chemokines & growth factors

8/8/2019 Chemokines & Growth Factors

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chemokines( diapedesis) (60~ 70 aa )

: ( Cys) 40 : chemokines receptors G actin

lamellipodia integrin

histamine allergic inflammation, lymphocyte homing, lymphoid organdevelopment, lymphoid organ homostasis, angiogenesis


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Chemokiney 19 ( leukocytes)

diapedesis Elias Metschnikoff ( 1908 ( engulf )

diapedesis ( ) ( host defense)

y Chemokines actinlamellipodia integrin

histamineeotaxin lymphocyte homing, lymphoid

organ development, lymphoid organ homostasis, angiogenesis )

y

classical chemoattractants chemokines

y

1. IL-8 was the key neutrophil -chemoattractant factor,2 . CC L2 (MCP -1 ) was the key monocyte attractant, and 3. CC L11 (eotaxin) was identified later as the major eosinophil attractant .


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Chemokines


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Chemokine receptor(1) ligand (yellow); (2) ligand r edundant or shar ed (r ed); (3) ligand orp han - r ecep tor s (p urp le); (4) chemokine non-signaling- -r ecep tor s (blue). chemokine r ecep tor ligand( ).

chemokine r ecep tor antagonists ( ); (5) r egulated byvMIP-II(r epr esented in a smaller f ont size to indicate those r ecep tor s atwhich it exhibits only p ar tial antagonism). Vir ally encoded chemokines that ar e agonists at indicated

r ecep tor s ( gr een).A bbr eviations:y BC A-1/ CXCL 13, B-cell-attr actingchemokine 1; y CT ACK / CCL 27, cutaneous T -cell attr acting chemokine; y DARC , Duff y antigen r ecep tor f or chemokines; y ELC / CCL 19, Ep stein B arr vir us-induced r ecep tor ligandchemokine; y ENA78/ CXCL 5, ep ithelial cell-der ived neutr ohil-activating factor 78;y GC P-2/ CXCL6 , gr anulocyte chemoattr actant pr otein 2; y HCC -1/ CCL 14, hemof iltr ate CC chemokine 1; y HC R, human chemokine r ecep tor ; y IL -8/ CXCL 8, inter leukin 8;y IP-10/ CXCL 10, inter f er on-inducible pr otein 10; y I-T AC / CXCL 11, inter f er on-inducible T -cell a chemoattr actant; y L ARC / CCL 20, liver and activation-r elated chemokine;y MC P-1/ CCL 2, monocyte chemoattr actant pr otein 1; y MDC / CCL 22, macr op hage-der ived chemokine; MIG/ CXCL 9, y monocyte-induced by K-inter f er on; y MIPF-1/ CCL 23, myeloid pr ogenitor inhibitor y factor 1; y NAP-2/ CXCL 7, neutr op hil-activating p ep tide 2; y RANT ES/ CCL 5, r egulation on activation of nor mal T cells expr essed and secr eted;y SDF-1/ CXCL 12, str omal cell-der ived factor 1; y SLC / CCL 21, second lymp hoid tissue chemokine; y T ARC / CCL 17, T cell and activation-r elated chemokine; y

T ECK / CCL 25, thymus expr essed chemokine; y vMIP-II, vir ally encoded macr op hage inf lammator y pr otein II.


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Chemokines


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Chemokine receptor


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Chemokiney classical chemoattractants : fM LP ( N -formyl -methionyl -leucyl - phenylalanine, f -Met -leu - phe),

C5a, leukotriene B4 PAF ( platelet -activating factor) chemoattractants phagocytic cells neutrophils, monocyteclassical chemoattractants innate immunity

y neutrophil chemoattractants IL-8 1978 40 18 ( 60 -130 aa. ) ,

chemokines

y Chemokine chemoattractant cytokine199 2 I nternational Symposium of Chemotactic Cytokines

Chemokine classical chemoattractant chemokine phagocytic cells dendritic cells T

lymphocytes stromal cell progenitor B cell chemotaxisI nnate nonspecific immunity phagocytic cells Adaptive

acquired, specific immunity lymphocytes dendritic cells

y chemokine innate adaptive immunity innate adaptive immunity chemokines

chemokines ( host defense) chemokineschemokine

y 199 6 chemokine CXCR4 receptor CD4 H IV gp120 H IV chemokine chemokine receptor


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Chemokinesy Chemokines 4 cysteine, N Cys 1

