kirkwood, Duncan Humphries, Sarah R. Walmsley, Calum C. Bain, Anna M. Hoy, David H. Dockrell, MRC Centre for Regenerative Medicine and Multiple Sclerosis Society/University of Edinburgh Centre for Translational Research, Centre for Inflammation Research, The Queen’s Medical Research Institute, Edinburgh, UK., University of Edinburgh, Cancer Research UK Beatson Institute [Glasgow], Centre d'Immunologie de Marseille - Luminy (CIML), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre d'Immunophénomique (CIPHE), and Institute of Cancer Sciences, University of Glasgow
- Subjects
Male ,health care facilities, manpower, and services ,[SDV]Life Sciences [q-bio] ,Immunology ,education ,Biology ,Article ,Pneumococcal Infections ,Mice ,Macrophages, Alveolar ,medicine ,Animals ,Humans ,Imprinting (psychology) ,Transcription factor ,health care economics and organizations ,Early Growth Response Protein 2 ,Lung ,General Medicine ,Tissue repair ,respiratory system ,Phenotype ,Cell biology ,medicine.anatomical_structure ,Streptococcus pneumoniae ,Alveolar macrophage ,Female ,Homeostasis ,Respiratory tract - Abstract
Alveolar macrophages are the most abundant macrophages in the healthy lung where they play key roles in homeostasis and immune surveillance against air-borne pathogens. Tissue-specific differentiation and survival of alveolar macrophages relies on niche-derived factors, such as granulocyte-macrophage colony stimulating factor 2 (GM-CSF) and transforming growth factor beta (TGF-). However, the nature of the downstream molecular pathways that regulate the identity and function of alveolar macrophages and their response to injury remains poorly understood. Here, we identify that the transcriptional factor EGR2 is an evolutionarily conserved feature of lung alveolar macrophages and show that cell-intrinsic EGR2 is indispensable for the tissue-specific identity of alveolar macrophages. Mechanistically, we show that EGR2 is driven by TGF- and GM-CSF in a PPAR--dependent manner to control alveolar macrophage differentiation. Functionally, EGR2 was dispensable for regulation of lipids in the airways, but crucial for the effective handling of the respiratory pathogen Streptococcus pneumoniae. Finally, we show that EGR2 is required for repopulation of the alveolar niche following sterile, bleomycin-induced lung injury and demonstrate that EGR2-dependent, monocyte-derived alveolar macrophages are vital for effective tissue repair following injury. Collectively, we demonstrate that EGR2 is an indispensable component of the transcriptional network controlling the identity and function of alveolar macrophages in health and disease.
- Published
- 2021
36. Laparoscopic Surgery: A New Technique to Induce Endometriosis in a Mouse Model
- Author
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M. Mercedes Binda, Daniëlle Peterse, Amelie Fassbender, Joris Vriens, Thomas D'Hooghe, Arne Vanhie, Dorien F. O, and Philippa T. K. Saunders
- Subjects
Laparoscopic surgery ,medicine.medical_specialty ,medicine.medical_treatment ,Endometriosis ,Uterus ,Reproductive medicine ,Estrous Cycle ,Pilot Projects ,Mice ,03 medical and health sciences ,Peritoneal cavity ,0302 clinical medicine ,Laparotomy ,medicine ,Animals ,Prospective Studies ,Laparoscopy ,Prospective cohort study ,Peritoneal Cavity ,Mice, Inbred BALB C ,030219 obstetrics & reproductive medicine ,medicine.diagnostic_test ,business.industry ,General surgery ,Obstetrics and Gynecology ,medicine.disease ,Surgery ,Disease Models, Animal ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Female ,business - Abstract
This prospective pilot study was designed to induce endometriosis in a mouse model using laparoscopy, a less invasive and more precise approach than laparotomy. We aimed to achieve a peritoneal implant rate of at least 50% by varying both duration of anesthesia and intra-abdominal insufflation pressure.Female BALB/cANnCrl mice in metestrus or diestrus were used as donors (n = 5) or recipients (n = 20) of uterine transplant tissue. Each recipient mouse was laparoscopically inoculated with 10 uterine pieces (range: 10-12) from donor mice into the abdominal cavity. Before starting the study, recipient mice were randomly assigned to 4 groups with variable duration of anesthesia (ketamine/xylazine or pentobarbital) and variable intra-abdominal pressure (5 or 15 mm Hg). One week after laparoscopy, endometriosis incidence and peritoneal implant take rate were documented visually during laparotomy. The retrieved lesions were histologically analyzed.Laparoscopic inoculation of uterine pieces in recipient mice resulted in an endometriosis incidence of 100% (20/20 animals) and an individual peritoneal implant take rate of 60% (121/206), ranging from 17% (2/12) till 83% (10/12), without differences between the 4 subgroups, and with a histological confirmation rate of 92% (58/63).To the best our knowledge, this is the first report showing that endometriosis can be induced by laparoscopic surgery in rodents, with a 100% incidence and a median peritoneal implant take rate of 60%. This laparoscopic model offers important advantages over traditional laparotomy models that are limited by surgery-associated trauma and/or adhesion formation.
