mechanisms of resistance to jak inhibitors - imedex, llc · tyrosine kinase inhibitors adapted from...

Mechanisms of resistance to JAK inhibitors

L. Knoops

1 : Resistance to tyrosine kinase inhibition in cancer

MYT1Wee1Wee1B

PIK3R4

SgK493

VRK1VRK2

Bub1BubR1

CK1!2

CK1"

CK1"2

CK1#

CK1$

CK1!1CK1!3

PRPKHaspin

SCYL2SCYL1SCYL3

SgK196

SgK396Slob

VRK3

MSK1

p70S6Kp70S6K%

RSK3

PKC&PKC'

PKN3

RSK4

PKC#PKC(

RSK1/p90RSKRSK2

PKC!

PKC)PKC$

PKC"PKC%

GRK4

GRK5

TLK2

PLK1

PLK4

PLK3

PLK2

MRCK%

MRCK"

GRK6

RHODK/GRK1

AurA/Aur2

TLK1

AurB/Aur1AurC/Aur3

DMPK

ROCK1ROCK2

NDR1NDR2

DMPK2

YANK2YANK3

YANK1

SgK494

PDK1

BARK1/GRK2BARK2/GRK3

GRK7

MAST3MASTL

MAST2

MAST4 MAST1

PKN1/PRK1PKN2/PRK2

MSK2

LATS1LATS2

Akt3/PKB!

SGK2

Akt1/PKB"Akt2/PKB%

SGK1SGK3

CRIK

PKA"PKA%

PRKYPRKX

PKG2PKG1

PKA!

CAMKK1CAMKK2

ULK1

ULK2

BIKE

AAK1

SBK

RSKL2RSKL1

IKK"

IKK%IKK$

TBK1

SgK069SgK110

GAK

MPSK1

MAP2K5

MAP2K7

OSR1STLK3

STLK5STLK6

MKK3/MAP2K3MKK6/MAP2K6SEK1/MAP2K4

TAO3

PAK3

PAK6MEK1/MAP2K1

MEK2/MAP2K2

LOKSLK

COTNIK

GCN2~b

TAO2TAO1

PAK1

PAK2 PAK4PAK5

PBK

CDK8 CDK11

ERK7

ERK3ERK4

CDKL5

GSK3"GSK3% ICK

MAK

CCRK

CDK9

CDK7

NLK

CDKL1CDKL4

CDKL2CDKL3

CHEDCRK7

CDK4CDK6

CDK10

PITSLRE

ERK5

JNK2

CDK5

ERK1

ERK2

JNK1JNK3

p38"p38%

p38#p38!

CDC2/CDK1

PCTAIRE3

PFTAIRE1PFTAIRE2

CDK2 CDK3

PCTAIRE1

PCTAIRE2

CK2"2CK2"1

MOK

SgK071

RNAseL

CLIK1LCLIK1

TTKKIS

IRE1IRE2 TBCK

HRI

GCN2PKRPERK/PEK

CDC7

MAP3K4

KHS1KHS2

NRK/ZC4

MYO3A

MYO3B

MST1MST2

MST4

TNIK/ZC2

HGK/ZC1MINK/ZC3

MAP3K8

MEKK6/MAP3K6

RIPK3

LIMK1LIMK2

TESK1TESK2

ALK1ALK2

TGF%R2

MISR2BMPR2

RIPK2

HH498TAK1ILK

ARAF

KSR KSR2

BMPR1ABMPR1B

ALK7

ActR2ActR2B

ANKRD3 SgK288

ZAK BRAFRAF1

ALK4TGF%R1

DLKLZK

MLK2

MLK4

MLK1

MLK3

LRRK1LRRK2

SgK496

RIPK1

WNK3WNK1

WNK2NRBP1NRBP2

WNK4MEKK1/MAP3K1

MEKK2/MAP3K2MEKK3/MAP3K3

ASK/MAP3K5MAP3K7

MLKL

SgK307SgK424

IRAK4

HSER

CYGD

CYGF

IRAK2

ANP"

ANP%

IRAK1IRAK3

ULK3

ULK4Fused

MELK

NIM1SNRK

SSTK

TSSK3

TSSK1

AMPK"1AMPK"2

BRSK1BRSK2

NuaK1NuaK2

MARK4

QSK

MARK3

QIKSIK

MARK1MARK2

CaMK1%

CaMK1!

RSK3~b

CHK2

PKD2

HUNK

DCAMKL3

CASK

MAPKAPK5VACAMKL

PSKH1PSKH2

MAPKAPK2MAPKAPK3

MSK1~bMSK2~b

RSK4~b

TSSK2

TSSK4

Nek6Nek7

Nek2

Nek8Nek9

Nek11

Nek4

Nek3

Nek1

Nek5

Nek10

STK33

MNK1MNK2PhK!1

PhK!2

PKD3 DCAMKL1DCAMKL2

CaMK4

CaMK2"CaMK2#

CaMK2%CaMK2!