F-strands Cys1 Cys-3 Cys2 Cys4 -S -S - Cys-2 Cys420 -30 E-helix C chemokines

( 1 ) (2) 2 Cysteines :( 1 ). CXC ( Cys1- X -Cys3 ) chemokine: IL- 8 75 CXCR1-5

IL-8, GCP -2; IL-8, GROE / F / K( growth-regulated oncogene ), NAP -2 ( neutrophil -activating peptide2 ), ENA78 ( epithelial -cell derived neutrophil activator ), GCP -2 ( granulocyte chemotactic protein-2 ); I P 10 ( interferon-inducible protein ), M I G ( monokine induced by interferon-K ); SDF -1 ( stromal cell - derived factor );BCA-1 / BLC

(2). CC ( Cys2 -Cys4 ) chemokine : F-chemokine RANTES ( Regulated upon activation, normal T cell expressed and secreted ), M I P 1E , M I P 1 F ( macrophage inflammatory protein )H IV T 10 CCR1-8 RANTES, M I P -1E , MCP -2 , MCP -3

( monocyte chemoattractant protein ) ; MCP -1-4; Eotaxin, eotaxin-2, RANTES, MCP -2,3,4; TARC ( thymusand activation-regulated chemokine ), RANTES, M I P -1E ,, M I P -1; RANTES, M I P -1E , F; L ARC / M I P -3E / exodus; E LC / M I P -3 F; 1-30 9

** eotaxin : a powerful attractant of eosinophils during asthma, atopic dermatitis, nasal polyps, ulcerative colitis; itsreceptor CCR3 is present in RBC, basophils & a subset of T lymphocytes with TH2 helper

** I P 10, Mig are selective chemokines induced by I FN -Kfor IL-2 -activated T lymphocytes.** BCA-1 / BLC is a selective chemokine for B lymphocyte

** CCR5 is expressed preferentially in TH 1 cells, where CCR3 & CCR4 seem to be characteristic of TH2 cells

** Lymphocye-chemotactic protein : RANTES, M I P -1E , F chemokines ( 1 ) (2)

(3). CX3C : mucin N , CX3CR1 Fractalkine / Neurotacin(4). C : lymphotactin ( K-chemokine )


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1. CXC 2 e.g. IL-8 ---

-

2 . CC ( ) )---MCP -1( -1 ) ( IL-6 E )

T LPS, IL-1, TNF, C5a, LTB4,I NF T

3. C ( ) )---lymphotactin- lymphotactin-

4. CX3C ---Fractalkine

B : B-lymphocyte; Bs: basophil; DC : dendritic cell; Eo : eosinophil; Mc: mast cell; Mo : monocyte; NK : natural killer cell;

No : neutrophil;LEC : lymphatic endothelial cell; RBC : red blood cell; T: T -lymphocyte;V EC : vascular endothelial cell .


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chemokines


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y CC chemokines1. DC -CK -1 or PARC : Dendritic cell -derived chemokine-1 or pulmonary and activation-regulated chemokine2 . E LC : EBI1-ligand chemokine3. L ARC :Liver and activation-regulated chemokine4. MCP -1 : Monocyte chemotactic protein-15 . MDC : Macrophage-derived chemokine6 . M I P -1E : Macrophage inflammatory protein-1E7 . S LC : Secondary lymphoid tissue chemokine8 . RANTES : Regulated on activation, normal T cell expressed and secreted

9. TARC : Thymus and activation-regulated chemokine10 . TECK : Thymus-expressed chemokine

y CXC chemokines1. BCA-1 : B-cell -attracting chemokine-12 . GROE : Growth-related oncogene-E3. IL-8 : I nterleukin 8 4. I P -10 : I nterferon-c -inducible protein

5 . Nap-2 : Neutrophil -activating protein-2 6 . M I G : Monokine induced by interferon-K7 . PF -4 : Platelet factor -48 . SDF -1 : Stromal cell -derived factor -1


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Chemokiney Eotaxin, eotaxin-2, eotaxin-3, RANT ES

macr op hage chemoattr actant pr otein-4 (MC P-4) CCR3

.

1. Eotaxin Ab

y MCP -1 CCR-2 master cell


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Chemokine receptor & cancer

CXCR4 antagonist AMD3100 inhibited the growth of intracranialglioblastoma and medulloblastoma xenografts, and increasedtumour cell apoptosis within 24 hours. Therefore,CXCR4 is apotential therapeutic target in human cancer


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Chemokines can act as growth / survival factors for cancer cell

y CXCL 12 stimulates pr olif er ation and/o r sur vival of CXC R4-expr essing cancer cells, including glioma, melanoma, ovar ian, pr ostate, small cell lung, r enal and thyr oid cancer.

