PHYSIOLOGICAL ROLE OF PROGESTERONE IN THE LUTEAL PHASE AND THE ENDOMETRIUM

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PHYSIOLOGICAL ROLE OF

PROGESTERONE IN THE LUTEAL PHASE AND THE ENDOMETRIUM

Professor IOANNIS E. MESSINIS

MD, PhD (Aberdeen, UK), FRCOG (UK)

Department of Obs/Gynae University of Thessaly

Larissa, Greece

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DISCLOSURE

Nothing to disclose

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•  Discuss the role of LH in corpus luteum formation

•  Describe the endocrine role (feedback) of progesterone in the luteal phase

•  Summarize the paracrine actions of

progesterone within the corpus luteum

•  Describe the progesterone actions in the endometrium preparing for receptivity

Learning Objectives

At the conclusion of this presentation, the participant should be able to:

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Figure 1.

-14-12-10 -8 -6 -4 -2 +2 +4 +6 +8+10+12+14 -14-12-10 -8 -6 -4 -2 +2 +4 +6 +8+10+12+14 0

20 40 60 80 100

LH (IU/I)

-14-12-10 -8 -6 -4 -2 +2 +4 +6 +8+10+12+14 -14-12-10 -8 -6 -4 -2 +2 +4 +6 +8+10+12+14 0

100 200 300

Estradiol (pg/ml)

-14-12-10 -8 -6 -4 -2 +2 +4 +6 +8+10+12+14 -14-12-10 -8 -6 -4 -2 +2 +4 +6 +8+10+12+14 0

5 10 15 20 25

Cycle days

FSH (IU/I)

-14-12-10 -8 -6 -4 -2 +2 +4 +6 +8+10+12+14 -14-12-10 -8 -6 -4 -2 +2 +4 +6 +8+10+12+14 0

2 4 6 8 10 12

Cycle days

Progesterone (ng/ml)

100 80 60 40 20 0 25 20 15 10 0 5

300 200 100 0 12 10 8 6 4 2 0 14

10 6

2 -2 -6 -10

-14 -14 -10 -6 -2 2 6 10 14 LH

IU/L

FSH IU/L

E2

pg/ml

P4

ng/ml

Cycle days Cycle days

Dafopoulos et al., 2009 Fertil. Steril.

92,1389-94

NORMAL CYCLE

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FSH

CLASS 5 CLASS 6 CLASS 7 CLASS 8 INTERCYCLE

MIDCYCLE

10-12 16 18-20 mm

GROWTH OF THE DOMINANT FOLLICLE

Messinis, 2008 In: Infertility and Assisted Reproduction, Cambridge UP, pp. 10-24

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80 60 40 20 1200

800 400

FSH LH E2

pmol/l

mIU/ml

0 16 8

0

-36 0 24 36 72 HOURS

Progesterone nmol/l

0 100

NORMAL LH SURGE

Messinis, 1989 PhD thesis (UK)

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•  Luteinization

•  Resumption of oocyte meiosis

•  Follicle rupture

•  Corpus luteum formation

LH SURGE INDUCES:

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Granulosa cells

Theca cells

Small luteal cells

Large luteal cells

FORMATION OF CORPUS LUTEUM FOLLOWING THE LH SURGE

Retamales et al., 1994 Hum. Reprod. 9, 591-96

(Human) Progesterone

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CORPUS LUTEUM FUNCTION

growth regression

LH pulses

LH receptors

Early Mid Late

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Fas Caspase-8

Caspase-3 Apoptosis

Bcl-2/Bax Caspase-9

Survivin TNFα or IFN-γ

Fas-ligand Macrophage

HCG

HCG RESCUES CORPUS LUTEUM

Sugino & Okuda, 2007, J. Reprod. Dev. 53, 977-986 LUTEAL CELL

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PROGESTERONE ROLE IN THE LUTEAL PHASE

•  Endocrine (feedback mechanisms)

•  Local in corpus luteum (luteotrophic)

•  Endometrium

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PROGESTERONE ROLE IN THE LUTEAL PHASE

•  Endocrine (feedback mechanisms)

•   Local in corpus luteum (luteotropic)

•   Endometrium

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O CONTROL Δ E2

E2+P4

Messinis et al., 2002 Hum. Reprod.

