cikk-referáló Génátírás szabályozása: transzkripciós mechanizmusok Dr. Marcsek Zoltán
Génexpresszió Courtesy The Protein Society
eukaryoták promóter elemei • a promóter elemei többé-kevésbé jól definiáltak gyakran hiányzik általában van • a transzkripció start helyétől különböző távolságra vannak
egy tipikus promóter
az eukaryota transzkripció szabályozása
szabályozás transzkripciós faktorokkal • egy transzkripciós faktor (vagy transzkripciós faktorok egy csoportja) egy géncsoport expresszióját szabályozhatja (pl. heat shock fehérjék, G 1/S restrikciós pont fehérjék, stb) • egy gén átírását egy vagy több, egymástól független transzkripciós faktor szabályozhatja (pl. metallothionine) • magas expresszió kiváltásához gyakran több, különböző 'response' elemhez kötő transzkripciós faktor lehet szükséges
Nuclear Factor-κB: Factor-κB 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.
NFk. B mi az, mit csinál, miért jó, ha ismerjük? transzkripciós faktor
NFk. B • Baltimore & Sen fedezték fel mint egy B-sejt-specifikus magi proteint amely az immunoglobulin könnyű láncának meghatározott szekvenciájához kötődik és a gén átírását serkenti (enhancer) (Cell 47: 921 -928, 1986) 1986 • az NF B a Rel transzkripciós faktor családba tartozik amely nagyszámú, az immunválasszal, a gyulladással, a sejtosztódással és apoptózissal összefüggő gén expresszióját szabályozza • az emlős sejtekben öt eltérő NFk. B (rel) alegységet ismerünk amelyek kb. 300 nagyon konzervált aminosat tartalmaznak • a citoplazmában az NFk. B inaktív formában több, különböző kombinációban található (heterodimer p 50/p 65 (Rel A) és az inhibítor I B alegység)
Family of NF B and Inhibitory B (I B) proteins Nat. Rev. Cancer 2: 301 -310, 2002
p 105 (contains p 50 NFk. B 1) • Name: p 105 (contains p 50 subunit, a. k. a. NFk. B 1) • Descr: Once in the nucleus initiates transcription of hundreds of genes • Length: 968
p 100 (contains p 52 NFk. B 2) • Name: p 100 (contains p 52 subunit, a. k. a. NFk. B 2) • Descr: Once in the nucleus initiates transcription of hundreds of genes • Length: 899
Ik. B • Name: Inhibitor of NF kappa. B beta • Length: 356 • Consists mostly of ankyrin repeats that serve inhibitory function
Ik. B • Name: Ikappa. B epsilon • Length: 361 • Consists mostly of ankyrin repeats that serve inhibitory function
IKK Name: IKappa. B kinase alpha Descr: Forms complexes with IKK Length: 745 aa Conserved Serine/Threonine protein kinase Domain detected at 10 – 300 aa.
IKK • Name: IKappa. B kinase beta • Descr: Forms complexes with IKK and destroys Ik. B’s • Length: 756 Similarly to IKK a slightly less conserved Serine/Threonine protein kinase Domain detected at 10 - 290 aa.
IKK • Name: IKappa. B kinase gamma • Length: 487 aa Contains Chromosome segregation ATPases domain
az NF B aktiválása • NF B a citoplazmában található inaktív formában mint az I B inhibítorához kötött p 50/65 heterodimer • extracelluláris indukciós hatásra az I B -kináz komplex (IKK) aktiválódik és foszforilálja az I B -t (szerin 32 és 36) amely az I B lizin 21 és lizin 22 ubiquin-kötését okozza • ez az I B alegység lebotását eredményezi (26 S proteasome) • az inhibitor alegység lebomlása a p 50/p 65 komplex aktivációját eredményezi: az NFk. B a citoplazmából a sejtmagba kerül (transzlokáció) • a sejtmagban az NFk. B transzkripciós faktor a ‘consensus szekvenciához’ (5’-GGGACTTTC-3’) kötődik és aktiválja a specifikus gének expresszióját
Aktiváló Stimulus (kfl) Ik. B-kináz/NEMO/IKAP komplex I B p 50 p 65 P p 65 gyulladási-/immunstb. fehérjék I B degradáció m. RNS CITOPLAZMA p 50 NUKLEUSZ p 65 transzkripció transzlokáció p 50 p 65 Target genes
Factors that induce NF B induce Reactive Oxygen Species (ROS) Leukemia 16: 1053 -1068, 2002
Free Radic Biol Med 25: 346 -361
kérdés: mely szignálok aktiválják az NFk. B aktivitást?