Semin Cancer Biol 2004;14:171-9.

y CXCL 1, CXCL 2, CXCL 3 ar e not expr essed bynor mal melanocytes but ar e upr egulatedin melanoma and stimulate the gr owth of melanoma cells in vitroand in vivo.

J Leukoc Biol 2002;72:9-18.

y CCL 20 pr omotes pr olif er ation of CC R6 -expr essing p ancr eatic cells.Int J Cancer 1999;81:6 50-7.

y CXCL 1 and CXCL 8 also stimulate pr olif er ation of p ancr eatic cancer cell lines.Pancreas2000;21:52-6 .

y CXCL 8 can act as an autocr ine gr owth factor f or human ovar ian cancer cells.O ncol Res2000;12:97-106 .


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Chemokines involved in adhesion / invasion incancer cell

y CXCL 12 modulates the expr ession and function of cell sur face integr in molecules, and in tur npr omotes adhesion toVC AM-1, f ibr onectin, andcollagen.O ncogene22:8093-101, 03, & 24:446 2-71, 05.

y CXCL12 induces metallopr otease (MMP) synthesisin diff er ent cell typ es, including r ha bdomyosar coma,

p ancr eatic and pr ostate cancer. MMPs ar e involvedin the degr adation of extr acellular matr ix in the surr ounding nor mal tissue and known to mediate cancer invasion and metastases.


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Chemokines regulate angiogenicityy CXC chemokines r egulate angiogenesis either p ositively or negatively

dep ending on the EL R motif . Angiogenic chemokines include CXCL 1, 2, 3, 5, 6 , 7 and 8. CXCL 12 lacking the EL R motif can also exer t angiogenic activity.

y Elevated levels of CXCL 5 and CXCL 8 ar e detected in non-small cell lungcancer and corr elated with the vascular ity. Dep letion of these chemokines invivor esulted in decr eased angiogenesis and decr eased tumor size and metastasis.

y CXCL 12 pr omotes blood vessel f or mation (to induce endothelial cell migr ationand pr olif er ation and to incr easeVEGF r elease by endothelial cells).

Cytokine Growth Factor Rev 2005;16 :593-6 09.


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CXCR4 & CCR7 in tumor

CXCR4 expression is associatedwith metastasis in in nasopharyngealcarcinoma, osteosarcoma, non-smallcell lung cancer, colorectal or prostate cancer.CXCR4 expression predicts LNmetastasis in breast cancer.

CXCR4 expression is associatedwith poor clinical outcome inesophageal cancer and melanoma.

CCR7 expression is associated withmetastasis in esophageal, gastric,head and neck, and non-small celllung cancer.CCR7 expression correlates with,and can be used to predict,metastasis in breast and colorectalcancer patients.Patients with oral and pharyngealsquamous cell carcinomas thatexpress CCR7 have lower survivalrates.


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Dendritic cell Discovered by Ralph Steinman (TheRockefeller University, New York)


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D entritic cell


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Chemokine T Helper T lymphocyte ( TH 1 ) CCR5 1 TH2 CCR3, CCR4


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LPS

y

LPS CC chemokine monocyte

1. LPS MCP -1, M I P -1E, MCP -3

2 . LPS ( CCR2> CCR5= CCR1 )


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Chemokiney Chemokine receptor 7 ( E- ) G

199 6 Gordon Conference on Chemotactic Cytokines chemokine receptor receptor ligand receptor

y IL-8 receptor chemokine receptor IL-8 CXC chemokine CXCR1 ( CXC ) CCRCC MCP -1 18 chemokine receptor

5 CXC chemokine receptor (CXCR) CXCR1 - CXCR5 11 CC chemokine

receptor (CCR) CCR1 -CCR

11; CX3CR

1 XCR

1 Chemokine receptor orphan receptors

y chemokine chemokine () eotaxin CCR3

T H 2 T ) CCR3eotaxin, eotaxin-2, RANTES, MCP -2,3,4

y :

Anti -IL-8 Ab : lung reperfusion injury, Urate-induced arthritisM I P -2 Ab : rat glomerulonephritisMCP -1 Ab : Cutaneous delayed hypersensitivity M I P -1E & RANTES Ab : Mouse allergic airway inflammationCC -chemokine antagonist : murine models of arthritis


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. growth factor

TGF - ( transforming growth factor - , - )EGF ( epidermal growth factor, )FGF ( fibroblast growth factor, )NGF ( nerve growth factor, )

PDGF ( platelet -derived growth factor, )I nsulinI nsulin-like growth factor V EGF ( vascular endothelial cell growth factor, )

:1.