- Published
- 2016
37. Androgens and androgen receptor: Above and beyond
- Author
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Iain J. McEwan, Douglas A Gibson, and Philippa T. K. Saunders
- Subjects
Male ,0301 basic medicine ,medicine.medical_specialty ,Drug resistance ,Biochemistry ,Mice ,03 medical and health sciences ,Endocrinology ,Internal medicine ,medicine ,Animals ,Humans ,Neoplasm ,Receptor ,Molecular Biology ,business.industry ,medicine.disease ,Androgen Metabolism ,Androgen receptor ,030104 developmental biology ,Drug Resistance, Neoplasm ,Receptors, Androgen ,Androgens ,Female ,business - Published
- 2018
38. Profiling the expression and function of ER46 in human endometrial tissues and uterine NK cells
- Author
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Frances Collins, Hilary O. D. Critchley, Philippa T. K. Saunders, Cristina Bajo-Santos, Arantza Esnal-Zufiaurre, and Douglas A Gibson
- Subjects
0303 health sciences ,education.field_of_study ,Stromal cell ,Decidua ,Population ,Motility ,Biology ,Endometrium ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,Live cell imaging ,030220 oncology & carcinogenesis ,embryonic structures ,medicine ,Immunohistochemistry ,Receptor ,education ,030304 developmental biology - Abstract
Study questionDoes the oestrogen receptor isoform, ER46, contribute to regulation of endometrial function?Summary answerER46 is expressed in endometrial tissues during the proliferative and secretory phases and is the predominant ERα isoform in first trimester decidua. ER46 is abundantly expressed in uterine NK (uNK) cells and localised to the cell membrane. Activation of ER46 regulates the function of human uNK cells by increasing cell motility.What is known alreadyOestrogens acting via their cognate receptors are essential regulators of endometrial function and play key roles in establishment of pregnancy. ER46 is a 46kDa truncated isoform of full length ERα (ER66, encoded by ESR1) that contains both ligand and DNA binding domains. Expression of ER46 in human endometrium has not been investigated previously. ER46 is located at the cell membrane of peripheral blood leukocytes and mediates rapid responses to oestrogens. UNK cells are a phenotypically distinct (CD56brightCD16-) population of tissue-resident immune cells that regulate vascular remodelling within the endometrium and decidua. We have shown that oestrogens stimulate rapid increases in uNK cell motility. Previous characterisation of uNK cells suggests they are ER66-negative but expression of ER46 has not been characterised. We hypothesise that uNK cells express ER46 and that rapid responses to oestrogens are mediated via this receptor.Study design, size, durationThis laboratory-based study used primary human endometrial (n=24) and decidual tissue biopsies (n=30) as well as uNK cells which were freshly isolated from first trimester human decidua (n=18).Participants/materials, setting, methodsPrimary human endometrial and first trimester decidual tissue biopsies were collected using methods approved by the local institutional ethics committee (LREC/05/51104/12 and LREC/10/51402/59). The expression of oestrogen receptors (ER66, ER46 and ERβ) was assessed by qPCR, western blot and immunohistochemistry. Uterine Natural Killer (uNK) cells were isolated from first trimester human decidua by magnetic bead sorting. Cell motility of uNK cells was measured by live cell imaging: cells were treated with oestradiol (E2)-BSA (10nM equivalent), the ERβ-selective agonist 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN; 10nM) or vehicle control (DMSO).Main results and the role of chanceER46 was detected in proliferative and secretory phase tissues and was the predominant ERα isoform in first trimester decidua samples. Immunohistochemistry revealed ER46 was co-localised with ER66 in cell nuclei during the proliferative phase but detected in both the cytoplasm and cell membrane of stromal cells in the secretory phase and in decidua. Triple immunofluorescence staining of decidua tissues identified expression of ER46 in the cell membrane of CD56-positive uNK cells which were otherwise ER66-negative. Profiling