html5-img
1 / 50

in vivo animal model studies in biological science

in vivo animal model studies in biological science. Cancer 2. Neuroscience. Cancer research. 2. Neuroscience. Lung Cancer. Cure rate for all patients: 15%. Male. Female. EGFR Expression in NSCLC. Tumours showing high EGFR expression.

maja
Télécharger la présentation

in vivo animal model studies in biological science

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. in vivo animal model studies in biological science • Cancer • 2. Neuroscience • Cancer research 2. Neuroscience

  2. Lung Cancer • Cure rate for all patients: 15% Male Female

  3. EGFR Expression in NSCLC Tumours showinghigh EGFR expression High expression associated with Poor outcome NSCLC 60-80%

  4. EGFR signaling pathways HER HER g PLC PI3K PKC PDK-1 Ras Sos AKT PTEN Grb2 Raf p70 S6K Shc Bad/Bcl2 JAK Mek GSK3 STAT Erk Myc p27 KIP1 E2F Jun/Fos Sp1 FKHR-L1 PEA3 Elk1 CyclinD1 Citri A, et al. Exp Cell Res 2003

  5. Molecular Targeting of EGF Receptor Cetuximab Gefitinib (IRESSA) Erlotinib (Tarceva)

  6. Response to EGFR-TKI in NSCLC patients Initial diagnosis 2 months after gefitinib 9 months later Yu CJ. (2005) PloS Med

  7. Primary activating mutations are mainly found in EGFR tyrosine kinase domain exons 18-21 EGF Ligand Binding TM Tyrosine Kinase L858R (~40%) S768I (2%) R776C (2%) G719A/C (5%) L861Q (4%) 719 768 776 858 861 GxGxxG 18 K 19 20 R L 21 R 22 23 24 E746-A750 766-768 Deletions (~45%) Insertions (3%)

  8. Modeling EGFR-TKI responses in pre-clinical model system PC9, HCC827 TKI Response(+) Response(-) ( Acquired resistance ) EGFR Mutation(+) TKI Response(-) Primary mutation(s) NSCLC TKI Response(+) EGFR Mutation(-) TKI Response(-)

  9. Clinical response to EGFR-TKI in NSCLC patients Initial diagnosis 2 months after gefitinib 9 months later Yu CJ. (2005) PloS Med

  10. Mechanisms of the Acquired Resistance to EGFR tyrosine kinase inhibitors in NSCLC (2008) Unknown Mechanisms T790M secondary mutation MET amplification

  11. Modeling EGFR-TKI responses in pre-clinical model system PC9, HCC827 TKI Response(+) Response(-) ( Acquired resistance ) EGFR Mutation(+) TKI Response(-) Primary mutation(s) NSCLC TKI Response(+) EGFR Mutation(-) TKI Response(-)

  12. Human cancer cell immunocompromised mouse xenograft model

  13. Erlotinib treatment in PC9 orthotopic lung cancer model vehicle treated erlotinib (50mg/kg/day) treated Before administration 1 week 2 weeks 3 weeks

  14. Data summary of erlotinib treatment in pre-clinical animal tumor xenograft model system

  15. Multi-cycle resistance test Inoculate previously obtained in vivo drug resistant cells to second animals and subject the animals to repeated drug cycle PC9TR PC9

  16. 1st generation EGFR-TKI gefitinib, erlotinb : reversible EGFR blocker 2nd generation EGFR-TKI e.g. pan-erbB blocker, multi-target EGFR blocker, irreversible EGFR blocker (BIBW2992)

  17. The Effect on Cell Viability of si-EGFR in PC-BR clones scram si-EGFR si-EGFR si-EGFR si-EGFR scram scram scram EGFR Actinin PC9 #1 #6 #10 EGFR dependent EGFR independent

  18. Sequencing in PC9 & BR1,6,10 BR#1 PC9 I T(790) L Q I T(790) L Q BR#6 BR#10 I T(790) L Q I T(790) L Q

  19. Protein expression and phosphorylation profile in PC9-BR clones #6 #1 #10 PC9 0 0.2 2 20 200 BIBW2992 for24H 0 0.2 2 20 200 0 0.2 2 20 200 0 0.2 2 20 200 p-EGFR EGFR p-Her3 p-MET MET p-STAT3 STAT3 p70 S6K p-AKT AKT p-ERK ERK BIM Actinin

  20. #1 #6 #10 PC BIBW2992 treatment in inoculated mouse in vivo

  21. invivo response to BIBW2992 (25mg/kg)

  22. Primary activating mutations are mainly found in EGFR tyrosine kinase domain exons 18-21 EGF Ligand Binding TM Tyrosine Kinase L858R (~40%) S768I (2%) R776C (2%) G719A/C (5%) L861Q (4%) 719 768 776 858 861 GxGxxG 18 K 19 20 R L 21 R 22 23 24 E746-A750 766-768 Deletions (~45%) Insertions (3%)

  23. EGFR Mutations Gefitinib Responders 8/9 Non-responders 0/7 p= 0.00075 Lynch et al, NEJM 2004

  24. Construction of transgenic mouse model

  25. Mouse EGFR non-small cell lung cancer transgenic mouse model CCSP rtTA Wong et al. (2006) Cancer Cell

  26. Tet-inducible mutant EGFR expression in mouse lung Wong et al. (2006) Cancer Cell

  27. EGFR mutation is oncogenic Wong et al. (2006) Cancer Cell

  28. EGFR expression is required for the maintenance of tumor Wong et al. (2006) Cancer Cell

  29. Wong et al. (2006) Cancer Cell

  30. Lung cancer originated from mutant EGFR respond to various EGFR inhibitors Wong et al. (2006) Cancer Cell

  31. Transgenic mutant EGFR animal model study 1. mutant EGFR is oncogenic 2. continued expression of EGFR is required for the maintenance of tumor 3. mutant EGFR is a therapeutic target

  32. Factors controlling tumorigenesis Tumor suppressor genes Oncogenes metastasis Immune Stroma Angiogenesis WT cells

  33. Utility of genetically-engineered mouse models of cancer early detection Genetically engineered mouse prevention Progression analysis Tumor development chemotherapy

  34. Genetic engineering of mouse genome : knock-out and knock-in via homologous recombination

  35. Embryonic stem cell culture

  36. Homologous recombinant ES cell selection and blastocyst injection

  37. Generation of chimera mouse

  38. Confirmation of germ line transmission and generation of knock-out(in) mouse

  39. Conditional activation of p53 Advantages: p53 Native promoters Temporal, spatial Regulation of gene expression unbiased

  40. Conditional knock-out system

  41. P53 LSL/LSL is a phenocopy of p53-/- Cre-recombinase-Oestrogen-Receptor-T2

  42. p53 reactivation mouse model study with conditional gene expression mouse • p53 inactivation is required for the maintenance of p53 mutant tumors • p53 gene delivery or other ways to reactivate p53 in p53 mutant tumor • could be a therapeutic option

More Related