Table 1. A list of inducers of NF-k. B activation Cytokines TNFα , TNFβ RANKL, TRAl. L CD 40 ligand CD 30 ligand TWEAK THANK BAFF LTβ IL-1, IL-2, IL-4 IL-8, IL-12, IL-15 IL-17, IL-18 CD 11 b/CD 18 ligand CD 2 ligand CD 35 ligand CD 3 ligand CD 28 ligand CD 43 ligand CD 4 ligand CD 66 a ligand Ll. F S 100 B Agonist antibodies to AT 1 receptor Ig. M ligand Beta-D-glucan ligand Ig. G 2 a Flt-1 ligand Ly 6 A/E 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) Prostratin Oncogenes Chemical Agents Ras Myc Bcr-abl All-trans retinoic acid RET/PTC 3 fusion oncoprotein S 100 B Serum Sulphatide TGF-α TGF-β L-NMA, Ethanol, Monensin Nicotine, Adriamycin 2 -Deoxyglucose Anisomycin Benzyl isothiocyanate Bisperoxovanadium phosphotyrosyl Brefeldin A, Cadmium Calyculin A, Colchicine Calcium lonophore plus phorbol ester Ceramide-beta-galactose 2 -chloroethyl 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 Phorbol ester, Podophy/lotoxin Polychlorinated biophenyl-77 Prostratin, Pyrogallol Staurosporine, Tunicamycin Transglutaminase 2 WF 10, WY-14 643, CFTR, GPR 56 Hematopoietic progenitor kinase 1 Ig heavy chain, MHC Class I
Table 1. A list of inducers of NF-k. B activation (ctd) activation Viruses Viral products Bacteria and fungi Bacterial/fungal products Adenovirus Cytomegalovirus EBV Hammarskjöld 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 Adenovírus 5: E 1 A Adenovirus: E 3/19 K African swine fever virus: IAP Antibody to dengue virus nonstructural protein 1 CMV: ie 1 EBNA-2 EBV: LMP 1 HBV: HBx HBV: LHBs HBV: MHBst HCV: Core protein HVS 13 HVS: Stp. C HIV-1: gp 160 HIV-1: Nef HIV-1: p 9 (9 aa peptide) HIV-1: Tat HTLV-I: Tax HTLV-II: Tax Influenza vírus: hemagglutinin KSHV: K 1 Parvovirus B 19: NSI Respiratory syncytial virus: M 2 -1 SV 40 Anaplasma phagocytophilum Angiostrongylus 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 Listeria 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 f. Iexneri Staphy/lococcus aureus Streptococcus Streptomyces californicus Trichomonas vaginalis Ureaplasma urealyticum Yersinia enterocolitica LPS Wogonin Enterotoxin GIycosylphosphatidylinositols Fumonisin B 1 Staphylococcus enterotoxins A and B Apicularen A Cp. G Cytolysin Diphosphoryl lipid A Exotoxin 8 Lipoprotein Lp. K Lipoteichoic acid Membrane lipoproteins (Mycoplasma fermentans, Mycoplasma penetrans) Mannoproteins Muramyl peptides Mycobacterium species lipoarabinomannan Pic. A, Plc. B Porin 1 B Toxic shock syndrome toxin 1 Eukariotic parasietis Eimeria tenella and necatrix Leishmania Phospholipomannan Pneumocystis Theileria parva Trypanoplasma borreli
Table 1. A list of inducers of NF-k. B activation (ctd) activation Physiologic (stress) cond. Asthma, Rheumatoid arthritis Crohn's disease/ulcerative colitis Corneal epidemic keratoconjunctivitis Coronary 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, fo. Ga. I) 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 Environmental hazards Chemotherapeutic drugs UV A, B and C radiation Bile duct ligation Cyclic mechanical muscle strain Exercice γ 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 Cigarette smoke Arsenic Polychlorinated biphenyl Benzo[a]pyrene diol epoxide Crocidolite asbestos fibers Dicamba Diesel exhaust particles Gliadin House dust mite Dust particles Fear-potentiated startie response Lead chromate Noise Olly fly ash Silica partlcles Wood smoke Zymosan Oxldative stress Glutathione