Prenatal glucocorticoids exposure and fetal adrenal developmental programming

Journal Pre-proof Prenatal glucocorticoids exposure and fetal adrenal developmental programming Yawen Chen, Zheng He, Guanghui Chen, Min Liu, Hui Wang

PII:

S0300-483X(19)30265-3

DOI:

https://doi.org/10.1016/j.tox.2019.152308

Reference:

TOX 152308

To appear in:

Toxicology

Received Date:

24 June 2019

Revised Date:

25 August 2019

Accepted Date:

7 October 2019

Please cite this article as: Chen Y, He Z, Chen G, Liu M, Wang H, Prenatal glucocorticoids exposure and fetal adrenal developmental programming, Toxicology (2019), doi: https://doi.org/10.1016/j.tox.2019.152308

This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. © 2019 Published by Elsevier.

Title page Prenatal glucocorticoids exposure and fetal adrenal developmental programming

Yawen Chena, Zheng Heb, Guanghui Chena, Min Liua, Hui Wanga,b,*

Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan 430071, China

b

Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan 430071, China

*

Corresponding author information:

ro of

a

Prof. Hui Wang, M.D., Ph.D. Department of Pharmacology

-p

School of Basic Medical Science of Wuhan University Wuhan, 430071, Hubei Province, China

re

Telephone: +86-13627232557

lP

E-mail: [email protected]

na

Abstract: Clinically, we apply synthetic glucocorticoids to treat fetal and maternal diseases, such as premature labor and autoimmune diseases. Although its clinical efficacy is positive, the fetus will be exposed

ur

to exogenous synthetic glucocorticoids. Prenatal adverse environments (such as xenobiotics exposure, malnutrition, infection, hypoxia and stress) can cause fetuses overexposure to excessive endogenous maternal

Jo

glucocorticoids. The level of glucocorticoids is the key to fetal tissue maturation and postnatal fate. A large number of studies have found that prenatal glucocorticoids exposure can lead to fetal adrenal dysplasia and dysfunction, continuing after birth and even into adulthood. As the core organ of fetal-originated adult diseases, fetal adrenal dysplasia is closely related to the susceptibility and occurrence of multiple chronic diseases, and there are also obvious gender differences. However, its intrauterine programming mechanisms have not been fully elucidated. This review summarizes recent advances in prenatal glucocorticoids exposure 1

and fetal adrenal developmental programming alterations, which is of great significance for explaining adrenal developmental toxicity and the intrauterine origin of fetal-originated adult diseases.

Keywords: Glucocorticoids; Adrenal; Intrauterine programming; Epigenetic mechanism; Glucocorticoid insulin like growth 1 axis; Gender differences.

Abbreviations: DOHaD, Developmental Origins of Health and Disease; IUGR, intrauterine growth retardation; HPA, hypothalamus-pituitary-adrenal; sGC, Synthetic glucocorticoids; WHO, World Health

ro of

Organization; 11β-HSDs, 11β-hydroxysteroid dehydrogenases; ZG, zona glomerulosa; ZF, zona fasciculata; ZR, zona reticularis; IGF1, insulin-like growth factor 1; TGFβ, transforming growth factor β; cAMP, cyclic adenosine monophosphate; SF1, steroidogenic factor 1; Dax 1, dosage-sensitive sex reversal, adrenal

-p

hypoplasia congenita, X-linked-1; TCF21,Transcription factors 21; GC-IGF1, glucocorticoids-insulin-like growth factor 1; GR, glucocorticoids receptor; C/EBP , C/EBP homologous protein ; MR,

re

mineralocorticoid receptors; PNC, parvocellular neuroendocrine; PVN, paraventricular nucleus; CRH, corticotropin-releasing hormone; AVP, arginine vasopressin; ACTH, adrenocorticotropic hormone; StAR,

lP

steroidogenic acute regulatory protein; P450scc, cytochrome P450 cholesterol side chain cleavage; PI3K, phosphorylated phosphatidylinositol-3-hydroxykinase; GRE, glucocorticoid response element; HDAC7,

na

histone deacetylase 7.

1 Introduction

ur

Contents

Jo

2 Status of prenatal glucocorticoids exposure 2.1 Clinical application of synthetic glucocorticoids during pregnancy 2.2 Maternal glucocorticoids over-exposure caused by prenatal adverse environment 2.3 Placental glucocorticoids barrier and 11β-hydroxysteroid dehydrogenases system 3 Prenatal glucocorticoids exposure-induced adrenal dysplasia 3.1 Adrenal morphological development abnormality 2

3.2 Adrenal functional development abnormality 4 Intrauterine programming mechanism of glucocorticoids-induced adrenal dysfunction 4.1 Low-functional programming of adrenal de novo steroidogenesis 4.2 Developmental programming alterations of glucocorticoid-insulin-like growth 1 axis 4.3 Epigenetic modification changes and adrenal developmental programming 5 Prenatal glucocorticoids exposure and gender differences of adrenal development 6 Conclusions and prospects

ro of

1 Introduction

In the past three decades, scholars have carried out a large number of studies into the relationship

between prenatal adverse environments, fetal low birth weight and adult chronic diseases, and proposed a new

-p

concept of Developmental Origins of Health and Disease (DOHaD). Fetal programming has become an

important theory to understand the programming of lifetime health (Fleming et al., 2018). Glucocorticoids

re

have some vital roles during the time of intrauterine and neonatal development. Not only is it essential for normal maturation close to term, it also acts as an important signal of environmental compromise earlier in

lP

gestation. The glucocorticoids-triggered switch from tissue accretion to differentiation improves offspring fitness by maximizing the chances of the fetus surviving into adulthood. Prenatally, early activation of this

na

switch ensures that fetal growth is commensurate with the nutrient supply in utero and that fetal tissues are sufficiently mature to function ex utero when delivery occurs (Fowden et al., 2016). However, the fetus is

ur

particularly susceptible to internal and external environmental stimuli, many of which can alter the level of glucocorticoids and individual developmental trajectories. It subsequently increases susceptibility to adult

Jo

diseases. Prenatal glucocorticoids exposure can lead to intrauterine growth retardation (IUGR), appearing as low birth weight and fetal multiple organ dysfunctions. Accumulated studies have shown that IUGR caused fetal distress, neonatal asphyxia and perinatal death, and that the harm could last after birth. Eventually, it results in susceptibility to chronic diseases in adulthood (Bremer, 2010; Moisiadis and Matthews, 2014a, b). As the core organ of fetal-originated adult diseases, the adrenal gland is not only the terminal effector organ of the hypothalamus-pituitary-adrenal (HPA) axis, but also the earliest and fastest-growing organ. It can 3

synthesize multiple steroid hormones, which is important for normal gestation and fetal growth (Busada and Cidlowski, 2017). The development of the adrenal gland and the level of fetal glucocorticoids not only determine the maturation of fetal tissue, but also affect its fate after birth (Ishimoto and Jaffe, 2011; Reynolds, 2013). In this review, we summarize the advance in adrenal development and intrauterine programming mechanism induced by prenatal glucocorticoids exposure in offspring. It can provide the experimental and theoretical basis for the effective evaluation of the harmful factors during pregnancy, and explore the early prevention of fetal-originated adult diseases.

ro of

2 Status of prenatal glucocorticoids exposure

Synthetic glucocorticoids (sGC) are applied to treat fetal and maternal diseases during pregnancy, such as premature labor and autoimmune diseases. Although its clinical efficacy is positive, the fetus is exposed to

-p

sGC. Meanwhile, fetus will be over-exposed to endogenous maternal glucocorticoids, which is induced by prenatal adverse environments.

re

2.1 Clinical application of synthetic glucocorticoids during pregnancy

Clinically, sGC such as dexamethasone and betamethasone, help pregnant women relieve nausea and

lP

vomiting, prevents placenta previa, and improves pregnancy outcomes such as habitual abortion (Pattanittum et al., 2008; Practice, 2011; Roberts et al., 2017). The World Health Organization (WHO) reported that the

na

prevalence of prenatal sGC treatment for premature labor in 22-36 weeks of pregnancy is 54% in the maternal and child health survey data of 359 institutions in 29 countries, and even up to 91% in some countries (Vogel

ur

et al., 2014). Although the clinical application of sGC during pregnancy is extensive and effective, studies found the neonatal weight of infants treated by sGC is much lower than normal individuals, especially in

