1. dia

1
cikk-referáló Génátírás szabályozása
transzkripciós mechanizmusok Dr. Marcsek
Zoltán
2
(No Transcript)
3
(No Transcript)
4
eukaryoták promóter elemei

• a promóter elemei többé-kevésbé jól definiáltak
• a transzkripció start helyétol különbözo
  távolságra vannak

gyakran hiányzik
általában van
5
egy tipikus promóter


6
az eukaryota transzkripció szabályozása
7
szabályozás transzkripciós faktorokkal

• egy transzkripciós faktor (vagy transzkripciós
  faktorok egy csoportja) egy géncsoport
  expresszióját szabályozhatja (pl. heat shock
  fehérjék, G1/S restrikciós pont fehérjék, stb)
• egy gén átírását egy vagy több, egymástól
  független transzkripciós faktor szabályozhatja
  (pl. metallothionine)
• magas expresszió kiváltásához gyakran több,
  különbözo 'response' elemhez köto transzkripciós
  faktor lehet szükséges

8
Nuclear Factor-?B A Holy Grail in Cancer
Prevention and Therapy Bharat B. Aggarwal,
Gautam Sethi, Asha Nair, Haruyo
Ichikawa Cytokine Res. Lab, Dept. Exp.
Therapeutics, Univ. Texas M.D. Anderson Cancer
Center, Houston, Tx invited review Current
Signal Transduction and Therapeutics 2006.
9
NFkB mi az, mit csinál, miért jó, ha
ismerjük? transzkripciós faktor
10
NFkB

• Baltimore Sen fedezték fel mint egy
  B-sejt-specifikus magi proteint amely az
  immunoglobulin k könnyu láncának meghatározott
  szekvenciájához kötodik és a gén átírását
  serkenti (enhancer) (Cell 47921-928, 1986)
• az NFkB a Rel transzkripciós faktor családba
  tartozik amely nagyszámú, az immunválasszal, a
  gyulladással, a sejtosztódással és apoptózissal
  összefüggo gén expresszióját szabályozza
• az emlos sejtekben öt eltéro NFkB (rel)
  alegységet ismerünk amelyek kb. 300 nagyon
  konzervált aminosat tartalmaznak
• a citoplazmában az NFkB inaktív formában több,
  különbözo kombinációban található (heterodimer
  p50/p65 (Rel A) és az inhibítor IkBa alegység)

11
Family of NFkB and Inhibitory kB (IkB) proteins
Nat.Rev.Cancer 2301-310, 2002
12
p105 (contains p50 NFkB1)

• Name p105 (contains p50 subunit, a.k.a. NFkB1)
• Descr Once in the nucleus initiates
  transcription of hundreds of genes
• Length 968

13
p100 (contains p52 NFkB2)

• Name p100 (contains p52 subunit, a.k.a. NFkB2)
• Descr Once in the nucleus initiates
  transcription of hundreds of genes
• Length 899

14
IkB?

• Name Inhibitor of NF kappaB beta
• Length 356
• Consists mostly of ankyrin repeats that serve
  inhibitory function

15
IkB?

• Name IkappaB epsilon
• Length 361
• Consists mostly of ankyrin repeats that serve
  inhibitory function

16
IKK?
Name IKappaB kinase alpha Descr Forms
complexes with IKK? Length 745 aa Conserved
Serine/Threonine protein kinase Domain detected
at 10 300 aa.
17
IKK?

• Name IKappaB kinase beta
• Descr Forms complexes with IKK? and destroys
  IkBs
• Length 756
• Similarly to IKK? a slightly less conserved
  Serine/Threonine protein kinase Domain detected
  at 10 - 290 aa.

18
IKK?

• Name IKappaB kinase gamma
• Length 487 aa
• Contains Chromosome segregation ATPases domain

19
az NFkB aktiválása

• NFkB a citoplazmában található inaktív formában
  mint az IkBa inhibítorához kötött p50/65
  heterodimer
• extracelluláris indukciós hatásra az IkBa-kináz
  komplex (IKK) aktiválódik és foszforilálja az
  IkBa-t (szerin 32 és 36) amely az IkBa lizin21 és
  lizin22 ubiquin-kötését okozza
• ez az IkBa alegység lebotását eredményezi (26S
  proteasome)
• az inhibitor alegység lebomlása a p50/p65 komplex
  aktivációját eredményezi az NFkB a citoplazmából
  a sejtmagba kerül (transzlokáció)
• a sejtmagban az NFkB transzkripciós faktor a
  consensus szekvenciához (5-GGGACTTTC-3)
  kötodik és aktiválja a specifikus gének
  expresszióját

20
Aktiváló Stimulus (kfl)
gyulladási-/immun- stb. fehérjék
IkB-kináz/NEMO/IKAP komplex
P
IkBa
IkBa
IkBa
p65
p65
p50
p50
degradáció
mRNS
CITOPLAZMA
p65
p50
NUKLEUSZ
transzkripció
p65
p50
transzlokáció
Target genes
21
Factors that induce NFkB
Reactive Oxygen Species (ROS)
Leukemia 161053-1068, 2002
22
Free Radic Biol Med 25346-361
23
kérdés mely szignálok aktiválják az NFkB
aktivitást?
24
Table 1. A list of inducers of NF-kB activation

