From Cancer Genomics to Cancer Treatment : from hope to reality Yusuke Nakamura Human Genome Center Institute of Medical Science The University of Tokyo
International HapMap International HapMap Consortium Consortium October 27, 2005 Construction of public Construction of public database for genetic database for genetic variations in human variations in human
Country Genotyping Center %Genome Chromosome Platform Japan RIKEN 24.3% 5, 11, 14, 15, 16, 17, 19 Third Wave Invader Wellcome Trust Sanger UK 23.7% 1, 6, 10, 13, 20 Illumina BeadArray Institute We (The SNP Research Center in RIKEN) We (The SNP Research Center in RIKEN) McGill Univ. / Genome Canada 10.1% 2, 4p Illumina BeadArray Quebec Innovation Centre contributed the largest SNP data (24.3%) contributed the largest SNP data (24.3%) Sequenom MassExtend, China Chinese HapMap Consortium 9.5% 3, 8p, 21 Illumina BeadArray among the SNP typing centers among the SNP typing centers Illumina 16.1% 8q, 9, 18q, 22, X Illumina BeadArray in this paper in this paper Broad Institute of Harvard and Sequenom MassExtend, 9.7% 4q, 7q, 18p, Y, mtDNA MIT Illumina BeadArray USA Baylor College of Medicine 4.6% 12 ParAllele MIP PerkinElmer AcycloPrime- UCSF / Washington Univ. 2.0% 7p FP High-denstity Perlegen Sciences All oligonucleotide array
Biobank Japan Sample Collection Biobank Japan Sample Collection at February 29, 2008 at February 29, 2008 Hyperlipidemia 42,354 Prostate cancer 5,839 Leukemia 1,597 Diabetes 39,982 Periodontitis 5,652 Esophageal cancer 1,461 Cataract 19,070 Pollinosis 5,572 Cervical caner 1,405 Cerebral infarction 16,012 Glaucoma 5,257 Hepatitis B 1,391 Arrhythmia 15,440 Lung cancer 4,845 Uterine corpus cancer 1,189 Stable angina pectoris 14,855 Unstable angina pectoris 4,161 Nephrotic syndrome 1,038 We have genotyped more than 15,000 patients We have genotyped more than 15,000 patients Myocardial Infarction 12,956 Rheumatoid arthritis 4,155 Ovarian cancer 976 Bronchial asthma 8,657 Atopic dermatitis 2,967 Tuberculosis 894 at 250K-550K SNPs at 250K-550K SNPs Cardiac failure 7,438 COPD 2,797 Keloid 814 Breast cancer 7,349 Cerebral aneurysm 2,735 ALS 788 Colorectal cancer 6,957 Arteriosclerotic obliterans 2,609 ILD 780 A total of 6-billion data-points. A total of 6-billion data-points. Gastric cancer 6,869 Liver cirrhosis 2,494 Drug-induced hypersensitivity 598 Urinary stone 6,605 Liver cancer 2,452 Pancreatic cancer 531 Osteoporosis 6,412 Hyperthyroidism 2,320 CCC 503 Myoma uteri 5,988 Epilepsy 2,250 Febrile seizures 475 1,827 Total 5,962 Endometriosis Hepatitis C 295,278 IC asked 235,841 patients IC asked 235,841 patients Total Cases 295,278 cases Total Cases 295,278 cases IC obtained 201,805 patients IC obtained 201,805 patients Withdrawn 172 individuals （ Withdrawn 172 individuals （ 85.6%) 85.6%)
Genes isolated through genome-wide association studies in RIKEN and University of Tokyo Myocardial Infarction LTA Nature Genetics 2002 LGALS2 Nature 2004 PSMA6 Nature Genetics 2006 Rheumatoid Arthritis PADI4 Nature Genetics 2003 SLC22A4 Nature Genetics 2003 FcRH3 Nature Genetics 2005 Diabetic nephropathySLC12A3 Diabetes 2003 ELMO1 Diabetes 2005 (Diabetes) KCNQ1 Nature Genetics 2008 IgA nephropathy SEL-L,-E Am J Hum Genet 2002 Osteoarthritis Asporin Nature Genetics 2005 Human Mol. Gen 2005 Calmodulin 1 GDF5 Nature Genetics 2007 DVWA Nature Genetics 2008 Disc herniation CILP Nature Genetics 2005 Brain Infarction PRKCH1 Nature Genetics 2007 Kawasaki disease ITPKC Nature Genetics 2008 Crohn disease TNSF15 Human Mol. Gen 2005 Colon cancer multipel genes Nature Genetics 2008 Lung fibrosis TERT JMG 2008 Hapmap Nature 2003 Nature 2005 Nature 2007
From molecular targets to anti- -cancer drugs cancer drugs From molecular targets to anti From molecular targets to anti-cancer drugs Antibody SMC Peptide Dominant-negative Target Target peptide Molecules vaccine Molecules Anti-sense DNA Cell Gene siRNA Therapy Therapy
From molecular targets to anti- -cancer drugs cancer drugs From molecular targets to anti From molecular targets to anti-cancer drugs Antibody SMC Peptide Dominant-negative Target Target peptide Molecules vaccine Molecules Anti-sense DNA Cell Gene siRNA Therapy Therapy