Hydrogen peroxide Catalase Nitric oxide Ozone Peroxiynitrite Butyl peroxide Pervanadate Reoxygenation Cisplatin, Paclitaxel Doxorubicin, Tamoxifen Vinblastine, Vincristine Etoposide Ara-C Bryostatin-1 Camptothecin Daunomycin Daunorubicin Mitoxantrone Gemcitabine Therapeutic drugs Celecoxib, AZT, Ciprofibrate Bleomycin ABR-25757 SN 38 SA 981, WR 1065 Ad. AMP, Anthralin Cycloprodigiosin Dacarbasine Orengedeokuto Diazoxide 5, 6 -dimethylxanthenone-4 -acetic acid Flavone-8 -acetic acid Haloperidol Cunbi-Boshin. Hangam-Tang Methamphetamine Oltipraz Phenobarbital Protocatechuic acid
Table 1. A list of inducers of NF-k. B activation (ctd) Fatty acids Apoptotlc Mediators Palmitate Leukotriene B 4 Free fatty acids Llnolelc acid Oleic acid Saturated fattx acids Lysophosphatldlc acid Anti-Fas/Apo-1 PARP Physiological Mediators Alloxan Angiotensin II Bradykinin Fibrinogen Metals Hemoglobin Lithium Titanium and copper implants Hyaluronan Thioredoxin Nickel sulfate Kainan acid Iron L-Glutamate Modified Proteins Manganase Melanin LDL Nickel Violacein AGEs Chromium Vitamin D 3 Amyloid protein fragment Cobalt Adesonine Glycosylated Aluminium Activin A oxyhemaglobin Albumin Maleylated bovine serum Natural products Allergin albumin Areca nut extract Acrp 30/adiponectin Neurotrophin receptor Baicalin Adrenomedullin proteolytic fragments Beta-carotene Aeginetia indica L (AIL)b-A Nonamyloid beta Quercetin Bronchoalveolar lavage fluid component of Quinolinic acid Brusatol Alzheimer's disease Safflower polysaccharides Homocysteine ATM Sanglifehrin A Cryptdins Deoxicholic acid Heat shock protein 12 -hydroxyeicosatrienoic acid HSP 60 6 -Hydroxydopamine HSP 71 Human peptide transporter 1 LTPLyso PCNeuomelanin NS-398 Osteopontin PAF PCSC Prolactin PIF 12 -Lipoxygenase Thrombin Streptozotocin Substance P Potassium Bum patient serum Anaphyloxin C 3 a and C 5 a Angiotensin II type 1 receptoractivating antibodies Antiphospholipid antibodies Basic Calcium Phosphate crystals C 2 -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 S 100 B Sleep deprivation TUDCA Tuberous sclerosis complex Uremic toxins, VLDL
Constitutive NF-k. B Signaling Module Input signal I K P I B P NF B I B NF B NLS NF B I B Degraded Cytoplasm I B NF B Negative feedback I B gene NF B IL-6 gene I B NF B
kérdés: mely szignálutak vesznek részt az NFk. B aktivitásában?
Table 2. protein kinases implicated in NF-k. B activation Kinase Ligand IKKα IKKβ IKKγ IKKi/IKKε NIK NAK PI-3 K Akt 1 HPK-1 PKC-α PKC-β PKC-τ PKC-δ PKC-ε a. PKC MEKK 1/2 MEKK 3 TPL-2/Cot kinase pp 90 rsk Raf-1 kinase PKR TAK 1/MEKK MLK 3 PAK 1 BTK JAK 2 PKA IRAK-1 IRAK-2 IRAK-M P 56 Ick Syk Proinflammatory cytokines (eg TNF, IL-1, IL 6), LPS, Ik. B TNF, PMA, Ik. B TNF, CD 95, and IL-1 Phorbol esters, growth factors IL-1 TNF, TCR/CD 28 Unknown TPA, TNF, Bimp 1, Bcl 1 O/MAL T 1 Ig. M receptor, Bimp 1, Bc. I 10/MAL T 1 CD 3 -CD 28 (TCR/CD 28), Bimp 1, Bcl 1 O/MAL T 1 TNFα Phorbol ester, Bimp 1, Bcl 1 O/ MAL T 1 TNF, ras p 21 Proinflammatory cytokines TNF, other proinflammatory cytokines TNF Pseu omonas aeruglnosa Serum ~rowth factors, phorbol ester, and PTK oncogenes ds. RNA XIAP, TGF-β CD 3/CD 28 p 21, LPS B-cell antigen receptor Erythropoietin IL-1, LPS, many others IL-1, LPs IL-1, LPS Ceramide TNF, H 2 O 2 min. 32 ismert kináz
kérdés: mely betegségekben mutatható ki abberáns (állandóan magas) NFk. B aktivitás? többet tudunk, ha ismerjük a rendszer hibáit. . .