• Cytokines
• TNFa , TNFß
• RANKL, TRAlL
• CD40 ligand
• CD30 ligand
• TWEAK
• THANK
• BAFF
• LTß
• IL-1, IL-2, IL-4
• IL-8, IL-12, IL-15
• IL-17, IL-18
• CD11b/CD18 ligand
• CD2 ligand
• CD35 ligand
• CD3 ligand
• CD28 ligand
• CD43 ligand
• CD4 ligand

• Growth factors and hormones
• GMCSF
• M-CSF
• NGF
• EGF
• Insulin-like growth factor 1
• Insulin
• Hepatocyte growth factor
• Gastrin
• PEDF
• Platelet-derived growth factor
• Bone morphogenic protein 2
• Bone morphogenic protein 4
• Cortical releasing hormone
• Follicle-stimulating hormone
• Mullerian inhibiting substance
• PAF
• Plant steroids (diosgenin,
• hecogenin, tigogenin)

Oncogenes Ras Myc Bcr-abl All-trans retinoic
acid RET/PTC3 fusion oncoprotein
S100B Serum Sulphatide TGF-a TGF-ß
Chemical Agents L-NMA, Ethanol,
Monensin Nicotine, Adriamycin 2-Deoxyglucose Aniso
mycin Benzyl isothiocyanate Bisperoxovanadium
phosphotyrosyl Brefeldin A, Cadmium Calyculin A,
Colchicine Calcium lonophore plus phorbol
ester Ceramide-beta-galactose 2-chloroethyl ethyl
sulfide Con A, Cycloheximide Cyclopiazonic
acid Diquat, 2,4-Dinitrofluorobenzene Ferrocene,
Forskolin Gadolinium chloride Glass fibers Sodium
butyrate trichostatin A Malondialdehyde MDMA, MEN
17055 N-methyl-D-aspartate Mycophenolic
acid Nafenopin, N-Nitrosomorphine Nocodazol,
Okadaic acid Peplomycin, Phytohemagglutinin Phorbo
l ester, Podophy/lotoxin Polychlorinated
biophenyl-77 Prostratin, Pyrogallol Staurosporine,
Tunicamycin Transglutaminase 2 WF10, WY-14 643,
CFTR, GPR56 Hematopoietic progenitor kinase 1 Ig
heavy chain, MHC Class I
25
Table 1. A list of inducers of NF-kB activation
(ctd)
Viruses Adenovirus Cytomegalovirus EBV Hammarskjöl
d and hepatitis B virus HVS Human herpes virus
6 HIV-1 Herpes simplex virus-1 HTLV-1 Influenza
virus Measies virus Moloney murine leukemia
virus Newcastle disease virus Respiratory
syncytial virus Rhinovirus SARS
coronavirus Sendai paramyxovirus Sindbis
virus Vaccinia virus akara West Nile flavavirus
Viral products Adenovírus 5 E1A Adenovirus
E3/19K African swine fever virus
IAP Antibody to dengue virus nonstructural
protein 1 CMV ie1 EBNA-2 EBV LMP1 HBV HBx HBV
LHBs HBV MHBst HCV Core protein HVS13 HVS
StpC HIV-1 gp160 HIV-1 Nef HIV-1 p9 (9 aa
peptide) HIV-1Tat HTLV-I Tax HTLV-II
Tax Influenza vírus hemagglutinin KSHV
K1 Parvovirus B19 NSI Respiratory syncytial
virus M2-1 SV40
Bacteria and fungi Anaplasma phagocytophilum Angio
strongylus cantonensis Bacteroides forsythus
Bartonel/a henselae Bordetel/a
pertussis Borrelia burgdorferi Chlamydia
pneumoniae Cryptococcus neoformans Ehrlichia
chaffeensis Enteropathogenic Escherichia
coli Fusobacterium nucleatum Gardnerella
vaginalis Helicobacter pylori Lactobacilli Lister
ia monocytogenes Mycoplasma fermentans
Mycobacteria tuberculosis Neisseria gonorrhoeae
Neisseria meningitidis Porphyromonas gingivalis
Prevote/la intermedia Pseudomonas aerogenosa
Rhodococcus equi Rickettsia rickettsii Salmonella
