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,JapanASCO-COSA-CSCO-ESMO-JCOG International SymposiumCSCO Annual Meeting 2008Shanghai,ChinaDisclosure of COIResear
2、ch 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 Consultant or advisort role:none Expert
3、Testimony:none 000000000Todays talkClinical practice of NHL Registration system for WHO classification of NHL Standard chemotherapy in NHL and HL RCHOP in DLBCL Resistance to rituximabLive-cell confocal fluorescence microscopy CDC ADCC Relationship CDC and response to rituximab and prognosis Mutatio
4、n of CD20 and CDCClinical practice of Non-Hodgkins Lymphoma(B-cell lymphoma)in CIHOld,but an important prognostic marker sIL-2R,and CD5 in RCHOP treatment2400160080001.0.9.8.7.6.5.4.3.2.10.0DLBCL:treatment result by CHOPRituximab CHOP n=83:76%R-CHOPn=92:97%P=0.0002 0.05%:3-year PFSDays from treatmen
5、tOver-all survival rate87 cases treated by CHOP and 141 cases treated by RCHOP Were analized in DLBCL.RCHOP was superior to CHOP in bothRCHOP is superior to CHOP,and sIL-2R is still good marker For prognosis.Mechanisms of action of rituximabproliferationblockADCCCDCapoptosisNK,M,PMNFcRC1qC2-C9CD20CD
6、20+lymphomarituximabResearch methodology Prediction of response and prognosisEstablishment of clinically applicable bio-imagingCancer cells from the patients Ultra super speedySensitivity test Live-cell bio-imagingConfocal fluoresencePrediction of response to rituximab by imagingResponse to rituxima
7、b in lymphoma cells from the patientsProbably ineffectiveProbably effective10min10minThe principle of imaging-based CDC susceptibility assayDiagnostic details of patients evaluated for CDC susceptibilityCorrelation between CDC and clinical response DLBCLFLchemotherapywith rituximabchemotherapyw/o ri
8、tuximabP=0.0023P=0.00067Reproducible ADCC assayKHYG-1LTRFcgr3a(158V)LTRLTRFcgr3a(158F)LTRIRESIRESZsGreenZsGreenKHYG-1/mock-ZsGreenKHYG-1/158V-ZsGreenKHYG-1/158F-ZsGrreenNK leukemia cell line CD3-,CD5-,CD7+,CD16-,CD56+,TCR-ZsGreen,rituximab,PIADCC assayKHYG-1/mockKHYG-1/158VKHYG-1/158FLDH release ass
9、ay Target:Ramos E/T:1 Co-culture:4hr51Cr release assay Target:Ramos E/T:1 Co-culture:4hrEffect of serum on ADCC activitySerum(-)Serum(+)Rituximab(-)Rituximab(+)Ramos vs KHYG-1/FcRIIIa(158V)in heat-inactivate serumEffects of IgG on ADCC activityDaudi vs.KHYG-1/FcRIIIaRituximab:0.1g/mL Co-culture:for
10、4hrDaudi vs.KHYG-1/FcRIIIaRituximab:0.1g/mL Co-culture:for 4hr IgG(12mg/mL)Effect of complement on ADCC activityDaudi vs.KHYG-1/FcRIIIaRituximab:0-100g/mL Co-culture:for 4hr serum/heat-inactivated serum:50%(v/v)SummaryDepletion of peripheral B cellscomplementConsumption of CSerum IgGCDCADCCC2-C9ritu
11、ximabC1qNK,M,PMNCD20FcRMutations of C-terminal region of CD20 molecule predict CD20 expression and time to progression after rituximab in non-Hodgkins lymphomaYasuhito Terui,Yuji Mishima,Natsuhiko Sugimura,Kiyotsugu Kojima,Takuma Sakurai,Yuko Mishima,Ryoko Kuniyoshi,Akiko Rokudai,Masahiro Yokoyama,K
12、engo Takeuchi,Chie Watanabe,Shunji Takahashi,Yoshinori Ito,and Kiyohiko Hatake Department of Medical Oncology and Hematology,Cancer Institute Hospital,Japanese Foundation for Cancer Research;Division of Clinical Chemotherapy,Cancer Chemotherapy Center,Japanese Foundation for Cancer Research;Olympus
13、Bio-imaging Laboratory,Cancer Chemotherapy Center,Japanese Foundation for Cancer Research;Department of Pathology,Cancer 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
14、Methods1.Since June 2002 to November 2004,retrospective analysis was performed in CIH(evaluable 50 cases of NHL)2.We performed flow cytometry(CD20 antigen)in each fresh lymphoma cells from the patients3.Mutation analysis of CD20Structure of CD20 mutationCD20 negative DLBCL lymphoma transformed durin