:1. G protein receptor tyrosine kinase (EGF, FGF, PDGF,

I nsulin,I nsulin-like growth factor )


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PDGF platelets, endothelial cells, placenta

promotes proliferation of connective tissue, glial and smooth muscle cells

two different protein chains form 3distinct dimer forms; AA, AB and BB

EGF submaxillary gland, Brunnersgland

promotes proliferation of mesenchymal, glial and epithelial cells

TGF -E common in transformed cells may be important for normal wound healing related to EGF

FGF wide range of cells; protein isassociated with the ECM

promotes proliferation of many cells; inhibits some stemcells; induces mesoderm to form in early embryos

at least 19 family members, 4 distinct receptors

NGF promotes neurite outgrowth and neural cell survival several related proteins first identified as proto-oncogenes; trkA (trackA),trkB, trkC

Erythropoietin kidney promotes proliferation and differentiation of erythrocytes

TGF - F activated TH 1 cells (T -helper)and natural killer (NK) cellsanti

-inflammatory (suppresses cytokine production and class II MHC expression), promotes wound healing,

inhibits macrophage and lymphocyte proliferationat least 100 different family members

I GF -I primarily liver promotes prol iferation of many cel l typesrelated to I GF -II and proinsulin, alsocalled Somatomedin C

I GF -II variety of cells promotes proliferation of many cell types primarily of fetal origin related to

I GF -I and proinsulin


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1. Platelet -Derived Growth Factor ( PDGF, )

y Ligand 1. domain : 5 ( I g )y :1. 2 A B ( dimeric) ( homodimers , AA

BB) ( heterodimers , AB).

2 .

3. c -Sis proto-oncogene PDGF A chainy :1. tyrosine kinase PDGF ,

( autophosphorylation ). 2 .

y

:1. EPDGF (mesenchymal)2 . , P I . 3. proto-oncogene ( Fos, Jun Myc) . 4. TGF -b PDGF TGF -b .


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2 . Epidermal growth factor ( , EGF )

y :1. tyrosine kinase EGF

2. kinase( autophosphorylation ).

3. Neu proto-oncogene EGF ( homologue).

y :1. EGF ( mesoderm)

( ectoderm) ,keratinocytes fibroblasts.

2 ..

3. proto-oncogene ( Fos, Jun Myc)

. 4. .


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EGFR ( ErbB HER)y EGFR RTKs

1. EGFR (ErbB1 ) ErbB2 (HER2 / neu ErbB2) ErbB3 (HER3TK ) ErbB4 (HER4) Kinase C

2 . Ligand : EGF, TGF E ( transforming growth factor ), HB-EGF ( hep ar in- binding EGF), amp hir egulin, betacellulin, ep ir egulin ; neuroregulin( NRG )

ErbB3, B4 ligand, 2:2

3. domain ligand monomer (homodimer) (heterodimer);hetero-oligomer

4. EGFR 170kD,1186aay

1. EGF receptor Gs ( F receptor) Gi Go . 2 . domain (Tyrosine kinase) (phosphorylation)

signal transduction pathway ( Ras-Raf

-MEK

-MAPK pathway;P I 3K - Akt pathway, SAPK pathway, STAT pathway) substrate ,

3.ligand -receptor complex

4. ErbB2 ligand


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E GFR tyrosine kinase pathway1. The Ras / Raf mitogen-activated protein (MAP)

kinase :y EGFR recruits the guanine-nucleotide exchange

factor via the Grb2 and Shc adapter proteins,activating Ras through guanosine triphosphateexchange.

y affect cell proliferation, tumor invasion, and metastasis

2 . P I 3K / Akt y activated by the recruitment of the adaptor protein

Gab1 to phosphorylated EGFR. Activation leads toantiapoptotic signaling through nuclear factor -OBand the generation of H2O2, blocking the inhibitory tyrosine protein phosphatase that terminatesEGFR activation

3. Jak2 / STAT3 pathwaysy The activation of Jak2 / STAT3 is mediated by the

cytoplasmic TK c -Scr and plays a critical role intumor resistance to chemotherapy


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E GFR tyrosine kinase


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EGFRy

(EGFR)y tyrosine kinase (malignancies)

(a) retroviral tyrosine kinase proto-oncogene ( ); (b)( oncogenic t tyrosine kinase ( e.g., Bcr - Abl in Philadelphia

chromosomepositive leukemias); (c) tyrosine kinases ( stromal tumorsK I T); (d) tyrosine kinase ( solid tumors EGFR ).