Jo

multi-course treatment. Besides, low birth weight is closely related to the occurrence of chronic diseases in adulthood (Lahti et al., 2015). Animal experiments have also confirmed that prenatal dexamethasone exposure caused low birth weight, developmental toxicity of multiple organs and related diseases in adulthood (Dodic et al., 1998; Long et al., 2012). 2.2 Maternal glucocorticoids over-exposure caused by prenatal adverse environment Adverse environments during pregnancy (such as xenobiotics exposure, malnutrition, infection, hypoxia 4

and stress) could elevate the level of maternal glucocorticoids (Buss et al., 2012; Correia-Branco et al., 2015; Cuffe et al., 2014; Lesage et al., 2001; Pawluski et al., 2012). Maternal malnutrition during pregnancy increased the level of intrauterine maternal cortisol and the occurrence of IUGR, and it was shown that cortisol could program offspring susceptibility to cardiovascular disease in adulthood in a cohort study (Drake et al., 2012). Further, exposure to nicotine has been associated with increased maternal glucocorticoids and alterations in offspring HPA stress response in animal and human studies (Varvarigou et al., 2006; Xu et al., 2012a). And prenatal glucocorticoids programed an adult psychiatric disorder, namely, nicotine dependence, among daughters over a 40 year prospective study (Stroud et al., 2014). Raised maternal cortisol was

ro of

associated with maternal anxiety through karyotype analysis of maternal plasma and amniotic fluid samples, and there is a positive correlation between maternal cortisol and fetal cortisol, its IUGR as well as HPA axis in functional disorders (Sarkar et al., 2008). Besides, our lab also confirmed that prenatal xenobiotics (such as

-p

caffeine, nicotine, and ethanol) and food restriction could cause fetal rats to overexpose to maternal

glucocorticoids, and this was accompanied by IUGR and multiple organ dysplasia (Chen et al., 2007; He et

re

al., 2017b; Liang et al., 2011; Xu et al., 2012c). Moreover, recent experimental work has demonstrated that air pollutants can act as stressors and induce endocrine stress responses. Exposure of rats to concentrated ambient

lP

particles activated stress centers in the brain and increased circulating levels of corticosterone (Sirivelu et al., 2006). In utero exposure to ultrafine particles significantly increased the serum cortisol levels in the dams and

na

fetuses, also influenced the susceptibility to cardiovascular disease in adulthood (Morales-Rubio et al., 2019). In all, it may be a universal phenomenon that prenatal adverse environments induce over-exposure to maternal

ur

glucocorticoids in fetuses.

2.3 Placental glucocorticoids barrier and 11β-hydroxysteroid dehydrogenases system

Jo

The placenta is an important organ for the development of the fetus. Placental transport of maternal glucocorticoids is the main source of glucocorticoids in the fetal circulation (Murphy et al., 2006). Environmental perturbation in pregnancy can increase fetal glucocorticoid exposure by altering placental transfer of glucocorticoids. Both exogenous and endogenous glucocorticoids can cross the placental barrier by metabolism, and glucocorticoids metabolism in the placenta is associated with the expression and activity of 11β-hydroxysteroid dehydrogenases (11β-HSDs), including 11β-HSD1 and 11β-HSD2 (McNeil et al., 2007). 5

Generally, glucocorticoids are activated by 11β-HSD1 and inactivated by 11β-HSD2 (Chapman et al., 2013). 11β-HSD2 converts cortisol (in humans) or corticosterone (in rodents) into their inactive forms, cortisone or 11-dehydrocorticosterone (Yang, 1997). 11β-HSD2 is responsible for inactively metabolizing glucocorticoids by oxidation, thereby effectively regulating the level of active glucocorticoids in the cells. Therefore, 11 HSD2 is an important regulatory factor to protect the fetus from excessive maternal glucocorticoids (Salvante et al., 2017). Maternal vitamin D deficiency in a mouse model of pregnancy elevates circulating maternal glucocorticoids as well as adrenal weight. Vitamin D deficiency also reduces placental 11β-HSD2 expression

ro of

(Yates et al., 2017). Adverse environments during pregnancy can inhibit placental 11 -HSD2 expression, and open the placental glucocorticoids barrier, so the fetus is overexposed to maternal glucocorticoids (Yu et al., 2018; Zhou et al., 2018a). Further, as placental endothelial cells are glucocorticoid targets (Kipmen-Korgun et al., 2012), overexposure to maternal glucocorticoids during pregnancy may have adverse effects on placental

-p

angiogenesis mechanisms (Ozmen et al., 2017), thereby affecting oxygen and nutrient availability to support fetal development. Hence, maternal glucocorticoid excess may impact disease risk in offspring through

re

impacting placental function (Fowden et al., 2015). Eventually, this high level of glucocorticoids in utero

lP

leads to IUGR, appearing as low birth weight and multiple organ dysplasia (Fig. 1).

3 Prenatal glucocorticoids exposure-induced adrenal dysplasia

na

The nature and severity of glucocorticoids effects seem to be influenced by the stressors intervening during pregnancy. As a matter of fact, many scientific reports support the idea that fetal adrenal development

ur

is characterized by sensitive periods or developmental windows when it is more vulnerable to stressors. 3.1 Adrenal morphological development abnormality

Jo

The landmark study of human adrenal cortex development began in the 1960s. Research has basically revealed the physiological development of the adrenal gland, as shown in Table 1. However, is the adrenal development under prenatal glucocorticoids over-exposure consistent with physiological trajectories? A significant reduction in fetal adrenal volume and delayed fetal growth appeared when pregnant women were treated with sGC, and the adrenal gland of IUGR children was smaller than that of normal individuals’ (Iijima et al., 2010; Karsli et al., 2017). These results suggest that glucocorticoids over-exposure during pregnancy 6

inhibits the development of the fetal adrenal gland in human. The adrenal development of rodents is different from that of humans, but its basic process and related important genes are similar (Mazilu and McCabe, 2011), as shown in Table 1. Much research has shown that prenatal xenobiotics (such as caffeine, ethanol, nicotine) exposure and food restriction could induce the fetal over-exposure to maternal glucocorticoids, and the fetal adrenal cortex got thinner, but it was close to normal in adulthood (Chen et al., 2018; Chen et al., 2007; He et al., 2017b; Huang et al., 2015). Similarly, when the mother rats were exposed to dexamethasone during pregnancy, the volume of the fetal adrenal gland decreased, as did the zona glomerulosa (ZG), zona fasciculata (ZF), and zona reticularis (ZR), but they tended

ro of

to be normal in six months after birth (Waddell et al., 2010). Studies have found that stress during pregnancy can increase the ratio of adrenal cortex/medullary after birth in offspring rats (Liaudat et al., 2015). All these results suggest that prenatal glucocorticoids exposure inhibited the morphological development of the fetal

-p

adrenal gland, but the cortex quickly grew after birth, even close to normal in adulthood. Table 1. Adrenal physiologically morphological development. Adrenal morphogenesis

re

Species Gestational age / Age The 4th week

Emerging of AGP: characterized by SF1 expression.

(Bandiera et al., 2013)

The 8th week

New adrenocortical cells emerge between the

(Goto et al., 2006)

lP

Human

References

na

capsule and ZF, forming the definitive zone (DZ) that later develops into the adult cortex. DZ is

ur

composed of SF1-positive, densely packed basophilic cells arranged in a narrow band of cellular

Jo

clusters.

The 9th week

Mesenchymal cells of the glomerular envelope

(Ross and Louw,

encapsulate the adrenal embryonic base to form the

2015)

adrenal capsule. The 16th-20th

The fetal zone is characterized by typical steroid

(Mesiano and Jaffe,

week

hormone-secreting cells. The permanent zone is

1993)

7

characterized by typical proliferating cells. It forms a transitional zone which has the characteristics of adult adrenal ZG. The 30th week

The transition zone is in the form of an adult ZF.

(Mesiano and Jaffe, 1997)

Neonatal period

Regression of ZF: ZF cells undergo intense apoptosis. Adrenarche: ZR forms and begins to produce adrenal androgens.

(Xing et al., 2015)

ro of

Pubertal period

Homeostasis: All zones established. Stem/progenitor cells replenish cortex centripetally. The 9th day

The development of the adrenal cortex begins with

(Hatano et al., 1996)

-p

Rat

epithelial cells.

re

the migration of urogenital corpus callosum

Adrenal cortical cells begin to differentiate.

(Mitani et al., 2003)

The 15th day

The adrenal medulla forms.

(Schulte et al., 2007)

The 18th day

The undifferentiated zone does not express steroidal

(Mitani et al., 1999)

lP

The 10.5th day

na

synthase. Adrenal cortical function may be a stem cell belt.

Retention of XZ: XZ transiently activate FAdE and

(Mitani, 2014)

ur

After birth

Jo

start to express 20αHSD. Regression of XZ: XZ disappears male at puberty, female at first pregnancy.

AGP, adrenogenital primordium; SF1, steroidogenic factor 1; ZF, zona fasciculata; DZ, definitive zone; ZG, zona glomerulosa; ZR, zona reticularis; XZ, X-zone; FAdE, fetal adrenal enhancer; 20α-HSD, 20αhydroxysteroid dehydrogenase.