Table 3. association of NF-k. B activation with diseases Cancer Cardiovascular diseases Other diseases 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 Atherosclerosis Cardiac hypertrophy Heart failure Hypercholesterolemia 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 Diabetes mellitus 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
kérdés: mely gének aktiválódnak az NFk. B hatására? mi a hiba okozója (fenotípus)
Table 4. list of genes regulated by NF-k. B Cytokines Receptors TNF-α, TNF-β IL-1 -α, IL-1β IFN-α, IFN-γ TRAl. L, M-CSF RANTES FAS ligand Lymphotoxin α, β G-CSF, GM-CSF IL- 2, IL- 6, IL -8, IL- 9, IL-10, IL-11, IL-12, IL-13, IL-15 NK 4, GCP-2 (CXCL 6) Gro α, Gro γ, Gro-1 ICOS IL-1 receptor antagonist IP-10, KC, BMP-2 CCL 15, CCL 22, CCL 28, CCL 5 CD 40 ligand CINC-1, CXCL 11 ENA-78 (CXCL 5) Eotaxin Erythropoietin MCP-1/JE, MIP-1 α β MIP-2, MIP-3 α, Mob-1 Neutrophil- activating peptide 78 NK-1 R Stem cell factor Angiotensinogen TCA 3 TFF 3 TSP-1 TSP-2 beta 2 microglobulin µ-opioid receptor 2 A A 1 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 γ 4, Ig Cγ 1, Ig ε heavy chain Ig к light chain, invariant chain II Lox-1, Mdr 1 MHC class (HLA-B 7) MHC class I (H-2 Kb) Neuropeptode YY 1 - receptor NMDA receptor subunint NR-1 NMDA receptor subunit Nod 2 Polymeric Ig receptor RAF receptor 1, RAGE T -cell receptor β chain T -cell receptor CD 3γ TNF-Receptor, p 75/80 DR 4, DR 5 Early response genes p 62, p 22/RPG 1, B 94 Egr-1, TIEG Transcription factors NF-КB 1, NF-КB 2, c-Rel, Rel. B Ik. Bα c-fos, p 53, c-myb, Jun. B E 2 F 3 a, Elf 3 , ELYS ETR 1001 IRF-1, IRF-2, IRF-4, IRF-7 Mail, Nurr 1, Stat 5 a, WT 1 Metastasis ICAM-1, VCAM-1 ELAM-1, E-selectin Endoglin, Fibronectin Mad. CAM-1, P-selectin Tenascin-C, DC-SIGN MMP-3, MMP-9, CXCR 4 KA 11/CD 82, u. PA Antiapoptosis Bcl-2, Bcl-XL, Bfl 1/A 1 TRAF-1, TRAF-2, TRAF 6 c-FLIP, IAPs Survivin, A 20, Bax Caspase-11 FAP-1, Fas-ligand IEX-1 L, Nr 13 Angiogenesis VEGF Cell proliferation COX-2, c-myc Cell cycle Cyclin D 1, P 21, GADD 45 Cyclin D 2, Cyclin D 3 Viruses HIV-1 EBV wp promoter HBV pregenomic promoter HSV, SV-40 Adenovirus (E 3 region) Avian leukosis virus Bovine leukemia virus Cytomegalovirus JC virus, HPV 16, SIV Proteins involved in antigen presentation Tapasin LMP 2 TAP 1 Complement B Complement receptor 2 Complement component 3