. y EGFR

ErbB2 EGFR(proliferation) (apoptosis) (de-differentiation)

(angiogenesis) (metastasis)y EGFR ,

(survival)y EGF EGF distal convoluted tubules thick

ascending limbs of Henel loop.y EGFR basal cells ErbB2 ErbB3 ErbB4 superficial layer

urothelial tumor EGFR , EGFR-ErbB3 ErbB3-ErbB4high grade tumor tumor invasion

y ErbB2, EGFR-ErbB2, ErbB2 -ErbB3 Grade 2 EGFR-ErbB2 -ErbB3 EGFR-ErbB2 second recurrence ErbB2 -ErbB3


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Protein kinase

Overexpression of growth factor receptorsEGFR,HER2,HER3,HER4 in breast, ovary, lung, head & neck and glioblastomaFGFR in lung, ovary and breastPDGFR in glioblastomaIGF-1R in solid tumors

Overexpression of growth factorsTGF- E in carcinoma overexpressing the EGFRPDGF-BB expression in glioblastomaVEGF expression for neo-vascularization (angiogenesis andmetastasis)

Al tered protein kinase leve ls and/or activitiesBcr-Abl in CML (95%), ALL (15%)c-Met in renal carcinomac-Kit in gastric cancers (GIST)Cancers with Onc Ras (about 30^50%) have deregulated PKC,Raf-kinase (bladder, colon, lung, breast)Deregulationof cell cycle by CdKs


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EGFRy ErbB2 Trastuzumab ( Herceptin ) 98 EGFR

ErbB2 ( Her2 / neu ) ( 30% )

y EGFR / ErbB1 Cetuximab (C -225) (Erbitux) ir inotecanckEGFR ( 30% )

y : Gefitinib ( I ressa) Erlotinib (Tarceva)Gefitinib ( I ressa, ZD1839 : EGFR EGFR ATP bindingdomain tyrosine kinase ( tyrosine kinase domain

)1. EGFR

I ressa 12 .5 200mg / kgDC I S

2 . EGFR mutation ( 10 %) I ressa

3. EGFR EGFR-ErbB2 y :

1. ErbB2 ErbB2 ErbB2

2 . ErbB2 cDNA DNA IL2 -MBT2 cellsCD4+ T lymphocytes anti -neu antibody

3. ErbB2 ErbB2 ErbB2 - -12 cDNA - ErbB2 IL-12 DNA

vaccine t DNA ,


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E GFR kinase inhibitor:imatinib, dasatinib,nilotinib,erlotinib ( Nov,04 ) and gefitinib ( May,03)

y Mutations involving the BRAF gene mutation : 2 -3% of nonsmall -cell lung cancer (NSC LC),y KRAS proto-oncogene mutation : 30% of adenocarcinomas.y The structural similarity between Abl and EGFR TKs is fairly high because the threonine-to-

isoleucine mutation at codon 315 corresponds to the threonine-to-methionine mutation at codon 7 90 (T7 90M) in EGFR.

y a single mutation (T315 I ) deep within the ATP -binding pocket of the Abl TK that confers a high degree of resistance to imatinib, dasatinib, and nilotinib as a result of

steric hindrance caused by the replacement of threonine with the bulkier isoleucineresidue


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tyrosine kinasey The Philadelphia (Ph) chromosome is a foreshortened chromosome 22 resulting from

an exchange between the long arms of chromosomes9 and 22 .y The translocation t( 9;22) results in the juxtaposition of 3' DNA sequences derived

from the ABL proto-oncogene on chromosome9 with 5' sequences of the breakpoint cluster region(BCR) gene on chromosome 22, forming a fusion gene, BCRABL. ( Thereciprocal formation of the ABL-BCR fusion gene on chromosome9q+ is not depicted . )

y BCR- ABL produces a chimeric messenger RNA (not shown) from which a fusion BCR- ABL oncoprotein is translated . The length of the BCR- ABL protein varies and isdetermined by the breakpoint within the BCR gene

y Chronic - phase CM L is driven by the constitutively active BCR- ABL tyrosine kinase

protein, which activates multiple pathways, leading to the malignant expansion of myeloid cells through the stimulation of mitosis, the disruption of cytoadherence and regulatory control by stromal cells, and the inhibition of apoptosis.