dublin Salmonella typhimurium Shigella fIexneri
Staphy/lococcus aureus Streptococcus Streptomyce
s californicus Trichomonas vaginalis Ureaplasma
urealyticum Yersinia enterocolitica
Bacterial/fungal products LPS Wogonin Enterotoxin
GIycosylphosphatidylinositols Fumonisin
B1 Staphylococcus enterotoxins A and B Apicularen
A CpG Cytolysin Diphosphoryl lipid A Exotoxin 8
Lipoprotein LpK Lipoprotein LpK Lipoteichoic
acid Membrane lipoproteins (Mycoplasma
fermentans, Mycoplasma penetrans) Mannoproteins Mu
ramyl peptides Mycobacterium species
lipoarabinomannan PicA, PlcB Porin 1B Toxic
shock syndrome toxin 1 Eukariotic
parasietis Eimeria tenella and necatrix
Leishmania Phospholipomannan Pneumocystis Theile
ria parva Trypanoplasma borreli
26
Table 1. A list of inducers of NF-kB activation
(ctd)
Physiologic (stress) cond. Asthma, Rheumatoid
arthritis Crohn's disease/ulcerative
colitis Corneal epidemic keratoconjunctivitis Coro
nary artery bypass Hypercholesterolemia
Hyperglycemia Hyperhomocysteinemia
Hyperosmotic shock, Shear stress Hypoxia,
Reoxygenation Acute jung injury Acute respiratory
distress syndrome Adhesion, Hemorrhage Angina
pectoris Antiphospholipid antibodies
Appendicitis Depolarization Ischemia (transient,
foGaI) Ischemic preconditioning Liver
regeneration Biotin deficiency Butter and walnut
diet Cecalligatin and puncture Mechanical
ventilation Memory retrieval Middie cerebral
artery occlusion Muscle disuse, Muscular
dystrophy Neuronal firing Overventilation
pancreatitis Proteinuria Senescence Neuroonal
trimethyltin injury Uniaxial cyclic cell
stretching T -cell selection
Physical stress UV A, B and C radiation Bile
duct ligation Cyclic mechanical muscle
strain Exercice ? radiation Heavy ion
irradiation High-fat diet Laminar shear
stress Dietary lipid hydroperoxide PPME
photosensitization Mechanical lung
ventilation Obesity Wounding combined with
He-Ne irradiation Wounding combened with
thermal irradiathion Oxldative
stress Glutathione Hydrogen peroxide Catalase
Nitric oxide Ozone Peroxiynitrite Butyl
peroxide Pervanadate Reoxygenation
Environmental hazards Cigarette
smoke Arsenic Polychlorinated biphenyl
Benzoapyrene diol epoxide Crocidolite
asbestos fibers Dicamba Diesel exhaust
particles Gliadin House dust mite Dust
particles Fear-potentiated startie
response Lead Lead chromate Noise Olly fly
ash Silica partlcles Wood smoke Zymosan
Chemotherapeutic drugs Cisplatin,
Paclitaxel Doxorubicin, Tamoxifen Vinblastine,
Vincristine Etoposide Ara-C Bryostatin-1 Camptothe
cin Daunomycin Daunorubicin Mitoxantrone Gemcitabi
ne Therapeutic drugs Celecoxib, AZT,
Ciprofibrate Bleomycin ABR-25757 SN38 SA 981,
WR1065 AdAMP, Anthralin Cycloprodigiosin Dacarbasi
ne Orengedeokuto Diazoxide 5,6-dimethylxanthenone-
4-acetic acid Flavone-8-acetic acid Haloperidol Cu
nbi-BoshinHangam-Tang Methamphetamine Oltipraz Phe
nobarbital Protocatechuic acid
27
Table 1. A list of inducers of NF-kB activation
(ctd)
Fatty acids

