Regulation of Cell Function by FGFR3 Pavel Krejci Limb development: At the beginning there was retinoic acid… DNA binding domain derived from the prototype gene called transcription factor T. Limb identity factors Tbx4 (hindlimb) and Tbx5 (forelimb) Later came TBX Development 2003;130:2741-51. Wild-type Fgf10-/- FGF10 proliferace mesodermu růst končetinového pupene ……..followed by FGF10 Wild-type Fgf10-/- 4 receptors: FGFR1-4 22 ligands: FGF1-23 ……..followed by more FGF G. Martin x 109 = Pattern and Shape vs. Growth and Proportions Resting cartilage Bone Growing cartilage Age Resting cartilage Growing cartilage FGFR3 Fgfr3+/- Fgfr3+/- Fgfr3-/- Tail Fgfr3-/- Cell 1996, 84(6):911-21. Achondroplasia Thanatophoric Dysplasia FGFR3-related skeletal dysplasias Nat Genet 1995, 9:321-8.. - short long bones - brachydactyly - macrocephaly - low nasal bridge - spinal stenosis - temporal lobe malformations FGFR3-related skeletal dysplasia Hypochondroplasia Achondroplasia SADDAN Thanatophoric Dysplasia STATURE AC TM TK I II III FGF binds here FGF4 FGF4 wild-type Skeleton: hypochondroplasia, achondroplasia, thanatophoric dysplasia, SADDAN, Muenke syndrome Skin: epidermal nevi, seborrhaeic keratosis, acanthosis nigricans Cancer: multiple myeloma, bladder cancer, seminoma, other. Exp Cell Res. 2004;297:152-64. J Cell Sci. 2005; 118: 5089-100. J Biol Chem. 2007 ;282:2929-36. Pediatr Res. 2007; 61(3):267-72. Invest New Drugs 2007; 25:391-95. PLoS One 2008; 3:e3961. J Cell Sci 2008; 121:272-81. Cell Signal 2009; 21:151-60. Hum Mol Genet. 2009; 18:227-40. J Biol Chem 2010; 285:20644-53. Bone 2010; 47:102-10. Leukemia 2011; 25:538-50. Human Mutation 2012; 33:29-41. Am J Hum Genet. 2012;90(3):550-7. PLoS One. 2012;7(4):e35826. J Clin Invest. 2012;122(6):2153-64. PLoS One. 2014;9(1):e86470. Mutat Res Rev. 2014 Jan-Mar;759:40-8. healthy TD resting proliferating hypertrophic bone FGF2 (h) FGF2 concentration (ng/ml) FGFR3 inhibits chondrocyte proliferation by arresting their cell cycle in G1 phase Exp Cell Res 2004, 297:152-64. Lower expression levels of FGFR3 than FGFR1 but all zones express same amounts. control FGF2 FGFR3 alters the cartilage-like phenotype of chondrocytes FGF2 (h) M C 0.5 1 4 8 12 24 48 72 aggrecan collagen II collagen I GAPDH proMMP9 matureMMP9 intermediate matureMMP2 proMMP13 matureMMP13 proMMP2 mature MMP3/10 pro MMP3/10 J Cell Sci. 2005; 118: 5089-100. Lower expression levels of FGFR3 than FGFR1 but all zones express same amounts. FGFR3 causes premature senescence in chondrocytes SA-b-gal control FGF2 FGF2 control Bone 2010; 47:102-10. C1 C2 1’ 5’ 10’ 30’ 1h 2h 4h 8h FGF2 P-Erk1/2 Erk1/2 P-p38 p38 P-PLCg1 PLCg1 .001 .01 .1 1 10 100 Inhibitor concentration (mM) U0126 80 60 40 20 0 Ras RasN17 - F1 F10 F100 FGFR3 arrests chondrocyte growth via RAS-ERK MAP kinase pathway Exp Cell Res 2004, 297:152-64. Lower expression levels of FGFR3 than FGFR1 but all zones express same amounts. STAT3-YFP DAPI merge DIC/merge control IL6 30m FGF2 48h FGF2 2h IL6 30m FGF2 48h IL11, LIF Cish Socs1 Socs3 FGF2 IL6 IL6RA Shp2 gp130 STAT3 STAT3 nucleus FGFR3 Frs2 Chronic FGFR3 activation inhibits cytokine/STAT signaling in chondrocytes Cell Signal 2009;21:151-60. Fgfr3Ach Kazuwa Nakao Nature Medicine 10, 80-86 (2004). CNP wild-type Fgfr3Ach/CNP CNP wild-type wild-type C-natriuretic peptide (CNP) FGFR3 Ras CNP cGMP PKG Raf-1 MEK Erk NP-R FRS2 STOP FGF2 Growth arrest Matrix degradation J Cell Sci. 2005; 118: 5089-100. J Biol Chem 2010; 285:20644-53. NF449 Leukemia 2011. 25:538-50. An external file that holds a picture, illustration, etc. Object name is nihms332356f3.jpg Human Mutation 2012; 33:29-41.