of isolated uNK cells confirmed expression ER46 and localised ER46 protein to the cell membrane. Functional analysis of isolated uNK cells using live cell imaging demonstrated that activation of ER46 with E2-BSA significantly increased uNK cell motility.Limitations, reasons for cautionExpression patterns in endometrial tissue was only determined using samples from proliferative and secretory phases. Assessment of first trimester decidua samples was from a range of gestational ages which may have precluded insights into gestation specific changes in these tissues. Our results are based on in vitro responses of primary human cells and we cannot be certain that similar mechanisms occur in situ.Wider implications of the findingsE2 is an essential regulator of reproductive competence. This study provides the first evidence for expression of ER46 in human endometrium and decidua of early pregnancy. We describe a mechanism for regulating the function of human uNK cells via expression of ER46 and demonstrate that selective targeting with E2-BSA regulates uNK cell motility. These novel findings identify a role for ER46 in human endometrium and provide unique insight into the importance of membrane-initiated signalling in modulating the impact of E2 on uNK cell function in women.Study funding/competing interest(s)These studies were supported by MRC Programme Grants G1100356/1 and MR/N024524/1 to PTKS. HODC was supported by MRC grant G1002033.
- Published
- 2019
39. Selective androgen receptor modulators (SARMs) have specific impacts on the mouse uterus
- Author
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Olympia Kelepouri, Elisabeth O’Flaherty, Douglas A Gibson, Arantza Esnal-Zuffiaure, Ioannis Simitsidellis, and Philippa T. K. Saunders
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Stromal cell ,DHT ,medicine.drug_class ,Endocrinology, Diabetes and Metabolism ,Gene Expression ,030209 endocrinology & metabolism ,androgen ,Endometrium ,Aminophenols ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Endocrinology ,GTx-024 ,Internal medicine ,Acetamides ,medicine ,Humans ,Anilides ,endometrium ,Andarine ,Cell Proliferation ,Danazol ,uterus ,business.industry ,Research ,Dihydrotestosterone ,Epithelial Cells ,SARM ,Androgen ,Androgen receptor ,Ostarine ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Enobosarm ,Receptors, Androgen ,Female ,business ,medicine.drug - Abstract
Selective androgen receptor modulators (SARMs) have been proposed as therapeutics for women suffering from breast cancer, muscle wasting or urinary incontinence. The androgen receptor (AR) is expressed in the uterus but the impact of SARMs on the function of this organ is unknown. We used a mouse model to compare the impact of SARMs (GTx-007/Andarine®, GTx-024/Enobosarm®), Danazol (a synthetic androstane steroid) and dihydrotestosterone (DHT) on tissue architecture, cell proliferation and gene expression. Ovariectomised mice were treated daily for 7 days with compound or vehicle control (VC). Uterine morphometric characteristics were quantified using high-throughput image analysis (StrataQuest; TissueGnostics), protein and gene expression were evaluated by immunohistochemistry and RT-qPCR, respectively. Treatment with GTx-024, Danazol or DHT induced significant increases in body weight, uterine weight and the surface area of the endometrial stromal and epithelial compartments compared to VC. Treatment with GTx-007 had no impact on these parameters. GTx-024, Danazol and DHT all significantly increased the percentage of Ki67-positive cells in the stroma, but only GTx-024 had an impact on epithelial cell proliferation. GTx-007 significantly increased uterine expression of Wnt4 and Wnt7a, whereas GTx-024 and Danazol decreased their expression. In summary, the impact of GTx-024 and Danazol on uterine cells mirrored that of DHT, whereas GTx-007 had minimal impact on the tested parameters. This study has identified endpoints that have revealed differences in the effects of SARMs on uterine tissue and provides a template for preclinical studies comparing the impact of compounds targeting the AR on endometrial function.