Table 4. list of genes regulated by NF-k. B (ctd) NF-k. B Kinases PI 3 K PIK 3 PKC 8 MAP 4 K 1 P 450 CYP 2 C 11 CYP 2 E 1 Oxidative stress XOD GST Mn SOD INOS Enzymes γ-GCS 11 b. GSD 2 12 -LOX 5 -LOX ABC transporters ADH ARFRP 1 Aromatase BACE Catepsin B and ceramide glycosyl transferase Collagenase 1 CRAD 1 and 2 Dihydrodiol dehydrogenase DT –diaphorase Enzymes (ctd) Miscellaneous Gelatinase B GSTP 1 -1 Guanylyl cyclase α H+-K+-ATPase a 2 Heparanase HO-1 Hyaluronan synthase lodothyronine deiodinase Lysozyme MKP-1 N- Acethylglucosaminyl transferase 1 NGAL NQO 1 PDE 7 A 1 PGESL-PGDS Phospholipase A 2 Phospholipase C δ 1 PIM-1 PTGIS RACK 1 REV 3 Seprin 2 A SNARK TERT Transglutaminase α-1 acid glycoprotein a 1 -Antitrypsin, a 2(1)-Collagen α-Fetprotein AMH Apolipoprotein C III β-Amyloid Biglycan Caveolin-1 Claudin-2 Clone 156 Clone 330 Clone 68 Connexin 32 Epsilon-globin Factor VIII Gadd 45β Galectin 3 GIF GS 3686 HMG-14 K 15 keratin K 3 keratin K 6 keratin Laminin B 2 chain m. CGM 3 Mts 1 MUC-2 Mx 1 Neutrophil gelatinaseassosiated lipocalin Nl. F 1 P 11 PAI-1 Pax 8 PCBD Perforin PGK PSA RICK S 100 A 6 (calcyclin) Spergen-1 Syndecan-4 TFPI-2 Transferrin Urokin 16 UBE 2 M UCP-2 Vimentin Wilms' tumor suppressor gene
kérdés: mivel gátolható az abberáns NFk. B aktivitás?
Table 5. list of agents that inhibit NF-KB activation Anti-inflammatory agents Aspirin, Indomethacin Diclofenac, Naproxen Dexamethasone Phenylbutazone Tamoxifen, Sulfasalazine Sulindac, Ibuprofen Leflunomide Trichodion Acetaminophen Gabexate mesilate Glucosamine sulfate Petrosaspongiolide M Dexanabinol, BSASM Cloricromene, Rolipram Pranlukast Immu. Inosupressive agents Cyclosporin, LF 15 -0195 15 -Deoxyspergualin 6(5 H)-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 K 1 L, HIV-1 Vpu protein Canine distemper virus Antioxidants ADT, Apocynin, Benidipine BHA, Carvedilol Catechol derivatives DHEAS, DMDTC DMSO, Disulfiram Ebselen, EPC-K 1 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 HIV-1 protease inhibitors Nelfinavir Ritonavir Saquinavir ACE inhibitor Antidiabetes agents Ouinadril Adiponectin Glimepiride Celecoxib Tyrosine phosphatase inhibitors COX-2 inhibitor Interferon inducible protein p 202 a Diamides Pervanadate Phenylarsine oxide Methanesulfonilid e antiarthritis inhibitor Serine protease inhibitor T -614 Pefabloc Phosphathidylcholine phospholipase C inhibitor D 609 PTK inhibitor Herbimycin A Farnesyl protein transferase inhibitor SCH 66336