RSK1~b

RSK2~b

PKD1

Trb1

Trb3

Trb2

SgK495

DRAK1

DAPK1DAPK2

caMLCK

SgK085

DAPK3

DRAK2skMLCK

smMLCK

TTN

TradTrio

Obscn

SPEG

Obscn~bSPEG~b

Pim1

Pim3Pim2

CHK1

PASKLkb1

SuRTK106JAK1~bTyk2~b

JAK2~bJAK3~b

MOS

Lmr1Lmr2

Lmr3

EphA2

Etk/BMXBTK

ITK

BLK

Fgr

EphA8

TECTXK

Lck

FynHCKLyn

Src

Yes

EphA7

EphB4

EphA6

EphB3

EphA4

EphB1

EphB2

EphA3

EphA5

EphA10

EphB6

Fer

FRK

BrkSrm

EphA1

AblArg

CSKCTK

FLT4

CCK4/PTK7

PDGFR"PDGFR%

AxlMer

FGFR1FGFR4

FLT1KDR Fms/CSFR

Kit

ALKLTK

IGF1RInsR

DDR1DDR2

MuSK

TRKA

FGFR2 FGFR3

TRKB

TRKC

EGFR HER2

Tyro3/Sky

MetRon

Ret

FLT3

Ros

IRR

ROR1ROR2

Tnk1HER3 JAK3

PYK2ZAP70

RYKTIE1 HER4

Ack JAK1Tyk2

JAK2

Syk

TIE2

FAK

Fes

HPK1

GCKMST3

YSK1

SgK223SgK269

PINK1

TTBK1TTBK2

CaMK1"CaMK1#

PRP4

SRPK2

HIPK4

CLK3

CLK4

CLK2CLK1

MSSK1

SRPK1

DYRK1ADYRK1B

DYRK4

HIPK3

DYRK2DYRK3

HIPK1HIPK2

TKLTK

STE

CK1

CMGC

AGC

CAMK

THE

KinomeKinomeHuman

Authors Gerard Manning, David B. Whyte, Ricardo Martinez, Sucha Sudarsanam, Sugen Inc., South San Francisco, CA, USA; Tony Hunter, Salk Institute, La Jolla, CA, USA. This poster accompanies the paper “The Protein Kinase Complement of the Human Genome” (Manning et al., Science, 6 December 2002).Science coordinator L. Bryan RayDesign and production Anne Ashley, David Comb, Michael Melnick, C. Faber Smith, J. White, David M. Tompkins, Marcus SpieglerCopyeditor Harry Jach Sponsored by Cell Signaling Technology, Inc. and Sugen, Inc.

© 2002 Science, a publication of the American Association for the Advancement of Science

ABC1

Alpha

Brd

PDHK

PIKK

RIO

AlphaK1AlphaK2

ChaK2ChaK1

EEF2KAlphaK3

Brd2Brd3

Brd4BrdT

PDHK2PDHK3PDHK1

PDHK4BCKDK

RIOK3RIOK1

RIOK2

ATMATR

SMG1

FRAPDNAPK

TRRAP

TIF1TIF1%

TIF1!TIF1"

ADCK1ADCK5

ADCK3ADCK4ADCK2

ATYPICAL PROTEIN KINASES

� his phylogenetic tree depicts the relationships between members of the complete �� superfamily of human protein kinases. Protein kinases constitute one of the largest �� human gene families and are key regulators of cell function. The 518 human protein kinases control protein activity by catalyzing the addition of a negatively charged phosphate group to other proteins. Protein kinases modulate a wide variety of biological processes, especially those that carry signals from the cell membrane to intracellular targets and coordinate complex biological functions. � Most protein kinases belong to a single superfamily of enzymes whose catalytic domains are related in sequence and structure. The main diagram illustrates the similarity between the protein sequences of these catalytic domains. Each kinase is at the tip of a branch, and the similarity between various kinases is inversely related to the distance between their positions on the tree diagram. Most kinases fall into small families of highly related sequences, and most

families are part of larger groups. The seven major groups are labeled and colored distinctly. Other kinases are shown in the center of the tree, colored gray. The relationships shown on the tree can be used to predict protein substrates and biological function for many of the over 100 uncharacterized kinases presented here. � The inset diagram shows trees for seven atypical protein kinase families. These proteins have verified or strongly predicted kinase activity, but have little or no sequence similarity to members of the protein kinase superfamily. A further eight atypical protein kinases in small families of one or two genes are not shown.

