OVERVIEW

Biology of Estrogen &

Progesterone Receptors

Benita S. Katzenellenbogen
Swanlund Professor of Physiology,
Cell and Structural Biology
University of Illinois and College of Medicine
 Important Issues
• Roles of ER and ER
• Distinct Functions of PR-A and PR-B
• Selective Ligands (SERMs and SPRMs)
• Coregulators
• Interrelationships Between Estrogen and
Progestin Hormone-Receptor Pathways
• Tissue Selectivity in Estrogen and
Progestin Actions
Target Tissues in Females & Males

Also:
Colon (& Intestine)
Bladder & Urogenital Tract
Lung

Korach, 2001
Estrogen Receptor in Breast Cancer

Predicts:
• Improved Disease-Free Survival

• Response to Tamoxifen/Endocrine
Therapy
 New York Times, April 7, 1998, p. A1

Researchers Find the First Drug

Known to Prevent Breast Cancer

Tamoxifen Helps Women, but Has Side Effects

 Tissue Selectivity

- Ideal Profile for HRT -

• Antagonist - Breast, Uterus

• Agonist - Bone, Brain, Colon,
Lipid Profile, Vasomotor,
Cardiovascular
 TRIPARTITE PHARMACOLOGY
MEDIATED BY NUCLEAR HORMONE RECEPTORS
– Underlies Tissue Selectivity –

“Interactions”
“Actions”

Ligands
• Receptor
• Effectors

• natural • subtypes • DNA response TRANSCRIPTION

• synthetic • isoforms elements OTHER Actions
• environmental • splice variants • Co-regulators
activators
repressors
• Other TFs • REs
 Receptor Actions in Cells
Growth Peptides and
Estrogen Factors Neurotransmitters
E GF

E R

E Second Messengers
and
Signaling Protein Kinase Cascades

EREs
R R Receptor
E E

Co-Regulators
TAFs
B
TBP Transcription
A F
TATA
pol II

(B. Katzenellenbogen et al. Recent Prog. Horm. Res.,2000)

Human Estrogen Receptors  and 
AF-1 DNA Ligand / AF-2
A/B C D E F

1 180 263 302 552 595

ER N- -C

1 144 227 255 504 530

ER N- -C
(18) (97) (30) (59) (18)

• Different tissue/cell distributions

• Different affinity for ligands
• Different gene activations
Exquisite Precision in Receptor Regulation

Small Changes Major Changes

in in
Ligand Structure Biological Character

Different Pharmacology
ERER
At
Different Ligands
Different Target Genes
 Ligands for Estrogen Receptors
Novel ER/ER
Estrogens Known SERMs
Selective Ligands
OH
N
O

OH
OH
N N

CN
HO OH HO
HO
CH3
PPT (Pyrazole) DPN (Nitrile)
Estradiol Tamoxifen
ER Agonist ER Agonist
O
O N
N
OH
Et OH Me
O
N
OH N
HO S OH
HO HO Et HO

Diethylstilbestrol Raloxifene R,R-THC MPP

ER Agonist & ER Antagonist
Droloxifene, Idoxifene,
Toremifene, GW5638, ER Antagonist
EM652, Cp-336156, others
 SERM Action

(B. & J. Katzenellenbogen, Science, 295: 2380, 2002)

Structures of Diethylstilbestrol-ER LBD
and Hydroxytamoxifen-ER LBD

Diethylstilbestrol Hydroxytamoxifen Greene, 4/99

 PPT, an ER Specific Ligand, Prevents
Ovariectomy-Induced Loss of Bone Mineral Density

800
Total
* * *
Bone Mineral Density

700 Trabecular
600
*
(mg/cm3)

500 * *
400
300
200
100
0
Sham Ovx PPT (2mg/kg) 17beta-estradiol
(6ug/kg)

* significantly greater than ovx (p < 0.01) (Harris et al. and Katzenellenbogens,
Endocrinology, 2002)
 PPT, an ER Specific Ligand,
Prevents Hot Flush in a Rat Model
6
Temperature Increase

5
(degrees +/- SEM)

0
Vehicle Ethinyl PPT
Estradiol (Harris et al. and Katzenellenbogens,
Endocrinology 2002)
 Gene
Regulation
Estrogen Effects on Cell Cytoarchitecture

From Vic et al., Cancer Res., 1982

Estrogen Effects in Mammary Gland
and Breast Cancer

Cell Cycle Regulation --

 Proliferation  Apoptosis
Cytoskeletal Organization
Signal Transduction
 Estrogen Receptor Beta

• A “dampener” of ER activity?