Table 5. list of agents that inhibit NF-KB activation (ctd) activation Protease inhibitor Nafamostat mesilate PPARγ ligand Pioglitazone Heat shock proteins HSP 70, HSP 72 Statins Atorvastatin Erbstatin Fluvastatin Lovastatin Metals Chromium Cadmium Gold Lead Mercury Zink Arsenic Titanium IKB-like protein Encoded by ASFV Acetylation inhibitor HDAC Others β-Amyloid protein, β –Catenin γ-Glutamylcysteine 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 APCO 576, Arsenite Auranofin, Avr. A protein AZT, Bcl-Ab 1, Benfotiamine Biliverdin Biotinylated isopanepoxydone Bisphenol A, BMT Bovine serum albumin BZLF 1, C 5 a Carbon monoxide Cyclopentones CYL-19 s and CYL-26 z CYLD, Desloratadine Disulfiram, Dobutamine Da 65 -79, DTD E 3330, E-73, Ecabet sodium Hydroquinone, Hypochlorite Inhaled isobutil nitrite Jesterone dimer, KT-90 APC Losartin, Calcitriol, Ca. MKK Campthothecin, Caprofin Decoy oligonucleotides Dehydroxymethylepoxy quinomicin Diethyldithiocarbamic acid Diethylmaleate Diltiazem, DMF, Epoxyquinol Erythromycin Estrogen (E 2) Ethacrynic acid Ethyl 2 -[(3 -methyl-2, 5 dioxo(3 -pyrrolinyl)) Ethyl pyruvate, Evans blue Fibrates, Fish oil feeding Flunixin meglumine Flurbiprofen, Fox 1 j G-120 3032, Gangliosides Gemfibrozil, Ghrelin GILZ, Glucocorticoids Hydroquinone, HSCO Human breast milk Hycrochloride Hyperosmolarity Hypothermia, IMD-O 354 Intravenous immunoglobulin Jesterone, JSH-21, JSH-23 Kaposi's sarcoma-associated herpesvirus K 1 protein Ketamine, KL-1156, LMB Macro/ide antibiotics MAST 205, MEB, Melatonin Mercaptopyrazine Mesalamine, Methotrexate Monochloramine Monomethylfumarate MTA, Murr 1, MX 781, EM, Myxoma Virus MNF N-(p-coumaroyl) serotonin Neurofibromatosis-2 protein nicotinamide 3 - aminobenzamide Nilvadipine, Nitrosoglutathione Nitrosylcobalamin mild hypothermia, N-Octylcaffeate N-Oleoyldopamine NPM-ALK oncoprotein, NRF Nucling, ox-LDL and HNE OXPAPC, p. B, p 53, PACAP PAN 1, Panepoxidone, PBN PDTC, PEDF, Pentoxifylline Photo oxidative stress PIAS, Pirfenidone, PPM-1 B Probiotics Pg J 2, Pg A 1, Pg E 2, PSK Psychosine, PTEN , PTX-B Pyrithione, Quinazolines pyrrolidinedithiocarbamic -acid Rapamycin, Rebamipide Rel. A peptides, Retinoic acid receptor-related orphan receptor-alpha
Table 5. list of agents that inhibit NF-KB activation (ctd) activation Others (ctd) Ribavirin Rifamides , Ritonavir RKIP Rocaglamide Saline Selenomethionine Siah 2, Si. RNA SLPI, SOCS 1 Sodium salicylate SUN C 8079 Surfactant protein A Survanta TAT -SR-IКBα Tetrathiomobdate Thalidomide THI 52 Thiopental TNAP Tom 1 Tranilast Triflusal Triglyceride-rich lipoproteins Troglitazone Tyrphostin Ursodeoxycholic acid Uteroglobin Vasoactive intestinal peptide vl. RF 3 Yop. J ZAS 3 protein ZUD protein
kérdés: hol találhatók olyan anyagok, amelyek csökentik a kóros NFk. B aktivitást?
