J. Coen NetelenbosJ. Coen Netelenbos

8 oktober 2008, IWO, Utrecht8 oktober 2008, IWO, Utrecht

Wnt Signaling SystemWnt Signaling System*Wat moeten we weten?*Wat moeten we weten?*Nieuwe medicijnen tegen *Nieuwe medicijnen tegen Osteoporose Osteoporose

Wnt signalingWnt signaling

WWg (wingless)g (wingless)

IntInt(egration)(egration)

The Wnt Homepage: http://www.stanford.edu/~rnusse/wntwindow.html o.l.v.Roel Nusse de ontdekker 1982 en naamgever Wnt in 1992

The Wnt Homepage: http://www.stanford.edu/~rnusse/wntwindow.html o.l.v.Roel Nusse de ontdekker 1982 en naamgever Wnt in 1992

plasterk

Nature 425, 633-637 Nature 425, 633-637 (9 October 2003)(9 October 2003)The Wnt/   -catenin pathway The Wnt/   -catenin pathway regulates cardiac valve regulates cardiac valve formationformationAdam F. L. Hurlstone……. Adam F. L. Hurlstone……. Ronald Ronald H. A. PlasterkH. A. Plasterk11 and and Hans CleversHans Clevers11

Wnt pathway componentsWnt pathway components ` `

Wnt &Wnt &

Wnt & heart diseaseWnt & heart disease

• Wound healing response after myocardial infarction in rats Wound healing response after myocardial infarction in rats and mice (and mice (Cardiovascular Research 2002 55(1):16-24)Cardiovascular Research 2002 55(1):16-24)

• Cardiac Hypertrophy and the Wnt Pathway Cardiac Hypertrophy and the Wnt Pathway (Cingolani OH et al. (Hypertension. (Cingolani OH et al. (Hypertension.

2007;49:427.)2007;49:427.)

.

Boyden LM et al. N Engl J Med 2002;346:1513-21Boyden LM et al. N Engl J Med 2002;346:1513-21

High bone density due to a gain-of-function mutation High bone density due to a gain-of-function mutation in in LDL-Receptor Related Protein 5 (LRP5)LDL-Receptor Related Protein 5 (LRP5)

Wnt pathway speelt rol Wnt pathway speelt rol bij stimulatie humane bij stimulatie humane osteoblasten met LRP5 osteoblasten met LRP5 als co-receptor!als co-receptor!

Gong Y et al. Cell 2001;107:513-23Gong Y et al. Cell 2001;107:513-23

OsteoPorosis-PseudoGlioma syndrome (OPPG) due to OsteoPorosis-PseudoGlioma syndrome (OPPG) due to a loss-of-function mutation in LDL-Receptor Related a loss-of-function mutation in LDL-Receptor Related

Protein 5 (LRP5) Protein 5 (LRP5)

Wnt Signaling by Low-Density Lipoprotein Wnt Signaling by Low-Density Lipoprotein Receptor-Related Protein 5 (LRP5),Receptor-Related Protein 5 (LRP5),

a coreceptor in Wnt signaling system a coreceptor in Wnt signaling system

LRP5(-/-) KO muis: LRP5(-/-) KO muis: osteoporose en osteoporose en pseudoglioom (OPPG) pseudoglioom (OPPG) LRP5 gain of function LRP5 gain of function muation: hoge bot massa muation: hoge bot massa

Central Role of Canonical Wnt SignalingCentral Role of Canonical Wnt Signaling

Khosla S et al. JCI 2008;118:421-8

List target genes of Wnt/List target genes of Wnt/ßß-catenin signaling-catenin signaling

sept 2008sept 2008

Regulation Canonical Wnt SignalingRegulation Canonical Wnt Signaling

Piters E et al. Arch Biochem Biophysics 2008;473:112-6

Regulation Canonical Wnt Signaling (A)Regulation Canonical Wnt Signaling (A)

Piters E et al. Arch Biochem Biophysics 2008;473:112-6

Frizzeld baconFrizzeld bacon

Low density Low density lipoprotein lipoprotein Receptor-Receptor-Related Protein Related Protein =coreceptor=coreceptor