Palmitate Leukotriene B4 Free fatty acids
Llnolelc acid Oleic acid Saturated fattx
acids Lysophosphatldlc acid Modified Proteins
LDL AGEs Amyloid protein fragment
Glycosylated oxyhemaglobin Maleylated
bovine serum albumin Neurotrophin receptor
proteolytic fragments Nonamyloid beta
component of Alzheimer's disease ATM Heat
shock protein HSP60 HSP71
Apoptotlc Mediators Anti-Fas/Apo-1 PARP Metals L
ithium Titanium and copper implants Nickel
sulfate Iron Manganase Nickel Chromium Cobalt Alum
inium Natural products Areca nut
extract Baicalin Beta-carotene Quercetin Quinolini
c acid Safflower polysaccharides Sanglifehrin A
Physiological Mediators Alloxan Angiotensin
II Bradykinin Fibrinogen Hemoglobin Hyaluronan Thi
oredoxin Kainan acid L-Glutamate Melanin Violacein
Vitamin D3 Adesonine Activin A Albumin Allergin A
crp30/adiponectin Adrenomedullin Aeginetia indica
L (AIL)b-A Bronchoalveolar lavage
fluid Brusatol Homocysteine Cryptdins Deoxicholic
acid 12-hydroxyeicosatrienoic acid 6-Hydroxydopami
ne Human peptide transporter 1 LTPLyso
PCNeuomelanin NS-398 Osteopontin PAF PCSC Prolacti
n
PIF 12-Lipoxygenase Thrombin Streptozotocin Substa
nce P Potassium Bum patient serum Anaphyloxin C3a
and C5a Angiotensin II type 1 receptoractivating
antibodies Antiphospholipid antibodies Basic Calci
um Phosphate crystals C2-ceramide
cerulein Chelidonium majus extract Collagen
latice Collagen type 1 Cysteinyl
leukotrines Des-Arg 10-kallidin Mixed meal
ingestion Monosudium urate crystals Neutrophil
elastase Phellinus linteus proteoglycan Platelet
type arachidonate Polysachcharides of Poria
cacos Regulatory RNA S100B Sleep
deprivation TUDCA Tuberous sclerosis
complex Uremic toxins, VLDL
28
Constitutive NF-kB Signaling Module
Input signal
IkK
IkK
I?B
NF
?B
NLS
I?B Degraded
Cytoplasm
IL-6 gene
Nucleus
29
kérdés mely szignálutak vesznek részt az
NFkB aktivitásában?
30
Table 2. protein kinases implicated in NF-kB
activation
min. 32 ismert kináz
31
kérdés mely betegségekben mutatható ki
abberáns (állandóan magas) NFkB
aktivitás? többet tudunk, ha ismerjük a
rendszer hibáit...
32
Table 3. association of NF-kB activation with
diseases
Cancer B-cell lymphoma Breast cancer Hodgkin's
disease Liver cancer Mantle cell
lymphoma Multiple myeloma Non-Hodgkin's
lymphoma Ovarian cancer Prostate cancer T -cell
lymphoma Thyroid cancer Bladdercancer Colon
cancer Esophagealcancer Head and neck squamous
cell carcinoma Laryngealcancer Lung
cancer Pancreatic cancer Pharyngealcancer Renal
carcinoma Acute lymphoblastic leukemia Adult
T-cell leukemia Cervical cancer Nasopharyngeal
carcinoma Melanoma
Cardiovascular diseases Atherosclerosis Cardiac
hypertrophy Heart failure Hypercholesterolemia Di
abetes mellitus Type I Type II Inflammatory
diseases Alzheimer's disease Arthritis Asthma Bone
resorption Chronic obstructive pulmonary
disease (COPD) Crohn's disease Huntington's
disease Inflammatory bowel disease Inflammatory
response syndrome Multiple sclerosis Ocular
allergy Parkinson's disease
Other diseases Aging AIDS Catabolic
disorders Ectodermal dysplasia Gut
diseases Headache Helicobacter pylori-associated
gastritis Inontinentia pigmenti
Ischemia/reperfusion injury Systemic lupus
erythematosus Muscular dystrophy Neuropathologic
diseases Psychosocial stress diseases Renal
diseases Sepsis Skin diseases Sleep apnea Viral
infection
33
kérdés mely gének aktiválódnak az NFkB
hatására? mi a hiba okozója (fenotípus)
34
Table 4. list of genes regulated by NF-kB
Cytokines TNF-a, TNF-ß IL-1-a, IL-1ß IFN-a,
IFN-? TRAlL, M-CSF RANTES FAS ligand Lymphotoxin
a, ß G-CSF, GM-CSF IL- 2, IL- 6, IL -8, IL- 9,
IL-10, IL-11, IL-12, IL-13, IL-15 NK4, GCP-2
(CXCL6) Gro a, Gro ?, Gro-1 ICOS IL-1 receptor
