Sex-specific disease aetiology from developmental steroid insults: mechanistic understanding and biomarker development towards disease prevention.
Lead Research Organisation:
Edinburgh Napier University
Department Name: School of Applied Science
Abstract
We are living and working longer, but capitalising on this means enjoying extended 'healthspan' as well as extended lifespan. Risks of developing many adult illnesses are set down as we develop in the womb, hence the environment we experience during fetal life needs to be 'just right'. Understanding how the fetal environment colours development helps decision making during pregnancy to ensure best health chances for our children.
Hormones drive development. Steroid hormones direct numerous processes, including correct male or female development. Unfortunately, imbalance in sex steroids during development can have lifelong health consequences. Numerous clinical conditions associated with altered early life steroid exposure have effects that are 'silent' until later in life. Some of these consequences are sex-specific, likely due to differing hormonal requirements of male or female development. There are also many chemicals in our environment that can reach a developing fetus. Some of these compounds behave like steroids, or affect natural steroid actions; these are 'endocrine disrupting compounds'.
The sex steroids androgens and oestrogens play critical male and female-specific roles in development. Common conditions such as Polycystic Ovary Syndrome (PCOS) have origins of incorrect sex steroid exposure during development. Endocrine disrupting compounds commonly mimic, or alter, effects of these sex steroids. If we knew what the effects of the incorrect sex steroid exposure were during fetal life, and could connect this information to adult disease development, then we could predict what the effects of chemical exposures, and clinical conditions such as PCOS would have on the next generation. By identifying biomarkers in adult life that are legacies of incorrect fetal steroid exposure, we could identify those individuals at risk of developing associated illness. We could then design treatments to prevent disease prior to its occurrence in at risk individuals, and thereby protect such people from having 'healthspans' shorter than their lifespans.
Whilst investigating origins of PCOS we developed sheep models of this human condition by altering the sex steroid environment that developing lambs experience. This leads to development of insulin resistance, obesity, altered pancreatic and liver function in adulthood. Here we have chosen to study the liver since it responds to, deactivates and transforms steroids. Our work indicated that the liver was affected in a health-relevant manner by our altered fetal steroid exposure studies.
Recent technological advances will let us build 'maps' of effects of altered steroid exposure during fetal life on development and adult function of liver, specific to males and females. As we already know some adult health consequences of altered fetal steroid exposure we can now discover the reasons behind them. Once we know the genes and proteins affected, we can examine the possibility of some alterations in the liver being detectable in the circulation. This will give us blood tests to see if someone has been incorrectly exposed to altered steroid signalling during their early life, and then we can consider how to help such individuals prior to them getting ill. All sheep samples required are already collected from previous scientific investigations, saving time, money, and reducing animal use in research. We know the sheep samples we propose for study are affected in terms of human-relevant health, so these samples hold answers to how these effects occur. We will reveal the consequences of altered sex steroid exposure during development, how such alterations affect adult disease risk, develop blood tests to identify during early life those at increased risk in later life, and provide information regarding possible treatment routes.
Hormones drive development. Steroid hormones direct numerous processes, including correct male or female development. Unfortunately, imbalance in sex steroids during development can have lifelong health consequences. Numerous clinical conditions associated with altered early life steroid exposure have effects that are 'silent' until later in life. Some of these consequences are sex-specific, likely due to differing hormonal requirements of male or female development. There are also many chemicals in our environment that can reach a developing fetus. Some of these compounds behave like steroids, or affect natural steroid actions; these are 'endocrine disrupting compounds'.
The sex steroids androgens and oestrogens play critical male and female-specific roles in development. Common conditions such as Polycystic Ovary Syndrome (PCOS) have origins of incorrect sex steroid exposure during development. Endocrine disrupting compounds commonly mimic, or alter, effects of these sex steroids. If we knew what the effects of the incorrect sex steroid exposure were during fetal life, and could connect this information to adult disease development, then we could predict what the effects of chemical exposures, and clinical conditions such as PCOS would have on the next generation. By identifying biomarkers in adult life that are legacies of incorrect fetal steroid exposure, we could identify those individuals at risk of developing associated illness. We could then design treatments to prevent disease prior to its occurrence in at risk individuals, and thereby protect such people from having 'healthspans' shorter than their lifespans.
Whilst investigating origins of PCOS we developed sheep models of this human condition by altering the sex steroid environment that developing lambs experience. This leads to development of insulin resistance, obesity, altered pancreatic and liver function in adulthood. Here we have chosen to study the liver since it responds to, deactivates and transforms steroids. Our work indicated that the liver was affected in a health-relevant manner by our altered fetal steroid exposure studies.