- Published
- 2019
40. Macrophage-derived insulin-like growth factor-1 is a key neurotrophic and nerve-sensitizing factor in pain associated with endometriosis
- Author
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Alexandra Sarginson, Andrew W Horne, Chloe Hogg, Ashley Dorning, Philippa T. K. Saunders, Rachel Forster, Erin Greaves, and Atanaska N. Velichkova
- Subjects
0301 basic medicine ,Linsitinib ,medicine.medical_treatment ,Neurogenesis ,Endometriosis ,Pain ,Biochemistry ,Cell Line ,Receptor, IGF Type 1 ,03 medical and health sciences ,Insulin-like growth factor ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Nerve Fibers ,Genetics ,medicine ,Macrophage ,Animals ,Humans ,Insulin-Like Growth Factor I ,Molecular Biology ,biology ,business.industry ,Pelvic pain ,Macrophages ,medicine.disease ,Pathophysiology ,3. Good health ,Mice, Inbred C57BL ,Editorial Commentary ,030104 developmental biology ,chemistry ,Hyperalgesia ,Cancer research ,biology.protein ,Female ,medicine.symptom ,business ,030217 neurology & neurosurgery ,Biotechnology ,Neurotrophin - Abstract
Endometriosis is a common incurable inflammatory disorder that is associated with debilitating pelvic pain in women. Macrophages are central to the pathophysiology of endometriosis: they dictate the growth and vascularization of endometriosis lesions and more recently have been shown to promote lesion innervation. The aim of this study was to determine the mechanistic role of macrophages in producing pain associated with endometriosis. Herein, we show that macrophage depletion in a mouse model of endometriosis can reverse abnormal changes in pain behavior. We identified that disease‐modified macrophages exhibit increased expression of IGF‐1 in an in vitro model of endometriosis‐associated macrophages and confirmed expression by lesion‐resident macrophages in mice and women. Concentrations of IGF‐1 were elevated in peritoneal fluid from women with endometriosis and positively correlate with their pain scores. Mechanistically, we demonstrate that macrophage‐derived IGF‐1 promotes sprouting neurogenesis and nerve sensitization in vitro. Finally, we show that the Igf‐1 receptor inhibitor linsitinib reverses the pain behavior observed in mice with endometriosis. Our data support a role for macrophage‐derived IGF‐1 as a key neurotrophic and sensitizing factor in endometriosis, and we propose that therapies that modify macrophage phenotype may be attractive therapeutic options for the treatment of women with endometriosis‐associated pain.—Forster, R., Sarginson, A., Velichkova, A., Hogg, C., Doming, A., Home, A. W., Saunders, P. T. K., Greaves, E. Macrophage‐derived insulin‐like growth factor‐1 is a key neurotrophic and nerve‐sensitizing factor in pain associated with endometriosis. FASEB J. 33, 11210–11222 (2019). www.fasebj.org
- Published
- 2019
41. Repurposing simvastatin as a therapy for preterm labor: evidence from preclinical models
- Author
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Adriano G. Rossi, Ashley K Boyle, Jane E. Norman, Philippa T. K. Saunders, and Sara F. Rinaldi
- Subjects
0301 basic medicine ,Lipopolysaccharides ,Simvastatin ,Geranylgeranyl pyrophosphate ,Farnesyl pyrophosphate ,Inflammation ,CCL2 ,Pharmacology ,Biochemistry ,Proinflammatory cytokine ,statins ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,Uterine Contraction ,0302 clinical medicine ,Obstetric Labor, Premature ,Pregnancy ,Genetics ,Medicine ,Animals ,Humans ,Molecular Biology ,Cells, Cultured ,business.industry ,Anticholesteremic Agents ,Research ,Drug Repositioning ,preterm birth ,contraction ,Muscle, Smooth ,3. Good health ,CXCL1 ,Disease Models, Animal ,030104 developmental biology ,chemistry ,Myometrium ,Female ,medicine.symptom ,business ,Vascular smooth muscle contraction ,030217 neurology & neurosurgery ,Biotechnology ,medicine.drug ,Signal Transduction - Abstract