antagonist IP-10, KC, BMP-2 CCL 15, CCL 22, CCL
28, CCL 5 CD40 ligand CINC-1, CXCL 11 ENA-78
(CXCL5) Eotaxin Erythropoietin MCP-1/JE, MIP-1
a ß MIP-2, MIP-3 a, Mob-1 Neutrophil- activating
peptide 78 NK-1R Stem cell factor Angiotensinogen
TCA3 TFF3 TSP-1 TSP-2
Receptors beta2 microglobulin µ-opioid
receptor 2A A1 Adenosine receptor
Amiloride-sensitive Na channel Androgen receptor
B 7.1 Bradykinin B-1 receptor BRL-1, CCR 5, CCR
7, CD 137 CD 154, CD 23, CD 40, CD 48 CD 69,
CD 83, CD 95 EGFR Gal 1 receptor,
GFBP-2 Glucocorticoid receptor IGFBP-1 IL-1
receptor antagonist IL-2 receptor a-chain Ig ?4,
Ig C?1, Ig e heavy chain Ig ? light chain,
invariant chain II Lox-1, Mdr 1 MHC class
(HLA-B7) MHC class I (H-2Kb) Neuropeptode YY1-
receptor NMDA receptor subunint NR-1 NMDA
receptor subunit Nod 2 Polymeric Ig receptor RAF
receptor 1,RAGE T -cell receptor ß chain T -cell
receptor CD3? TNF-Receptor, p75/80 DR4, DR5
Angiogenesis VEGF Cell proliferation COX-2,
c-myc Cell cycle Cyclin D1, P21, GADD 45 Cyclin
D2, Cyclin D3 Viruses HIV-1 EBV wp promoter HBV
pregenomic promoter HSV, SV-40 Adenovirus (E3
region) Avian leukosis virus Bovine leukemia
virus Cytomegalovirus JC virus, HPV 16,
SIV Proteins involved in antigen presentation
Tapasin LMP2 TAP 1 Complement B Complement
receptor 2 Complement component 3
Early response genes p62, p22/RPG1, B 94 Egr-1,
TIEG Transcription factors NF-?B1, NF-?B2,
c-Rel, RelB IkBa c-fos, p53, c-myb, JunB E2F3a,
Elf3 , ELYS ETR 1001 IRF-1, IRF-2, IRF-4,
IRF-7 Mail, Nurr1, Stat5a, WT1 Metastasis
ICAM-1, VCAM-1 ELAM-1, E-selectin Endoglin,
Fibronectin MadCAM-1, P-selectin Tenascin-C,
DC-SIGN MMP-3, MMP-9, CXCR4 KA11/CD82, uPA
Antiapoptosis Bcl-2, Bcl-XL, Bfl1/A1 TRAF-1,
TRAF-2, TRAF6 c-FLIP, IAPs Survivin, A20,
Bax Caspase-11 FAP-1, Fas-ligand IEX-1 L, Nr13
35
Table 4. list of genes regulated by NF-kB (ctd)
Kinases PI3K PIK3 PKC8 MAP 4K1 P450 CYP2C11 CYP2E
1 Oxidative stress XOD GST Mn SOD
INOS Enzymes ?-GCS 11bGSD2 12-LOX 5-LOX ABC
transporters ADH ARFRP1 Aromatase BACE Catepsin
B and ceramide glycosyl transferase
Collagenase 1 CRAD1 and 2 Dihydrodiol
dehydrogenase DT diaphorase
Enzymes (ctd) Gelatinase B GSTP1-1 Guanylyl
cyclase a H-K-ATPase a2 Heparanase HO-1 Hyaluro
nan synthase lodothyronine deiodinase Lysozyme MK
P-1 N- Acethylglucosaminyl transferase
1 NGAL NQO1 PDE 7A1 PGES\L-PGDS Phospholipase
A2 Phospholipase C d1 PIM-1 PTGIS RACK1 REV3 Sepri
n 2A SNARK TERT Transglutaminase
Miscellaneous a-1 acid glycoprotein
a1-Antitrypsin, a2(1)-Collagen a-Fetprotein AMH A
polipoprotein C III ß-Amyloid Biglycan Caveolin-1
Claudin-2 Clone 156 Clone 330 Clone 68 Connexin
32 Epsilon-globin Factor VIII Gadd45ß Galectin
3 GIF GS 3686 HMG-14 K15 keratin K3 keratin K3
keratin K6 keratin
Laminin B2 chain mCGM3 Mts1 MUC-2 Mx1 Neutrophil
gelatinaseassosiated lipocalin NlF
1 P11 PAI-1 Pax 8 PCBD Perforin PGK PSA RICK S100A
6 (calcyclin) Spergen-1 Syndecan-4 TFPI-2 Transf
errin Urokin 16 UBE 2M UCP-2 Vimentin Wilms'
tumor suppressor gene
36
kérdés mivel gátolható az abberáns
NFkB aktivitás?
37
Table 5. list of agents that inhibit NF-KB
activation
Anti-inflammatory agents Aspirin,
Indomethacin Diclofenac, Naproxen Dexamethasone Ph
enylbutazone Tamoxifen, Sulfasalazine Sulindac,
Ibuprofen Leflunomide Trichodion Acetaminophen
Gabexate mesilate Glucosamine sulfate
Petrosaspongiolide M Dexanabinol,
BSASM Cloricromene, Rolipram Pranlukast ImmuInosu
pressive agents Cyclosporin, LF15-0195 15-Deoxyspe
rgualin 6(5H)-Phenanthridinone and
benzamide Chemopreventive agents Carboplatin,
Vesnarinone Cycloprodigiosin (- )-Cycloepoxydon
Campthothecin, Gemcitabine Levamisole, Epoxomicin
Angiogenesis 2-Methoxyestradiol TNP-470 Cell
proliferation 6-Aminoquinazoline derivatives