y Differentiation and maturation of the leukemic clone are relatively intact in chronic - phaseCM L, but BCR- ABL is also thought to promote genomic instability, leading to secondary mutations and to the blast phase. I matinib mesylate inhibits the tyrosine kinase activity of the BCRABL oncoprotein, thus blocking the leukemogenic effects of the Phchromosome. Dimerization and activation of the normal c -kit receptor by its ligand stem-cell factor are shown in Panel B.

y

The proto-oncogene c kit encodes a transmembrane tyrosine kinase receptor located on the long arm of chromosome 4 (4q11 q12). I n gastrointestinal stromal tumors, in-frame deletions and point mutations in c -kit produce ligand -independent constitutiveactivation of c -kit (Panel C). Mutations of c -kit in the juxtamembrane domain ingastrointestinal stromal tumors (exon11 ) are found in approximately 60 percent of cases . Mutations also occur in the extracellular domain (exon9 ) and in the more distal phosphokinase domain (exon13). ATP denotes adenosine triphosphate, ADP adenosinediphosphate, and P phosphate.


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Receptor -coupled tyrosine kinase ( RTK )y 90 y RTK : homodimer ( insulin ,I nsulin-like

growth factor ) (autophosphorylation)tyrosine

: , ( epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet -derived growth factor (PDGF), vascular endothelial growthfactor ( V EGF), nerve growth factor (NGF),Epo.

: monomer dimer : ,

SH2 domains : p-Tyr moiety C 1-6 ,PTK (Src kinases),

protein tyrosine phosphatase (PTP, Shp2), phospholipase C activity (P LC K ), Ras-GAP

PTB domains : p-Tyr moiety N 3-5 SH3 domains : proline-rich sequence moiety PXXP WW domain : proline-rich motif PXPX Pleckstrin homology (PH) domains : Ptd I ns(4,5)P2 P I 3K FY V E domains: Ptd I ns-3-P PDZ domains : target protein C


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RTKs


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RTK

(A) PKB (also known as Akt) . Ptd I ns(3,4,5)P3 generated in response to growth factor stimulation serves as a binding site for the PH domains of PDK 1 and PKB. Membrane translocation is accompanied by releaseof an autoinhibition leading to activation of PDK 1 and PKBkinase activities. Full activation of PKB requires phosphorylationby PDK 1 (and also byPDK2?). Activated PKB phosphorylates avariety of target proteins that prevent apoptotic death and regulate various metabolic processes.

(B) Binding of the SH2 domains of p85,the regulatory subunit of P I-3 kinase to pTyr sites on activated receptors releases an autoinhibitory constraint that stimulatesthe catalytic domain (p110). P I-3 kinase catalyzes the phosphorylation of the 39 positions of the inositol ring of Ptd I ns(4)P and Ptd I ns(4,5)P2 to generate Ptd I ns(3,4)P2 and Ptd I ns(3,4,5)P3, respectively .

(C) tyrosine phosphorylation Binding of the SH2 domains of P LC Kto pTyr sites in activated receptors facilitatestyrosine phosphorylation of P LC Kas well as membranetranslocation; a process mediated in part by binding of the PH domain to P I-3 kinase products. Tyrosine phosphorylation isessential for P LCg activation leading to hydrolysis of Ptd I ns(4,5)P2 and the generation of the two second messengers I ns( 1,4,5)P3 and diacyglycosol .


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RTK

0

10

20

30

40

50

60

70

80

90

(A) RTK .For example, in addition to activation of the MAP kinasesignaling cascade, Ras activates P I-3 kinase and Cdc42 . Stimulation of P I-3 kinase leads to activation of PDK 1 and PKB,two kinases that regulate various metabolic processes and prevent apoptotic death. I n addition, P I-3 kinase activationstimulates generation of hydrogen peroxide which in turnoxidizes and blocks the action of an inhibitory protein tyrosine phosphatase (PTP) . The signaling cassettes presented in thefigure regulate the activity of multiple cytoplasmic targets. However, the Ras / MAP, STAT, JNK, and P I-3 kinase signaling pathways also regulate the activity of transcriptional factors by phosphorylation and by other mechanisms.