Preterm birth (PTB), the leading cause of neonatal morbidity and mortality, urgently requires novel therapeutic agents. Spontaneous PTB, resulting from preterm labor, is commonly caused by intrauterine infection/ inflammation. Statins are well‐established, cholesterol‐lowering drugs that can reduce inflammation and inhibit vascular smooth muscle contraction. We show that simvastatin reduced the incidence of PTB in a validated intrauterine LPS‐induced PTB mouse model, decreased uterine proinflammatory mRNA concentrations (IL‐6, Cxcl1, and Ccl2), and reduced serum IL‐6 concentration. In human myometrial cells, simvastatin reduced proinflammatory mediator mRNA and protein expression (IL‐6 and IL‐8) and increased anti‐inflammatory cytokine mRNA expression (IL‐10 and IL‐13). Critically, simvastatin inhibited myometrial cell contraction, basally and during inflammation, and reduced phosphorylated myosin light chain concentration. Supplementation with mevalonate and geranylgeranyl pyrophosphate, but not farnesyl pyrophosphate, abolished these anticontractile effects, indicating that the Rho/Rho‐associated protein kinase pathway is critically involved. Thus, simvastatin reduces PTB incidence in mice, inhibits myometrial contractions, and exhibits key anti‐inflammatory effects, providing a rationale for investigation into the repurposing of statins to treat preterm labor in women.—Boyle, A. K., Rinaldi, S. F., Rossi, A. G., Saunders, P. T. K., Norman, J. E. Repurposing simvastatin as a therapy for preterm labor: evidence from preclinical models. FASEB J. 33, 2743–2758 (2019). www.fasebj.org
- Published
- 2019
42. Androgen modulation of mouse uterus: a tissue-based bioassay for testing endogenous and synthetic androgen receptor modulators (SARMs)
- Author
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Olympia Kelepouri, Douglas A Gibson, Philippa T. K. Saunders, Ioannis Simitsidellis, and Arantza Esnal-Zufiaurre
- Subjects
Androgen receptor ,Mouse Uterus ,Chemistry ,medicine.drug_class ,medicine ,Bioassay ,Endogeny ,Androgen ,Cell biology - Published
- 2018
43. Endometrial Intracrinology: Oestrogens, Androgens and Endometrial Disorders
- Author
-
Ioannis Simitsidellis, Douglas A Gibson, Philippa T. K. Saunders, and Frances Collins
- Subjects
aromatase ,endometriosis ,medicine.medical_specialty ,media_common.quotation_subject ,medicine.medical_treatment ,oestradiol ,Endometrium ,sulfatase ,lcsh:Chemistry ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,medicine ,Aromatase ,Receptor ,lcsh:QH301-705.5 ,Menstrual cycle ,Testosterone ,030304 developmental biology ,media_common ,0303 health sciences ,030219 obstetrics & reproductive medicine ,biology ,business.industry ,Endometrial cancer ,decidualisation ,nursing_health_studies ,medicine.disease ,3. Good health ,Androgen receptor ,Steroid hormone ,medicine.anatomical_structure ,Endocrinology ,dehydroepiandrosterone (DHEA) ,lcsh:Biology (General) ,lcsh:QD1-999 ,testosterone ,endometrial cancer ,biology.protein ,business - Abstract
Peripheral tissue metabolism of steroids (intracrinology) is now accepted as a key way in which tissues, such as the endometrium, can utilize inactive steroids present in the blood to respond to local physiological demands and ‘fine-tune’ the activation or inhibition of steroid hormone receptor-dependent processes. Expression of enzymes that play a critical role in activation and inactivation of bioactive oestrogens (E1, E2) and androgens (A4, T, DHT), as well as expression of steroid hormone receptors, has been detected in endometrial tissues and cells recovered during the menstrual cycle. There is robust evidence that increased expression of aromatase is important for creating a local microenvironment that can support a pregnancy. Measurement of intra-tissue concentrations of steroids using liquid chromatography–tandem mass spectrometry has been important in advancing our understanding of a role for androgens in the endometrium acting both as active ligands for the androgen receptor and as substrates for oestrogen biosynthesis. The emergence of intracrinology, associated with disordered expression of key enzymes such as aromatase, in the aetiology of common women’s health disorders such as endometriosis and endometrial cancer has prompted renewed interest in development of drugs targeting these pathways opening up new opportunities for targeted therapies and precision medicine.  