Raxofelast Virus K1L, HIV-1 Vpu protein Canine
distemper virus Antioxidants ADT, Apocynin,
Benidipine BHA, Carvedilol Catechol
derivatives DHEAS, DMDTC DMSO, Disulfiram Ebselen,
EPC-K1 21 Ethyl pyruvate Ethylene glycol
tetraacetic acid Iron tetrakis, Mn-SOD Melatonin,
NAL, NDGA Orthophenanthroline Hydroquinone and
tert-butyl hydroquinone Ref-1, Tempol,
Tepoxaline N-Acetylcysteine Reactive oxygen
species Nitric oxide
Antibacterial agents Fosfomycin 5-Aminosalicylic
acid Clarithromycin Rifampicin Roxithromycin Ant
idiabetes agents Adiponectin Glimepiride Tyrosine
phosphatase inhibitors Diamides Pervanadate Pheny
larsine oxide Serine protease inhibitor Pefabloc
Phosphathidyl-choline - phospholipase C
inhibitor D609
HIV-1 protease inhibitors Nelfinavir Ritonavir Sa
quinavir ACE inhibitor Ouinadril COX-2
inhibitor Celecoxib Interferon inducible
protein p202a Methanesulfonilide antiarthritis
inhibitor T -614 PTK inhibitor Herbimycin
A Farnesyl protein transferase inhibitor
SCH66336
38
Table 5. list of agents that inhibit NF-KB
activation (ctd)
Others ß-Amyloid protein, ß Catenin ?-Glutamylcys
teine synthetase 1,2,3,4,6-Penta-O-galloyl-
beta-D-glucose 1,2,4- Thiadiazolidine derivatives
15-Deoxy-prostaglandin J(2) 17 -Allylamino-17-
demethoxygeldanamycin 2-Acetylaminofluorene 3,
4,5- Trimethoxy-4'- fluorochalcone 4-Hydroxy-
2 -nonenal 5'-Methylthioadenosine Acrolein,
Adenosine AMP-activated protein kinase Amrinone,
Anandamide Angiopoietin-1, ANP Antihrombin,
Antithrombin III APCO576, Arsenite Auranofin,
AvrA protein AZT, Bcl-Ab1, Benfotiamine Biliverdin
Biotinylated isopanepoxydone Bisphenol A,
BMT Bovine serum albumin BZLF1, C5a Carbon
monoxide Cyclopentones CYL-19s and CYL-26z CYLD,
Desloratadine Disulfiram, Dobutamine Da 65-79,
DTD
Protease inhibitor Nafamostat mesilate PPAR?
ligand Pioglitazone Heat shock proteins HSP70,
HSP72 Statins Atorvastatin Erbstatin Fluvastatin
Lovastatin Metals Chromium Cadmium
Gold Lead Mercury Zink Arsenic Titanium
IKB-like protein Encoded by ASFV Acetylation
inhibitor HDAC
E3330, E-73, Ecabet sodium Hydroquinone,
Hypochlorite Inhaled isobutil nitrite Jesterone
dimer, KT-90 APC Losartin, Calcitriol,
CaMKK Campthothecin, Caprofin Decoy
oligonucleotides Dehydroxymethylepoxy
quinomicin Diethyldithiocarbamic acid
Diethylmaleate Diltiazem, DMF,
Epoxyquinol Erythromycin Estrogen (E2) Ethacrynic
acid Ethyl 2-(3-methyl-2,5-
dioxo(3-pyrrolinyl)) Ethyl pyruvate, Evans
blue Fibrates, Fish oil feeding Flunixin
meglumine Flurbiprofen, Fox1j G-120 3032,
Gangliosides Gemfibrozil, Ghrelin GILZ,
Glucocorticoids Hydroquinone, HSCO Human breast
milk Hycrochloride Hyperosmolarity Hypothermia,
IMD-O354 Intravenous immunoglobulin Jesterone,
JSH-21, JSH-23 Kaposi's sarcoma-associated
herpesvirus K1 protein
Ketamine, KL-1156, LMB Macro/ide
antibiotics MAST205, MEB, Melatonin Mercaptopyrazi
ne Mesalamine, Methotrexate Monochloramine Monomet
hylfumarate MTA, Murr1, MX781, EM, Myxoma Virus
MNF N-(p-coumaroyl) serotonin Neurofibromatosis-
2 protein nicotinamide 3- aminobenzamide Nilv
adipine, Nitrosoglutathione Nitrosylcobalamin
mild hypothermia, N-Octylcaffeate N-Oleoyldopamine
NPM-ALK oncoprotein, NRF Nucling, ox-LDL and
HNE OXPAPC, pB, p53, PACAP PAN1, Panepoxidone,
PBN PDTC, PEDF, Pentoxifylline Photo oxidative
stress PIAS, Pirfenidone, PPM-1B Probiotics Pg
J2, Pg A1, Pg E2, PSK Psychosine, PTEN ,
PTX-B Pyrithione, Quinazolines pyrrolidinedithioca
rbamic -acid Rapamycin, Rebamipide RelA
peptides, Retinoic acid receptor-related orphan
receptor-alpha
39
Table 5. list of agents that inhibit NF-KB
activation (ctd)
Others (ctd) Ribavirin Rifamides
,Ritonavir RKIP Rocaglamide Saline Selenomethionin
e Siah2, SiRNA SLPI, SOCS1 Sodium salicylate SUN