Recent technological advances will let us build 'maps' of effects of altered steroid exposure during fetal life on development and adult function of liver, specific to males and females. As we already know some adult health consequences of altered fetal steroid exposure we can now discover the reasons behind them. Once we know the genes and proteins affected, we can examine the possibility of some alterations in the liver being detectable in the circulation. This will give us blood tests to see if someone has been incorrectly exposed to altered steroid signalling during their early life, and then we can consider how to help such individuals prior to them getting ill. All sheep samples required are already collected from previous scientific investigations, saving time, money, and reducing animal use in research. We know the sheep samples we propose for study are affected in terms of human-relevant health, so these samples hold answers to how these effects occur. We will reveal the consequences of altered sex steroid exposure during development, how such alterations affect adult disease risk, develop blood tests to identify during early life those at increased risk in later life, and provide information regarding possible treatment routes.
Technical Summary
Fetal development, including sex development, is modulated by steroids. Incorrect steroid exposure in utero predisposes to altered postnatal disease risk. If we understand the postnatal health legacies that are consequences of excess steroid exposure in utero, we can advise upon issues such as limiting exposure to exogenous compounds that alter steroid signalling, and develop amelioration strategies where exposure is due to underlying medical conditions.
RESEARCH PLAN
We will interrogate, using Next Generation Sequencing and shotgun proteomics, pre-existing sets of ovine tissues, where excess sex steroid signalling was experimentally driven during fetal life to model human PCOS. Due to its modulatory role in steroid exposure during development and postnatal life, the liver is our focus. We capitalise on previous investment in large animal modelling, and require no additional animal experimentation. We add translation to humans by interfacing this work with ongoing work examining the human fetal liver.
DELIVERABLE 1 We will identify hepatic differences in postnatal life attributable to developmental androgenisation in sheep. We will examine the outcomes of steroidal excess during development on male and female postnatal hepatic function. This work will identify postnatal legacies, and fetal antecedents of, disrupted steroid signalling in utero, in a sex-specific context.
DELIVERABLE 2 We will use health relevant indices collected in vivo, to delineate 'effect' from 'adverse effect'. We will utilise human data sets to provide translational confidence between interventional ovine models and observational human studies.
DELIVERABLE 3 We will identify postnatal biomarkers of specific fetal steroid excesses (androgen or oestrogen) in offspring from birth to adulthood that are relevant to health, and deliver mechanistic insights to inform future therapeutic strategies.
RESEARCH PLAN
We will interrogate, using Next Generation Sequencing and shotgun proteomics, pre-existing sets of ovine tissues, where excess sex steroid signalling was experimentally driven during fetal life to model human PCOS. Due to its modulatory role in steroid exposure during development and postnatal life, the liver is our focus. We capitalise on previous investment in large animal modelling, and require no additional animal experimentation. We add translation to humans by interfacing this work with ongoing work examining the human fetal liver.
DELIVERABLE 1 We will identify hepatic differences in postnatal life attributable to developmental androgenisation in sheep. We will examine the outcomes of steroidal excess during development on male and female postnatal hepatic function. This work will identify postnatal legacies, and fetal antecedents of, disrupted steroid signalling in utero, in a sex-specific context.
DELIVERABLE 2 We will use health relevant indices collected in vivo, to delineate 'effect' from 'adverse effect'. We will utilise human data sets to provide translational confidence between interventional ovine models and observational human studies.
DELIVERABLE 3 We will identify postnatal biomarkers of specific fetal steroid excesses (androgen or oestrogen) in offspring from birth to adulthood that are relevant to health, and deliver mechanistic insights to inform future therapeutic strategies.
Planned Impact
The advances made in this project will impact upon, and benefit, research disciplines, spanning basic developmental biology, fetal origins of adult disease and medical research. Medical communities, and individuals will all benefit from the new knowledge generated. We recognise the need for increased understanding of the mechanisms of health programming to occur in tandem with improved identification of at risk individuals and this proposal is driven by the unmet clinical needs of both of these ambitions.
