ReACH Registr achondroplazie
csen
 
 

Výzkum

Head of laboratory

Pavel Krejci, PhD

Office

Masaryk University, Faculty of Medicine, Department of Biology, Room A3/246
Kamenice 5, 62500 Brno, Czech Republic 

E-mail

krejcip@med.muni.cz

Phone

+420-549495395

Masaryk University page

Our research on FGFR3-related skeletal dysplasia

The major focus of our research for over 15 years has been to dissect the mechanisms of fibroblast growth factor (FGF) signaling. We are interested in pathological FGF-receptor (FGFR) signaling in disease, namely the skeletal disorders caused by activating mutations in FGFR3 (hypochondroplasia, achondroplasia, thanatophoric dysplasia, SADDAN). Our studies encompass many different areas of the FGF field including expression of FGF ligands in vivo, mechanisms of FGF/FGFR-mediated regulation of cell function, molecular mechanisms of FGFR signal transduction, biochemistry of FGFR kinase activation, development of FGFR inhibitors, and others. In our research, we actively collaborate with other scientists worldwide, including those from Norway (Dr. A. Wiedlocha, Oslo University), USA (Dr. D. Krakow, University of California Los Angeles; Dr. K. Hristova, John Hopkins University, Baltimore) and the Czech Republic (Dr. L. Trantirek, Masaryk University CEITEC, Brno; Dr. P. Konik, University of South Bohemia, Ceske Budejovice; Dr. M. Buchtova, Masaryk University).

The current lab projects related to achondroplasia address three important and poorly understood areas of FGFR3 function in chondrocytes: (1) the composition of signaling complexes proximal to activated FGFR3 at the cell membrane, (2) the nature of molecular effectors utilized by FGFR3 signaling to mediate specific cell functions, and (3) the mechanisms by which FGFR3 regulates chondrocyte differentiation. In addition to understanding the basic mechanics of FGFR3 signal transduction, we also focus on development of novel ways to target FGFR3 therapeutically. In the past, we characterized one novel small chemical inhibitor of FGFR3, as well as the mechanism of inhibition of FGFR3 signaling by a biological pathway, i.e. the C-natriuretic peptide (CNP) signaling. The latter work provided the basis for our collaboration with Biomarin Pharmaceuticals (Novato, CA) on development of a CNP analogue as a treatment for achondroplasia. The work on therapeutic targeting of FGFR3 never ceases, and we currently have several projects addressing novel avenues of FGFR3 inhibition.

Methods

Krejci lab operates extensive toolkit to study FGFR signaling at biochemical, molecular and cellular level. Please click at the animation below to see our main chondrocyte in vitro model used to interrogate the FGFR3 signaling in vitro. We also employ up-to-date proteomics and phosphoproteomics to identify and characterize novel members of FGFR3 signal transduction pathways. Through our collaborations, we validate in vitro findings using several ex vivo and in vivo models to limb development such as mouse limb bud micromass cultures, limb explant cultures and chick limb morphogenesis model.

We are open to collaboration on any aspect of FGFR signaling in achondroplasia or other FGFR-related pathologies, please contact P. Krejci at krejcip@med.muni.cz.

Inhibition of chondrocyte growth by FGFR3 movie

Lab members

Postdocs

Michaela Kunova Bosakova

Bohumil Fafilek

 

Physician

Martin Pesl

 

Postgradual students

Iva Gudernova

Alexandru Nita 

Marketa Tomanova

Neha Basheer 

Sara P. Abraham

Tomas Gregor

 

Lab technicians

Jitka Krenova

Vladimira Hykelova

Selected publications

(See PubMed for complete publication list)

Reviews

  • Krejci P. (2014) The paradox of FGFR3 signaling in skeletal dysplasia: why chondrocytes growth arrest while other cells over proliferate. Mutat Res Rev Mutat Res.  759:40-8.
  • Foldynova-Trantirkova S, Wilcox WR, Krejci P. (2012) Sixteen years and counting: the current understanding of fibroblast growth factor receptor 3 (FGFR3) signaling in skeletal dysplasias. Hum Mutat. 33:29-41.
  • Pejchalova K, Krejci P, Wilcox WR. (2007) C-natriuretic peptide: an important regulator of cartilage. Mol Genet Metab. 92:210-5.