(B ) RTK I n several cases the activity of RTKs can be negatively regulated by ligand antagonists or by hetero-oligomerizationwith naturally occurring dominant interfering receptor variants. The PTK activity of EGFR is attenuated by PKC -induced

phosphorylation at the juxtamembrane region. Dephosphorylation of key regulatory pTyr residues by proteintyrosine phosphatases (PTP) may inhibit kinase activity or eliminate docking sites. An important mechanism for signal termination is via receptor endocytosis and degradation. Theoncogenic protein Cbl binds to pTyr sites in activated RTKs viaits SH2 -like domain. The RI NG finger domain of Cbl functionsas a ubiquitin-ligase leading to receptor ubiquitination and degradation by the proteosome.


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Tyrosine kinase


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E GFR ( erb B )

The EGFR (erbB) Family

Membrane

Extracellular

Intracellular

Cysteine-richReceptor domain

K

EGFTGF-E

AmphiregulinF-cellulinHB-EGF

Epiregulin

Tyrosine kinasedomain

erbB4

HER4

erbB3

HER3

erbB1

HER1EGFR

erbB2

HER2neu

Ligands

K

No specificligands -

often acts asdimer partner

K

Heregulins

K

NRG2NRG3

HeregulinsF-cellulin

www.egfr-info.com

: RTK dimer . I nsulin I nsulin-like growth factor heterotetrameric ( 2 E sulfhydryl 2 E I RS adaptor protein ( ) 2 F tyrosine kinase. Anticancer drugs as Tarceva directly target the EGFR. EGFR positive patients have shown animpressive 60% response rate .


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E GF pathway


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E GFR overexpression inhibitors inchemotherapy

y Gef itinib & Er lotinib f or lung cancery C etuxima b f or colon cancery

Monoclone A b :C

etuxima b, p anitumuma b, zalutumuma b, nimotuzuma b, matuzuma by Small molecule kinaseinhibitor s : Gef itinib, er lotinib

and lap atinib (mixed EGFR and ERBB 2 inhibitor )


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3. Fibroblast Growth Factors Receptor ( , FGFRs)

y

1. 2 12 . 3 FGF FGFR, heparin

heparan-sulfated proteoglycans FGFR 2 :2 ( ) FGF 1 ( acidic -FGF, aFGF) FGF2 ( basic -FGF, bFGF ).

3. neurotrophic factor

4. Kaposis FGF K -FGF proto-oncogene.

5 . I nt -1 I nt -2 . I nt -2 FGF y :1. FGFR1 - FGFR4. tyrosine kinase, ( autophosphorylation ). 2 . (achondroplasia)

FGFR3 glycine arginine(craniosynostosis ) FGFR1, FGFR2 FGFR3 FGFR2

cysteine tyrosine Pfeiffer Crouzon syndrome3. Flg proto-oncogene FGFR4. FGFR1


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Affected Receptor Syndrome Phenotypes

FGFR1 Pfeiffer broad first digits, hypertelorism

FGFR2 Apert mid-face hypoplasia, fusion of digits

FGFR2 Beare-Stevenson mid-face hypoplasia, corrugated skin

FGFR2 Crouzon mid-face hypoplasia, ocular proptosis

FGFR2 Jackson-Weiss mid-face hypoplasia, foot anamolies

FGFR2 Pfeiffer same as for FGFR1 mutations

FGFR3 Crouzon mid-face hypoplasia, acanthosis nigricans, ocular proptosis

FGFR3 Non-syndromatic craniosynostosis digit defects, hearing loss


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4-1. Transforming growth factor - ( - )

: 100 TGF - F1~4 activin A, B , inhibin A,B, Mullerian inhibiting substance(M I S) bone morphogenetic protein (BMP)

T B ( ): serine / threonine kinase

y T y B T y

NK IL-

2 L AK y M

y

y TGF - Fy immunoglobulin hematopoiesis, myogenesis, adipogenesis

adrenal steroidogenesis. y TGF -b activin A

f h f ( )


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4-2 . Transforming growth factor -E ( -E )

y : 1. macrophages and

keratinocytes (and possibly other epithelial cells)2. keratinocyte

( autocrine )

3. TGF -E EGF 4. :y TGF F ( )

1 ). cyclin- dependent kinases , 2 ). myc transcription factor, 3).