T

Mapping ProceduresThe main dendrogram shows the sequence similarity between protein kinase

domains, derived from public sequences and gene-prediction methods detailed in

Manning et al. (Science, 6 December 2002). Domains were defined by hidden

Markov model profile analysis and multiple sequence alignment. The initial

branching pattern was built from a neighbor-joining tree derived from a ClustalW

protein sequence alignment of the domains. This was extensively modified by

reference to other alignment and tree-building methods (hmmalign and parsimony

trees) and by extensive pairwise sequence alignment of kinase domains. The

curved layout was created manually. Many branch lengths are semiquantitative, but

the branching pattern is more informative than any single automatic method. The

atypical kinase trees were generated automatically by ClustalW alignment of full-

length protein sequences followed by neighbor-joining tree building. Unpublished

kinases are named where possible according to family nomenclature. Some

divergent kinases retain a numerical SgK (Sugen kinase) accession number. The

second domains of dual-domain kinases are named with a “~b” suffix. Detailed

subtrees and sequence alignments of individual groups and families, and

comparative genomic trees are available at http://www.kinase.com. Information on

regulation and substrates of many of these kinases is available at

http://www.cellsignal.com.

Group namesAGC Containing PKA, PKG, PKC families; CAMK Calcium/calmodulin-dependent protein kinase; CK1 Casein kinase 1; CMGC Containing CDK, MAPK, GSK3, CLK families; STE Homologs of yeast Sterile 7, Sterile 11, Sterile 20 kinases; TK Tyrosine kinase; TKL Tyrosine kinase–like.

Kinase names(A selective list includes those cases in which the full name is more informative than the abbreviation or acronym shown on the tree. Other full names and synonyms are available at http://www.kinase.com.) ActR Activin receptor; ALK (TK group) Anaplastic lymphoma kinase; ALK (TKL group) Activin-like receptor kinase; AMPK Adenosine monophosphate–activated protein kinase; Aur Aurora; BARK !-adrenergic receptor kinase; BLK B lymphocyte tyrosine kinase; BMPR Bone morphogeneic protein receptor; BMX Bone marrow tyrosine kinase gene in chromosome X; BRD Bromodomain kinase; BRSK Brain-selective kinase; CaMK Calcium/calmodulin-dependent protein kinase; CAMKK CaMK kinase; CCK-4 Colon carcinoma kinase–4; CDK Cyclin-dependent kinase; CDKL Cyclin-dependent kinase–like; CK Casein kinase; CLK Cdc2-like kinase; CSFR Colony-stimulating factor receptor; DAPK Death-associated protein kinase; DCAMKL Doublecortin- and CaMK-like; DDR Discoidin domain receptor; DMPK Dystrophia myotonica protein kinase; DNAPK DNA-activated protein kinase; DRAK DAPK-related apoptosis-inducing kinase; DYRK Dual-specificity tyrosine phosphorylation–regulated kinase; EEF2K Eukaryotic elongation factor–2 kinase; EGFR Epidermal growth factor receptor; Eph Ephrin receptor; ERK Extracellular signal–regulated kinase; FAK Focal adhesion kinase; FGFR Fibroblast growth factor receptor; FRK Fos-regulatory kinase; GRK G protein–coupled receptor kinase; GSK Glycogen synthase kinase; HIPK Homeodomain-interacting protein kinase; IKK I-"B kinase; ILK Integrin-linked kinase; InsR Insulin receptor; IRAK Interleukin-1 receptor–associated kinase; IRE Inositol-requiring; IRR Insulin receptor–related; JAK Janus kinase; JNK c-Jun NH2-terminal kinase; KSR Kinase suppressor of Ras; LATS Large tumor suppressor; LIMK Lim domain–containing kinase; LMR Lemur kinase; LRRK Leucine rich–repeat kinase; MAP2K Mitogen-activated protein kinase kinase; MAP3K Mitogen-activated protein kinase kinase kinase; MAPK Mitogen-activated protein kinase; MAPKAPK MAPK–activated protein kinase; MARK Microtubule-associated protein/microtubule affinity–regulating kinase; MAST Microtubule-associated serine-threonine kinase; MLCK Myosin light chain kinase; MLK Mixed lineage kinase; MNK MAPK-interacting kinase; MRCK Myotonic dystrophy–related CDC42-binding kinase; MSK Mitogen- and stress-activated protein kinase; MuSK Muscle-specific kinase; NDR Nuclear, Dbf2-related kinase; NIK Nuclear factor "B–inducing kinase; PAK p21-activated kinase; PDGFR Platelet-derived growth factor receptor; PDHK Pyruvate dehydrogenase kinase; PDK Phosphoinositide-dependent kinase; PhK Phosphorylase kinase; PIKK Phosphatidylinositol 3-kinase–related kinase; PKA Protein kinase A; PKB Protein kinase B; PKC Protein kinase C; PKD Protein kinase D; PKG Protein kinase G; PKN Protein kinase N; PKR Protein kinase, double-stranded RNA–dependent; PRK Protein kinase C–related kinase; PSKH Protein serine kinase H; RIPK Receptor-interacting protein kinase; ROCK Rho-associated, coiled-coil–containing kinase; ROR Regeneron orphan receptor; RSK Ribosomal protein S6 kinase; RSKL RSK-like; SgK Sugen kinase; SGK Serum- and glucocorticoid-regulated kinase; SRPK Serine-arginine splicing factor protein kinase; SYK Spleen tyrosine kinase; TAK Transforming growth factor–!–activated kinase; TEC Tyrosine kinase expressed in hepatocellular carcinoma; TESK Testis-specific kinase; TGF%R Transforming growth factor–! receptor; TIE Tyrosine kinase with immunoglobulin and EGF repeats; TIF1 Transcriptional intermediary factor 1; TLK Tousled-like kinase; TSSK Testis-specific serine