Prenatal steroids affect metabolic development and maturation, and increasing evidence links the prenatal steroid environment to altered lifetime risk of metabolic disease development. This research will map what the effects are during development, and in postnatal life, of in utero sex steroid excess, focussing on the liver as a loci of such effects with relevance to development and lifelong health. We will deliver all information in a sex-specific context, since the outcomes of prenatal steroid alterations are predicated upon sex-specific function, itself driven by endogenous sex steroid exposure. Thus the work entailed will also deliver basic biological information regarding hepatic function differences between males and females that are underwritten by steroids during fetal development. This may have implications beyond the current focus of the project, for example in terms of organ transplantation considerations. By understanding the consequences of in utero steroid excesses (clinical or chemical in origin), and relationships to development of disease in adulthood, then knowledge of the systems affected and how they are affected will have implications for preventative and amelioration therapy approaches. We will identify biomarkers of altered development due to incorrect sex steroid exposures, thus placing us in a position to begin to identify at risk individuals earlier than when first signs of postnatal disease development are evident, with congruent increased understanding of the systems affected and consequences of the early life insult. Collectively, these aspects are all relevant and of benefit to both medical practitioners and researchers.
Our previously collected samples are tied to health indices measured in vivo, and so our proposal will be powered to delineate effects from adverse effects, and will be translated to humans from the ovine models used via inclusion of relevant human samples throughout all analyses.
In summary, we will deliver health relevant biomarkers discernible in fetal life, health relevant biomarkers in adult life caused by fetal steroidal alterations, and the mechanisms of action of excess sex steroids during development which will define the consequences of exposure and direct interventional strategy. Collectively, the knowledge gleaned from these studies will positively contribute to and impact upon future lifelong human health.
Prenatal steroids affect metabolic development and maturation, and increasing evidence links the prenatal steroid environment to altered lifetime risk of metabolic disease development. This research will map what the effects are during development, and in postnatal life, of in utero sex steroid excess, focussing on the liver as a loci of such effects with relevance to development and lifelong health. We will deliver all information in a sex-specific context, since the outcomes of prenatal steroid alterations are predicated upon sex-specific function, itself driven by endogenous sex steroid exposure. Thus the work entailed will also deliver basic biological information regarding hepatic function differences between males and females that are underwritten by steroids during fetal development. This may have implications beyond the current focus of the project, for example in terms of organ transplantation considerations. By understanding the consequences of in utero steroid excesses (clinical or chemical in origin), and relationships to development of disease in adulthood, then knowledge of the systems affected and how they are affected will have implications for preventative and amelioration therapy approaches. We will identify biomarkers of altered development due to incorrect sex steroid exposures, thus placing us in a position to begin to identify at risk individuals earlier than when first signs of postnatal disease development are evident, with congruent increased understanding of the systems affected and consequences of the early life insult. Collectively, these aspects are all relevant and of benefit to both medical practitioners and researchers.
Our previously collected samples are tied to health indices measured in vivo, and so our proposal will be powered to delineate effects from adverse effects, and will be translated to humans from the ovine models used via inclusion of relevant human samples throughout all analyses.
In summary, we will deliver health relevant biomarkers discernible in fetal life, health relevant biomarkers in adult life caused by fetal steroidal alterations, and the mechanisms of action of excess sex steroids during development which will define the consequences of exposure and direct interventional strategy. Collectively, the knowledge gleaned from these studies will positively contribute to and impact upon future lifelong human health.
Organisations
Publications
Siemienowicz KJ
(2022)
Hepatic Mitochondrial Dysfunction and Risk of Liver Disease in an Ovine Model of "PCOS Males".
in Biomedicines
Johnston ZC
(2018)
The human fetal adrenal produces cortisol but no detectable aldosterone throughout the second trimester.
in BMC medicine
Siemienowicz KJ
(2021)
Aberrant subcutaneous adipogenesis precedes adult metabolic dysfunction in an ovine model of polycystic ovary syndrome (PCOS).
in Molecular and cellular endocrinology
Siemienowicz KJ
(2021)
Pubertal FGF21 deficit is central in the metabolic pathophysiology of an ovine model of polycystic ovary syndrome.
in Molecular and cellular endocrinology
Walker N
(2019)
Nutrient transporter expression in both the placenta and fetal liver are affected by maternal smoking.
in Placenta
Siemienowicz KJ
(2019)
Fetal androgen exposure is a determinant of adult male metabolic health.
in Scientific reports
Siemienowicz K
(2020)
Early pregnancy maternal progesterone administration alters pituitary and testis function and steroid profile in male fetuses
in Scientific Reports
Bongaerts E
(2022)
Maternal exposure to ambient black carbon particles and their presence in maternal and fetal circulation and organs: an analysis of two independent population-based observational studies.