8

3.2 Adrenal functional development abnormality The fetal adrenal gland secretes steroid hormones from early embryonic period (McNutt and Jones, 1970). The adrenal cortex consists of three concentric zones, ZG, ZF and ZR, which secrete mineralocorticoids, glucocorticoids, and sex hormones, respectively. The adrenal gland acts as the terminal effector organ of the HPA axis, and its development is mainly regulated by local factors before birth and the HPA axis after birth. Some signaling pathways, including insulin-like growth factor 1 (IGF1), transforming growth factor β (TGFβ) and the cyclic adenosine monophosphate (cAMP) signaling pathway, also have an important regulatory role in the proliferation and differentiation of the adrenal gland (Hofland and de Jong,

ro of

2012; Pitetti et al., 2013). Moreover, some important transcriptional factors are also involved in the regulation of adrenal development, e.g., steroidogenic factor 1 (SF1), dosage-sensitive sex reversal, adrenal hypoplasia congenita, X-linked-1 (Dax 1) and Transcription factors 21 (TCF21) (Lee et al., 2011; Val and Swain, 2010;

-p

Wood et al., 2013).

Clinical cases indicated that high-dose methylprednisolone or prednisolone in a pregnant woman

re

inhibited fetal adrenal steroidogenesis (Homar et al., 2008; Kurtoglu et al., 2011). Prenatal betamethasone exposure suppressed infants' HPA axis response to a stressor, resulting in an impaired ability of the adrenal

lP

gland to produce cortisol after birth (Davis et al., 2004). Pregnant women are treated with dexamethasone to promote fetal lung maturation, but inappropriate courses and doses could cause low adrenal function in IUGR

na

neonates (Ng et al., 1999; Ng et al., 1997). Besides, IUGR neonates presented low levels of cortisol three days postnatal (Khan et al., 2011). Furthermore, when IUGR newborns grew to puberty, the ability to excrete

ur

cortisol was impaired, which resulted in adrenal insufficiency and chronic disease in adulthood (Jensen et al., 2007; Meuwese et al., 2010).

Jo

Study has shown that maternal betamethasone administration results in adrenal suppression in adult sheep (Sloboda et al., 2007). In addition, high levels of glucocorticoids induced by prenatal caffeine exposure inhibited the levels of endogenous corticosterone, aldosterone and the expression of adrenal steroidal synthases through IGF1 signaling pathway in male offspring rats (Chen et al., 2018). Meanwhile, a similar dysfunction was found in a rat model of maternal glucocorticoids over-exposure by prenatal ethanol exposure (Huang et al., 2015). These results suggest that prenatal glucocorticoids exposure inhibited adrenal 9

steroidogenesis and continued after birth.

4 Intrauterine programming mechanism of glucocorticoids-induced adrenal dysfunction Glucocorticoids are critical in the physiological development of fetuses. The effects of glucocorticoidsmediated intrauterine programming are evident in fetal development, even in adult diseases (Moisiadis and Matthews, 2014a, b). Growing evidence indicates that glucocorticoids can program adrenal development throughout life, including low-functional programming of adrenal de novo steroidogenesis, glucocorticoidsinsulin-like growth factor 1 (GC-IGF1) axis programming and epigenetic modification programming, etc (He

ro of

et al., 2017a; Ping et al., 2014; Xu et al., 2012b). If the offspring have not adapted to glucocorticoids-induced phenotypic changes, this will lead to susceptibility to chronic disease in adulthood (Fowden et al., 2016; Gluckman and Hanson, 2004; Kim et al., 2015).

-p

4.1 Low-functional programming of adrenal de novo steroidogenesis

The synthetic function of the adrenal gland is an important index to evaluate adrenal development, and

re

acute stress at a critical stage of adrenal development may determine steroid synthesis and secretion in adult life (Quinn et al., 2014). Our lab has confirmed that high levels of glucocorticoids could inhibit expression of

lP

steroidogenic enzymes in vivo and in vitro, which is associated with the activated glucocorticoids-activation system, mainly including the 11 -HSD1/11 -HSD2 ratio, glucocorticoids receptor (GR) and C/EBP (C/EBP ) expression (He et al., 2019b; Huang et al., 2015). Furthermore, the inhibited

na

homologous protein

adrenal function in offspring with prenatal caffeine exposure and a post-weaning high-fat diet may principally

ur

be the result of intrauterine programming of glucocorticoids. At the same time, the adrenal glucocorticoidsactivation system showed a significant inactivation, mainly including a decreased 11β-HSD1/11β-HSD2 ratio

Jo

and decreased mineralocorticoid receptor (MR) expression. The de novo synthesis pathway presented a lowfunctional programming alteration, and extended to a lifetime after birth, and even intergenerational effects existed (He et al., 2019b; Luo et al., 2014). The HPA axis can actively participate in physiological functions, such as regulating fetal growth and development, glucocorticoids biosynthesis, and adrenal differentiation and maturation (Wood and Walker, 2015). The activity of the HPA axis is mainly adjusted to the function of parvocellular neuroendocrine (PNC) 10

in the paraventricular nucleus (PVN) of the hypothalamus. After stress, the hypothalamic PNC secretes corticotropin-releasing hormone (CRH) and arginine vasopressin (AVP) to initiate the body’s stress response. CRH and AVP rapidly stimulate the secretion of adrenocorticotropic hormone (ACTH) in the pituitary, which further increases the secretion of glucocorticoids in the adrenal gland. Inversely, the high level of glucocorticoids can dynamically regulate the HPA axis by negative feedback, thereby maintaining the balance of adrenal function (Herman et al., 2012; Tasker and Herman, 2011). Prenatal glucocorticoids exposure tends to alter the developmental programming of the HPA axis, mainly presenting as low basic activity after birth (Charmandari et al., 2012; Glover et al., 2010). The study showed that the levels of ACTH and cortisol were

ro of

lower than baseline in the pig model of dexamethasone exposure during late pregnancy, which suggested that the function of the HPA axis is inhibited, accompanied by adrenal dysplasia (Schiffner et al., 2017). Our

laboratory also found some similar phenomenona suggesting that maternal corticosterone overexposure due to

-p

various xenobiotics (such as caffeine, nicotine, ethanol) exposure could suppress the expression of

hypothalamic CRH and AVP, adrenal steroidogenic acute regulatory protein (StAR) and cytochrome P450

re

cholesterol side chain cleavage (P450scc), as well as decrease the production of endogenous corticosterone (Chen et al., 2007; Liang et al., 2011; Xu et al., 2012b; Xu et al., 2012c). Finally, the adrenal gland is

lP

hypersensitive to acute and chronic stress and subsequently susceptible to disease. Over all, prenatal glucocorticoids exposure inhibits adrenal steroidogenesis by directedly activating

na

adrenal glucocorticoids-activation system or indirectly inhibiting function development of HPA axis in utero, which causes low basic activity of the HPA axis after birth. Eventually, it makes offspring adrenal

stress (Fig. 2).

ur

hypersensitive to postnatal adverse environments and increases susceptibility to adult disease with chronic

Jo

4.2 Developmental programming alterations of glucocorticoid-insulin-like growth 1 axis IGF1 is known to be a major endocrinal regulator of adaptive response to the internal and external

environment changes of early development (Klammt et al., 2008). IGF1 plays an important role in proliferation, differentiation, migration and survival, and even regulates tissue growth, reproduction, lifespan (Fowden, 2003). The fetal circulation of IGF1 is mainly from the fetal liver and its level is directly related to embryonic or fetal development in utero (Coppola et al., 2009). Studies have confirmed that IGF1 and its 11

receptor, IGF1R, are highly expressed in the adrenal cortex, which can regulate the expression of SF1 and steroidal synthases through the phosphorylated phosphatidylinositol-3-hydroxykinase (PI3K) / protein kinase pathway (Ito et al., 1997; Pitetti et al., 2013; Sirianni et al., 2007). Changes in fetal circulation glucocorticoids concentrations due to the intrauterine environments can cause changes in the circulation of IGF1 concentrations (Fowden, 2003). We found that prenatal xenobiotics (e.g. caffeine, ethanol and nicotine) exposure and food restriction induced high levels of serum corticosterone in fetal and newborn rats, while the levels of IGF1 in serum and multiple tissues (e.g. adrenal, liver and bone) were decreased. The level of serum corticosterone is gradually reduced after birth, but the level of IGF1 in serum and multiple tissues were

ro of

gradually increased (He et al., 2019a; He et al., 2017a; He et al., 2017b; Huang et al., 2015; Pei et al., 2019; Shangguan et al., 2018; Tan et al., 2018; Zhou et al., 2018b). Interestingly, a post-weaning high-fat diet can further decrease the level of serum corticosterone after birth and increase the level of serum IGF1 (Xu et al.,

-p

2013; Zhang et al., 2016). This negative relationship between the serum glucocorticoids and tissue IGF1

levels suggests that a high level of maternal glucocorticoids in utero can program the tissue IGF1 signaling

re

pathway, which we named “intrauterine programming of GC-IGF1 axis”. In addition, “intrauterine programming of GC-IGF1 axis” is also involved in compensatory adrenal growth after birth. “Compensatory

lP

adrenal growth” has been used to describe the phenomenon of adapting to or compensating for a physiological or pathological condition by strengthening the adrenal function, which may lead to metabolic diseases by

na

elevating glucocorticoid levels. The compensatory adrenal growth response is tightly controlled by both humoral and neuronal factors that have been elucidated in increasing detail over recent decades (Sfikakis et

ur

al., 2008).