SIGNAL TRANSDUCTIONKNOWLEDGE ENVIRONMENT

WWW.STKE.ORG


WWW.STKE.ORG

Tyrosine kinases

www.stke.org

Tyrosine Kinase

inhibitors

Tyrosine Kinase

inhibitors

Tyrosine kinase inhibitors

Adapted from Nagar et al, Cancer Res, 2002 cABL kinase domain cristal strucure with imatinib

Tyrosine kinase inhibitors in cancer

time TKI

Mut

ant

Clon

e W

T

Resistance

Resistant clone

MYT1Wee1Wee1B

PIK3R4

SgK493

VRK1VRK2

Bub1BubR1

CK1!2

CK1"

CK1"2

CK1#

CK1$

CK1!1CK1!3

PRPKHaspin

SCYL2SCYL1SCYL3

SgK196

SgK396Slob

VRK3

MSK1

p70S6Kp70S6K%

RSK3

PKC&PKC'

PKN3

RSK4

PKC#PKC(

RSK1/p90RSKRSK2

PKC!

PKC)PKC$

PKC"PKC%

GRK4

GRK5

TLK2

PLK1

PLK4

PLK3

PLK2

MRCK%

MRCK"

GRK6

RHODK/GRK1

AurA/Aur2

TLK1

AurB/Aur1AurC/Aur3

DMPK

ROCK1ROCK2

NDR1NDR2

DMPK2

YANK2YANK3

YANK1

SgK494

PDK1

BARK1/GRK2BARK2/GRK3

GRK7

MAST3MASTL

MAST2

MAST4 MAST1

PKN1/PRK1PKN2/PRK2

MSK2

LATS1LATS2

Akt3/PKB!

SGK2

Akt1/PKB"Akt2/PKB%

SGK1SGK3

CRIK

PKA"PKA%

PRKYPRKX

PKG2PKG1

PKA!

CAMKK1CAMKK2

ULK1

ULK2

BIKE

AAK1

SBK

RSKL2RSKL1

IKK"

IKK%IKK$

TBK1

SgK069SgK110

GAK

MPSK1

MAP2K5

MAP2K7

OSR1STLK3

STLK5STLK6

MKK3/MAP2K3MKK6/MAP2K6SEK1/MAP2K4

TAO3

PAK3

PAK6MEK1/MAP2K1

MEK2/MAP2K2

LOKSLK

COTNIK

GCN2~b

TAO2TAO1

PAK1

PAK2 PAK4PAK5

PBK

CDK8 CDK11

ERK7

ERK3ERK4

CDKL5

GSK3"GSK3% ICK

MAK

CCRK

CDK9

CDK7

NLK

CDKL1CDKL4

CDKL2CDKL3

CHEDCRK7

CDK4CDK6

CDK10

PITSLRE

ERK5

JNK2

CDK5

ERK1

ERK2

JNK1JNK3

p38"p38%

p38#p38!

CDC2/CDK1

PCTAIRE3

PFTAIRE1PFTAIRE2

CDK2 CDK3

PCTAIRE1

PCTAIRE2

CK2"2CK2"1

MOK

SgK071

RNAseL

CLIK1LCLIK1

TTKKIS

IRE1IRE2 TBCK

HRI

GCN2PKRPERK/PEK

CDC7

MAP3K4

KHS1KHS2

NRK/ZC4

MYO3A

MYO3B

MST1MST2

MST4

TNIK/ZC2

HGK/ZC1MINK/ZC3

MAP3K8

MEKK6/MAP3K6

RIPK3

LIMK1LIMK2

TESK1TESK2

ALK1ALK2

TGF%R2

MISR2BMPR2

RIPK2

HH498TAK1ILK

ARAF

KSR KSR2

BMPR1ABMPR1B

ALK7

ActR2ActR2B

ANKRD3 SgK288

ZAK BRAFRAF1

ALK4TGF%R1

DLKLZK

MLK2

MLK4

MLK1

MLK3

LRRK1LRRK2

SgK496

RIPK1

WNK3WNK1

WNK2NRBP1NRBP2

WNK4MEKK1/MAP3K1

MEKK2/MAP3K2MEKK3/MAP3K3

ASK/MAP3K5MAP3K7

MLKL

SgK307SgK424

IRAK4

HSER

CYGD

CYGF

IRAK2

ANP"