in The Lancet. Planetary health
Description | Can the human fetal plasma proteome be used to monitor "exposure" to increased maternal body mass index during fetal life and predict resulting childhood diseases? |
Amount | £11,935 (GBP) |
Funding ID | 17/034 |
Organisation | NHS Grampian |
Sector | Public |
Country | United Kingdom |
Start | 04/2018 |
End | 03/2019 |
Description | Discovery of retinoic acid receptor control of stress granules as a treatment for neurodegenerative diseases |
Amount | £96,875 (GBP) |
Funding ID | CT-21-18 |
Organisation | The Cunningham Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 10/2022 |
End | 09/2025 |
Description | Female Reproductive toxicity of Endocrine disrupting chemicals (EDCs): a human evidence-based screening and Identification Approach (FREIA) |
Amount | € 5,216,904 (EUR) |
Funding ID | 825100 |
Organisation | European Commission |
Sector | Public |
Country | European Union (EU) |
Start | 01/2019 |
End | 12/2023 |
Description | Towards a novel treatment targeting obesity in women with polycystic ovary syndrome (PCOS) |
Amount | £379,663 (GBP) |
Funding ID | MR/W015439/1 |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 12/2021 |
End | 03/2024 |
Description | Towards a novel treatment targeting obesity in women with polycystic ovary syndrome (PCOS) |
Amount | £379,664 (GBP) |
Funding ID | MR/W015439/1 |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 12/2021 |
End | 03/2024 |
Title | Establishment of the 10-year SAFeR (Scottish Advanced Fetal Research) study |
Description | This was an aim of MRC MR/L010011/1 award. The collection of human fetal material has been approved by the NHS North of Scotland Research Ethics Committee (REC 15/NS/0123 "Normal development of the human fetus and the influences and mechanisms by which that development occurs and is perturbed") which includes the collection of an extensive range of fetal tissues and maternal data. Women seeking elective terminations of normal pregnancy between 7 and 20 weeks of gestation aree recruited with full written informed consent by clinical staff working independently at hospitals at both Aberdeen and Glasgow. Permission has been granted for collection of up to 1,600 fetuses over 10 years and this will represent an unprecedented step change in the power of studies of the human fetus. One cautionary note is that Glasgow NHS R&D have insisted on a separate ethics application resulting in ongoing delays so that collection in Glasgow has not started yet. In Aberdeen the ethics application started in January 2015 with final approval obtained in December 2015. Aberdeen collection started in May 2016 once governance and sponsorship processes were completed. |
Type Of Material | Biological samples |
Year Produced | 2016 |
Provided To Others? | Yes |
Impact | In Aberdeen, as of March 2018, 180 fetuses and their placentae have been collected. First contribution of this collection to publication was: The human fetal adrenal produces cortisol but no detectable aldosterone throughout the second trimester. Johnston ZC, Bellingham M, Filis P, Soffientini U, Hough D, Bhattacharya S, Simard M, Hammond GL, King P, O'Shaughnessy PJ, Fowler PA. BMC Med. 2018 Feb 12;16(1):23. The study has contributed to 3 grant applications (to Wellcome Trust, Wellbeing of Women and Kidney Research UK) and funded Arthritis Research UK, BBSRC EASTBIO DTP and MSCA-ITN projects. |
Title | Human fetal liver and placenta collection |
Description | As part of MRC MR/L010011/1 and added to during MRC MR/P011535/1, we have accumulated over 260 human fetal liver and placenta pairs. RNA, DNA, protein and steroid fractions have been extracted. This collection is growing as part of the SAFeR study. |
Type Of Material | Database/Collection of data |
Year Produced | 2017 |
Provided To Others? | Yes |
Impact | The collection has been used to validate/extend MRC MR/L010011/1 findings and form part of the major publications in preparation. |
Title | Human fetal plasma proteome |
Description | MRC MR/L010011/1 and added to during MRC MR/P011535/1. Shot-gun proteomics data on 60 second trimester human fetal plasma samples, categorised according to fetal age, fetal sex and maternal smoking status. >300 proteins quantified. The data have been uploaded to PRIDE and will remain confidential until the major publication is in press. |
Type Of Material | Database/Collection of data |
Year Produced | 2018 |
Provided To Others? | No |
Impact | Major publications from MRC MR/L010011/1 and MRC MR/P011535/1 are in progress |
URL | https://www.ebi.ac.uk/pride/archive/ |
Title | Liver 80 RNAseq |
Description | RNA-seq data on 80 second trimester human fetal livers, categorised according to fetal age, fetal sex and maternal smoking status. 13,599 genes quantified. The data have been uploaded to ArrayExpress (E-MTAB-6103) and will remain confidential until the major publications from MRC MR/L010011/1 are in press. |