C8079 Surfactant protein A Survanta TAT
-SR-I?Ba Tetrathiomobdate Thalidomide THI
52 Thiopental TNAP Tom1 Tranilast Triflusal Trigly
ceride-rich lipoproteins Troglitazone Tyrphostin
Ursodeoxycholic acid Uteroglobin Vasoactive
intestinal peptide vlRF3 YopJ ZAS3 protein ZUD
protein
40
kérdés hol találhatók olyan anyagok,
amelyek csökentik a kóros NFkB aktivitást?
41
Table 6. natural products that suppress NF-kB
activation
Lupeol Luteolin Magnolol ?-Mangostin Manassantis M
angiferin Nicotine Oleandrin Onconase Panduratin
A Palthenolide PC-SPES Phenethylisothiocyanate Pho
mol Phytic acid Phallacidin Piceatonnol Piperine Q
ingkailing Shuanghuanglian Quercetin Quercetin Res
iniferatoxin Resveratrol Rhein Rocaglamide Rotenon
e Sanggenon C Sanguinarine Saucerneol Sauchinone S
esamin Sesquiterpene lactones Silymarin Sinomenine
a-Lipoic acid a-Pinene a-Tocopherol a-Torphryl
succinate ß-Lapachone 1-Acetoxychavic
ole acetate 20(S)-Protopanaxatriol 23-Hydroxyursol
ic acid 7-Amino-4-methy- coumarin Acetyl-bosw
ellic acids Allyl-cysteine Mentoflavone Andalusol
Andrographolide Anethole Apigenin Artemisia
sylvatca Artemisinin Ataxanthin Astragaloside
IV Aucubin Avicin Baicalein Bambara groundnut Bee
venon Benzyl isothiocynate Betulinic
acid Bis-eugenol Bupleurum fruticosum
phenylpropanoids Cacospongionolide
B Calagualine Cancer bush
Epoxyquinone A Falcarindol Flavenoids Flavopiridol
Fungal gliotoxin Ganoderma lucidum
polysaccharides Garcinol Genistein Galdenamycin Ge
nipin Ginkgolide B Glabridin Glitoxin Glossogyne
tenuifolia Glycyrrhizin Guggulsterone Helenarin He
matein Herbal compound861 Hydroxyethyl
starch Hypericin 10Z-hymenialdisine Hypoestoxide I
ndirubin-3-oxime IRFI 042 Isomallotochromanol
isomallotochromene Isorhapontigenin Kahweol
Quinic acid Kamebakurin Lacidipine L-Ascorbic
acid Lazaroids
CAPE Capsaicin Capsiate Camosol Catalposide Cinnam
aldehyde/2-methoxycinnamaldehyde Cycloepoxydon Cel
astrol Cepharantin Cytochalasin
D Conophyline Cheongyeolsaseuplang Chromene
derivatives Chicory root Chitosan Curcumin Cycloli
nteinone Danshenshu DDC Dehydroascorbic
acid Diarylheptanoid Dibenzylbutyrolactone
lignans Diferoxamine Digitoxin Dihydroisoeugenol D
ihydrolipoic acid DIM/I3C Ebselen EGCG Emodin Ent-
kaurene diterpenoids Ergothioneine Evodiamine
Solona nigrum L Sphondin Staurosporine Tansinones
Tetrandine Theaflavin Trilinolein Triptolide Ursol
ic acid Vitamin C Wedalolactone Withaferin
A Wogonin Wortmannin Yakuchinone A and
B Zerumbone
42
Table 6. natural products that suppress NF-kB
activation
Others 1,2,4-Thiadiazolidine derivatives Aged
garlic extract, Apple juice Artemisia capillaris
extract Blueberry and berry mix Black raspberry
extracts Catsclaw bark, Extract of the stem
bark of Mangifera indica L Extracts of Ochna
macroccalyx Fomes fomentarius methanol
extracts Fructus benincasae recens extract Ginkgo
biloba extract Harpagophytum procumbens
extracts Kochia scoparia fruit, Omega 3 fatty
acids Phyllanthus amarus extracts Platycodi radix
extract, Red wine Sophorae radix extract Stinging
nettle plant extrats Tanacetum larvatum
extract Uncaria tomentosum plant
extract Trichomonas vaginalis Xanthium strumarium
L Yucca schidigera extract
43
kérdés praktikusan - mely kis-molekula-
tömegu természetes vagy szintetikus anyagok
csökkentik az NFkB aktivitását?
44
Table 7. proteins and peptides that inhibit NF-kB
activation
Interleukin IL-4 IL-11 IL-10 IL-13 Interferon IF
N-a
Peptides TRAF6 inhibitory peptide NEMO binding
peptide p65 binding peptide Boronic acid
peptide NLS cell permeable peptides Peptide
nucleic acid-DNA decoys Peptide YY TIRAP
inhibitor peptide
Growth factors and hormones VEGF Hepatocyte
growth factor MSH HB-EGF Chorionic gonadotropin

