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 Cons
3、ultant or advisort role: none Expert Testimony: none000000000,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 res
4、ponse 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 PF
5、S,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,apo
6、ptosis,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 fromthe patients,Ultra super speedy Sensitivity test,Live-cell bio-imaging,Confocal fluoresence,Prediction of respon
7、se 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 respon
8、se,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-, CD
9、56+, TCR-,ZsGreen, rituximab, PI,ADCC assay,KHYG-1/mock,KHYG-1/158V,KHYG-1/158F,LDH release assayTarget: RamosE/T: 1Co-culture: 4hr,51Cr release assayTarget: RamosE/T: 1Co-culture: 4hr,Effect of serum on ADCC activity,Serum (-),Serum (+),Rituximab (-),Rituximab (+),Ramos vs KHYG-1/FcRIIIa (158V) in
10、heat-inactivate serum,Effects of IgG on ADCC activity,Daudi vs. KHYG-1/FcRIIIaRituximab: 0.1g/mL Co-culture: for 4hr,Daudi vs. KHYG-1/FcRIIIaRituximab: 0.1g/mL Co-culture: for 4hrIgG (12mg/mL),Effect of complement on ADCC activity,Daudi vs. KHYG-1/FcRIIIaRituximab: 0-100g/mL Co-culture: for 4hrserum
11、 / 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-Hodgkins lymphoma
12、 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, Cancer Institute Ho
13、spital, 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 Institute Hospi
14、tal, 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,MethodsSince June 2002 to November 2004, retrospective analysis was performed in CIH (evaluable 50 cases of NHL)2. We performed
15、 flow cytometry (CD20 antigen) in each fresh lymphoma cells from the patients3. 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 cases with biopsy
16、after PD,Histopathology treatment cases Biopsy after PDMALT R 3 1R-CHOP like 11R-CHOPRTx 1R-VP-16 1 1FL R 1R-CHOP like 5 2R-CHOPRTx 1DLBCL R-CHOP 21 2RTxR-CHOP 1 1CLL R 1 1R-CHOP like 1SLL R 1Lymphoplasmacytic R-CHOP like 1Mantle CHOPRTR 1 1,total 50 9,CD20 mutation site region casesC-terminal delet
17、ion C-terminal cytoplasmic 4(truncation) 2. Extracellular 2nd extracellular 1 TM 4th transmembrane 14. 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/WT,K562/ TM,K562/ CD-1,K562/ CD-2,K56
18、2/ 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 treatment in both normal and CD20 mutated ca
19、ses,Progression-free survival after R-containing chemo in normal and C-terminal deletion group,n=4,n=40,ConclusionC-terminal deletion mutation is related to loss of CD20 and shortened PFS,Other MoAbs,B-cell lymphoma CD20 Rituximab、GA101、HumaxCD20Zevalin CD22 CMC-544T-cell lymphoma CD4 HumaxCD4 CCR4
20、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 lymphoma,IGF1R,Receptor tyrosine kinase q chro
21、mosome A.A., single polypeptidecleaved 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,KMS-BM,KMS-PE,IM-9,IM-9 VR,IGF1R,IGF1R,-ac
22、tin,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 expression changes on Jurkat cells,0.1%
23、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% Glucose,0 1 2 3 4,0 1 2 3 4,IGF1R,IGF1R
24、,-actin,Bortezomib 20nM addition,Day,IGF1R molecular targeting drugs,MoAbs CP-751871 IMC-A12 Em164 MK-0646 R1507 AMG-479 Low molecular weight drugsAEW541 INSM18Rinfabate,Carboplatin And Paclitaxel With Or Without CP-751, 871 (An IGF-1R Inhibitor) For Advanced NSCLC Of Squamous,Large Cell And Adenosq
25、uamous Carcinoma Histology 2. Phase 3 Trial of CP-751,871 And Erlotinib in Refractory Lung Cancer3Phase 2 Study of CP-751,871 in Combination With Docetaxel and Prednisone in HRPCStudy Of CP-751,871 In Combination With Exemestane In Postmenopausal Women With Hormone Receptor Positive Advanced Breast
26、Cancer5. Study Of CP-751,871 In Patients With Ewings Sarcoma Family Of Tumors6. Study Using CP-751,871 In Patients With Stage IV Colorectal Cancer That Has Not Responded To Previous Anti-Cancer Treatments7. Phase 2 Trial Of CP-751,871 And Docetaxel In Advanced Breast Cancer8. Combination Study of CP
27、-751,871 with Paclitaxel and Carboplatin in Advanced Lung Cancer9. Study of anti-IGF-IR CP-751,871 in patients with solid tumorsStudy of CP-751,871 in Combination With Cisplatin and Gemcitabine in Chemotherapy-naive Patients WithAdvanced Non-Small Cell Lung Cancer11. Study of CP-751,871 in combinati
28、on with carboplatin and Paclitaxel in Advanced Lung Cancer12. 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 Type 1 ReceptorMonoclonal Antibody CP-751,871 in Patients
29、 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 expression 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,