15、g R-CHOP treatmentM Raji PT K562 CD20Flow cytometric analysis RT-PCRSequence analysisSummary of 50 cases with biopsy after PDHistopathology treatment cases Biopsy after PDMALT 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 1CLL R 1 1 R-C
16、HOP like 1SLL R 1Lymphoplasmacytic R-CHOP like 1 Mantle CHOPRTR 1 1 total 50 9 CD20 mutation site region cases1.C-terminal deletion C-terminal cytoplasmic 4 (truncation)2.Extracellular 2nd extracellular 1 3.TM 4th transmembrane 14.Early termination N-terminal cytoplasmic 5CD20 mutation in 11/50 NHL0
17、20406080100120140160p=0.0434p=0.0035CD20 expression in mutated lymphoma cells C-terminal deletion and its structureK562K562/WTK562/TMK562/CD-1K562/CD-2K562/CD-337 kDa20 kDa25 kDaFull lengthC-terminal deletionCD20 expression in K562 cells with C-terminal deleteion Surface CD20Western blot analysis us
18、ing N-terminal antibody for CD20 antigen normal C-terminal deletion groupCR#(%)19(49)1(25)Non-CR#(%)21(51)3(75)計 40 4Response to R-containing treatment in both normal and CD20 mutated cases0.00.20.40.60.81.00510152025303540Months after rituximab-treatmentNon-mutationC-terminaldeletionProportionp=0.0
19、481Progression-free survival after R-containing chemo in normal and C-terminal deletion groupn=4n=40ConclusionC-terminal deletion mutation is related to loss of CD20 and shortened PFS Other MoAbsB-cell lymphoma CD20 Rituximab、GA101、HumaxCD20 Zevalin CD22CMC-544T-cell lymphoma CD4 HumaxCD4 CCR4 Anti-
20、CCR4 AbB-cell：82.7 T/NK-cell：10.3 HL：7.0 B-cell lymphoma n=414T/NK-cell lymphoma n=53Overall survival rateCHOP n=24:58%Other therapy n=10:57%Overall survival ratePTCLClinical aspects of T-cell lymphoam in CIHB-cell lymphoma&T/K lymphomaIGF1RReceptor tyrosine kinaseq chromosomeA.A.,single polypeptide
21、cleaved by protease and subunit and subunit makes heterodimer and expressed on the surface chain is degraded by ubiquitin/proteasomeExpressed on T cells（CDCD）Homology with IR,IGF1R expression on cell linesJurkatTALLCCRFKMS-BMKMS-PEIM-9IM-9 VRIGF1RIGF1R-actinJurkatK562RajiDaudiKYSEBALL-1ARHSKWIGF1R e
22、xpression on the cell surfaceIGF1RIGF1R-actinThe effect of Glucose on Jurkat cell proliferationWith bortezomibHours in culture0.1%Glucose2.0%Glucosex 104 cells/mlThe effect of glucose for IGF1R expression changes on Jurkat cells0.1%Glucose2.0%Glucose0 1 2 3 40 1 2 3 4IGF1RIGF1R-actinDayThe effect of
23、 bortezomib on Jurkat cellsBortezomib(10nM)Hours in culturex 104 cells/ml0.1%Glucose2.0%GlucoseIGF1R expression after addition of bortezomib on Jurkat cells0.1%Glucose2.0%Glucose0 1 2 3 40 1 2 3 4IGF1RIGF1R-actinBortezomib 20nM additionDayIGF1R molecular targeting drugs MoAbs CP-751871 IMC-A12 Em164
24、 MK-0646 R1507 AMG-479 Low molecular weight drugs AEW541 INSM18 RinfabateCarboplatin And Paclitaxel With Or Without CP-751,871(An IGF-1R Inhibitor)For Advanced NSCLC Of Squamous,Large Cell And Adenosquamous Carcinoma Histology2.Phase 3 Trial of CP-751,871 And Erlotinib in Refractory Lung Cancer3Phas
25、e 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 Cancer5.Study Of CP-751,871 In Patients With Ewings Sarcoma Family Of Tumors6.Study Using CP-751,871 I
26、n 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-751,871 with Paclitaxel and Carboplatin in Advanced Lung Cancer9.Study of anti-IGF-IR CP-751,871 in patie
27、nts with solid tumorsStudy of CP-751,871 in Combination With Cisplatin and Gemcitabine in Chemotherapy-nai ive Patients WithAdvanced Non-Small Cell Lung Cancer11.Study of CP-751,871 in combination with carboplatin and Paclitaxel in Advanced Lung Cancer12.Phase I Pilot Study of Neoadjuvant CP-751,871
28、 in Women with operable early breast cancer 13.Phase I,Pharmacokinetic and Pharmacodynamic Study of the AntiInsulinlike Growth Factor Type 1 Receptor Monoclonal Antibody CP-751,871 in Patients With Multiple MyelomaAnti-IGF1R Ab clinical trialsSummary Rituximab resistance of CDC can be explained by t
29、he 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