Researchers found that high levels of serum corticosterone during pregnancy could increase 11β-HSD1

Jo

expression and decrease 11β-HSD2 expression, leading to increased 11β-HSD1/11β-HSD2 expression ratio. Meanwhile, GR expression as well as C/EBPα expression and C/EBPα / C/EBPβ expression ratios were increased, but the expression of the adrenal IGF1 signal pathway was suppressed. After birth, in the case of lower serum corticosterone induced by prenatal glucocorticoids exposure, the adrenal glucocorticoidactivation system was inhibited, and this manifested as decreased 11β-HSD1 expression and increased 11βHSD2 expression. Meanwhile, adrenal IGF1 signal pathway expression was increased. Hence, it caused 12

compensatory adrenal growth, then the level of glucocorticoids gradually increased. Thereby, it increased the susceptibilities of metabolic syndrome and related metabolic diseases. These suggest that there is a good negative correlation between the level of serum glucocorticoids and the expression of the adrenal IGF1 signaling pathway due to glucocorticoids exposure during pregnancy (He et al., 2019b). How does the glucocorticoid-activation system mediate the “GC-IGF1 axis programming alteration”? We verified that glucocorticoids could increase 11β-HSD1 and decrease 11β-HSD2 expression to enhance the local glucocorticoids concentration in adrenal glands, and the latter promoted GR expression and its nucleus translocation. After transferring into the nuclear, GR interactions with C/EBPα could inhibit the expression of

ro of

the IGF1 signaling pathway and its downstream steroidogenesis (such as StAR and P450scc), eventually leading to steroid synthesis damage of adrenocortical cells. These results suggest that the glucocorticoidactivation system might participate in the “GC-IGF1 axis programming alteration” in the adrenal of the

-p

prenatal glucocorticoids exposure offspring (He et al., 2019b) [Fig. 3].

4.3 Epigenetic modification changes and adrenal developmental programming

re

Epigenetic mechanisms mediate the influence of adverse environmental factors in early life on long-term metabolic health, which induces persistent changes in gene expression and organ functions (Tobi et al., 2018).

lP

These changes can be influenced by environment, forming a specific phenotype that may be adaptive to individual growth and development. However, they are also influenced by stress response. The effects of

na

stress may be a result of cumulative stress or a mismatch between the environment experienced early in life versus conditions much later. These effects, including associated epigenetic modifications, are potentially

ur

reversible (Buschdorf and Meaney, 2015). Therefore, the dynamic levels of glucocorticoids caused by prenatal adverse environment and postnatal chronic stress may cause epigenetic modification of genes.

Jo

Glucocorticoids can program adrenal development during the early period by epigenetic modifications, such as DNA hypermethylation and histone deacetylation (Liu et al., 2016; Yang et al., 2011). It is known that glucocorticoids can regulate downstream gene expression by binding to GR. The GR binds the genome either directly via DNA-sequence-specific interactions with a glucocorticoid response element (GRE) or, more often, indirectly via tethering to other transcription factors such as the AP-1 family (Gertz et al., 2013), and then, it can recruit related epigenetic enzymes (such as CBP/p300) (Kassel and Herrlich, 2007). Studies have found 13

that overexposure of fetal rats to maternal glucocorticoids could result in abnormal DNA methylation and histone acetylation of the SF1 promoter region, thereby inhibiting the fetal adrenal steroidogenesis (Ping et al., 2014). Recently, our lab has found that prenatal dexamethasone exposure increased the level of serum dexamethasone in fetal rats, which recruited histone deacetylase 7 (HDAC7) through GR to reduce the level of histone acetylation in the StAR promoter region, and then inhibited testicular steroid synthase in utero and this continued after birth (Liu et al., 2018). Moreover, we found similar phenomenon in the adrenal glands of the same rats. Interestingly, and this abnormal adrenal development could inherite to the F3 generation, and it

ro of

may be related to DNA methylation of the imprinted gene in germ cells (unpublished data). In addition, other research has found that prenatal betamethasone exposure induced adrenal total DNA methylation and related gene expression in guinea pigs, which could continue to adulthood or even the F2 generation (Crudo et al.,

-p

2012). These results suggest that prenatal glucocorticoids exposure can cause epigenetic modification changes of adrenal developmental genes through the GR recruitment to epigenetic enzymes, which alters the

re

intrauterine programming of adrenal development in offspring, and even affects multiple generations.

lP

5 Prenatal glucocorticoids exposure and gender differences of adrenal development The same environmental exposure during pregnancy has different effects on the occurrence, process and

na

outcome of chronic diseases in male and female offspring (van Abeelen et al., 2011; Waddell and McCarthy, 2012). The adrenal gland is an important target for glucocorticoids intrauterine programing effects, and acute

ur

stress at a critical stage of adrenal development may determine steroid synthesis and secretion in adult life. The outcome also depends on the sex of the fetus (Quinn et al., 2014). There are obvious gender differences in

Jo

adrenal function and sensitivity change in offspring induced by prenatal glucocorticoids exposure (Arnetz et al., 2015; Pinheiro et al., 2011). Our study found that there were some obvious gender differences in the glucocorticoids-activation system and IGF1 signaling pathway in the offspring, which was induced by prenatal caffeine exposure under normal diet and post-weaning high-fat diet conditions, especially in females, and there was even disorder after chronic stress (He et al., 2017a). Glucocorticoids secreted by the adrenal gland are an important metabolic regulator. Metabolic syndrome is a typical disease caused by metabolic 14

abnormality. It comprises of obesity, hypertension, dyslipidemia, and insulin resistance, which shares many similar characteristics with Cushing’s syndrome, caused by excess systemic glucocorticoids, thereby glucocorticoids effects might play a role in the development of metabolic syndrome (Goodwin and Geller, 2012). Moreover, a large number of studies indicate that there are significant gender differences in many metabolic diseases, such as nonalcoholic fatty liver, cardiovascular diseases, hypertension, etc. (Ayonrinde et al., 2011; Dasinger and Alexander, 2016; O'Regan et al., 2004; Ojeda et al., 2014). It suggests that the gender differences in glucocorticoids levels caused by adrenal dysplasia mediates the different risk of metabolic

ro of

diseases in males and females.

6 Conclusions and prospects

In conclusion, prenatal glucocorticoids exposure can cause abnormal morphological and functional

-p

development of the adrenal gland. This organ is the synthetic and secretory organ of glucocorticoids, and adrenal dysplasia in utero causes potential or persistent harm to individual development, including the

re

occurrence of IUGR and susceptibility to chronic diseases in adulthood. The adrenal gland acts as a core organ of the susceptibility to fetal-originated adult diseases, and we summarize two intrauterine programming

lP

mechanisms of fetal adult-originated diseases (Fig. 4). On the one hand, prenatal glucocorticoids exposure inhibits fetal adrenal de novo steroidogenesis and the basic activity of the HPA axis, leading to lasting low

na

steroidogenesis and adrenal hypersensitivity after birth, which may mediate the offspring’s susceptibility to chronic diseases in adulthood. On the other hand, the glucocorticoid-activation system inhibits fetal adrenal

ur

development in utero and caused compensatory adrenal growth after birth through the intrauterine programming alteration of GC-IGF1 axis. Eventually, the postnatal morphological and functional

Jo

compensations of the adrenal gland increased the offspring’s susceptibility to chronic diseases in adulthood, and there are also appears obvious gender differences. Among them, epigenetic modification mediates adrenal developmental changes from intrauterine to postnatal days, even in multiple generations. Therefore, we can target the epigenetic markers of fetal adrenal developmental toxicity, and carry out evaluation and screen for adverse environmental factors. Furthermore, based on the intrauterine programming mechanisms of adrenal dysplasia caused by maternal glucocorticoids over-exposure, we can explore early-warning techniques and 15

improve prevention strategies.

Conflict of interest The authors declare that there is no conflict of interest.

ro of

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 81673524, 81701463, 81430089), the National Key Research and Development Program of China (2017YFC1001300)

Jo

ur

na

lP

re

-p

and Hubei Province Health and Family Planning Scientific Research Project (No. WJ2017C0003).