CatenaCatena

T-Cell-specific T-Cell-specific transcription transcription

FactorFactor

•Proliferatie Proliferatie •DifferentiatieDifferentiatie•ApoptosisApoptosis

Dvl

DishevelledDishevelled protein (Dvl)protein (Dvl)

Osteoblasten

Regulation Canonical Wnt Signaling (A)Regulation Canonical Wnt Signaling (A)

DvlGlycogeen Glycogeen Synthetase KinaseSynthetase Kinase

LithiumLithium&&Small moleculesSmall moleculesmet zelfde effectmet zelfde effectop bot bij OVX-op bot bij OVX-rat als PTH!rat als PTH!

•Proliferatie Proliferatie •DifferentiatieDifferentiatie•ApoptosisApoptosis

Osteoblasten

Regulation Canonical Wnt Signaling (B)Regulation Canonical Wnt Signaling (B)

Piters E et al. Arch Biochem Biophysics 2008;473:112-6

Dickkopf Dickkopf (Dkk)(Dkk)

Dkk-1 InhibitorDkk-1 Inhibitor(mAb) met (mAb) met zelfde effectzelfde effectop bot bij OVX-op bot bij OVX-muis als PTH!muis als PTH!

Regulation Canonical Wnt Signaling (B)Regulation Canonical Wnt Signaling (B)

Piters E et al. Arch Biochem Biophysics 2008;473:112-6

Dickkopf Dickkopf (Dkk)(Dkk)

Kringle Kringle containing containing transmembrane transmembrane protein; protein; KremenKremen(Krm)(Krm)

* * SFRP KO (-/-) muisSFRP KO (-/-) muis::O-blast ontwikkeling en functieO-blast ontwikkeling en functieApoptosis O-blast en O-cyteApoptosis O-blast en O-cyte

* * rat met GIOP:rat met GIOP:SFRP-1 expressie SFRP-1 expressie

Secreted Frizzled Related Protein-1 (sFRP-1) is Secreted Frizzled Related Protein-1 (sFRP-1) is inhibitor of Wnt andinhibitor of Wnt and

Key Regulator Survival O-blast & O-cyteKey Regulator Survival O-blast & O-cyte

Sclerostin and Wnt Signaling Sclerostin and Wnt Signaling

Buchem diseaseBuchem disease &&Sclerostosis met Sclerostosis met mutaties SOST gene, mutaties SOST gene, waardoor tekort aan waardoor tekort aan sclerostin sclerostin toename botvormingtoename botvorming

First Case (1955)First Case (1955)

Antilichamen tegenAntilichamen tegenSclerostin (Scl-Ab)Sclerostin (Scl-Ab)

Phase 1 studyPhase 1 study: : BMD , BFR , BRRBMD , BFR , BRR =

Weekly s.c. antibody to sclerostin elevates BMD Weekly s.c. antibody to sclerostin elevates BMD reversibly in OVX rats (1211 ASBMR 2008)reversibly in OVX rats (1211 ASBMR 2008)

Ab-Scerostin

Piters E et al. Arch Biochem Biophysics 2008;473:112-6

LRP5 in MechanotransductionLRP5 in Mechanotransduction

Sawakami K et al. J Biol Chem 2006;281:23698-711 Sawakami K et al. J Biol Chem 2006;281:23698-711

Anabolic effects PTHAnabolic effects PTH

Khosla S et al. JCI 2008;118:421-8

PTHR1 Signaling in OsteocytesPTHR1 Signaling in Osteocytes

O’Brien CA et al. Plos ONE 2008;3:e 2942

Bij LRP5-/- muisBij LRP5-/- muisgeen effect van geen effect van intermitt. PTH op HBMintermitt. PTH op HBM

Bij LRP5-/- transgene Bij LRP5-/- transgene PTHR1 muis: geen PTHR1 muis: geen verhoogde BM maar verhoogde BM maar wel verhoogde wel verhoogde remodeling dus andereremodeling dus andererouteroute

Bij PTHR1- (gain of Bij PTHR1- (gain of function) muis: function) muis: verhoogde botmassaverhoogde botmassa

Wnt signaling: eating bone or adding it!Wnt signaling: eating bone or adding it!