ANP%

IRAK1IRAK3

ULK3

ULK4Fused

MELK

NIM1SNRK

SSTK

TSSK3

TSSK1

AMPK"1AMPK"2

BRSK1BRSK2

NuaK1NuaK2

MARK4

QSK

MARK3

QIKSIK

MARK1MARK2

CaMK1%

CaMK1!

RSK3~b

CHK2

PKD2

HUNK

DCAMKL3

CASK

MAPKAPK5VACAMKL

PSKH1PSKH2

MAPKAPK2MAPKAPK3

MSK1~bMSK2~b

RSK4~b

TSSK2

TSSK4

Nek6Nek7

Nek2

Nek8Nek9

Nek11

Nek4

Nek3

Nek1

Nek5

Nek10

STK33

MNK1MNK2PhK!1

PhK!2

PKD3 DCAMKL1DCAMKL2

CaMK4

CaMK2"CaMK2#

CaMK2%CaMK2!

RSK1~b

RSK2~b

PKD1

Trb1

Trb3

Trb2

SgK495

DRAK1

DAPK1DAPK2

caMLCK

SgK085

DAPK3

DRAK2skMLCK

smMLCK

TTN

TradTrio

Obscn

SPEG

Obscn~bSPEG~b

Pim1

Pim3Pim2

CHK1

PASKLkb1

SuRTK106JAK1~bTyk2~b

JAK2~bJAK3~b

MOS

Lmr1Lmr2

Lmr3

EphA2

Etk/BMXBTK

ITK

BLK

Fgr

EphA8

TECTXK

Lck

FynHCKLyn

Src

Yes

EphA7

EphB4

EphA6

EphB3

EphA4

EphB1

EphB2

EphA3

EphA5

EphA10

EphB6

Fer

FRK

BrkSrm

EphA1

AblArg

CSKCTK

FLT4

CCK4/PTK7

PDGFR"PDGFR%

AxlMer

FGFR1FGFR4

FLT1KDR Fms/CSFR

Kit

ALKLTK

IGF1RInsR

DDR1DDR2

MuSK

TRKA

FGFR2 FGFR3

TRKB

TRKC

EGFR HER2

Tyro3/Sky

MetRon

Ret

FLT3

Ros

IRR

ROR1ROR2

Tnk1HER3 JAK3

PYK2ZAP70

RYKTIE1 HER4

Ack JAK1Tyk2

JAK2

Syk

TIE2

FAK

Fes

HPK1

GCKMST3

YSK1

SgK223SgK269

PINK1

TTBK1TTBK2

CaMK1"CaMK1#

PRP4

SRPK2

HIPK4

CLK3

CLK4

CLK2CLK1

MSSK1

SRPK1

DYRK1ADYRK1B

DYRK4

HIPK3

DYRK2DYRK3

HIPK1HIPK2

TKLTK

STE

CK1

CMGC

AGC

CAMK

THE

KinomeKinomeHuman

Authors Gerard Manning, David B. Whyte, Ricardo Martinez, Sucha Sudarsanam, Sugen Inc., South San Francisco, CA, USA; Tony Hunter, Salk Institute, La Jolla, CA, USA. This poster accompanies the paper “The Protein Kinase Complement of the Human Genome” (Manning et al., Science, 6 December 2002).Science coordinator L. Bryan RayDesign and production Anne Ashley, David Comb, Michael Melnick, C. Faber Smith, J. White, David M. Tompkins, Marcus SpieglerCopyeditor Harry Jach Sponsored by Cell Signaling Technology, Inc. and Sugen, Inc.

© 2002 Science, a publication of the American Association for the Advancement of Science

ABC1

Alpha

Brd

PDHK

PIKK

RIO

AlphaK1AlphaK2

ChaK2ChaK1

EEF2KAlphaK3

Brd2Brd3

Brd4BrdT

PDHK2PDHK3PDHK1

PDHK4BCKDK

RIOK3RIOK1

RIOK2

ATMATR

SMG1

FRAPDNAPK

TRRAP

TIF1TIF1%

TIF1!TIF1"