Table 6. natural products that suppress NF-k. B activation α-Lipoic acid α-Pinene α-Tocopherol α-Torphryl succinate β-Lapachone 1’-Acetoxychavicole acetate 20(S)-Protopanaxatriol 23 -Hydroxyursolic acid 7 -Amino-4 -methy coumarin Acetyl-boswellic 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 CAPE Capsaicin Capsiate Camosol Catalposide Cinnamaldehyde/2 methoxycinnamaldehyde Cycloepoxydon Celastrol Cepharantin Cytochalasin D Conophyline Cheongyeolsaseuplang Chromene derivatives Chicory root Chitosan Curcumin Cyclolinteinone Danshenshu DDC Dehydroascorbic acid Diarylheptanoid Dibenzylbutyrolactone lignans Diferoxamine Digitoxin Dihydroisoeugenol Dihydrolipoic acid DIM/I 3 C Ebselen EGCG Emodin Ent-kaurene diterpenoids Ergothioneine Evodiamine Epoxyquinone A Falcarindol Flavenoids Flavopiridol Fungal gliotoxin Ganoderma lucidum polysaccharides Garcinol Genistein Galdenamycin Genipin Ginkgolide B Glabridin Glitoxin Glossogyne tenuifolia Glycyrrhizin Guggulsterone Helenarin Hematein Herbal compound 861 Hydroxyethyl starch Hypericin 10 Z-hymenialdisine Hypoestoxide Indirubin-3’-oxime IRFI 042 Isomallotochromanol isomallotochromene Isorhapontigenin Kahweol Quinic acid Kamebakurin Lacidipine L-Ascorbic acid Lazaroids Lupeol Luteolin Magnolol γ-Mangostin Manassantis Mangiferin Nicotine Oleandrin Onconase Panduratin A Palthenolide PC-SPES Phenethylisothiocyanate Phomol Phytic acid Phallacidin Piceatonnol Piperine Qingkailing Shuanglian Quercetin Resiniferatoxin Resveratrol Rhein Rocaglamide Rotenone Sanggenon C Sanguinarine Saucerneol Sauchinone Sesamin Sesquiterpene lactones Silymarin Sinomenine Solona nigrum L Sphondin Staurosporine Tansinones Tetrandine Theaflavin Trilinolein Triptolide Ursolic acid Vitamin C Wedalolactone Withaferin A Wogonin Wortmannin Yakuchinone A and B Zerumbone
Table 6. natural products that suppress NF-k. B activation Others 1, 2, 4 -Thiadiazolidine derivatives Aged garlic extract, Apple juice Artemisia capillaris extract Blueberry and berry mix Black raspberry extracts Cat’sclaw 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
kérdés: praktikusan - mely kis-molekulatömegű természetes vagy szintetikus anyagok csökkentik az NFk. B aktivitását?
Table 7. proteins and peptides that inhibit NF-k. B activation Interleukin IL-4 IL-11 IL-10 IL-13 Interferon Growth factors and hormones VEGF Hepatocyte growth factor MSH HB-EGF Chorionic gonadotropin IFN-α Peptides TRAF 6 inhibitory peptide NEMO binding peptide p 65 binding peptide Boronic acid peptide NLS cell permeable peptides Peptide nucleic acid-DNA decoys Peptide YY TIRAP inhibitor peptide Table 8. synthetic compounds that inhibit NF-k. B activation IKK inhibitors BAY-11 -7082 BAY-11 -7085 BMS-345541 SPC 839 AS 602868 PS 1145 MNL 120 SC-514 Polyubiquitination inhibitor Ro 106 -9920 Ubiquitin Ligase Inhibitors Z-LLL, Z-LLn. V Proteosome inhibitors ALLn. L PS-341 Lactacystine, β-lactone SN-50 FK 506 Ro 106 -9920 PS 5190 TLCK , TPCK Cyclosporin A DCIC Deoxyspergualin DFP LLM BTEE MG 115 MG 120 APNE P 42 MAPK inhibitor AG 126 P 38 MAPK inhibitor SB 203580 PI-3 K-kinase inhibitor LY 294002 PKC inhibitor RO 31 -8220 MEK 1/2 inhibitor UO 126 Protein kinase inhibitor A 771726
kérdés: mely pontokon lehet/kell szerkezeti változtatást létrehozni (pl. rekombináns technikákkal) az NFk. B aktivitásának specifikus csökkentéséhez?
Table 9. inhibition of NF-k. B through the critical cystein residue IKKα/β Cys 179 IKKβ Cys 59 IKKγ Cys 417 P 65 Cys 38 p 50 Cys 62
kérdés: milyen célmolekulákat tervezzünk az NFk. B aktivitásának csökkentsére? a szignálút melyik szakaszára?
kérdés: az aktivációs mechanizmus gátlásával változik-e az NFk. B aktivitása? gátló stimulusok használhatók-e? hát az Ik. B gátlása?
az NF-k. B gátlás kehetőségei 1. Aktiváló Stimulus (kfl) gyulladási-/immunstb. fehérjék Ik. B kináz /NEMO/IKAP I B p 50 p 65 P p 65 I B degradáció m. RNS inaktív forma marad CITOPLAZMA p 50 NUKLEUSZ p 65 transzkripció transzlokáció p 50 p 65 Target genes
kérdés: az Ik. B kináz ativitásának vagy aktiválásának gátlásával lehet csökkenteni az NFk. B aktivitását? mi az ubiquitináció gátlás hatása?