- Published
- 2018
44. A Role for Androgens in Epithelial Proliferation and Formation of Glands in the Mouse Uterus
- Author
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Douglas A Gibson, Philippa T. K. Saunders, Ioannis Simitsidellis, Arantza Esnal-Zufiaurre, and Fiona L. Cousins
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Stromal cell ,medicine.drug_class ,Gene Expression ,Biology ,Endometrium ,Mice ,03 medical and health sciences ,Endocrinology ,Internal medicine ,WNT4 ,medicine ,Animals ,Original Research ,Cell Proliferation ,Cell Cycle ,Dihydrotestosterone ,Epithelial Cells ,Androgen ,Androgen receptor ,030104 developmental biology ,medicine.anatomical_structure ,WNT7A ,Reproduction-Development ,Receptors, Androgen ,Androgens ,Female ,Immunostaining ,medicine.drug - Abstract
The endometrium consists of stromal and epithelial compartments (luminal and glandular) with distinct functions in the regulation of uterine homeostasis. Ovarian sex steroids, namely 17β-estradiol and progesterone, play essential roles in modulating uterine cell proliferation, stromal-epithelial cross-talk and differentiation in preparation for pregnancy. The effect of androgens on uterine function remains poorly understood. The current study investigated the effect of the non-aromatizable androgen dihydrotestosterone (DHT) on mouse endometrial function. Ovx female mice were given a single sc injection (short treatment) or 7 daily injections (long treatment) of vehicle alone (5% ethanol, 0.4% methylcellulose) or vehicle with the addition of 0.2 mg DHT (n=8/group) and a single injection of bromodeoxyuridine 2 hours prior to tissue recovery. Treatment with DHT increased uterine weight, the area of the endometrial compartment and immunoexpression of the androgen receptor in the luminal and glandular epithelium. Treatment-dependent proliferation of epithelial cells was identified by immunostaining for MKi67 and bromodeoxyuridine. Real-time PCR identified significant DHT-dependent changes in the concentrations of mRNAs encoded by genes implicated in the regulation of the cell cycle (Wee1, Ccnd1, Rb1) and stromal-epithelial interactions (Wnt4, Wnt5a, Wnt7a, Cdh1, Vcl, Igf1, Prl8, Prlr) as well as a striking effect on the number of endometrial glands. This study has revealed a novel role for androgens in regulating uterine function with an effect on the glandular compartment of the endometrium. This previously unrecognized role for androgens has implications for our understanding of the role of androgens in regulation of endometrial function and fertility in women.
- Published
- 2016
45. Cortisol regulates the paracrine action of macrophages by inducing vasoactive gene expression in endometrial cells
- Author
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Philippa T. K. Saunders, Hilary O. D. Critchley, Jacqueline A. Maybin, Gregory M. Armstrong, Erin Greaves, and Uma Thiruchelvam
- Subjects
0301 basic medicine ,endocrine system ,medicine.medical_specialty ,Hydrocortisone ,Angiogenesis ,Adipose tissue macrophages ,Immunology ,Inflammation ,Translational & Clinical Immunology ,Vascular Remodeling ,progesterone ,Biology ,Endometrium ,angiogenesis ,03 medical and health sciences ,Paracrine signalling ,Receptors, Glucocorticoid ,0302 clinical medicine ,estradiol ,Internal medicine ,Paracrine Communication ,glucocorticoid receptor ,medicine ,Humans ,Immunology and Allergy ,Macrophage ,Interleukin 8 ,Menstrual Cycle ,030219 obstetrics & reproductive medicine ,Macrophages ,Endothelial Cells ,Cell Biology ,CXCL2 ,3. Good health ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,Gene Expression Regulation ,Angiogenesis Inducing Agents ,Female ,medicine.symptom ,hormones, hormone substitutes, and hormone antagonists - Abstract
Cortisol regulation of macrophages induces angiogenic gene expression in endometrial cells at menstruation., The human endometrium undergoes inflammation and tissue repair during menstruation. We hypothesized that the local availability of bioactive glucocorticoids plays an important role in immune cell–vascular cell interactions in endometrium during tissue repair at menstruation, acting either directly or indirectly via tissue resident macrophages. We sought to determine whether endometrial macrophages are direct targets for glucocorticoids; whether cortisol-treated macrophages have a paracrine effect on angiogenic gene expression by endometrial endothelial cells; and whether endometrial macrophages express angiogenic factors. Human endometrium (n = 41) was collected with ethical approval and subject consent. Donor peripheral blood monocyte-derived macrophages were treated with estradiol, progesterone, or cortisol. The effect of peripheral blood monocyte-derived macrophage secretory products on the expression of angiogenic RNAs by endothelial cells was examined. Immunofluorescence was used to examine localization in macrophages and other endometrial cell types across the menstrual cycle. Endometrial macrophages express the glucocorticoid receptor. In vitro culture with supernatants from cortisol-treated peripheral blood monocyte-derived macrophages resulted in altered endometrial endothelial cell expression of the angiogenic genes, CXCL2, CXCL8, CTGF, and VEGFC. These data highlight the importance of local cortisol in regulating paracrine actions of macrophages in the endometrium. CXCL2 and CXCL8 were detected in endometrial macrophages in situ. The expression of these factors was highest in the endometrium during the menstrual phase, consistent with these factors having a role in endometrial repair. Our data have indicated that activation of macrophages with glucocorticoids might have paracrine effects by increasing angiogenic factor expression by endometrial endothelial cells. This might reflect possible roles for macrophages in endometrial repair of the vascular bed after menstruation.