Isolation of molecular targets for cancer treatment Isolation of molecular targets for cancer treatment using clinical materials using clinical materials cDNA microarray consisting of 32,000 genes Expression profiles of Comparison of expression Comparison of expression Expression profiles of 30 normal human profiles of cancer and profiles of cancer and 30 normal human tissues corresponding normal tissues tissues corresponding normal tissues Selection of novel molecular targets Diagnosis Treatment Diagnosis Treatment Tumor Marker Small molecular compound Tumor Marker Small molecular compound Prediction to Monoclonal Antibody Prediction to Monoclonal Antibody chemosensitivity Peptide Vaccine chemosensitivity Peptide Vaccine siRNA siRNA
cDNA Microarray system cDNA Microarray system H i g h - d e n s i t y s p o t t i n g C a n c e r N o r m a l 2 0 0 0 0 s p o t s / g l a s s t i s s u e t i s s u e Ｌ ＭＭ Ｌ ＭＭ C a n c e r c e l l s N o r m a l d u c t a l c e l l s T 7 - b a s e d R N A A m p l i f i c a t i o n T 7 - b a s e d R N A A m p l i f i c a t i o n （ 2 - r o u n d s ） （ 2 - r o u n d s ） a R N A a R N A l a b e l i n g l a b e l i n g C C y y 5 5 C C y y 3 3 c o - h y b r i d i z a t i o n
Number of clinical samples Number of clinical samples analyzed by cDNA microarray analyzed by cDNA microarray Tissue Number Tissue Number Lung 126 Bile duct 45 Breast 135 Uterus 44 Soft Tissue 101 ALL 25 AML 87 Kidney 25 Colon 78 Endometriosis 23 CML 84 Liver 20 Ovary 59 Pancreas 20 54 Melanoma 20 Malignant lymphoma Prostate 54 Thyroid 20 Stomach 51 Neuroblastoma 16 Bladder 55 Testis 13 Eshophagus 45 TOTAL 1200
Criteria for selection of candidate targets for drug development (S) Small molecular compound (P) Peptide vaccine (R) siRNA (A) Antibody 1. Genes which were highly over-expressed in a large 1. Genes which were highly over-expressed in a large proportion of clinical cancer samples examined proportion of clinical cancer samples examined (S, P, R, A) (S, P, R, A) 2. Genes which were expressed in none of important vital 2. Genes which were expressed in none of important vital organs; ideally not expressed in any organs (S, P, R, A) organs; ideally not expressed in any organs (S, P, R, A) 3. Genes whose expressions are essential for cell survival 3. Genes whose expressions are essential for cell survival (S, R, A?) (S, R, A?) 4. Cytoplasmic membrane protein, Secreted protein (A) 4. Cytoplasmic membrane protein, Secreted protein (A)
Development of anti-FZD10 antibody therapy Development of anti-FZD10 antibody therapy for Synovial Sarcoma for Synovial Sarcoma Chikako Fukukawa Chikako Fukukawa Satoshi Nagayama Satoshi Nagayama Toyomasa Katagiri Toyomasa Katagiri
Frizzled Homologue 10 (FZD10) Frizzled Homologue 10 (FZD10) RT-PCR C SS S S M MFH LMS Synovial Sarcoma MPNST cell lines L 1 2 3 4 5 6 7 8 9 10 1112 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 FZD10 b2MG SS Northern Blotting Bone marrow Srug. CL Pancreas Frizzled family (Wnt signal) HS-SY-2 Placenta YaFuSS Kidney SS582 SS487 Heart Brain Lung Liver Wnt? SS cell 9.5 7.5 4.4 2.37 1.35 SS ; Synovial Sarcoma
FZD10 Protein expression of FZD10 in colon cancer 03-24640 FZD10 expression in colon cancer 34 41 59 69 123 124 128 129 141 146 147 N T N T N T N T N T N T N T N T N T N Pr M N Pr M FZD10 β 2MG 149 150 151 153 154 155 201 N Pr N Pr M N Pr M N Pr M N Pr M N Pr N T SS FZD10 β 2MG 04-26192 04-25950 Paraffin slides EDTA buffer (pH9.0) 125 o C, 30sec α -FZD10 mAb 92-13 20 μ g/mL, 4 o C, O/N
Celler Immunization Monoclonal antibody recognizing a complex structure Cells (COS-7 etc.) transfection Ag pCAGGS-FZD10 (FL)-myc ･ His Immunization to mouse Monoclonal Antibody Hybrydoma
Antibody stayed at tumor lesion at 5 days after injection
Internalization of anti-FDZ10 antibody Internalization of anti-FZD10 antibody FZD10(+) FZD10(-) YaFuSS LoVo SYO-1 No Tx 92-13 93-22
In vivo effect of Y90-anti-FZD10 Antibody Day 0 Day 5 Day 9 Day 40 Day 54 90 Y-anti-FZD10 Day 33 Day 40 Day 0 Day 9 Day 54 90 Y-anti-FZD10 Day 9 Day 0 90 Y-CD20-Ab
In vivo effect of Y90-anti-FZD10 Antibody Balb-c/nu (male) / SYO-1 tumor 90 Y - DTPA - antibody 100uCi Intraveneous injection Single injection on Day0 7 Non-Labeled 92-13 (n=5) Non-treated (n=5) 6 Tumor Volume (cm 3 ) 5 4 3 2 90 Y - 92-13 (n=11) 1 0 0 5 10 15 20 25 30 35 40 Days 11 / 11 4 / 11 Injection
Development of cancer peptide vaccine Development of cancer peptide vaccine Development of cancer peptide vaccine and and and Construction of TR network in Japan Construction of TR network in Japan Construction of TR network in Japan
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