16

References Arnetz, L., Rajamand Ekberg, N., Brismar, K. and Alvarsson, M., 2015. Gender difference in adrenal sensitivity to ACTH is abolished in type 2 diabetes. Endocr Connect. 4, 92-99. Ayonrinde, O.T., Olynyk, J.K., Beilin, L.J., Mori, T.A., Pennell, C.E., de Klerk, N., Oddy, W.H., Shipman, P. and Adams, L.A., 2011. Gender-specific differences in adipose distribution and adipocytokines influence adolescent nonalcoholic fatty liver disease. Hepatology. 53, 800-809. Bandiera, R., Vidal, V.P., Motamedi, F.J., Clarkson, M., Sahut-Barnola, I., von Gise, A., Pu, W.T., Hohenstein, P., Martinez, A. and Schedl, A., 2013. WT1 maintains adrenal-gonadal primordium identity and marks a

ro of

population of AGP-like progenitors within the adrenal gland. Dev. Cell. 27, 5-18.

Bremer, A.A., 2010. Mechanism of a Metabolic Shift in Monkeys. Sci. Transl. Med. 2, 14ec14-14ec14.

Busada, J.T. and Cidlowski, J.A., 2017. Mechanisms of Glucocorticoid Action During Development. Curr.

-p

Top. Dev. Biol. 125, 147-170.

Buschdorf, J.P. and Meaney, M.J., 2015. Epigenetics/Programming in the HPA Axis. Comprehensive

re

Physiology. 6, 87-110.

Buss, C., Entringer, S. and Wadhwa, P.D., 2012. Fetal Programming of Brain Development: Intrauterine Stress

lP

and Susceptibility to Psychopathology. Science Signaling. 5, pt7-pt7.

Chapman, K., Holmes, M. and Seckl, J., 2013. 11beta-hydroxysteroid dehydrogenases: intracellular gate-

na

keepers of tissue glucocorticoid action. Physiol. Rev. 93, 1139-1206. Charmandari, E., Achermann, J.C., Carel, J.-C., Soder, O. and Chrousos, G.P., 2012. Stress Response and

ur

Child Health. Science Signaling. 5, mr1-mr1.

Chen, G., Yuan, C., Duan, F., Liu, Y., Zhang, J., He, Z., Huang, H., He, C. and Wang, H., 2018.

Jo

IGF1/MAPK/ERK signaling pathway-mediated programming alterations of adrenal cortex cell proliferation by prenatal caffeine exposure in male offspring rats. Toxicol. Appl. Pharmacol. 341, 64-76. Chen, M., Wang, T., Liao, Z.X., Pan, X.L., Feng, Y.H. and Wang, H., 2007. Nicotine-induced prenatal overexposure to maternal glucocorticoid and intrauterine growth retardation in rat. Exp. Toxicol. Pathol. 59, 245-251. Coppola, D., Ouban, A. and Gilbert-Barness, E., 2009. Expression of the insulin-like growth factor receptor 1 17

during human embryogenesis. Fetal Pediatr. Pathol. 28, 47-54. Correia-Branco, A., Keating, E. and Martel, F., 2015. Maternal undernutrition and fetal developmental programming of obesity: the glucocorticoid connection. Reprod. Sci. 22, 138-145. Crudo, A., Petropoulos, S., Moisiadis, V.G., Iqbal, M., Kostaki, A., Machnes, Z., Szyf, M. and Matthews, S.G., 2012. Prenatal synthetic glucocorticoid treatment changes DNA methylation states in male organ systems: multigenerational effects. Endocrinology. 153, 3269-3283. Cuffe, J.S., Walton, S.L., Singh, R.R., Spiers, J.G., Bielefeldt-Ohmann, H., Wilkinson, L., Little, M.H. and Moritz, K.M., 2014. Mid- to late term hypoxia in the mouse alters placental morphology, glucocorticoid

ro of

regulatory pathways and nutrient transporters in a sex-specific manner. J. Physiol. 592, 3127-3141. Dasinger, J.H. and Alexander, B.T., 2016. Gender differences in developmental programming of cardiovascular diseases. Clinical science. 130, 337-348.

-p

Davis, E.P., Townsend, E.L., Gunnar, M.R., Georgieff, M.K., Guiang, S.F., Ciffuentes, R.F. and Lussky, R.C.,

infants. Psychoneuroendocrinology. 29, 1028-1036.

re

2004. Effects of prenatal betamethasone exposure on regulation of stress physiology in healthy premature

Dodic, M., May, C.N., Wintour, E.M. and Coghlan, J.P., 1998. An early prenatal exposure to excess

lP

glucocorticoid leads to hypertensive offspring in sheep. Clinical science. 94, 149-155. Drake, A.J., McPherson, R.C., Godfrey, K.M., Cooper, C., Lillycrop, K.A., Hanson, M.A., Meehan, R.R.,

na

Seckl, J.R. and Reynolds, R.M., 2012. An unbalanced maternal diet in pregnancy associates with offspring epigenetic changes in genes controlling glucocorticoid action and foetal growth. Clin. Endocrinol. (Oxf.). 77,

ur

808-815.

Fleming, T.P., Watkins, A.J., Velazquez, M.A., Mathers, J.C., Prentice, A.M., Stephenson, J., Barker, M.,

Jo

Saffery, R., Yajnik, C.S., Eckert, J.J., Hanson, M.A., Forrester, T., Gluckman, P.D. and Godfrey, K.M., 2018. Origins of lifetime health around the time of conception: causes and consequences. Lancet. 391, 1842-1852. Fowden, A.L., 2003. The insulin-like growth factors and feto-placental growth. Placenta. 24, 803-812. Fowden, A.L., Forhead, A.J., Sferruzzi-Perri, A.N., Burton, G.J. and Vaughan, O.R., 2015. Review: Endocrine regulation of placental phenotype. Placenta. 36 Suppl 1, S50-59. Fowden, A.L., Valenzuela, O.A., Vaughan, O.R., Jellyman, J.K. and Forhead, A.J., 2016. Glucocorticoid 18

programming of intrauterine development. Domest. Anim. Endocrinol. 56 Suppl, S121-132. Gertz, J., Savic, D., Varley, K.E., Partridge, E.C., Safi, A., Jain, P., Cooper, G.M., Reddy, T.E., Crawford, G.E. and Myers, R.M., 2013. Distinct properties of cell-type-specific and shared transcription factor binding sites. Mol. Cell. 52, 25-36. Glover, V., O'Connor, T.G. and O'Donnell, K., 2010. Prenatal stress and the programming of the HPA axis. Neuroscience and biobehavioral reviews. 35, 17-22. Gluckman, P.D. and Hanson, M.A., 2004. Living with the Past: Evolution, Development, and Patterns of Disease. Science. 305, 1733-1736.

ro of

Goodwin, J.E. and Geller, D.S., 2012. Glucocorticoid-induced hypertension. Pediatr. Nephrol. 27, 1059-1066. Goto, M., Piper Hanley, K., Marcos, J., Wood, P.J., Wright, S., Postle, A.D., Cameron, I.T., Mason, J.I., Wilson, D.I. and Hanley, N.A., 2006. In humans, early cortisol biosynthesis provides a mechanism to

-p

safeguard female sexual development. J. Clin. Invest. 116, 953-960.

Hatano, O., Takakusu, A., Nomura, M. and Morohashi, K., 1996. Identical origin of adrenal cortex and gonad

re

revealed by expression profiles of Ad4BP/SF-1. Genes Cells. 1, 663-671.

He, H., Xiong, Y., Li, B., Zhu, Y., Chen, H., Ao, Y. and Wang, H., 2019a. Intrauterine programming of the

lP

glucocorticoid-insulin-like growth factor 1 (GC-IGF1) axis mediates glomerulosclerosis in female adult offspring rats induced by prenatal ethanol exposure. Toxicol. Lett. 311, 17-26.

na

He, Z., Lv, F., Ding, Y., Huang, H., Liu, L., Zhu, C., Lei, Y., Zhang, L., Si, C. and Wang, H., 2017a. High-fat diet and chronic stress aggravate adrenal function abnormality induced by prenatal caffeine exposure in male

ur

offspring rats. Sci. Rep. 7, 14825.