17, 299-303 FSH

IU/l

LH IU/l

E2

pmol/l

P4

nmol/l

30 15 500 250 0 10 5 0 18

9 0

0 1 2 3 4 5 6 7 Days after ovariectomy

NEGATIVE FEEDBACK

0 (Luteal phase)

OV

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FSH

Inhibin B

Inhibin A

E2 P4

INTERCYCLE RISE OF FSH

MENSES

Messinis 2006 Hum. Reprod.

Update 12, 557-571

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PROGESTERONE ROLE IN THE LUTEAL PHASE

•   Endocrine (feedback mechanisms)

•  Local in corpus luteum (luteotropic)

•   Endometrium

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1.5 1.0 0.5

0.0 EARLY MID LATE Progesterone receptor A/B protein (fmol/µg DNA)

PROGESTERONE RECEPTOR A/B (human corpus luteum)

Ottander et al., 2000

Biol. Reprod. 62, 655-63 a

a,b b

a,b P<0.05

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2.0 1.5 1.0 0.5 Caspase-3 mRNA level Arbitrary units 0.0

Control Onapristone

PROGESTERONE PREVENTS APOPTOSIS (luteotrophic action)

COWS (corpus luteum) Cultured mid-luteal cells

(Similar data with Fas mRNA)

P<0.01

Based on:

Okuda et al., 2004

Biol. Reprod. 71, 2065-71

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LH AND PROGESTERONE PREVENT CORPUS LUTEUM DEGRADATION

•  LH suppresses MMP-1 in the primate corpus luteum via progesterone

•  LH suppresses gelatinases, either at the mRNA (MMP-2) or protein (MMP-2 and -9) levels, perhaps in part via steroids,

including progesterone

•  LH promotes TIMP expression, perhaps via steroids, including progesterone.

Young & Stouffer, 2004 Biol. Reprod. 70, 244-50

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E2, P4 P4

CL CL

P4 HCG

Folli cle

ENDOCRINE AND PARACRINE REGULATION OF THE CORPUS

LUTEUM

LH surge

Pulsatile LH

Ovulation luteinization

Luteal

maintenance

Luteal Rescue

(early pregnancy)

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PROGESTERONE ROLE IN THE LUTEAL PHASE

•   Endocrine (feedback mechanisms)

•   Local in corpus luteum (luteotropic)

•  Endometrium

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FOLLICULAR PHASE

LUTEAL PHASE

Proliferative (Proliferation)

Secretory

(Differentiation) Follilce

Endome- trium

MENSTRUAL CYCLE

Shedding (Menstruation Regeneration)

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ENDOMETRIAL CHANGES

FOLLICULAR PHASE (Estrogen dominated)

•  Cellular proliferation in both the glandular

epithelium and stromal cells

LUTEAL PHASE

(Progesterone dominated)

•  First half

–  Decline in proliferation –  Secretory

differentiation

•  Second half

–  Proliferation in

epithelial cells is low –  Decidual changes

(high proliferation) in stroma

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ER1 PGR

Protein for cell division mRNAs

miRNAs

ER1 PGR

IGFBP1

Hbegf, BMP, Ovary

Estrogen &

Progesterone

Epithelium

Stroma

Ovarian steroids regulate the endometrium through cognate receptors (ESR1 or PGR)

Lessey, 2010

Biol. Reprod. 82, 653-655 _SC

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1 14 28 DAYS

Receptor pmol/mg DNA

P4

Ε2

RECEPTORS IN ENDOMETRIUM

(Epithelial cells)

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PROGESTERONE ACTS ON

•  The endometrial epithelial cells

•  The stroma

–  Cells –  Matrix

–  Blood vessels

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ROLE OF PROGESTERONE IN ENDOMETRIUM