ADCK1ADCK5

ADCK3ADCK4ADCK2

ATYPICAL PROTEIN KINASES

� his phylogenetic tree depicts the relationships between members of the complete �� superfamily of human protein kinases. Protein kinases constitute one of the largest �� human gene families and are key regulators of cell function. The 518 human protein kinases control protein activity by catalyzing the addition of a negatively charged phosphate group to other proteins. Protein kinases modulate a wide variety of biological processes, especially those that carry signals from the cell membrane to intracellular targets and coordinate complex biological functions. � Most protein kinases belong to a single superfamily of enzymes whose catalytic domains are related in sequence and structure. The main diagram illustrates the similarity between the protein sequences of these catalytic domains. Each kinase is at the tip of a branch, and the similarity between various kinases is inversely related to the distance between their positions on the tree diagram. Most kinases fall into small families of highly related sequences, and most

families are part of larger groups. The seven major groups are labeled and colored distinctly. Other kinases are shown in the center of the tree, colored gray. The relationships shown on the tree can be used to predict protein substrates and biological function for many of the over 100 uncharacterized kinases presented here. � The inset diagram shows trees for seven atypical protein kinase families. These proteins have verified or strongly predicted kinase activity, but have little or no sequence similarity to members of the protein kinase superfamily. A further eight atypical protein kinases in small families of one or two genes are not shown.

T

Mapping ProceduresThe main dendrogram shows the sequence similarity between protein kinase

domains, derived from public sequences and gene-prediction methods detailed in

Manning et al. (Science, 6 December 2002). Domains were defined by hidden

Markov model profile analysis and multiple sequence alignment. The initial

branching pattern was built from a neighbor-joining tree derived from a ClustalW

protein sequence alignment of the domains. This was extensively modified by

reference to other alignment and tree-building methods (hmmalign and parsimony

trees) and by extensive pairwise sequence alignment of kinase domains. The

curved layout was created manually. Many branch lengths are semiquantitative, but

the branching pattern is more informative than any single automatic method. The

atypical kinase trees were generated automatically by ClustalW alignment of full-

length protein sequences followed by neighbor-joining tree building. Unpublished

kinases are named where possible according to family nomenclature. Some

divergent kinases retain a numerical SgK (Sugen kinase) accession number. The

second domains of dual-domain kinases are named with a “~b” suffix. Detailed

subtrees and sequence alignments of individual groups and families, and

comparative genomic trees are available at http://www.kinase.com. Information on

regulation and substrates of many of these kinases is available at

http://www.cellsignal.com.

Group namesAGC Containing PKA, PKG, PKC families; CAMK Calcium/calmodulin-dependent protein kinase; CK1 Casein kinase 1; CMGC Containing CDK, MAPK, GSK3, CLK families; STE Homologs of yeast Sterile 7, Sterile 11, Sterile 20 kinases; TK Tyrosine kinase; TKL Tyrosine kinase–like.

Kinase names(A selective list includes those cases in which the full name is more informative than the abbreviation or acronym shown on the tree. Other full names and synonyms are available at http://www.kinase.com.) ActR Activin receptor; ALK (TK group) Anaplastic lymphoma kinase; ALK (TKL group) Activin-like receptor kinase; AMPK Adenosine monophosphate–activated protein kinase; Aur Aurora; BARK !-adrenergic receptor kinase; BLK B lymphocyte tyrosine kinase; BMPR Bone morphogeneic protein receptor; BMX Bone marrow tyrosine kinase gene in chromosome X; BRD Bromodomain kinase; BRSK Brain-selective kinase; CaMK Calcium/calmodulin-dependent protein kinase; CAMKK CaMK kinase; CCK-4 Colon carcinoma kinase–4; CDK Cyclin-dependent kinase; CDKL Cyclin-dependent kinase–like; CK Casein kinase; CLK Cdc2-like kinase; CSFR Colony-stimulating factor receptor; DAPK Death-associated protein kinase; DCAMKL Doublecortin- and CaMK-like; DDR Discoidin domain receptor; DMPK Dystrophia myotonica protein kinase; DNAPK DNA-activated protein kinase; DRAK DAPK-related apoptosis-inducing kinase; DYRK Dual-specificity tyrosine phosphorylation–regulated kinase; EEF2K Eukaryotic elongation factor–2 kinase; EGFR Epidermal growth factor receptor; Eph Ephrin receptor; ERK Extracellular signal–regulated kinase; FAK Focal adhesion kinase; FGFR Fibroblast growth factor receptor; FRK Fos-regulatory kinase; GRK G protein–coupled receptor kinase; GSK Glycogen synthase kinase; HIPK Homeodomain-interacting protein kinase; IKK I-"B kinase; ILK Integrin-linked kinase; InsR Insulin receptor; IRAK Interleukin-1 receptor–associated kinase; IRE Inositol-requiring; IRR Insulin receptor–related; JAK Janus kinase; JNK c-Jun NH2-terminal kinase; KSR Kinase suppressor of Ras; LATS Large tumor suppressor; LIMK Lim domain–containing kinase; LMR Lemur kinase; LRRK Leucine rich–repeat kinase; MAP2K Mitogen-activated protein kinase kinase; MAP3K Mitogen-activated protein kinase kinase kinase; MAPK Mitogen-activated protein kinase; MAPKAPK MAPK–activated protein kinase; MARK Microtubule-associated protein/microtubule affinity–regulating kinase; MAST Microtubule-associated serine-threonine kinase; MLCK Myosin light chain kinase; MLK Mixed lineage kinase; MNK MAPK-interacting kinase; MRCK Myotonic dystrophy–related CDC42-binding kinase; MSK Mitogen- and stress-activated protein kinase; MuSK Muscle-specific kinase; NDR Nuclear, Dbf2-related kinase; NIK Nuclear factor "B–inducing kinase; PAK p21-activated kinase; PDGFR Platelet-derived growth factor receptor; PDHK Pyruvate dehydrogenase kinase; PDK Phosphoinositide-dependent kinase; PhK Phosphorylase kinase; PIKK Phosphatidylinositol 3-kinase–related kinase; PKA Protein kinase A; PKB Protein kinase B; PKC Protein kinase C; PKD Protein kinase D; PKG Protein kinase G; PKN Protein kinase N; PKR Protein kinase, double-stranded RNA–dependent; PRK Protein kinase C–related kinase; PSKH Protein serine kinase H; RIPK Receptor-interacting protein kinase; ROCK Rho-associated, coiled-coil–containing kinase; ROR Regeneron orphan receptor; RSK Ribosomal protein S6 kinase; RSKL RSK-like; SgK Sugen kinase; SGK Serum- and glucocorticoid-regulated kinase; SRPK Serine-arginine splicing factor protein kinase; SYK Spleen tyrosine kinase; TAK Transforming growth factor–!–activated kinase; TEC Tyrosine kinase expressed in hepatocellular carcinoma; TESK Testis-specific kinase; TGF%R Transforming growth factor–! receptor; TIE Tyrosine kinase with immunoglobulin and EGF repeats; TIF1 Transcriptional intermediary factor 1; TLK Tousled-like kinase; TSSK Testis-specific serine