Mechanisms of FGFR3 and other FGF/FGFR signaling

  • Fafilek B, Balek L, Kunova Bosakova M, Varecha M, Nita A, Gregor T, Gudernova I, Krenova J, Ghosh S, Piskacek M, Jonatova L, Cernohorsky N, Zieba JT, Kostas M, Haugsten EM, Wesche J, Erneux C, Trantirek L, Krakow D, Krejci P. The inositol phosphatase SHIP2 enables sustained ERK activation downstream of FGF receptors by recruiting Src kinases to FGF-receptor. Science Signalling 2018;11(548).
  • Balek L, Nemec P , Konik P , Kunova Bosakova M, Varecha M, Gudernova I, Krakow D, Krejci P. Proteomic analyses of signalling complexes associated with receptor tyrosine kinase identify novel members of fibroblast growth factor receptor 3 interactome. Cellular Signaling 2018;42:144-154.
  • Kunova Bosakova M, Varecha M, Hampl M, Duran I, Buchtova M, Dosedelova H, Machat R, Xie Y, Ni Z, Martin JH, Chen L, Jansen G, Krakow D, Krejci P.  Regulation of ciliary function by fibroblast growth factor signaling identifies FGFR3-related disorders achondroplasia and thanatophoric dysplasia as ciliopathies. Hum Mol Genet. 2018;27(6):1093-1105.
  • Buchtova M, Chaloupkova R, Zakrzewska M, Vesela I, Cela P, Barathova J, Gudernova I, Zajickova R, Trantirek L, Martin J, Kostas M, Otlewski J, Damborsky J, Kozubik A, Wiedlocha A, Krejci P. (2015) Instability restricts signaling of multiple fibroblast growth factors. Cell Mol Life Sci. 72:2445-59.
  • Kant SG, Cervenkova I, Balek L, Trantirek L, Santen GW, de Vries MC, van Duyvenvoorde HA, van der Wielen MJ, Verkerk AJ, Uitterlinden A, Hannema SE, Wit JM, Oostdijk W, Krejci P, Losekoot M. (2015) A novel variant of FGFR3 causes proportionate short stature. Eur J Endocrinol. 172:763-70.
  • Buchtova M, Oralova V, Aklian A, Masek J, Vesela I, Ouyang Z, Obadalova T, Konecna Z, Spoustova T, Pospisilova T, Matula P, Varecha M, Balek L, Gudernova I, Jelinkova I, Duran I, Cervenkova I, Murakami S, Kozubik A, Dvorak P, Bryja V, Krejci P. (2015) Fibroblast growth factor and canonical WNT/β-catenin signaling cooperate in suppression of chondrocyte differentiation in experimental models of FGFR signaling in cartilage. Biochim Biophys Acta. 1852:839-50.
  • Salazar L, Kashiwada T, Krejci P, Meyer AN, Casale M, Hallowell M, Wilcox WR, Donoghue DJ, Thompson LM. (2014) Fibroblast growth factor receptor 3 interacts with and activates TGFβ-activated kinase 1 tyrosine phosphorylation and NFκB signaling in multiple myeloma and bladder cancer. PLoS One. 9:e86470.
  • Krejci P, Kunova M, Kubikova I, Tratirek L, Kozubik A, Dvorak P. (2013) Expression of FGF19 in human embryonic stem cells. Stem Cells. 31:2582-4.
  • Wang Y, Zhou X, Oberoi K, Phelps R, Couwenhoven R, Sun M, Rezza A, Holmes G, Percival CJ, Friedenthal J, Krejci P, Richtsmeier JT, Huso DL, Rendl M, Jabs EW. (2012) p38 Inhibition ameliorates skin and skull abnormalities in Fgfr2 Beare-Stevenson mice. J Clin Invest. 122:2153-64.