Goldring SR et al. Nat Med 2007:13:133Goldring SR et al. Nat Med 2007:13:133

Wnt/Wnt/ß-catenin signaling systeem remt ontwikkeling ß-catenin signaling systeem remt ontwikkeling van mesenchymale stamcel tot kraakbeen en vetcelvan mesenchymale stamcel tot kraakbeen en vetcel

Baron R et al. Endocrinology 2008:148:2635-43Baron R et al. Endocrinology 2008:148:2635-43

PPARPPARγγ remt osteoblastgenesis & remt osteoblastgenesis & bevordert osteoclastgenesisbevordert osteoclastgenesis

Takada I et al. IBMSBoneKEy 2008;5:258-61Takada I et al. IBMSBoneKEy 2008;5:258-61

ThiazolidinesThiazolidines

Role for SPPARMS:good for bone and fat?

RANKL in osteoclastogenesis in InflammationRANKL in osteoclastogenesis in Inflammation

Lorenzo J et al. Endocr Rev 2008;29:403-440Lorenzo J et al. Endocr Rev 2008;29:403-440

Wnt?

DKK-1 in Osteoarthritis (OA),Rheumatoid Arthritis DKK-1 in Osteoarthritis (OA),Rheumatoid Arthritis (RA) and Ankylosing Spondylitis (AS)(RA) and Ankylosing Spondylitis (AS)

Diarra D et al. Nat.Med 2007;13:156

Dickkopf-1 is a master regulator of joint remodeling

Effect anti-TNFEffect anti-TNFon DKK-1on DKK-1

Effect disease activityEffect disease activityon DKK-1 on DKK-1 in RA and ASin RA and AS

Synovial tissueSynovial tissue

RANKL & Wnt in Rheumatoid ArthritisRANKL & Wnt in Rheumatoid Arthritis

Diarra D et al. Nat.Med 2007;13:156

Dickkopf-1 is a master regulator of joint remodeling

normaal rheum.arthritis toepassen vantoepassen van

anti-DKK-1:

Role DDK1 Osteolysis in MyelomaRole DDK1 Osteolysis in Myeloma

Pinzone JJ et al. Blood 2008 sept on linePinzone JJ et al. Blood 2008 sept on line

KAHLER:KAHLER:* Bortezomib * Bortezomib remt DDK-1remt DDK-1* Antistoffen * Antistoffen tegen DDK1tegen DDK1 therapeutisch therapeutisch op bot- en op bot- en tumorlesietumorlesie•DenosumabDenosumab (Ab-RANKL)(Ab-RANKL) alleen effectiefalleen effectief op botlesiesop botlesies

Putative model involving Wnt/Putative model involving Wnt/ßcatenin in cancer cells;ßcatenin in cancer cells;Kremen as possible gatekeeperKremen as possible gatekeeper

Nakamura T et al. J Cell Mol Med 2008;12:391-408Nakamura T et al. J Cell Mol Med 2008;12:391-408

Dank voor uw aandacht

Putative model involving Wnt/Putative model involving Wnt/ßcatenin in cancer cellsßcatenin in cancer cells

Nakamura T et al. J Cell Mol Med 2008;12:391-408Nakamura T et al. J Cell Mol Med 2008;12:391-408

Wnt in Prostate CancerWnt in Prostate Cancer

Hall CL et al. JCB 2006;97:661-72Hall CL et al. JCB 2006;97:661-72

• osteolytic phaseosteolytic phase: mediated by RANKL, PTHRP, and : mediated by RANKL, PTHRP, and DKK-1DKK-1• transitional phasetransitional phase: expression Wnts and decrease : expression Wnts and decrease DKK-1DKK-1• osteoblastic phaseosteoblastic phase: VEGF, endothelin-1: VEGF, endothelin-1

Dickkopf-1 [ DDK1]Dickkopf-1 [ DDK1]

• Remmer van Wnt/catenin signaling systeem Remmer van Wnt/catenin signaling systeem