az NF-k. B gátlás kehetőségei 2. Aktiváló Stimulus (kfl) gyulladási-/immunstb. fehérjék I B kináz/NEMO/IKAP I B p 50 p 65 P p 65 I B degradáció m. RNS inaktív forma marad CITOPLAZMA p 50 NUKLEUSZ p 65 transzkripció transzlokáció p 50 p 65 Target genes
az NF-k. B gátlás kehetőségei 3. Aktiváló Stimulus (kfl) gyulladási-/immunstb. fehérjék I B kináz/NEMO/IKAP I B p 50 p 65 P p 65 I B degradáció m. RNS CITOPLAZMA p 50 NUKLEUSZ p 65 transzkripció transzlokáció p 50 p 65 Target genes
nem megoldott: - sejt-specificitás - alacsony toxicitás sok fejleszteni való van hátra
screen – ~220 e. Ft/kísérlet Human NFκB Signaling Pathway RT² Profiler™ PCR Array Functional Gene Grouping (more detailed information is provided in the gene table) Activation of NFκB pathway. Ligands and receptors: IL 1 B, IL 8, MYD 88, TLR 1, TLR 2, TLR 3, TLR 4, TLR 6, TLR 8, TNFRSF 10 A, TNFRSF 10 B. Membrane molecules: IRAK 1, IRAK 2. Kinases: CHUK, IKBKB, IKBKG, IRAK 1, IRAK 2. I-kappa-B kinase/NFkappa-B cascade: EDARADD, IKBKG, IRAK 2, STAT 1, TLR 8. Cytoplasmic sequestering/releasing of NF-kappa-B: BCL 3, IL 10, NALP 12, NFKBIA, TNFRSF 7, TNFSF 14. Transcription factors: IKBKB, IKBKG, IRAK 1, NFKBIA, RELA, STAT 1, TNF. Inflammatory response: IL 1 B, IL 8, IRAK 2, MYD 88, NFKB 1, TLR 2, TLR 3, TLR 4, TLR 6, TLR 8, TNF. Positive regulation of I-kappa-B kinase/NF-kappa-B cascade. Ligands and receptors: CD 40 (TNFRSF 5), F 2 R, FASLG (TNFSF 6), HTR 2 B, LTBR, SLC 20 A 1, TICAM 2, TNFRSF 1 A, TNFSF 10, TRIF. Membrane molecules: EDG 2, GJA 1, HMOX 1, RHOA. Kinases: IKBKE, RIPK 1, TBK 1. Other: BCL 10, BIRC 2, CARD 4, CASP 1, CASP 8, CFLAR, CTL 2, FADD, MALT 1, PPM 1 A, REL, RFP 2, TMED 4 (HNLF), TRADD. NFκB Responsive Genes. Acute phase response proteins: AGT, BF. Adhesion molecules: ICAM 1. Extracellular molecules: BF, CSF 2, CSF 3, IFNA 1, IFNB 1, IFNG, IL 6. Ligands and receptors: AGT, CCL 2, ICAM 1, LTA. Cytokines: CCL 2, CSF 3, IFNA 1, IFNB 1, IFNG, IL 6, IL 8, LTA, TNF. Inflammatory response: CCL 2. Other factors involved in NF-kappa-B pathway. Extracellular molecules: IL 1 A. Ligands and receptors: IL 1 R 1, RAF 1, TLR 7, TLR 9. Kinases: AKT 1, MAP 3 K 1, RAF 1. Transcription factors: ATF 1, EGR 1, ELK 1, FOS, JUN, NFKB 2, RELB, TNFAIP 3. Inflammatory response: FOS, IL 1 A, IL 1 R 1, TLR 7, TLR 9
köszönöm a figyelmet
(konstitutívan) aktív NFk. B-t expresszáló tumorok (pl. ) • • B cell lymphoma Hodgkin’s disease T-cell lymphoma Acute lymphoblastic leukemia • Breast • Liver • Thyroid • • Prostate Melanoma Head and neck SCC Colon Multiple myeloma Ovarian Bladder Lung Leukemia 16: 1053 -1068, 2002
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