  • Krejci P, Aklian A, Kaucka M, Sevcikova E, Prochazkova  J, Masek J, Mikolka P, Pospisilova T, Spoustova T, Weis M,  Paznekas W, Wolf J, Gutkind JS, Wilcox WR,  Kozubik A, Wang Jabs E, Bryja V, Salazar L, Vesela I, Balek L. (2012) Receptor tyrosine kinases activate canonical WNT/β-catenin signaling via MAP kinase/LRP6 pathway and direct β-catenin phosphorylation. PLoS ONE. 7:e35826
  • Merrill AE, Sarukhanov A, Krejci P, Idoni B, Camacho N, Estrada KD, Lyons KM, Deixler H, Robinson H, Chitayat D, Curry CJ, Lachman RS, Wilcox WR, Krakow D. (2012) Bent Bone Dysplasia-FGFR2 type, a Distinct Skeletal Disorder, Has Deficient Canonical FGF Signaling. Am J Hum Genet. 90:550-557.
  • Krejci P, Prochazkova J, Smutny J, Chlebova K, Lin P, Aklian A, Bryja V, Kozubik A, Wilcox WR. (2010) FGFR3 signaling induces a reversible senescence phenotype in chondrocytes similar to oncogene-induced premature senescence. Bone. 47:102-10.
  • Salazar L, Kashiwada T, Krejci P, Muchowski P, Donoghue D, Wilcox WR, Thompson LM. (2009) A novel interaction between fibroblast growth factor receptor 3 and the p85 subunit of phosphoinositide 3-kinase: activation-dependent regulation of ERK by p85 in multiple myeloma cells. Hum Mol Genet. 18:1951-61.      
  • Krejci P, Salazar L, Kashiwada TA, Chlebova K, Salasova A, Thompson LM, Bryja V, Kozubik A, Wilcox WR. (2008) Analysis of STAT1 activation by six FGFR3 mutants associated with skeletal dysplasia undermines dominant role of STAT1 in FGFR3 signaling in cartilage. PLoS One. 3:e3961.
  • Krejci P, Prochazkova J, Bryja V, Jelinkova P, Pejchalova K, Kozubik A, Thompson LM, Wilcox WR. (2009) Fibroblast growth factor inhibits interferon gamma-STAT1 and interleukin 6-STAT3 signaling in chondrocytes. Cell Signal. 21:151-60.
  • Matsushita T, Wilcox WR, Chan YY, Kawanami A, Bükülmez H, Balmes G, Krejci P, Mekikian PB, Otani K, Yamaura I, Warman ML, Givol D, Murakami S. (2009) FGFR3 promotes synchondrosis closure and fusion of ossification centers through the MAPK pathway. Hum Mol Genet. 18:227-40.
  • Krejci P, Salazar L, Goodridge HS, Kashiwada TA, Schibler MJ, Jelinkova P, Thompson LM, Wilcox WR. (2008) STAT1 and STAT3 do not participate in FGF-mediated growth arrest in chondrocytes. J Cell Sci. 121:272-81.
  • Krejci P, Masri B, Salazar L, Farrington-Rock C, Prats H, Thompson LM, Wilcox WR. (2007) Bisindolylmaleimide I suppresses fibroblast growth factor-mediated activation of Erk MAP kinase in chondrocytes by preventing Shp2 association with the Frs2 and Gab1 adaptor proteins. J Biol Chem. 282:2929-36.
  • Krejci P, Bryja V, Pachernik J, Hampl A., Pogue R, Mekikian P, Wilcox WR. (2004) FGF2 inhibits proliferation and alters the cartilage-like phenotype of RCS chondrocytes. Experimental Cell Research. 297: 152-164.