(bindt aan LRPF5) (bindt aan LRPF5) • Ontwikkeling skeletOntwikkeling skelet• Homeostasis skeletHomeostasis skelet• Hematopoeitische stamcel ?Hematopoeitische stamcel ?• Osteolytische metastasering Osteolytische metastasering • Betrokken bij - botverlies door oestrogeentekortBetrokken bij - botverlies door oestrogeentekort

- GIOP - GIOP

- erosieve artritis- erosieve artritis

- teratogene effecten thalidomide- teratogene effecten thalidomide

Role DDK1 in boneRole DDK1 in bone

Pinzone JJ et al. Blood 2008 sept on linePinzone JJ et al. Blood 2008 sept on line

19 Human Wnt Genes19 Human Wnt GenesLink toLink to Gene cards   SOURCE link HUGO link DiseaseDisease

WNT1  WNT1 WNT1  

WNT2  WNT2 WNT2  

WNT2B/13  WNT2B WNT2B  

WNT3   WNT3 Tetra-Amelia (Niemann 2004)

WNT3A   WNT3A  

WNT4     WNT4Mullerian-duct regression and virilization

(Biason-Lauber 2004) SERKAL syndrome (Mandel, 2008)

WNT5A  WNT5A WNT5A  

WNT5B       WNT5BAssociated with Susceptibility to type 2

diabetes (Kanazawa 2004)

WNT6       WNT6    

WNT7A  WNT7A WNT7A Fuhrmann syndrome

WNT7B     WNT7B

WNT8A     WNT8A  

WNT8B  WNT8B WNT8B  

WNT9A (previously WNT14)

 WNT9A WNT9A  

WNT9B Previously WNT15)

  WNT9B    

WNT10A   WNT10AOdonto-onycho-dermal dysplasia Adaimy

, 2007

WNT10B  WNT10B WNT10B

 Mutations in Obesity patients Christodoulides 2006

Split-Hand/Foot Malformation (Ugur 2008)

WNT11  WNT11 WNT11  

WNT16   WNT16

Human Diseases Caused by Abnormality Wnt SignalingHuman Diseases Caused by Abnormality Wnt Signaling

Nakamura T et al. J Cell Mol Med 2008;12:391-408Nakamura T et al. J Cell Mol Med 2008;12:391-408

Different Wnt signaling PathwaysDifferent Wnt signaling Pathways

Piters E et al. Arch Biochem Biophysics 2008;473:112-6

DDK1 & Wnt Signaling in MSC differentiationDDK1 & Wnt Signaling in MSC differentiation

Pinzone JJ et al. Blood 2008 sept on linePinzone JJ et al. Blood 2008 sept on line

Biology of the Osteoclast in Bone MetabolismBiology of the Osteoclast in Bone Metabolism

Deftos LJ. NEJM 2005;353:872-5Deftos LJ. NEJM 2005;353:872-5

DenosDenosuumab

OdanaOdanacatib

Freedom TrialFreedom Trial: : 3jaar bij 7868 PMP 3jaar bij 7868 PMP osteopor. vrouwen osteopor. vrouwen •wervelFx 68%wervelFx 68%•heupFx 40%heupFx 40%•nonVertFX 20%nonVertFX 20%

Wnt & heart diseaseWnt & heart disease• Wound healing response after myocardial infarction in rats and mice Wound healing response after myocardial infarction in rats and mice

((Cardiovascular Research 2002 55(1):16-24)Cardiovascular Research 2002 55(1):16-24)

• Cardiac Hypertrophy and the Wnt/Frizzled Pathway Cardiac Hypertrophy and the Wnt/Frizzled Pathway (Cingolani OH et al. (Hypertension. 2007;49:427.)(Cingolani OH et al. (Hypertension. 2007;49:427.)

• LRP6 mutation in a family with early coronary disease and metabolic LRP6 mutation in a family with early coronary disease and metabolic risk factors. risk factors. (Ani A et al. (Ani A et al. Science. 2007 Mar Science. 2007 Mar 2;315(5816):1278-82 )2;315(5816):1278-82 )

.