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1、“The molecular genetic development of hematologic malignancy in Japan”,Kiyohiko Hatake, M.D., Ph.D. Department of Medical Oncology and Hematology, Cancer Institute Hospital, Ariake, Koto, Tokyo, Japan,ASCO-COSA-CSCO-ESMO-JCOG International Symposium CSCO Annual Meeting 2008 Shanghai, China,Disclosur
2、e of COI,Research funding,Chugai/Roche, Kirin, BMS, Kyowa, Yakult, Weyth, Novartis, Pfizer, Taiho, Jansen, Otsuka, Bayer Employment or leadership position: none Stock ownership Takeda, Astellas, Daiichi-Sankyo,Donation,Pfizer, Kirin, Chugai/Roche, Yakult, Novartis, Jansen, Taiho, Honoraria: none Con
3、sultant or advisort role: none Expert Testimony: none 000000000,Todays talk,Clinical practice of NHL,Registration system for WHO classification of NHL Standard chemotherapy in NHL and HL RCHOP in DLBCL Resistance to rituximab,Live-cell confocal fluorescence microscopy,CDC ADCC Relationship CDC and r
4、esponse to rituximab and prognosis Mutation of CD20 and CDC,Clinical practice of Non-Hodgkins Lymphoma (B-cell lymphoma) in CIH,Old, but an important prognostic marker sIL-2R, and CD5 in RCHOP treatment,DLBCL : treatment result by CHOPRituximab,CHOP n=83: 76%,R-CHOP n=92: 97%,P=0.0002 0.05,%:3-year
5、PFS,Days from treatment,Over-all survival rate,87 cases treated by CHOP and 141 cases treated by RCHOP Were analized in DLBCL.,RCHOP was superior to CHOP in both,RCHOP is superior to CHOP, and sIL-2R is still good marker For prognosis.,Mechanisms of action of rituximab,proliferation block,ADCC,CDC,a
6、poptosis,NK, M, PMN,FcR,C1q,C2-C9,CD20,CD20+ lymphoma,rituximab,Research methodology,Prediction of response and prognosis,Establishment of clinically applicable bio-imaging,Cancer cells from the patients,Ultra super speedy Sensitivity test,Live-cell bio-imaging,Confocal fluoresence,Prediction of res
7、ponse to rituximab by imaging,Response to rituximab in lymphoma cells from the patients,Probably ineffective,Probably effective,10min,10min,The principle of imaging-based CDC susceptibility assay,Diagnostic details of patients evaluated for CDC susceptibility,Correlation between CDC and clinical res
8、ponse,DLBCL,FL,chemotherapy with rituximab,chemotherapy w/o rituximab,P=0.0023,P=0.00067,Reproducible ADCC assay,KHYG-1,LTR,Fcgr3a (158V),LTR,LTR,Fcgr3a (158F),LTR,IRES,IRES,ZsGreen,ZsGreen,KHYG-1/mock -ZsGreen,KHYG-1/158V -ZsGreen,KHYG-1/158F -ZsGrreen,NK leukemia cell line CD3-, CD5-, CD7+, CD16-,
9、 CD56+, TCR-,ZsGreen, rituximab, PI,ADCC assay,KHYG-1/mock,KHYG-1/158V,KHYG-1/158F,LDH release assay Target: Ramos E/T: 1 Co-culture: 4hr,51Cr release assay Target: Ramos E/T: 1 Co-culture: 4hr,Effect of serum on ADCC activity,Serum (-),Serum (+),Rituximab (-),Rituximab (+),Ramos vs KHYG-1/FcRIIIa (
10、158V) in heat-inactivate serum,Effects of IgG on ADCC activity,Daudi vs. KHYG-1/FcRIIIa Rituximab: 0.1g/mL Co-culture: for 4hr,Daudi vs. KHYG-1/FcRIIIa Rituximab: 0.1g/mL Co-culture: for 4hr IgG (12mg/mL),Effect of complement on ADCC activity,Daudi vs. KHYG-1/FcRIIIa Rituximab: 0-100g/mL Co-culture:
11、 for 4hr serum / heat-inactivated serum : 50%(v/v),Summary,Depletion of peripheral B cells,complement,Consumption of C,Serum IgG,CDC,ADCC,C2-C9,rituximab,C1q,NK, M, PMN,CD20,FcR,Mutations of C-terminal region of CD20 molecule predict CD20 expression and time to progression after rituximab in non-Hod
12、gkins lymphoma Yasuhito Terui, Yuji Mishima, Natsuhiko Sugimura, Kiyotsugu Kojima, Takuma Sakurai, Yuko Mishima, Ryoko Kuniyoshi, Akiko Rokudai, Masahiro Yokoyama, Kengo Takeuchi, Chie Watanabe, Shunji Takahashi, Yoshinori Ito, and Kiyohiko Hatake Department of Medical Oncology and Hematology, Cance
13、r Institute Hospital, Japanese Foundation for Cancer Research; Division of Clinical Chemotherapy, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research; Olympus Bio-imaging Laboratory, Cancer Chemotherapy Center, Japanese Foundation for Cancer Research; Department of Pathology, Cancer
14、Institute Hospital, Japanese Foundation for Cancer Research, Tokyo; Nutritional Science Laboratory, Morinaga Milk Co., Kanagawa, Japan.,Terui Y et al., abstracts in ASH 2006 and ASCO 2007,Methods Since June 2002 to November 2004, retrospective analysis was performed in CIH (evaluable 50 cases of NHL
15、) 2. We performed flow cytometry (CD20 antigen) in each fresh lymphoma cells from the patients 3. Mutation analysis of CD20,Structure of CD20 mutation,CD20 negative DLBCL lymphoma transformed during R-CHOP treatment,M Raji PT K562,CD20,Flow cytometric analysis,RT-PCR,Sequence analysis,Summary of 50
16、cases with biopsy after PD,Histopathology treatment cases Biopsy after PD MALT R 3 1 R-CHOP like 11 R-CHOPRTx 1 R-VP-16 1 1 FL R 1 R-CHOP like 5 2 R-CHOPRTx 1 DLBCL R-CHOP 21 2 RTxR-CHOP 1 1 CLL R 1 1 R-CHOP like 1 SLL R 1 Lymphoplasmacytic R-CHOP like 1 Mantle CHOPRTR 1 1,total 50 9,CD20 mutation s
17、ite region cases C-terminal deletion C-terminal cytoplasmic 4 (truncation) 2. Extracellular 2nd extracellular 1 TM 4th transmembrane 1 4. Early termination N-terminal cytoplasmic 5,CD20 mutation in 11/50 NHL,CD20 expression in mutated lymphoma cells,C-terminal deletion and its structure,K562,K562/ W
18、T,K562/ TM,K562/ CD-1,K562/ CD-2,K562/ CD-3,CD20 expression in K562 cells with C-terminal deleteion,Surface CD20,Western blot analysis using N-terminal antibody for CD20 antigen,normal C-terminal deletion group CR #(%) 19 (49) 1 (25) Non-CR #(%) 21 (51) 3 (75) 計 40 4,Response to R-containing treatme
19、nt in both normal and CD20 mutated cases,Progression-free survival after R-containing chemo in normal and C-terminal deletion group,n=4,n=40,Conclusion C-terminal deletion mutation is related to loss of CD20 and shortened PFS,Other MoAbs,B-cell lymphoma CD20 Rituximab、GA101、HumaxCD20 Zevalin CD22 CM
20、C-544 T-cell lymphoma CD4 HumaxCD4 CCR4 Anti-CCR4 Ab,B-cell ：82.7 ,T/NK-cell ：10.3 ,HL ：7.0 ,B-cell lymphoma n=414,T/NK-cell lymphoma n=53,Overall survival rate,CHOP n=24 : 58%,Other therapy n=10 : 57%,Overall survival rate,PTCL,Clinical aspects of T-cell lymphoam in CIH,B-cell lymphoma & T/K lympho