•  Prepares the uterine epithelium for receptivity

•  Antagonizes estrogen-induced

proliferation in uterine epithelium

•  Differentiates endometrial stromal

cells to a decidual phenotype

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DECIDUALIZATION

•  Morphological and biochemical

differentiation of endometrial stromal cells into decidual cells (from small stromal fibroblast into large

epithelioid decidual cells)

•  Influx of specialized uterine NK cells

•  Profound angiogenesis

•  (Critical role of PRA)

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PROGESTERONE-INDUCED ENDOMETRIAL PROTEINS

•  IGFBP-1 (Placental protein 12)

•  Glycodelin (Progesterone-associated endometrial protein or placental

protein 14)

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ENDOMETRIAL BIOMARKERS

•  LIF

•  Cytokines (IL-6, IL-11)

•  Growth factors (TGF-β, EGF, HB- EGF, IGF)

•  Integrins (α4β1, ανβ3)

•  Osteopontin

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PR

OPN Beta 3

Ovary with CL

HB-EGF

P4

Stromal cell

Epithelial cell BM

Lessey, 2003; Steroids, 68, 809-15

PROGESTERONE EFFECTS ON THE

ENDOMETRIUM (Paracrine mediators)

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Epithelium

Stroma

Wetendorf & DeMayo, 2012; Mol. Cell. Endocrinol. 357, 108-118 MUC1

ER ERK1/2

FGFR PR FGF

Ihh PR

Hand2

•Proliferation

•Decidualization

•Cell Survival

•Progesterone Sensitivity COUP-TFII

Bmp2

Wnt4

Prolifera- tion

?

Progesterone regulated paracrine signaling

model in endometrium at implantation

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80 60 40 20

0 P1 P2 S1 S2 S3 Ratio HOXA11/G3PDH Arbitrary Densitometry Units

HOXA 11 EXPRESSION IN HUMAN ENDOMETRIUM

P: Proliferative

S: Secretory Taylor et al., 1999

JCEM 84, 1129-35 HOXA 10 expression

shows a similar pattern

*

NH

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MARKERS ASSOCIATED WITH UTERINE RECEPTIVITY

•  Morphological features (histology, pinopodes)

•  Cellular adhesion molecules family (integrins, selectins, cadherins,

immunoglobulins, mucins)

•  Cytokines (LIF, IL-6, IL-1)

•  Prostaglandins

Achache & Revel, 2006

Hum. Reprod. Update, 12, 731-746

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LH+0

14 15 16 17 18 19 20 21 22 Days

LH+2 LH+7

WINDOW OF IMPLANTATION

Berg & Navot, 1992 Fertil. Steril.

58, 537-42

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Day 20

PINOPODES

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C P TGFβ1

P+TGFβ1 MMP2

MMP11

mRNA

Based on:

Itoh et al., 2012 JCEM 97,

E888-E897

REGULATION OF MMPs BY PROGESTERONE

Human endometrial

stromal cells _d

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4 3 2 1

0 C RU486

6hrs

RU486 24hrs MMP7

P<0.01

EXPRESSION IN HUMAN ENDOMETRIUM (P4 role)

Catalano et al., 2007

Mol. Hum. Reprod. 13, 641-54 RU486 was given in-vivo

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AFTER DECIDUALIZATION

•  …the endometrium becomes irreversibly dependent upon progesterone signalling

•  …falling levels of progesterone trigger a cascade of events that

results in proteolytic breakdown of the superficial endometrium, focal bleeding and cell death.

Brosens & Gellersen, 2006, 36, 389-98

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P4 declines

Stroma Perivascular

cells

↑Chemokines (eg. IL-8, MCP-1)

Leucocyte

influx ↑MMP activity

TGFβ2 Epithelium

Innate immune molecules (eg. NOD2) Tissue degradation

and repair

↑COX2, ↓PGDH

↑PG synthesis

(a) (b) (c)

CORPUS LUTEUM REGRESSES

King & Critchley, 2010

J. Steroid Biochem. Mol. Biol. 120, 116-26

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PHYSIOLOGICAL ROLE OF PROGESTERONE IN THE LUTEAL PHASE AND THE ENDOMETRIUM