- Published
- 2015
46. Supporting researchers in an era of team science
- Author
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Philippa T. K. Saunders
- Subjects
03 medical and health sciences ,Engineering ,0302 clinical medicine ,020205 medical informatics ,business.industry ,0202 electrical engineering, electronic engineering, information engineering ,Engineering ethics ,030212 general & internal medicine ,02 engineering and technology ,General Medicine ,business ,Team science - Published
- 2017
47. Endometrial Intracrinology: Oestrogens, Androgens and Endometrial Disorders
- Author
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Douglas A, Gibson, Ioannis, Simitsidellis, Frances, Collins, and Philippa T K, Saunders
- Subjects
Uterine Diseases ,aromatase ,endometriosis ,decidualisation ,Estrogens ,Dehydroepiandrosterone ,Review ,oestradiol ,sulfatase ,Endometrium ,dehydroepiandrosterone (DHEA) ,testosterone ,endometrial cancer ,Androgens ,Humans ,Female - Abstract
Peripheral tissue metabolism of steroids (intracrinology) is now accepted as a key way in which tissues, such as the endometrium, can utilise inactive steroids present in the blood to respond to local physiological demands and ‘fine-tune’ the activation or inhibition of steroid hormone receptor-dependent processes. Expression of enzymes that play a critical role in the activation and inactivation of bioactive oestrogens (E1, E2) and androgens (A4, T, DHT), as well as expression of steroid hormone receptors, has been detected in endometrial tissues and cells recovered during the menstrual cycle. There is robust evidence that increased expression of aromatase is important for creating a local microenvironment that can support a pregnancy. Measurement of intra-tissue concentrations of steroids using liquid chromatography–tandem mass spectrometry has been important in advancing our understanding of a role for androgens in the endometrium, acting both as active ligands for the androgen receptor and as substrates for oestrogen biosynthesis. The emergence of intracrinology, associated with disordered expression of key enzymes such as aromatase, in the aetiology of common women’s health disorders such as endometriosis and endometrial cancer has prompted renewed interest in the development of drugs targeting these pathways, opening up new opportunities for targeted therapies and precision medicine.
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- 2018
48. Effects of Boswellia serrata Roxb. and Curcuma longa L. in an In Vitro Intestinal Inflammation Model Using Immune Cells and Caco-2
- Author
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Bianca De Leo, Daniela Catanzaro, Elisabetta Miraldi, Monica Montopoli, Paolo Governa, Veronica Cocetta, Philippa T. K. Saunders, Marco Biagi, and Maddalena Marchi
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biology ,medicine.medical_treatment ,Pharmacology ,biology.organism_classification ,Intestinal epithelium ,Small intestine ,medicine.anatomical_structure ,Cytokine ,Immune system ,Mechanism of action ,Intestinal mucosa ,medicine ,Boswellia serrata ,Curcuma ,medicine.symptom - Abstract
Inflammatory bowel diseases, which consist of chronic inflammatory conditions of the colon and the small intestine, are considered a global disease of our modern society. Recently, the interest toward the use of herbal therapies for the management of inflammatory bowel diseases has increased because of their effectiveness and favourable safety profile, compared to conventional drugs. Boswellia serrata Roxb. and Curcuma longa L. are amongst the most promising herbal drugs, however, their clinical use in inflammatory bowel diseases is limited and little is known on their mechanism of action. The aim of this work was to investigate the effects of two phytochemically characterized extracts of B. serrata and C. longa in an in vitro model of intestinal inflammation. Their impact on cytokine release and reactive oxygen species production, as well as the maintenance of the intestinal barrier function and on intestinal mucosa immune cells infiltration, has been evaluated. The extracts showed a good protective effect on the intestinal epithelium at 1 µg/mL, with TEER values increasing by approximately 1.5 fold, compared to LPS-stimulated cells. C. longa showed an anti-inflammatory mechanism of action, reducing IL-8, TNF-α and IL-6 production by approximately 30%, 25% and 40%, respectively, compared to the inflammatory stimuli. B. serrata action was linked to its antioxidant effect, with ROS production being reduced by 25%, compared to H2O2-stimulated Caco-2 cells. C. longa and B. serrata resulted to be promising agents for the management of inflammatory bowel diseases by modulating in vitro parameters which have been identified in the clinical conditions.