WWW.STKE.ORG


WWW.STKE.ORG

Tyrosine kinases

Tyrosine Kinase

inhibitors

www.stke.org

Tyrosine Kinase

inhibitors

Resistance ?

JAK2 kinase domain

ABL kinase domain JAK2 kinase domain

cABL kinase domain cristal strucure with imatinib JAK2 kinase domain cristal structure with CP690 550

Adapted from Nagar et al, Cancer Res, 2002;Williams et al, J Mol Biol, 2009

Adapted from Giordanette F et al, Protein Eng, 2002

N C

FERM

PSEUDO- KINASE DOMAIN

KINASE DOMAIN

V617F

JAK Kinases

JAK2 V617F model

JAK2 JH1-JH2 domains model Adapted from Giordanetto et al, Protein Eng, 2002

P!Y

Y

Y

Y

Y

Y

Y

Y

Y

JAK JAK JAK Y

Y

Y

JAK Y

Y

Y

JAK Y

Y

Y

JAK P! P!

P!

P!

P!

P!

P!

STAT Y P! STAT Y P!

Cytokine

STAT Y P! STAT Y P!

Cytokine

Cytokine

Cytokine receptors signal transduction

JAK-STAT

Y

Y

Y

Y

P!

P!

P!

P!

STAT Y P! STAT Y P!

STAT Y P! STAT Y P!

RAS Y JAK Y JAK P! P!

Cytokine

JAK-STAT

MAPKinase

PI3Kinase


Y

Y

Y

JAK Y

Y

Y

JAK P!

P!

P!

P!

P!

P!

Cytokine


≈ 50 cytokine receptors

4 JAKs

Homo or heterodimers

JAK2 JAK2

ßc

ßc

IL-3, 5, GM-CSF

STAT5

JAK1 JAK2

Type II IFN

IFNγ

STAT1

JAK2

Homodimeric

EPO, TPO, GH, PRL

STAT5

JAK2

Cytokine receptors using JAK2

JAK1 JAK3

γc

IL-2, 4, 7, 9, 15, 21

STAT5 STAT3 STAT6

γc

JAK1 TYK2

gp130

IL-6, G-CSF, OSM

STAT3

gp130

JAK1 TYK2

Type I IFN

IFNa, ß

STAT1 STAT2

JAK1 JAK2

Type II IFN

IFNγ

STAT1

Cytokine receptors using JAK1


- JAK inhibitors are ATP-competitive inhibitors - JAK inhibitors are not imatinib! . Not specific for mutated vs WT JAKs . JAK inhibition will interfere with many cytokine receptors