He, Z., Lv, F., Ding, Y., Zhu, C., Huang, H., Zhang, L., Guo, Y. and Wang, H., 2017b. Insulin-like Growth

Jo

Factor 1 Mediates Adrenal Development Dysfunction in Offspring Rats Induced by Prenatal Food Restriction. Arch. Med. Res. 48, 488-497. He, Z., Zhang, J., Huang, H., Yuan, C., Zhu, C., Magdalou, J. and Wang, H., 2019b. Glucocorticoid-activation system mediated glucocorticoid-insulin-like growth factor 1 (GC-IGF1) axis programming alteration of adrenal dysfunction induced by prenatal caffeine exposure. Toxicol. Lett. 302, 7-17. Herman, J.P., McKlveen, J.M., Solomon, M.B., Carvalho-Netto, E. and Myers, B., 2012. Neural regulation of 19

the stress response: glucocorticoid feedback mechanisms. Braz. J. Med. Biol. Res. 45, 292-298. Hofland, J. and de Jong, F.H., 2012. Inhibins and activins: their roles in the adrenal gland and the development of adrenocortical tumors. Mol. Cell. Endocrinol. 359, 92-100. Homar, V., Grosek, S. and Battelino, T., 2008. High-dose methylprednisolone in a pregnant woman with Crohn's disease and adrenal suppression in her newborn. Neonatology. 94, 306-309. Huang, H., He, Z., Zhu, C., Liu, L., Kou, H., Shen, L. and Wang, H., 2015. Prenatal ethanol exposure-induced adrenal developmental abnormality of male offspring rats and its possible intrauterine programming mechanisms. Toxicology and applied pharmacology. 288, 84-94.

ro of

Iijima, S., Uga, N. and Ohzeki, T., 2010. Postnatal changes in adrenal size in very low-birth-weight infants: sonographic evaluation for the prediction of late-onset glucocorticoid-responsive circulatory collapse. American journal of perinatology. 27, 485-491.

-p

Ishimoto, H. and Jaffe, R.B., 2011. Development and function of the human fetal adrenal cortex: a key component in the feto-placental unit. Endocr. Rev. 32, 317-355.

re

Ito, M., Yu, R. and Jameson, J.L., 1997. DAX-1 inhibits SF-1-mediated transactivation via a carboxy-terminal domain that is deleted in adrenal hypoplasia congenita. Molecular and cellular biology. 17, 1476-1483.

lP

Jensen, R.B., Vielwerth, S., Larsen, T., Greisen, G., Veldhuis, J. and Juul, A., 2007. Pituitary-gonadal function in adolescent males born appropriate or small for gestational age with or without intrauterine growth

na

restriction. J. Clin. Endocrinol. Metab. 92, 1353-1357.

Karsli, T., Strickland, D., Livingston, J., Wu, Q., Mhanna, M.J. and Shekhawat, P.S., 2017. Assessment of

ur

neonatal adrenal size using high resolution 2D ultrasound and its correlation with birth demographics and clinical outcomes. 1-7.

Jo

Kassel, O. and Herrlich, P., 2007. Crosstalk between the glucocorticoid receptor and other transcription factors: molecular aspects. Molecular and cellular endocrinology. 275, 13-29. Khan, A.A., Rodriguez, A., Kaakinen, M., Pouta, A., Hartikainen, A.L. and Jarvelin, M.R., 2011. Does in utero exposure to synthetic glucocorticoids influence birthweight, head circumference and birth length? A systematic review of current evidence in humans. Paediatric and perinatal epidemiology. 25, 20-36. Kim, D.R., Bale, T.L. and Epperson, C.N., 2015. Prenatal programming of mental illness: current 20

understanding of relationship and mechanisms. Curr Psychiatry Rep. 17, 5. Kipmen-Korgun, D., Ozmen, A., Unek, G., Simsek, M., Demir, R. and Korgun, E.T., 2012. Triamcinolone upregulates GLUT 1 and GLUT 3 expression in cultured human placental endothelial cells. Cell Biochem. Funct. 30, 47-53. Klammt, J., Pfaffle, R., Werner, H. and Kiess, W., 2008. IGF signaling defects as causes of growth failure and IUGR. Trends Endocrinol. Metab. 19, 197-205. Kurtoglu, S., Sarici, D., Akin, M.A., Daar, G., Korkmaz, L. and Memur, S., 2011. Fetal adrenal suppression due to maternal corticosteroid use: case report. Journal of clinical research in pediatric endocrinology. 3, 160-

ro of

162.

Lahti, M., Eriksson, J.G., Heinonen, K., Kajantie, E., Lahti, J., Wahlbeck, K., Tuovinen, S., Pesonen, A.K., Mikkonen, M., Osmond, C., Barker, D.J. and Raikkonen, K., 2015. Late preterm birth, post-term birth, and

-p

abnormal fetal growth as risk factors for severe mental disorders from early to late adulthood. Psychol. Med. 45, 985-999.

re

Lee, F.Y., Faivre, E.J., Suzawa, M., Lontok, E., Ebert, D., Cai, F., Belsham, D.D. and Ingraham, H.A., 2011. Eliminating SF-1 (NR5A1) sumoylation in vivo results in ectopic hedgehog signaling and disruption of

lP

endocrine development. Dev. Cell. 21, 315-327.

Lesage, J., Blondeau, B., Grino, M., Breant, B. and Dupouy, J.P., 2001. Maternal undernutrition during late

na

gestation induces fetal overexposure to glucocorticoids and intrauterine growth retardation, and disturbs the hypothalamo-pituitary adrenal axis in the newborn rat. Endocrinology. 142, 1692-1702.

ur

Liang, G., Chen, M., Pan, X.L., Zheng, J. and Wang, H., 2011. Ethanol-induced inhibition of fetal hypothalamic-pituitary-adrenal axis due to prenatal overexposure to maternal glucocorticoid in mice. Exp.

Jo

Toxicol. Pathol. 63, 607-611.

Liaudat, A.C., Rodriguez, N., Chen, S., Romanini, M.C., Vivas, A., Rolando, A., Gauna, H. and Mayer, N., 2015. Adrenal response of male rats exposed to prenatal stress and early postnatal stimulation. Biotech. Histochem. 90, 432-438. Liu, L., Wang, J.F., Fan, J., Rao, Y.S., Liu, F., Yan, Y.E. and Wang, H., 2016. Nicotine Suppressed Fetal Adrenal StAR Expression via YY1 Mediated-Histone Deacetylation Modification Mechanism. International 21

journal of molecular sciences. 17. Liu, M., Chen, B., Pei, L., Zhang, Q., Zou, Y., Xiao, H., Zhou, J., Chen, L. and Wang, H., 2018. Decreased H3K9ac level of StAR mediated testicular dysplasia induced by prenatal dexamethasone exposure in male offspring rats. Toxicology. 408, 1-10. Long, N.M., Shasa, D.R., Ford, S.P. and Nathanielsz, P.W., 2012. Growth and insulin dynamics in two generations of female offspring of mothers receiving a single course of synthetic glucocorticoids. American journal of obstetrics and gynecology. 207, 203 e201-208. Luo, H., Deng, Z., Liu, L., Shen, L., Kou, H., He, Z., Ping, J., Xu, D., Ma, L., Chen, L. and Wang, H., 2014.

second generation rats. Toxicol. Appl. Pharmacol. 274, 383-392.

ro of

Prenatal caffeine ingestion induces transgenerational neuroendocrine metabolic programming alteration in

Mazilu, J.K. and McCabe, E.R., 2011. Moving toward personalized cell-based interventions for adrenal

proteins. Molecular genetics and metabolism. 104, 72-79.

-p

cortical disorders: part 1--Adrenal development and function, and roles of transcription factors and signaling

re

McNeil, C.J., Nwagwu, M.O., Finch, A.M., Page, K.R., Thain, A., McArdle, H.J. and Ashworth, C.J., 2007. Glucocorticoid exposure and tissue gene expression of 11beta HSD-1, 11beta HSD-2, and glucocorticoid

lP

receptor in a porcine model of differential fetal growth. Reproduction. 133, 653-661. McNutt, N.S. and Jones, A.L., 1970. Observations on the ultrastructure of cytodifferentiation in the human

na

fetal adrenal cortex. Laboratory investigation; a journal of technical methods and pathology. 22, 513-527. Mesiano, S. and Jaffe, R.B., 1993. Interaction of insulin-like growth factor-II and estradiol directs

ur

steroidogenesis in the human fetal adrenal toward dehydroepiandrosterone sulfate production. The Journal of clinical endocrinology and metabolism. 77, 754-758.

