CEDAM
School of Clinical & Experimental MedicineUniversity of Birmingham
Birmingham, United Kingdom
CEDAMCentre for Endocrinology, Diabetes and Metabolism
Wiebke Arlt
Adrenarche– nouveaux aspects de son
contrôle génétique
The Steroid Universe
DHEASDHEA
Oestradiol
Testosterone
Aldosterone
Androstenedione
Cortisol
Adrenarche
Seru
m D
HEA
-S (n
g/m
l)
1 10 20 30 40 50 60Age (yrs)
4000
2000
Serum DHEAS and Age
Age (years)
Korth-Schutz et al., JCE&M 1976
DHEAS Secretion during Adrenarche
Pubarche
Remer, Boye, Hartmann and Wudy , JCE&M 2005
Urinary Excretion of C19 Steroids from 3 to 19 years
Low birth weight
catch-upweight
gain
Premature adrenarche PCOS Metabolic
Syndrome
Early Onset Androgen Excess Premature Adrenarche and Adolescent PCOS
as a forerunner of adult metabolic disease
Idkowiak et al., Eur J Endocrinol 2011
Zona Glomerulosa
Aldosterone
Zona Fasciculata
DHEAZona Reticularis „Sex“
„Sugar“
„Salt“
Cortisol
CYP17A1
CYB5
HSD3B2
Pregnenolone
17Preg
Progesterone
17OHP
Cholesterol
Deoxycortico-sterone
11-Deoxycortisol
Cortisol
18OH-Cortico-sterone
AldosteroneCortico-sterone
DHEA Androstenedione Testosterone DHT
HSD3B2
HSD3B2 HSD17B3 SRD5A2
HSD3B2
CYP21A2
CYP21A2
CYP17A1
CYP17A1
CYP17A1
CYP17A1
CYP11A1
CYP11B1
CYP11B2 CYP11B2 CYP11B2
CortisoneHSD11B1
HSD11B2
Mineralocorticoids - SALTMineralocorticoid
precursors
Glucocorticoids- SUGAR
Androgens- SEX
Androgen precursors
Glucocorticoid precursors
CYP11B1
Adrenal Steroidogenesis
DHEA
DHEASSTSSULT
2A1
Estrone (E1)
17β-Estradiol (E2)
∆4-Andro-stenedione
Testosterone (T)
Dihydro-testosterone
(DHT)
Androstanediol glucuronide (ADG)
DHEA Metabolism and DHEA/DHEAS Interconversion
Adrenal Gland Peripheral target cell
17OH-Pregnenolone
CYP17A1
ACTH
Serum Hormones after 4 Months of DHEA 50 mg/d in Women with Adrenal Insufficiency (n=24)
DHEA
Andro-stene-dione
5α-Dihydro-
testo-sterone
ADG
Testo-sterone
DHEAS
Arlt et al., N Engl J Med
1999
DHEAS DHEA
STS
SULT2A1
Δ4-A‘dione
Testo
5α-DHT
E1
E2
Subjects• healthy young men (n=8, median age 25a)Protocol• frequent serum sampling 08:30 - 15:00 h• Day 1: Baseline• Day 2: 100 mg DHEA orally (09:00 h)• Day 3: 25 mg DHEAS i.v. (09:00 h)
Serum DHEA and DHEAS do not interconvert freely and continuously
Hammer F, Subtil S et al., JCE&M 2005
0
5
10
15
20
25
30
35
-60 0 60 120 180 240 300 360
Baseline25mg DHEAS i.v.
DH
EAS
(µm
ol/l)
Time (min)
25mg DHEAS i.v.
0
10
20
30
40
50
60
-60 0 60 120 180 240 300 360
25mg DHEAS i.v.