Type Of Material | Database/Collection of data |
Year Produced | 2017 |
Provided To Others? | No |
Impact | The major publications from MRC MR/L010011/1 are in preparation. Data mining is also supporting an EASTBIO PhD studentship and MSCA-ITN research. Outputs so far are part of two abstract at PPTOX VI 2018 international conference https://www.greengate.fo/pptox-conference |
URL | https://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-6103 |
Title | Liver 80 proteomics |
Description | Shot-gun proteomics data on 80 second trimester human fetal livers, categorised according to fetal age, fetal sex and maternal smoking status. 2,195 proteins quantified. The data have been uploaded to PRIDE and will remain confidential until the major publications from MRC MR/L010011/1 are in press. |
Type Of Material | Database/Collection of data |
Year Produced | 2017 |
Provided To Others? | No |
Impact | The major publications from MRC MR/L010011/1 are in preparation. Data mining is also supporting other publications and projects. |
URL | https://www.ebi.ac.uk/pride/archive/ |
Title | Ovine hepatic proteome/plasma proteome |
Description | Ovine postnatal male and female hepatic proteome and plasma proteome Ovine postnatal male and female hepatic and plasma proteome response to prenatal estrogenic and androgenic excess ProteomeXchange Consortium via the PRIDE140 partner repository with the dataset identifier PXD014050 |
Type Of Material | Database/Collection of data |
Year Produced | 2020 |
Provided To Others? | Yes |
Impact | Publication Sci Rep. 2019 Dec 27;9(1):20195. doi: 10.1038/s41598-019-56790-4 |
URL | https://www.ebi.ac.uk/pride/archive/projects/PXD014050 |
Title | Ovine hepatic transcriptome |
Description | Ovine postnatal hepatic transcriptome (RNAseq), male and female specific data sets. Ovine postnatal hepatic transcriptome (RNAseq), male and female specific responses to prenatal steroidal (androgenic and estrogenic) excess ArrayExpress data base, (under accession number E-MTAB-8032) |
Type Of Material | Database/Collection of data |
Year Produced | 2019 |
Provided To Others? | Yes |
Impact | Publication (Sci Rep. 2019 Dec 27;9(1):20195. doi: 10.1038/s41598-019-56790-4) |
URL | http://www.ebi.ac.uk/arrayexpress |
Description | Overweight or smoking mums cause 'worrying changes' to unborn children's thyroid |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | This was a University of Aberdeen press release that resulted some take up by on-line and traditional press. e.g. Smoking in pregnancy linked to 'worrying' thyroid changes in unborn baby: https://www.heraldscotland.com/news/16998982.smoking-in-pregnancy-linked-to-worrying-thyroid-changes-in-unborn-baby/ |
Year(s) Of Engagement Activity | 2018 |
URL | https://www.abdn.ac.uk/news/12358/ |
Description | Premature babies' low blood pressure puzzle explained |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | This was a University of Aberdeen press release that resulted some take up by on-line and traditional press. The University Social media unit also produced a very nice explainer : https://twitter.com/aberdeenuni/status/963697423167164422 |
Year(s) Of Engagement Activity | 2018 |
URL | https://www.abdn.ac.uk/news/11599/ |
Description | Public lecture |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | This lecture was an opportunity to engage the public in science in general, but also to introduce the concepts of prenatal programming, epigenetics, and how science is conducted. A turnout of approximately 200 people meant that the format was designed for a lay audience, and immediately after a Q and A session there was a reception (funded by Edinburgh Napier University) for further discussion. |
Year(s) Of Engagement Activity | 2018 |
Description | THE CONVERSATION Move over testosterone, another hormone is also vital for making boys - and it doesn't come from the testes |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | "Move over testosterone, another hormone is also vital for making boys - and it doesn't come from the testes" The Conversation 14/02/2019 between 14/02/.2019 and 13/03/2019 this article was read over 47,000 times |
Year(s) Of Engagement Activity | 2019 |
URL | https://theconversation.com/move-over-testosterone-another-hormone-is-also-vital-for-making-boys-and... |
Description | What makes a man? Testosterone only half the story, scientists say |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | This was a University of Aberdeen press release that resulted some take up by on-line and traditional press. Attention included other articles e.g. https://theconversation.com/you-need-more-than-just-testes-to-make-a-penis-111625 Some with further interview - eg NOVA at PBS Org USA, El Mundo Spain e.g. https://www.pbs.org/wgbh/nova/article/got-a-penis-you-might-have-moms-placenta-to-thank/ |
Year(s) Of Engagement Activity | 2019 |
URL | https://www.abdn.ac.uk/news/12707/ |