Jo

Mesiano, S. and Jaffe, R.B., 1997. Developmental and functional biology of the primate fetal adrenal cortex. Endocrine reviews. 18, 378-403. Meuwese, C.L., Euser, A.M., Ballieux, B.E., van Vliet, H.A., Finken, M.J., Walther, F.J., Dekker, F.W. and Wit, J.M., 2010. Growth-restricted preterm newborns are predisposed to functional adrenal hyperandrogenism in adult life. European journal of endocrinology. 163, 681-689. Mitani, F., 2014. Functional zonation of the rat adrenal cortex: the development and maintenance. Proc. Jpn. 22

Acad. Ser. B Phys. Biol. Sci. 90, 163-183. Mitani, F., Mukai, K., Miyamoto, H., Suematsu, M. and Ishimura, Y., 1999. Development of functional zonation in the rat adrenal cortex. Endocrinology. 140, 3342-3353. Mitani, F., Mukai, K., Miyamoto, H., Suematsu, M. and Ishimura, Y., 2003. The undifferentiated cell zone is a stem cell zone in adult rat adrenal cortex. Biochim. Biophys. Acta. 1619, 317-324. Moisiadis, V.G. and Matthews, S.G., 2014a. Glucocorticoids and fetal programming part 1: Outcomes. Nat. Rev. Endocrinol. 10, 391-402. Moisiadis, V.G. and Matthews, S.G., 2014b. Glucocorticoids and fetal programming part 2: Mechanisms. Nat.

ro of

Rev. Endocrinol. 10, 403-411.

Morales-Rubio, R.A., Alvarado-Cruz, I., Manzano-Leon, N., Andrade-Oliva, M.D., Uribe-Ramirez, M., Quintanilla-Vega, B., Osornio-Vargas, A. and De Vizcaya-Ruiz, A., 2019. In utero exposure to ultrafine

offspring results in altered blood pressure in adult mice.

-p

particles promotes placental stress-induced programming of renin-angiotensin system-related elements in the 16, 7.

re

Murphy, V.E., Smith, R., Giles, W.B. and Clifton, V.L., 2006. Endocrine regulation of human fetal growth: the role of the mother, placenta, and fetus. Endocr. Rev. 27, 141-169.

lP

Ng, P.C., Wong, G.W., Lam, C.W., Lee, C.H., Fok, T.F., Wong, M.Y. and Ma, K.C., 1999. Effect of multiple courses of antenatal corticosteroids on pituitary-adrenal function in preterm infants. Arch. Dis. Child. Fetal

na

Neonatal Ed. 80, F213-216.

Ng, P.C., Wong, G.W., Lam, C.W., Lee, C.H., Wong, M.Y., Fok, T.F., Wong, W. and Chan, D.C., 1997.

ur

Pituitary-adrenal response in preterm very low birth weight infants after treatment with antenatal corticosteroids. J. Clin. Endocrinol. Metab. 82, 3548-3552.

Jo

O'Regan, D., Kenyon, C.J., Seckl, J.R. and Holmes, M.C., 2004. Glucocorticoid exposure in late gestation in the rat permanently programs gender-specific differences in adult cardiovascular and metabolic physiology. Am. J. Physiol. Endocrinol. Metab. 287, E863-870. Ojeda, N.B., Intapad, S. and Alexander, B.T., 2014. Sex differences in the developmental programming of hypertension. Acta physiologica. 210, 307-316. Ozmen, A., Unek, G. and Korgun, E.T., 2017. Effect of glucocorticoids on mechanisms of placental 23

angiogenesis. Placenta. 52, 41-48. Pattanittum, P., Ewens, M.R., Laopaiboon, M., Lumbiganon, P., McDonald, S.J. and Crowther, C.A., 2008. Use of antenatal corticosteroids prior to preterm birth in four South East Asian countries within the SEAORCHID project. BMC pregnancy and childbirth. 8, 47. Pawluski, J.L., Brain, U.M., Underhill, C.M., Hammond, G.L. and Oberlander, T.F., 2012. Prenatal SSRI exposure alters neonatal corticosteroid binding globulin, infant cortisol levels, and emerging HPA function. Psychoneuroendocrinology. 37, 1019-1028. Pei, L.G., Zhang, Q., Yuan, C., Liu, M., Zou, Y.F., Lv, F., Luo, D.J., Zhong, S. and Wang, H., 2019. The GC-

ro of

IGF1 axis-mediated testicular dysplasia caused by prenatal caffeine exposure. J. Endocrinol. 242, M17-m32. Ping, J., Wang, J.F., Liu, L., Yan, Y.E., Liu, F., Lei, Y.Y. and Wang, H., 2014. Prenatal caffeine ingestion

induces aberrant DNA methylation and histone acetylation of steroidogenic factor 1 and inhibits fetal adrenal

-p

steroidogenesis. Toxicology. 321, 53-61.

Pinheiro, C.R., Oliveira, E., Trevenzoli, I.H., Manhaes, A.C., Santos-Silva, A.P., Younes-Rapozo, V., Claudio-

re

Neto, S., Santana, A.C., Nascimento-Saba, C.C., Moura, E.G. and Lisboa, P.C., 2011. Developmental plasticity in adrenal function and leptin production primed by nicotine exposure during lactation: gender

lP

differences in rats. Horm. Metab. Res. 43, 693-701.

Pitetti, J.L., Calvel, P., Romero, Y., Conne, B., Truong, V., Papaioannou, M.D., Schaad, O., Docquier, M.,

na

Herrera, P.L., Wilhelm, D. and Nef, S., 2013. Insulin and IGF1 receptors are essential for XX and XY gonadal differentiation and adrenal development in mice. PLoS Genet. 9, e1003160.

ur

Practice, A.C.o.O., 2011. ACOG Committee Opinion No. 475: antenatal corticosteroid therapy for fetal maturation. Obstetrics and gynecology. 117, 422-424.

Jo

Quinn, T.A., Ratnayake, U., Castillo-Melendez, M., Moritz, K.M., Dickinson, H. and Walker, D.W., 2014. Adrenal steroidogenesis following prenatal dexamethasone exposure in the spiny mouse. J. Endocrinol. 221, 347-362.

Reynolds, R.M., 2013. Glucocorticoid excess and the developmental origins of disease: two decades of testing the hypothesis--2012 Curt Richter Award Winner. Psychoneuroendocrinology. 38, 1-11. Roberts, D., Brown, J., Medley, N. and Dalziel, S.R., 2017. Antenatal corticosteroids for accelerating fetal 24

lung maturation for women at risk of preterm birth. The Cochrane database of systematic reviews. 3, Cd004454. Ross, I.L. and Louw, G.J., 2015. Embryological and molecular development of the adrenal glands. Clin. Anat. 28, 235-242. Salvante, K.G., Milano, K., Kliman, H.J. and Nepomnaschy, P.A., 2017. Placental 11 beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) expression very early during human pregnancy. J. Dev. Orig. Health Dis. 8, 149-154. Sarkar, P., Bergman, K., O'Connor, T.G. and Glover, V., 2008. Maternal antenatal anxiety and amniotic fluid

ro of

cortisol and testosterone: possible implications for foetal programming. J. Neuroendocrinol. 20, 489-496. Schiffner, R., Rodriguez-Gonzalez, G.L., Rakers, F. and Nistor, M., 2017. Effects of Late Gestational Fetal Exposure to Dexamethasone Administration on the Postnatal Hypothalamus-Pituitary-Adrenal Axis Response 18.

-p

to Hypoglycemia in Pigs.

Schulte, D.M., Shapiro, I., Reincke, M. and Beuschlein, F., 2007. Expression and spatio-temporal distribution

re

of differentiation and proliferation markers during mouse adrenal development. Gene expression patterns : GEP. 7, 72-81.

lP

Sfikakis, A., Pitychoutis, P.M., Antoniou, K., Bikas, N. and Papadopoulou-Daifoti, Z., 2008. The impact of the hormonal milieu and stress-associated hypothalamic dopaminergic activation on the rapid compensatory

na

adrenal growth in cycling female rats. Hormones (Athens, Greece). 7, 303-312. Shangguan, Y., Wen, Y., Tan, Y., Qin, J., Jiang, H., Magdalou, J., Chen, L. and Wang, H., 2018. Intrauterine

ur

Programming of Glucocorticoid-Insulin-Like Growth Factor-1 Axis-Mediated Developmental Origin of Osteoporosis Susceptibility in Female Offspring Rats with Prenatal Caffeine Exposure. Am. J. Pathol. 188,

Jo

2863-2876.

Sirianni, R., Chimento, A., Malivindi, R., Mazzitelli, I., Ando, S. and Pezzi, V., 2007. Insulin-like growth factor-I, regulating aromatase expression through steroidogenic factor 1, supports estrogen-dependent tumor Leydig cell proliferation. Cancer Res. 67, 8368-8377. Sirivelu, M.P., MohanKumar, S.M., Wagner, J.G., Harkema, J.R. and MohanKumar, P.S., 2006. Activation of the stress axis and neurochemical alterations in specific brain areas by concentrated ambient particle exposure 25

with concomitant allergic airway disease. Environ. Health Perspect. 114, 870-874. Sloboda, D.M., Moss, T.J., Li, S., Doherty, D., Nitsos, I., Challis, J.R. and Newnham, J.P., 2007. Prenatal betamethasone exposure results in pituitary-adrenal hyporesponsiveness in adult sheep. Am. J. Physiol. Endocrinol. Metab. 292, E61-70. Stroud, L.R., Papandonatos, G.D., Shenassa, E., Rodriguez, D., Niaura, R., LeWinn, K.Z., Lipsitt, L.P. and Buka, S.L., 2014. Prenatal glucocorticoids and maternal smoking during pregnancy independently program adult nicotine dependence in daughters: a 40-year prospective study. Biol. Psychiatry. 75, 47-55. Tan, Y., Lu, K., Li, J., Ni, Q., Zhao, Z., Magdalou, J., Chen, L. and Wang, H., 2018. Prenatal caffeine

of cartilage IGF-1 with histone acetylation. Toxicol. Lett. 295, 229-236.

ro of

exprosure increases adult female offspring rat's susceptibility to osteoarthritis via low-functional programming

Tasker, J.G. and Herman, J.P., 2011. Mechanisms of rapid glucocorticoid feedback inhibition of the

-p

hypothalamic-pituitary-adrenal axis. Stress. 14, 398-406.