21、ma,IGF1R,Receptor tyrosine kinase q chromosome A.A., single polypeptide cleaved by protease and subunit and subunit makes heterodimer and expressed on the surface chain is degraded by ubiquitin/proteasome Expressed on T cells（CDCD） Homology with IR, ,IGF1R expression on cell lines,Jurkat,TALL,CCRF,K
22、MS-BM,KMS-PE,IM-9,IM-9 VR,IGF1R,IGF1R,-actin,Jurkat,K562,Raji,Daudi,KYSE,BALL-1,ARH,SKW,IGF1R expression on the cell surface,IGF1R,IGF1R,-actin,The effect of Glucose on Jurkat cell proliferation With bortezomib,Hours in culture,0.1% Glucose,2.0% Glucose,x 104 cells/ml,The effect of glucose for IGF1R
23、 expression changes on Jurkat cells,0.1% Glucose,2.0% Glucose,0 1 2 3 4,0 1 2 3 4,IGF1R,IGF1R,-actin,Day,The effect of bortezomib on Jurkat cells,Bortezomib (10nM),Hours in culture,x 104 cells/ml,0.1% Glucose,2.0% Glucose,IGF1R expression after addition of bortezomib on Jurkat cells,0.1% Glucose,2.0
24、% Glucose,0 1 2 3 4,0 1 2 3 4,IGF1R,IGF1R,-actin,Bortezomib 20nM addition,Day,IGF1R molecular targeting drugs,MoAbs CP-751871 IMC-A12 Em164 MK-0646 R1507 AMG-479 Low molecular weight drugs AEW541 INSM18 Rinfabate,Carboplatin And Paclitaxel With Or Without CP-751, 871 (An IGF-1R Inhibitor) For Advanc
25、ed NSCLC Of Squamous, Large Cell And Adenosquamous Carcinoma Histology 2. Phase 3 Trial of CP-751,871 And Erlotinib in Refractory Lung Cancer 3Phase 2 Study of CP-751,871 in Combination With Docetaxel and Prednisone in HRPC Study Of CP-751,871 In Combination With Exemestane In Postmenopausal Women W
26、ith Hormone Receptor Positive Advanced Breast Cancer 5. Study Of CP-751,871 In Patients With Ewings Sarcoma Family Of Tumors 6. Study Using CP-751,871 In Patients With Stage IV Colorectal Cancer That Has Not Responded To Previous Anti-Cancer Treatments 7. Phase 2 Trial Of CP-751,871 And Docetaxel In
27、 Advanced Breast Cancer 8. Combination Study of CP-751,871 with Paclitaxel and Carboplatin in Advanced Lung Cancer 9. Study of anti-IGF-IR CP-751,871 in patients with solid tumors Study of CP-751,871 in Combination With Cisplatin and Gemcitabine in Chemotherapy-naive Patients With Advanced Non-Small
28、 Cell Lung Cancer 11. Study of CP-751,871 in combination with carboplatin and Paclitaxel in Advanced Lung Cancer 12. Phase I Pilot Study of Neoadjuvant CP-751,871 in Women with operable early breast cancer 13. Phase I, Pharmacokinetic and Pharmacodynamic Study of the AntiInsulinlike Growth Factor Ty
29、pe 1 Receptor Monoclonal Antibody CP-751,871 in Patients With Multiple Myeloma,Anti-IGF1R Ab clinical trials,Summary,Rituximab resistance of CDC can be explained by the expression of CD55, CD5, and CD20 mutation. CD20 mutated cases can be treated by ICE, or newer MoAbs for CD20 or CD22. IGFR-1 expre
30、ssion may be a next target.,Great thanks to:,Department of Medical Oncology and Hematology,Dr. Yasuhito Terui Dr. Yasuhiro Yokoyama Dr. Yuko Mishima Dr. Daisuke Ennishi Dr. Asai Data manager: Ms. Suitsu, Yamazaki, Yagou,Division of Clinical Chemotherapy and Olympus Bio-Imaging Labo,Dr. Yuji Mishima Dr. Satoshi Matsuzaka,