: fibronectin, collagens, proteoglycans

: collagenase, plasminogen activator

- TGF - F . c -Myc ( mick) proto-oncogene

Myc -binding elements .


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5. E rythropoietin ( , E po):

1. Epo ( 165 aa ) ( )

( erythropoiesis ) .

(erythroid progenitor cells ,erythroid burst -forming

colony -forming units )

erythrocyte colony -forming units ( 3-5 proerythroblasts,basophilic erythroblasts, polychromatophilic erythroblasts, orthochromatophilic erythroblasts (OEs)) OEs

reticulocyte .2 . Erythropoiesis :

pluripotent stem cells . :

1. Epo

2 . Epo 2 Epo receptorstransphosphorylation

JAK2 kinase.


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E po JAK-STAT


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6 . Insuliny insulin-dep endent lip olysis and inhibition of glucose oxidation in adip ocytes

thr ough G pr oteiny decr easing the expr ession of Gi2 in the liver and adip ose tissue r esults in

hyp er insulinemia, glucose intoler ance (decr eases the activation of glycogensynthase and antilip olytic activity ), and r esistance to insulin (decr eases glucose

tr ansp or ter GLUT

4 tr anslocation to the p lasma membr ane)y T he signal p athways : utilizeIRS-1/ IRS-2 and Shc as immediate downstr eam

adap tor s, leading to activation of the Ras, Raf , ERK kinases and PI-3 kinase p athways via insulin r ecep tor ( one of tyr osine kinase r ecep tor family )


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7 Insulin Like Growth FactorI II


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7 . I nsulin-Like Growth Factor -I , II ( , I GF -I , II )

y I GF -I ( somatomedin C) :1. I GF -I

somatomedin ( ) .2 . I GF -I autocrine paracrine3. : heterotetramer ( E2 F2 ) I GF -I receptor y activation of I GF -I receptor has been shown to inhibit adenylyl cyclase in a Gi2 -dependent manner and

activate Erk pathway via G subunitsy internalization of I GF -I receptor is necessary for activation of Erk cascadey tyrosine kinase , ( autophosphorylation )

.

y I GF -II 1. .2 . I GF -II .3. : mannose-6 - phosphate receptor

( mannose-6 - phosphate ) lysosomes .


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IGFy K ey downstr eam networ ks include the PI3K A KT TO R

system and the RAFMAPK systems. Activation of these p athways stimulates pr olif er ation and inhibits ap op tosis. For many cell typ es, the key eff ects of signalling downstr eam to

AKT

r elate to r egulation of cell sur vival and mRNA tr anslation, while the pr incip al eff ect of signalling downstr eam to RASinvolves r egulation of cellular pr olif er ation.

y 4EB P1 : eukar yotic tr anslation initiation factor 4E bindingpr otein 1;

y eIF4E : eukar yotic tr anslation initiation factor 4E; y ERK : extr acellular signal-r egulated kinase;y GRB 2 : gr owth-factor -r ecep tor - bound pr otein 2;y IRS1 : insulin-r ecep tor su bstr ate 1; y MAPK : mitogen-activated pr otein kinase; y MEK : mitogen-activated pr otein kinase kinase;y PI3K : p hosp hatidylinositol 3-kinase; y PIP : p hosp hatidylinositol; y PT EN : p hosp hatase and tensin homologue;y S6K : S6 kinase;y SHC : SRC -homology-2-domain tr ansf or ming pr otein; y SHP2 : p hosp hatidylinositol 3-kinase r egulator y su bunit; y SRF : ser um r esp onse factor ; y TO R : tar get of r ap amycin.


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y

diarthrodial joints of the hand and feet . y ( synovium) ( pannus)

y autoAbs & T cell ( CD4+ ) rheumatoid factor &( major histocompatibility complex, MHC ) in 3rd hypervariable ( or

susceptibility epitope ) region of DR F chain in 70 -74 aa- QKRAA or QRRAA- in humanleukocyte antigen ) ( 80 % )( chemotactic factor C5a ) .

y ( synovial fluid) : T -cell cytokine ( IL-2, I FN -K ) ;cytokines ( IL-1, 6, 10,18, TNF -E , GM -

CSF) ( cytokine network hyposis ) memoryT H 1 cell ( IL-17) ,chemokine receptor CCR5 , CXCR3 integrin E4 F1( delayed -type hypersensitivity) fibroblast activation, bone

destruction

chemokines & growth factors

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