2 : In vitro mechanisms of JAK inhibitor resistance

IL-‐9R signal transduc3on IL-9 R signal transduction

IL-9 R phe116 receptor

BaF3 phe116

IL-9

IL-9 responding

IL-9

Autonomous

/ /

JAK1 mutations

JAK1 overexpression

In vitro model of tumorigenesis

JAK1 activating mutations

JAK1 JH1-JH2 domains model


Mutated residues


V658F, L, I


Mutated residues



Mutated residues

Mutated residues in ALL

JAK1 JH1-JH2 domains model Flex et al., 2008, Asnafi et al., 2008, Jeong et al., 2008, Mullighan et al., 2009

JAK1 activating mutations : kinase domain

JAK1 kinase domain cristal structure with CP690 550 Adapted from Williams et al., J.Mol.Biol, 2009

JAK1 F958V is resistant to JAK inhibitor I

Western Blot Hornakova et al, Haematologica, 2011

BaF3 cells - H3 thymidine incorporation

JAK1 F958V is resistant to Ruxolitinib

Hornakova et al, Haematologica, 2011

0

20

40

60

80

100

120

0.001 0.01 0.1 1

Ruxolitinib (µM)

Prol

ifer

atio

n (%

of

cont

rol) M-RAS

Aut(F958V)

Aut(V658F)

Prolonged treatment with JAK inhibitor induces JAK1 overexpression

Quantitative PCR Hornakova et al, Haematologica, 2011

JAK1 and ABL1 hinge regions

Crystal structure of JAK1, JAK2 and ABL

P960T/S F958V/C/S/L (JAK inhibitors)

JAK1 925 KLIMEFLPSGSLKEYLPK-NKN 973 .:|.||:..|:|.:||.: |:.

ABL1 312 YIITEFMTYGNLLDYLRECNRQ 333 T315I/A/S (Imatinib) E316D F317L/I M318A G321V

Hornakova et al, Haematologica, 2011

0

20

40

60

80

100

120

0.001 0.01 0.1 1 10

Ruxolitinib (uM)

Prol

ifer

atio

n (in

% o

f co

ntro

l)

The JAK2 Y931C mutation induces resistance to ruxolitinib

BaF3 cells - H3 thymidine incorporation Hornakova et al, Haematologica, 2011

V617F

Y931C

V617F/Y931C

JAK2 kinase domain mutations resistant to JAK inhibitors

JAK2 kinase domain cristal structure with CP690 550 Adapted from Williams et al., J.Mol.Biol, 2009

Activating and resistant mutations

Resistant mutations

Y931C

G935R

E864A

Hornakova et al, Haematologica, 2011 Weigert et al, J Exp Med, 2012



2 : In vitro mechanisms of JAK inhibitor resistance - JAK inhibitors are able to kill JAK-dependent cells - JAK kinase domain mutation and overexpression can explain resistance

3 : Clinical resistance to JAK inhibitors in MF

% Cha

nge from

baseline at week 48

60

40

20

0

-‐20

-‐40

-‐60

-‐80

Primary endpoint

Ruxoli3nib BAT

80

Ruxoli3nib P < .0001

At Week 48

COMFORT-II : Percent change from baseline in spleen volume

Harrison C, ASH 2011, Abstract 279

Patient characteristics ?





- Clinical resistance exists – Mainly primary resistance (short follow-up)

4 : Molecular ‘resistance’ to JAK inhibitors in MF

Tyrosine kinase inhibitors in cancer

time TKI Resistance

Mut

ant

Clon

e W

T Resistant clone

JAK kinases inhibitors in PMF

time TKI

JAK2

V61

7F +

W

T JA

K2 V617F + W

T

Potential cause of JAK2 V617F persistance

0

20

40

60

80

100

120

0.001 0.01 0.1 1 10

Ruxolitinib (uM)

Prol

ifer

atio

n (in

% o

f co

ntro

l)

JAK2 V617F

M Ras

Is the JAK2 V617F MPN clone addicted to JAK-STAT signaling?

JAK kinases inhibitors in PMF

time TKI

JAK2

V61

7F +

W

T JA

K2 V617F + W

T

Potential cause of JAK2 V617F persistance

0

20

40

60

80

100

120

0.001 0.01 0.1 1 10

Ruxolitinib (uM)

Prol

ifer

atio

n (in

% o

f co

ntro

l)

JAK2 V617F

M Ras

Toxic dose JAK2 V617F persistance

Normal hematopoiesis







- No JAK kinase signaling addiction vs ‘effective’ dose too toxic

time TKI

JAK2

V61

7F +

W

T W

T

Better inhibitors ?

C. Hermans E. Van den neste L. Michaux M.C. Vekemans V. Havelange X. Poiré C. Lambert A. Ferrant

J.C. Renauld T. Hornakova L. Springuel A. Saumet

SN Constantinescu

Thanks

mechanisms of resistance to jak inhibitors - imedex, llc · tyrosine kinase inhibitors adapted from...

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