Table 8. synthetic compounds that inhibit NF-kB
activation
Proteosome inhibitors ALLnL PS-341 Lactacystine,
ß-lactone SN-50 FK506 Ro106-9920 PS5190 TLCK
,TPCK Cyclosporin A DCIC Deoxyspergualin DFP LLM B
TEE MG115 MG120 APNE
IKK inhibitors BAY-11-7082 BAY-11-7085
BMS-345541 SPC839 AS602868 PS1145 MNL
120 SC-514 Polyubiquitination inhibitor
Ro106-9920 Ubiquitin Ligase Inhibitors Z-LLL,
Z-LLnV
P42 MAPK inhibitor AG 126 P38 MAPK inhibitor
SB203580 PI-3K-kinase inhibitor LY294002 PKC
inhibitor RO31-8220 MEK1/2 inhibitor
UO126 Protein kinase inhibitor A771726
45
kérdés mely pontokon lehet/kell szerkeze
ti változtatást létrehozni (pl.
rekombináns technikákkal) az NFkB
aktivitásának specifikus csökkentéséhez?
46
Table 9. inhibition of NF-kB through the
critical cystein residue
IKKa/ß Cys179 IKKß Cys59 IKK?
Cys417 P65 Cys38 p50 Cys62
47
kérdés milyen célmolekulákat tervezzünk
az NFkB aktivitásának
csökkentsére? a szignálút melyik szakaszára?
48
kérdés az aktivációs mechanizmus
gátlásával változik-e az NFkB aktivitása?
gátló stimulusok használhatók-e? hát az
IkB gátlása?
49
az NF-kB gátlás kehetoségei 1.
Aktiváló Stimulus (kfl)
gyulladási-/immun- stb. fehérjék
IkB kináz /NEMO/IKAP
P
IkBa
IkBa
IkBa
p65
p65
p50
p50
degradáció
mRNS
inaktív forma marad
CITOPLAZMA
p65
p50
NUKLEUSZ
transzkripció
p65
p50
transzlokáció
Target genes
50
kérdés az IkB kináz ativitásának
vagy aktiválásának gátlásával lehet
csökkenteni az NFkB aktivitását? mi az
ubiquitináció gátlás hatása?
51
az NF-kB gátlás kehetoségei 2.
Aktiváló Stimulus (kfl)
gyulladási-/immun- stb. fehérjék
IkB kináz/NEMO/IKAP
P
IkBa
IkBa
IkBa
p65
p65
p50
p50
degradáció
mRNS
inaktív forma marad
CITOPLAZMA
p65
p50
NUKLEUSZ
transzkripció
p65
p50
transzlokáció
Target genes
52
az NF-kB gátlás kehetoségei 3.
Aktiváló Stimulus (kfl)
gyulladási-/immun- stb. fehérjék
IkB kináz/NEMO/IKAP
P
IkBa
IkBa
IkBa
p65
p65
p50
p50
degradáció
mRNS
CITOPLAZMA
p65
p50
NUKLEUSZ
transzkripció
p65
p50
transzlokáció
Target genes
53
nem megoldott - sejt-specificitás - alacsony
toxicitás sok fejleszteni való van hátra
54
screen 220eFt/kísérlet Human NF?B Signaling
Pathway RT² Profiler PCR Array
Functional Gene Grouping (more detailed
information is provided in the gene table)
Activation of NF?B pathwayLigands and receptors
IL1B, IL8, MYD88, TLR1, TLR2, TLR3, TLR4, TLR6,
TLR8, TNF, TNFRSF10A, TNFRSF10B. Membrane
molecules IRAK1, IRAK2. Kinases CHUK, IKBKB,
IKBKG, IRAK1, IRAK2. I-kappa-B kinase/NF-kappa-B
cascade EDARADD, IKBKG, IRAK2, STAT1, TLR8.
Cytoplasmic sequestering/releasing of NF-kappa-B
BCL3, IL10, NALP12, NFKBIA, TNFRSF7, TNFSF14.
Transcription factors IKBKB, IKBKG, IRAK1,
NFKB1, NFKBIA, RELA, STAT1, TNF. Inflammatory
response IL1B, IL8, IRAK2, MYD88, NFKB1, TLR1,
TLR2, TLR3, TLR4, TLR6, TLR8, TNF. Positive
regulation of I-kappa-B kinase/NF-kappa-B
cascadeLigands and receptors CD40 (TNFRSF5),
F2R, FASLG (TNFSF6), HTR2B, LTBR, SLC20A1,
TICAM2, TNFRSF1A, TNFSF10, TRIF. Membrane
molecules EDG2, GJA1, HMOX1, RHOA. Kinases
IKBKE, RIPK1, TBK1. Other BCL10, BIRC2, CARD4,
CASP1, CASP8, CFLAR, CTL2, FADD, MALT1, PPM1A,
REL, RFP2, TMED4 (HNLF), TRADD. NF?B Responsive
GenesAcute phase response proteins AGT, BF.
Adhesion molecules ICAM1. Extracellular
molecules BF, CSF2, CSF3, IFNA1, IFNB1, IFNG,
IL6. Ligands and receptors AGT, CCL2, ICAM1,
LTA. Cytokines CCL2, CSF2, CSF3, IFNA1, IFNB1,
IFNG, IL6, IL8, LTA, TNF. Inflammatory response
CCL2. Other factors involved in NF-kappa-B
pathwayExtracellular molecules IL1A. Ligands and
receptors IL1R1, RAF1, TLR7, TLR9. Kinases
AKT1, MAP3K1, RAF1. Transcription factors ATF1,
EGR1, ELK1, FOS, JUN, NFKB2, RELB, TNFAIP3.
Inflammatory response FOS, IL1A, IL1R1, TLR7,
TLR9  
55
köszönöm a figyelmet
56
(No Transcript)
57
(No Transcript)
58
(No Transcript)
59
(konstitutívan) aktív NFkB-t expresszáló
tumorok (pl.)

• Prostate
• Melanoma
• Head and neck SCC
• Colon
• Multiple myeloma
• Ovarian
• Bladder
• Lung

• B cell lymphoma
• Hodgkins disease
• T-cell lymphoma
• Acute lymphoblastic leukemia
• Breast
• Liver
• Thyroid

Leukemia 161053-1068, 2002
60
(No Transcript)
61
(No Transcript)
62
(No Transcript)
63
(No Transcript)
64
(No Transcript)
65
(No Transcript)
66
(No Transcript)