Tobi, E.W., Slieker, R.C., Luijk, R., Dekkers, K.F., Stein, A.D., Xu, K.M., Slagboom, P.E., van Zwet, E.W.,

re

Lumey, L.H. and Heijmans, B.T., 2018. DNA methylation as a mediator of the association between prenatal adversity and risk factors for metabolic disease in adulthood. Science Advances. 4.

lP

Val, P. and Swain, A., 2010. Gene dosage effects and transcriptional regulation of early mammalian adrenal cortex development. Molecular and cellular endocrinology. 323, 105-114.

na

van Abeelen, A.F., de Rooij, S.R., Osmond, C., Painter, R.C., Veenendaal, M.V., Bossuyt, P.M., Elias, S.G., Grobbee, D.E., van der Schouw, Y.T., Barker, D.J. and Roseboom, T.J., 2011. The sex-specific effects of

ur

famine on the association between placental size and later hypertension. Placenta. 32, 694-698. Varvarigou, A.A., Petsali, M., Vassilakos, P. and Beratis, N.G., 2006. Increased cortisol concentrations in the

Jo

cord blood of newborns whose mothers smoked during pregnancy. J. Perinat. Med. 34, 466-470. Vogel, J.P., Souza, J.P., Gulmezoglu, A.M., Mori, R., Lumbiganon, P., Qureshi, Z., Carroli, G., Laopaiboon, M., Fawole, B., Ganchimeg, T., Zhang, J., Torloni, M.R., Bohren, M. and Temmerman, M., 2014. Use of antenatal corticosteroids and tocolytic drugs in preterm births in 29 countries: an analysis of the WHO Multicountry Survey on Maternal and Newborn Health. Lancet. 384, 1869-1877. Waddell, B.J., Bollen, M., Wyrwoll, C.S., Mori, T.A. and Mark, P.J., 2010. Developmental programming of 26

adult adrenal structure and steroidogenesis: effects of fetal glucocorticoid excess and postnatal dietary omega3 fatty acids. J. Endocrinol. 205, 171-178. Waddell, J. and McCarthy, M.M., 2012. Sexual differentiation of the brain and ADHD: what is a sex difference in prevalence telling us? Current topics in behavioral neurosciences. 9, 341-360. Wood, C.E. and Walker, C.D., 2015. Fetal and Neonatal HPA Axis. Comprehensive Physiology. 6, 33-62. Wood, M.A., Acharya, A., Finco, I., Swonger, J.M., Elston, M.J., Tallquist, M.D. and Hammer, G.D., 2013. Fetal adrenal capsular cells serve as progenitor cells for steroidogenic and stromal adrenocortical cell lineages in M. musculus. Development. 140, 4522-4532.

ro of

Xing, Y., Lerario, A.M., Rainey, W. and Hammer, G.D., 2015. Development of adrenal cortex zonation. Endocrinol. Metab. Clin. North Am. 44, 243-274.

Xu, D., Liang, G., Yan, Y.E., He, W.W., Liu, Y.S., Chen, L.B., Magdalou, J. and Wang, H., 2012a. Nicotine-

-p

induced over-exposure to maternal glucocorticoid and activated glucocorticoid metabolism causes

hypothalamic-pituitary-adrenal axis-associated neuroendocrine metabolic alterations in fetal rats. Toxicol.

re

Lett. 209, 282-290.

Xu, D., Wu, Y., Liu, F., Liu, Y.S., Shen, L., Lei, Y.Y., Liu, J., Ping, J., Qin, J., Zhang, C., Chen, L.B.,

lP

Magdalou, J. and Wang, H., 2012b. A hypothalamic-pituitary-adrenal axis-associated neuroendocrine metabolic programmed alteration in offspring rats of IUGR induced by prenatal caffeine ingestion. Toxicol.

na

Appl. Pharmacol. 264, 395-403.

Xu, D., Xia, L.P., Shen, L., Lei, Y.Y., Liu, L., Zhang, L., Magdalou, J. and Wang, H., 2013. Prenatal nicotine

ur

exposure enhances the susceptibility to metabolic syndrome in adult offspring rats fed high-fat diet via alteration of HPA axis-associated neuroendocrine metabolic programming. Acta Pharmacol. Sin. 34, 1526-

Jo

1534.

Xu, D., Zhang, B., Liang, G., Ping, J., Kou, H., Li, X., Xiong, J., Hu, D., Chen, L., Magdalou, J. and Wang, H., 2012c. Caffeine-induced activated glucocorticoid metabolism in the hippocampus causes hypothalamicpituitary-adrenal axis inhibition in fetal rats. PloS one. 7, e44497. Yang, G., Zou, L.P., Wang, J. and Ding, Y.X., 2011. Epigenetic regulation of glucocorticoid receptor and infantile spasms. Medical hypotheses. 76, 187-189. 27

Yang, K., 1997. Placental 11 beta-hydroxysteroid dehydrogenase: barrier to maternal glucocorticoids. Rev. Reprod. 2, 129-132. Yates, N., Crew, R.C. and Wyrwoll, C.S., 2017. Vitamin D deficiency and impaired placental function: potential regulation by glucocorticoids? Reproduction (Cambridge, England). 153, R163-r171. Yu, L., Zhou, J., Zhang, G., Huang, W., Pei, L., Lv, F., Zhang, Y., Zhang, W. and Wang, H., 2018. cAMP/PKA/EGR1 signaling mediates the molecular mechanism of ethanol-induced inhibition of placental 11beta-HSD2 expression. Toxicol. Appl. Pharmacol. 352, 77-86. Zhang, L., Shen, L., Xu, D., Wang, L., Guo, Y., Liu, Z., Liu, Y., Liu, L., Magdalou, J., Chen, L. and Wang, H.,

liver disease is intrauterine programmed. Reprod. Toxicol. 65, 236-247.

ro of

2016. Increased susceptibility of prenatal food restricted offspring to high-fat diet-induced nonalcoholic fatty

Zhou, J., Liu, F., Yu, L., Xu, D., Li, B., Zhang, G., Huang, W., Li, L., Zhang, Y., Zhang, W. and Wang, H.,

-p

2018a. nAChRs-ERK1/2-Egr-1 signaling participates in the developmental toxicity of nicotine by epigenetically down-regulating placental 11beta-HSD2. Toxicol. Appl. Pharmacol. 344, 1-12.

re

Zhou, J., Zhu, C., Luo, H., Shen, L., Gong, J., Wu, Y., Magdalou, J., Chen, L., Guo, Y. and Wang, H., 2018b. Two intrauterine programming mechanisms of adult hypercholesterolemia induced by prenatal nicotine

Jo

ur

na

lP

exposure in male offspring rats. FASEB J., fj201800172R.

28

Figure legends Fig. 1. Prenatal glucocorticoids exposure altered 11 -HSD2 activity and opened glucocorticoid barrier in placenta. GCs, glucocorticoids; 11β-HSD1, 11β-hydroxysteroid dehydrogenases 1; 11β-HSD2, 11β-

na

lP

re

-p

ro of

hydroxysteroid dehydrogenases 2.

ur

Fig. 2. Prenatal maternal glucocorticoids over-exposure induces developmental programming alterations of hypothalamic-pituitary-adrenal (HPA) axis. CRH, corticotropin-releasing hormone; AVP,

Jo

arginine vasopressin; ATCH, adrenocorticotropic hormone.

29

ro of -p

re

Fig. 3. Prenatal maternal glucocorticoids over-exposure induces an intrauterine programming

Jo

ur

na

lP

alteration of in GC-IGF1 axis. GC, glucocorticoid; IGF1, insulin-like growth factor 1.

30

Fig. 4. Prenatal glucocorticoids exposure and fetal adrenal development programming. 11β-HSD2, 11β-

Jo

ur

na

lP

re

-p

ro of

hydroxysteroid dehydrogenases 2; IGF1, insulin-like growth factor 1.

31