DH
EA(n
mol
/l)Time (min)
=> Circulating DHEAS does not necessarily reflect circulating DHEA
DHEAS DHEA
STS
SULT2A1
Androstenedione
Testosterone
E1
E2
5α-DHT
Hammer F, Subtil S et al., J Clin Endocrinol Metab 2005Arlt W et al., J Clin Endocrinol Metab 2006
DHEAS DHEASULT2A1
8-year old girl • Premature pubarche at 6 yrs• Tanner PH4, B2• Height 125 cm, Weight 36.2 kg• Advanced bone age (12 yrs)Follow-up:• Menarche at 11 yrs• At 12 yrs Tanner PH5, B4• Height 139 cm, Weight 58.3 kg• Advanced bone age (16.5 yrs)•Significant hirsutism and acne•At 13 yrs secondary amenorrhoea
The Case
From Premature Pubarche to PCOSHormone Age 8 yrs Age 12 yrsACTH (pmol/L) 10.0 (2.2-13.2) 3.6 (2.2-13.2)Cortisol (nmol/L) 280 (190-550) 440 (190-550)17OHP (nmol/L) 1.80 (1.0-3.4)DHEAS (µmol/L) <0.40 (0.4-4.6) <0.40 (1.6-10.4)DHEA (nmol/L) 15.0 (1.3-18.0) 20.0 (2.0-22.0)
Androstenedione (nmol/L) 4.1 (0.1-1.4) 27.0 (<12.0)Testosterone (nmol/L) 1.2 (0.03-0.65) 2.1 (0.51-1.26)Dihydrotestosterone (nmol/L) 0.36 (0.05-0.25)LH (U/L) <0.2 (<0.05-0.7) 3.2 (<0.05-20.2)FSH (U/L) 2.0 (0.3-4.8) 5.2 (0.14-8.78)
SST (12 yrs) Baseline 30 min 60 minDHEA (nmol/L) 20.0 38.0 43.0DHEAS (µmol/L) <0.40 <0.40 <0.40
DHEA
DHEAS
SULT2A1
From Bedside to Bench:The DHEA/DHEAS Shuttle
PAP
PAPS
PAPS Synthase
APSATP+ SO4
ATP Sulfurylase
APS Kinase
+ ATP
Compound heterozygous mutations in PAPSS286 kb
6321 2 3 4 5 6 10 11 127B
1,359 6002,00149,186 3,760 752 561 120 5,912 5,512 13,7447 98
N APS KINASE C
44 194 244 6071 614
F
M
P
T48R R329X
c.143C>G c.985C>T
A
ATP SULFURYLASE
• Defects in APSK2 (=PAPSS2) are the cause of autosomal recessive spondyloepimetaphyseal dysplasia Pakistani type (SEMD). ul-Haque et al, Nat Genet 1998, 20:157-162
Spondyloepimetaphyseal dysplasia (SEMD) Pakistani type (OMIM +603005)
Ahmad et al., Am J Med Genet 1998, 78:468-473S475X
A
C D
T48RR329X
B
APS kinase domain
ATP sulfurylase
domain S475X
From Bedside to Bench: Mutant Protein Analysis
PAPS
3H DHEA 3H DHEAS
ATPSO42-
In vitro assessment of DHEA sulfation: Coupled SULT2A1/PAPSS2 assay
WT/ Mutant PAPSS2
WT SULT2A1
PAP
TLC
Viv Dhir
0
20
40
60
80
100
WT T48R R329X S475X
DH
EA
S s
ynth
esis
by
SU
LT2A
1 co
-in
cuba
ted
with
wild
-type
and
mut
ant
PAP
SS
2a (
% w
ild-ty
pe a
ctiv
ity)
WT T48R R329X S475X
100
80
60
40
20
0
WT
T48R
R32
9X
S475
X
Western Blot
Co-Incubation of bacterially expressed hSULT2A1 and wild-type or mutant PAPSS2a
PAPSS2 and Androgen Excess• Inactivating mutations in PAPSS2 in a girl with
premature pubarche and early onset polycystic ovary syndrome – novel monogenic cause of androgen excess
• Critical role of DHEA sulfation as a gatekeeper of human androgen synthesis – Impaired DHEA sulfation results in increased conversion of DHEA to androgens
N Engl J Med 2009, 360: 2310-2318.
Premature adrenarche
Polycystic ovary syndrome
(PCOS)
Metabolic Syndrome
DHEA Sulfation and Childhood Androgen Excess
Virdis R, Zampolli M, Ibáñez L, Ghizzoni L, Street ME, Vicens- Calvet E. II pubarca prematuro. Riv Ital Pediatr (IJP) 1993;19:569–579
PAPSS
DHEA sulfation and Androgen
Synthesis
SULT2A1
PAPSS2
DHEA
DHEAS
Sulfation during chondrocyte and
bone development
Sulfation of drugs and xenobiotics
(Hepatic Phase 2 Drug Metabolism)
PAPS Synthase and Human Sulfation
Vivek Dhir
Jan IdkowiakNils Krone
Jo McNelis
Edson Nogueira
Florian Schlereth
CEDAM
Sex Steroid Group Nicole Reisch
Angela TaylorHannah Ivison
Pushpa PatelDonna HolmesSilvia ParajesIan Rose
Fred Sweep Hedi Claahsen Jan Smeitink Kees Noordam Ron Smeets
Kees Nordam Hedi Clahhsen van der GrintenBill Rainey Enzo Lalli