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- 2018
49. Animal models of endometriosis: Replicating the aetiology and symptoms of the human disorder
- Author
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Ioannis Simitsidellis, Philippa T. K. Saunders, and Douglas A Gibson
- Subjects
0301 basic medicine ,Infertility ,Primates ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Endometriosis ,Psychological intervention ,Prodromal Symptoms ,Reproductive age ,Rodentia ,Pelvic Pain ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Medicine ,Animals ,Humans ,Intensive care medicine ,030219 obstetrics & reproductive medicine ,business.industry ,Chronic pain ,medicine.disease ,Key features ,Disease Models, Animal ,030104 developmental biology ,Preclinical testing ,Etiology ,Heterografts ,Female ,Chronic Pain ,business ,Infertility, Female - Abstract
Endometriosis is a chronic incurable disorder that affects 1 in 10 women of reproductive age: associated symptoms include chronic pain and infertility. The aetiology of endometriosis remains poorly understood but patients, clinicians and researchers are all in agreement that new non-surgical therapies are urgently needed to reduce the severity of symptoms. Preclinical testing of drugs requires the development and validation of models that recapitulate the key features of the disorder. In this review we describe the best-validated animal models (primate, rodent, xenograft) and their contributions to our understanding of the factors underpinning the development of symptoms. We consider the evidence that these models have provided the platform for identification of new therapeutic interventions and reflect on future directions for research and drug validation.
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- 2018
50. A pilot randomised double blind controlled trial of the efficacy of purified fatty acids for the treatment of women with endometriosis-associated pain (PurFECT):study protocol
- Author
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Ibtisam M. Abokhrais, Philippa T. K. Saunders, Fiona C. Denison, Ann Doust, Linda Williams, and Andrew W. Horne
- Subjects
Pilot trial ,lcsh:R5-920 ,Endometriosis ,Journal Article ,Purified fatty acids ,lcsh:Medicine (General) ,Chronic pelvic pain - Abstract
Background: Endometriosis affects 6-10% of women and is associated with debilitating pelvic pain. It costs the UK > £2.8 billion per year in loss of productivity. Endometriosis can be managed by surgical excision or medically by ovarian suppression. However, ~ 75% symptoms recur after surgery and available medical treatments have undesirable side effects and are contraceptive. Omega-3 purified fatty acids (PUFA) have been shown in animal models to reduce factors that are thought to lead to endometriosis-associated pain, have minimal side effects, and no effects on fertility. This paper presents a protocol for a two-arm, pilot parallel randomised controlled trial (RCT) which aims to inform the planning of a future multicentre trial to evaluate the efficacy of Omega-3 PUFA in the management of endometriosis-associated pain in women.Methods: The study will recruit women with endometriosis over a 12-month period in the National Health Service (NHS) Lothian, UK, and randomise them to 8 weeks of treatment with Omega-3 PUFA or comparator (olive oil). The primary objective is to assess recruitment and retention rates. The secondary objectives are to determine the effectiveness/acceptability to participants of the proposed methods of recruitment/randomisation/treatments/questionnaires, to inform the sample size calculation and to refine the research methodology for a future large randomised controlled trial. Response to treatment will be monitored by pain scores and questionnaires assessing physical and emotional function compared at baseline and 8 weeks.Discussion: We recognise that there may be potential difficulties in mounting a large randomised controlled trial for endometriosis to assess Omega-3 PUFA because they are a dietary supplement readily available over the counter and already used by women with endometriosis. We have therefore designed this pilot study to assess practical feasibility and following the 'Initiative on Methods, Measurement, and Pain Assessment in Clinical Trials' recommendations for the design of chronic pain trials.Trial registration: ISRCTN44202346.
- Published
- 2018
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