1. Regulation of cell function by FGFR3 receptor tyrosine kinase
Pavel Krejci
Medical Genetics Institute, Cedars-Sinai Medical Center, Los Angeles, CA
Institute of Experimental Biology, Masaryk University, Brno, Czech Republic

2. Fgfr3+/-
Fgfr3-/-
Fgfr3+/-
Fgfr3-/-
Tail
Cell 1996, 84(6):

3. FGFR3-related skeletal dysplasias
Achondroplasia
Thanatophoric Dysplasia
FGFR3-related skeletal dysplasias
Nat Genet 1995, 9:
- short long bones
- brachydactyly
- macrocephaly
- low nasal bridge
- spinal stenosis
- temporal lobe malformations

4. FGFR3-related skeletal dysplasia
STATURE AC TM TK I II
III
FGF binds here
Hypochondroplasia
Achondroplasia
SADDAN
Thanatophoric Dysplasia

6. Skeleton: hypochondroplasia, achondroplasia, thanatophoric dysplasia, SADDAN, Muenke syndrome
Skin: epidermal nevi, seborrhaeic keratosis, acanthosis nigricans
Cancer: multiple myeloma, bladder cancer, seminoma

7. Exp Cell Res. 2004;297:
J Cell Sci. 2005; 118:
J Biol Chem ;282:
Pediatr Res. 2007; 61(3):
Invest New Drugs 2007; 25:
PLoS One 2008; 3:e3961.
J Cell Sci 2008; 121:
Cell Signal 2009; 21:
Hum Mol Genet. 2009; 18:
J Biol Chem 2010; 285:
Bone 2010; 47:
Leukemia 2011; 25:
Human Mutation 2012; 33:29-41

8. healthy TD
resting
proliferating
hypertrophic
bone

9. FGFR3 inhibits chondrocyte proliferation by arresting their cell cycle in G1 phase
3H-thymidine (cpm x 103)
% of cells
Lower expression levels of FGFR3 than FGFR1 but all zones express same amounts.
FGF2 concentration (ng/ml)
FGF2 (h)
Exp Cell Res 2004, 297:

10. FGFR3 alters the cartilage-like phenotype of chondrocytes
control FGF2
proMMP9
matureMMP9
intermediate
matureMMP2
proMMP13
matureMMP13
control
FGF2
proMMP2
mature MMP3/10
pro MMP3/10
FGF2 (h)
M C
aggrecan
collagen II
collagen I
GAPDH
Lower expression levels of FGFR3 than FGFR1 but all zones express same amounts.
J Cell Sci. 2005; 118:

12. FGFR3 causes premature senescence in chondrocytes
SA-b-gal
control FGF2
FGF2
control
Bone 2010; 47:

13. FGFR3 arrests chondrocyte growth via RAS-ERK MAP kinase pathway
cell colonies/100 cells (%)
Ras
RasN17
F F10 F100
C1 C2 1’ 5’ 10’ 30’ 1h 2h 4h 8h
FGF2
P-Erk1/2
Erk1/2
P-p38
p38
P-PLCg1
PLCg1
Inhibitor concentration (mM)
U0126
cells/wellx103 80 60 40 20
Lower expression levels of FGFR3 than FGFR1 but all zones express same amounts.
Exp Cell Res 2004, 297:

15. FRS2 GAB1 SHC SHP2 GRB2 GRB2 SOS SOS J Biol Chem 2007; 282:2929-36.
FGF2
FGFR3
FRS2
GAB1 SHC
GRB2
SOS
SHP2
GRB2
SOS
Ras
Raf-1
MEK
Erk
J Biol Chem 2007; 282:

16. FGFR3
STAT1
CKI
Growth arrest
Skeletal dysplasia

17. ? FGFR3 associates with STAT1 and acts as STAT1-kinase in chondrocytes
CKI
FGFR3
STAT1
Growth arrest ?
FGFR3-wt
FGFR3-K650M
FGFR3-K508M
GFP
FGFR3
actin
IP: STAT1
FGFR3 WB
STAT1
FGFR3-wt
FGFR3-K650E
substrate: STAT
SU5402(mM) _ _ _ _
ATP _ _
P-Y701-STAT1
Skeletal dysplasia
STAT1
FGFR3

18. J Cell Sci 2008;121:

19. PLoS One 2008;3:e3961.

20. Chronic FGF stimulus inhibits cytokine/STAT signaling in chondrocytes
STAT3-YFP DAPI merge DIC/merge
IL6
control
IL11, LIF
FGFR3
IL6RA
Shp2
Frs2
FGF2 2h
gp130
FGF2 48h
Cish
Socs1
Socs3
STAT3
IL6 30m
FGF2 48h
nucleus
STAT3
IL6 30m
Cell Signal 2009;21:

21. 2001 2012 FGFR3 FGFR3 IL6, LIF, IL11, IFNg CNP SOCS1/3 ? PKC NPR-B
IL6, LIF, IL11, IFNg
FGFR3
FGFR3
CNP
SOCS1/3 ?
PKC
NPR-B
STAT1
Frs2, Gab1, SHC
STAT1/3
STAT1 ?
cGMP ?
CKI ?
Ras/Raf/MEK/Erk
PKG
proliferation
growth arrest
CKI MMP
growth arrest
matrix degradation

22. actin
FGF2: C1 15‘ 1h 3h 6h 12h 24h 48h 72h C2 C3
b-catenin
Wnt

25. submitted

26. C-natriuretic peptide (CNP)
wild-type
wild-type
Fgfr3Ach
CNP
Fgfr3Ach/CNP
Kazuwa Nakao
Nature Medicine 10, (2004).
wild-type
CNP

27. _ _ _ _ _ 10 100 200 500 pCPT-cGMP [mM] control control FGF2 CNP 50 4030
cells per well [x104]
control 20
FGF2 10
FGF2
_ _ _ _ _ CNP [mM]
_ _ _ _ _ pCPT-cGMP [mM]
control FGF2
control
CNP

28. STOP CNP NP-R J Cell Sci. 2005; 118: 5089-100. Growth arrest
FGFR3
Ras
CNP
cGMP
PKG
Raf-1
MEK
Erk
NP-R
FRS2
STOP
FGF2
Growth arrest
Matrix degradation
J Cell Sci. 2005; 118:

29. Tocriscreen™ Mini Library
A collection of 1120 biologically active compounds supplied as DMSO solutions.
Tocriscreen™ Mini Library

30. NF449
J Biol Chem 2010; 285:

31. Leukemia :

32. Cedars-Sinai Medical Center Los Angeles, California Betty Mekikian
Anie Aklian
UCI, Irvine, California
Leslie Thompson
Tamara Kashiwada
Lisa Salazar
UCLA, Los Angeles, California
Matthew Schibler
Masaryk University, Brno, Czech Republic
Jirina Prochazkova
Vita Bryja
Lukas Balek
Tereza Spoustova
Tereza Pospisilova
INSERM U589, Toulouse, France
Bernard Masri
Vincent Fontaine