• Some overlap with ER but also some

different gene activations
ER in Human Breast

(Roger et al., Cancer Res. 61:2537, 2001)

Gene Expression Profiling by DNA Microarrays
Differentiates Estrogen and SERM Activities

• Different Ligands Stimulate Somewhat Different Gene Sets

• Estrogen Stimulates Many Genes

• Tamoxifen also Stimulates Some of these Genes

• Raloxifene Stimulates a Few of these Genes

Comparative Patterns of Gene Stimulation by
Estradiol and SERMs (MCF-7 Cells) [n = 33]
A B
E2 Only (n = 17) E2 and TOT (n = 12)
Response

Response
C E2 ICI Ral TOT C E2 ICI Ral TOT

C
E2, Ral and TOT (n = 4)
Response

C E2 ICI Ral TOT

SERM Antagonism of Estradiol Stimulation
(MCF-7 Cells) [n = 35]
A B C
All 3 Antagonists (n = 12) ICI and Ral (n = 12) ICI only (n = 4)
Response

Response

Response
C E2 E2 E2 E2 C E2 E2 E2 E2 C E2 E2 E2 E2
+ + + + + + + + +
ICI RAL TOT ICI RAL TOT ICI RAL TOT

D E
Ral and TOT (n = 4) Ral only (n = 3)
Response

Response

C E2 E2 E2 E2 C E2 E2 E2 E2
+ + + + + +
ICI RAL TOT ICI RAL TOT
Progesterone
Receptor
 Ligands for Progesterone Receptor

Progestins Antiprogestins
OH
H 3C H
O
H
H3C CH3
N

O
Norethindrone OH
O [oral contraceptives] CH3
Progesterone
[natural] Et
O
Et O
H 3C RU486 (Mifepristone)
O
OAc
O H3C CH3
N
R5020
O (Promegestone)
CH3 H 3C OH
O
Medroxyprogesterone OH
Acetate (MPA) H Et
[HRT]
O
O ZK98299
ORG2058 (Onapristone)
 Human Progesterone Receptor:
A and B Forms
AF-3 AF-1 DBD AF-2 HBD hPR-B

AF-1 DBD AF-2 HBD hPR-A

• From single gene by alternate transcription

initiation (different promoters)

• Different activities
 Human Progesterone Receptor
Isoforms A and B

• Differential transcriptional activities

Cell type- and promoter-specific
Often PR-B > PR-A
• PR-A repressor of PR-B and ER signaling

• Present at unequal levels in breast tumors; some tissues

PR-A predominance as early event in carcinogenesis
 Interrelationships Between Estrogen and
Progestin Receptor Signaling Pathways

ER
–
PR Opposes + PR Enhances
ER Stimulation PR ER Stimulation

Uterus Breast

Other Tissues?
Women’s Health Initiative (WHI) Clinical Trial
Ended at 5.2 years, May 2002
[JAMA 288:321-333, 2002]

16,608 Healthy postmenopausal US women

Estrogen + Progestin
(CEE 0.625 mg/d) together daily (MPA 2.5 mg/d)
Would outcome be different if:
…estrogen only?
…different formulations, dosages?
…different estrogen(s)?
…different progestin?
Biology of Estrogens and Progestins
are Determined By:

• Ligand structure
• ER subtype ( or ) and PR isoform (A or B)
• Gene promoter responsive unit
• Character and balance of coactivators and
corepressors
 Take-Home Messages

1. Estrogens and progestins act through ER and PR.

2. Ligands can be:

Agonists-------SERMs/SPRMs-------Antagonists
(mixed agonists/antagonists)

3. Different ligands induce different receptor

conformations that determine all further interactions.
4. ER and PR work with partner proteins that modulate
their activity. Coregulator levels vary in different
tissues.
Ligand Receptor Coregulator Tissue Responses

5. Many interrelationships between ER and PR pathways:

• Progestins modulate estrogen action but also act
independently.
• Progestin effects on estrogen actions vary in
different tissues -
Mid-luteal when progesterone levels high --
– Breast: enhances mitotic activity
– Uterus: suppresses mitotic activity
 Questions that Remain
• What estrogen/SERM (± progestin/SPRM
combination) will give the best benefit/risk
for HRT in the diverse tissues in which
these hormones act ?

• Respective roles of ER and ER in

mediating the desired vs. undesired effects
of estrogen, and modulation of these
activities by progestins