Coordinator: Dr. Hsien-Ming Lee Coach Professor: Dr. Joseph J.-T. Huang Sit-in Professor: Dr. Su-Chang Lin. Presenter: Bagher Golzarroshan
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1 Coordinator: Dr. Hsien-Ming Lee Coach Professor: Dr. Joseph J.-T. Huang Sit-in Professor: Dr. Su-Chang Lin Presenter: Bagher Golzarroshan 1
2 Outline Introduction Cancer Apoptosis Apoptosis and disease Scope of this paper Identification of a Smac mimetic synergist SAR study and identification of Bioymifi Mechanism of Bioymifi Future work 2
3 1. UV radiation 2. Ionizing radiation cancer 1. Depyrimidination Cancer Hallmarks (Weinberg and Hanahan) 2. Depurination 1. DNA polymerases (proofreading) 2. Mismatch excision repair (MMR) 1. Oncogenes 2. Tumor suppressor genes 3. Suicide genes 4. DNA-repair genes
4 Apoptosis DR4 (TrailR1), DR5 (TrailR2) Fas-Associated protein with Death Domain inhibitor of apoptosis proteins
5 Apoptosis and disease Increased apoptosis: Alzheimer Parkinson Decreased or inhibited apoptosis: Lupus Cancer: BCL-2 Family proteins alterations (Over-expression) Apoptosome defects (Down-regulate Apaf1) Inhibitor of apoptosis proteins (IAP) inhibitors (Over-expression) Death receptor pathway defects Reduced expression (escape immune control) Toxicity of therapies targeting TNF and CD95 (therapy) Trail receptor pathway (not tissue toxic)
6 previous study Design and synthesize smac mimetic compound based on the IAP crystal structure that induces apoptosis synergistically with death receptor ligands such as TRAIL. Discover compound that promote cell death in synergy with a small-molecule mimetic of Smac through a highthroughput chemical screen for compounds.
7 Identification of a Smac mimetic synergist T98G human glioblastoma cells 7 A2C2
8 validate the chemical identity of A2C2? NMR and LC/MS tautomers 8 A2C2-induced cell death is attributable to the combined activity of compounds A2 and C2.
9 Bioymifi Structure-activity relationship (SAR) approach to search for a functional analog or derivative of A2C2 Single agent sensitivity Effect on different cancer cell lines focused further investigations on bioymifi-induced cell death
10 Bioymifi induces caspase-8 dependent apoptosis Poly (ADP-ribose) polymerase Bioymifi Fluorogenic activate induces assay caspase-8 dependent measured 8 and caspase-3 apoptosis 3activation Caspase-8 and the related extrinsic apoptotic pathway are essential
11 Bioymifi induces apoptosis via death receptor DR5 DR5 knockdown is the predominant rescued cells receptor from biymifi for TRAIL or A2C2-mediated in T98G cellsapoptosis
12 Bioymifi induces DR5-dependent proximal DISC as TRAIL does? -Fas-Associated protein with Death Domain -Tumor necrosis factor receptor type 1-associated DEATH domain protein -Receptor-interacting serine/threonine-protein Immunoprecipitated(IP) kinase 1 Use Treated TRADD caspase has by bioymifi: positive 8 antibody: FADD role FADD, in was TRAIL-induced rapidly TRADD recruited and apoptosis PIPK1 were DR5 complex present
13 Bioymifi-induced apoptosis is independent of TRAIL? This data indicated that Bioymifi acts on DR5 independently of TRAIL
14 Bioymifi directly binds DR5 Use recombinant DR5 and DR4 containing only ECD Bioymifi bound the ECD of DR5 with a K =2 µm but showed little bindind affinity to the DR4
15 Bioymifi promotes DR5 aggregation and activation Bioymifi DR5 knockdown induces decresed DR5 clustring the number (90% of and cells) intensity of the foci induced ECD by TRAIL Bioymifi domain induces simulated alone smaler the is sufficient DR5 clusters aggregation to for (30% DR5 but of aggregation cells) had little effect in response DR4 to ECD Bioymifi and A2C2 DR5 treated with Bioymifi and A2C2 aggregates
16 16
17 17
FEN1 DNA DNA DNA DNA DNA DNA DNA DNA DNA DT40 DNA RNA H3-K36. RNA H3-K36 Set2 RNA. Set2 DNA H3-K36 RNA H3-K36 H3-K36 TBS-I, -II, -III 3 TBS-I TBS-II
DNA DNA DNA DNA DNA RNA H3 36 Lys (K) H3-K36 H3-K36 24 DNA FEN1 DNA FEN1 DNA DNA 4 CAF1 DNA DNA DT40 DNA DNA RNA H3-K36 RNA H3-K36 Set2 Set2 RNA C RNA Set2 H3-K36 5 I 24 24 23 TBS-I, -II, -III 3 TBS-I
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