Drug development

  • Balek L, Buchtova M, Kunova Bosakova M, Varecha M, Foldynova-Trantirkova S, Gudernova I, Vesela I, Havlik J, Neburkova J, Turner S, Krzyscik MA, Zakrzewska M, Klimaschewski L, Claus P, Trantirek L, Cigler P, Krejci P.  Nanodiamonds as "artificial proteins": Regulation of a cell signalling system using low nanomolar solutions of inorganic nanocrystals. Biomaterials 2018;176:106-121.
  • Fafilek B, Hampl M, Ricankova N, Vesela I, Balek L, Kunova Bosakova M, Gudernova I, Varecha M, Buchtova M, Krejci P. (2017) Statins do not inhibit the FGFR signaling in chondrocytes. Osteoarthritis Cartilage 2017;25(9):1522-1530.
  • Balek L, Gudernova I, Vesela I, Hampl M, Oralova V, Kunova Bosakova M, Varecha M, Nemec P, Hall T, Abbadessa G, Hatch N, Buchtova M, Krejci P. ARQ 087 inhibits FGFR signaling and rescues aberrant cell proliferation and differentiation in experimental models of craniosynostoses and chondrodysplasias caused by activating mutations in FGFR1, FGFR2 and FGFR3. Bone 2017;105:57-66.
  • Gudernova I, Balek L, Varecha M, Kucerova Fialova J, Kunova Bosakova M, Fafilek B, Palusova V, Uldrijan S, Trantirek L, Krejci P. (2017) Inhibitor repurposing reveals ALK, LTK, FGFR, RET and TRK kinases as the targets of AZD1480. Oncotarget. 8:109319-109331.
  • Gudernova I, Foldynova-Trantirkova S, Ghannamova BE, Fafilek B, Varecha M, Balek L, Hruba E, Jonatova L, Jelinkova I, Kunova Bosakova M, Trantirek L, Mayer J, Krejci P. (2017) One reporter for in-cell activity profiling of majority of protein kinase oncogenes. Elife. 2017;6. pii: e21536
  • Gudernova I, Vesela I, Balek L, Buchtova M, Dosedelova H, Kunova M, Pivnicka J, Jelinkova I, Roubalova L, Kozubik A, Krejci P (2016). Multikinase activity of fibroblast growth factor receptor (FGFR) inhibitors SU5402, PD173074, AZD1480, AZD4547 and BGJ398 compromises the use of small chemicals targeting FGFR catalytic activity for therapy of short- stature syndromes. Hum Mol Genet. 25(1):9-23.
  • Wendt DJ, Dvorak-Ewell M, Bullens S, Lorget F, Bell SM, Peng J, Castillo S, Aoyagi-Scharber M, O'Neill CA, Krejci P, Wilcox WR, Rimoin DL, Bunting S. (2015) Neutral endopeptidase-resistant C-type natriuretic Peptide variant represents a new therapeutic approach for treatment of fibroblast growth factor receptor 3-related dwarfism. J Pharmacol Exp Ther. 353:132-49.
  • Scuto A, Krejci P, Popplewell L, Wu J, Wang Y, Kujawski M, Kowolik C, Xin H, Chen L, Wang Y, Kretzner L, Yu H, Wilcox WR, Yen Y, Forman S, Jove R. (2011) The novel JAK inhibitor AZD1480 blocks STAT3 and FGFR3 signaling, resulting in suppression of human myeloma cell growth and survival. Leukemia. 25:538-50.
  • Krejci P, Murakami S, Prochazkova J, Trantirek L, Chlebova K, Ouyang Z, Aklian A, Smutny J, Bryja V, Kozubik A, Wilcox WR. (2010) NF449 is a novel inhibitor of fibroblast growth factor receptor 3 (FGFR3) signaling active in chondrocytes and multiple myeloma cells. J Biol Chem. 285: 20644-53.
  • Krejci P, Pejchalova K, Wilcox WR. (2007) Simple, mammalian cell-based assay for identification of inhibitors of the Erk MAP kinase pathway. Investigational  New Drugs. 25: 391-395.
  • Krejci P, Masri B, Fontaine V, Mekikian PB, Weis M, Prats H, Wilcox WR. (2005) Interaction of fibroblast growth factor and C-natriuretic peptide signaling in regulation of chondrocyte proliferation and extracellular matrix homeostasis. J Cell Sci. 118: 5089-5100.

Contact us

Please contact Pavel Krejci (krejcip@med.muni.cz) for achondroplasia research-related questions and questions about how to collaborate with us, and Martin Pesl (mpesl@med.muni.cz) for clinical and patient-related questions and inquiries.

Links