QIAGEN (Suzhou) Translational Medicine Co., Ltd. Bringing Biomarkers and CDx to Precision Medicine

Transcription

1 QIAGEN (Suzhou) Translational Medicine Co., Ltd. Bringing Biomarkers and CDx to Precision Medicine 1

2 CONTENT Company Overview Our Capabilities & Expertise A Total Solution Provider for Precision Medicine 2

3 QIAGEN-World leader in Sample, Assay & Diagnostics Technologies Germantown, US Manchester, UK Hilden, Germany Shanghai/Suzhou Tokyo, Japan Copenhagen Stockholm Oslo Helsinki Los Angeles Mexico City Germantown/ Gaithersburg Toronto Manchester Venlo London Hamburg Duesseldorf Paris Vienna Milan Madrid Jesi Zurich Ankara Beijing Shenzhen Seoul Tokyo Shanghai Hong Kong > 500 core products with 2500 patents Consumable kits and instruments Kuala Lumpur Singapore > 500,000 customers worldwide Molecular Diagnostics Applied Testing Pharma Academia Sao Paolo Sales by Geography Asia RoW 2% 17% Americas 48% Brisbane Sydney Melbourne Melbourne Employees >5000 Europe 33% 33

4 QIAGEN (Suzhou) Translational Medicine Co., Ltd QIAGEN (Suzhou) is an innovative company that provides an unique integrated solution for Biomarker and CDx development and Clinical Testing for precision medicine & healthcare. PATHWAY TO BIOMARKER PATHWAY TO PRECISION MEDICINE Biomarkers Companion Diagnostics Clinical Testing Pathway To Biomarker Pathway To Companion Diagnostics Pathway To Precision Medicine 4

5 Our Business Model for Collaborations Rx pathway Drug Development Pre-clinical and Phase I Dx pathway Biomarker Development and Validation Drug Development Phase II and III Patient Stratifications for Clinical Trials FDA/CFDA Approval & Marketing Patient Screening for Therapy Preclinical Studies Clinical Trials Patients Testing PATHWAY TO BIOMARKER PATHWAY TO PRECISION MEDICINE 5

6 Our Industry-Leading Portfolio of CDx Selected co-development companion Diagnostics (CDx) Project Partner Indication Biomarker Status Eribitux (cetuximab) Vectibix (panitumumab) Gilotrif (afatinib) Iressa (gefitnib) Iressa (gefitnib) PF (dacomitinib) LY ) (Abemaciclib) Early-stage compound CO-1686 Colorectal cancer Colorectal cancer Lung cancer (NSCLC) Lung cancer (NSCLC) Lung cancer (NSCLC) Lung cancer (NSCLC) Lung cancer (NSCLC) KRAS KRAS EGFR EGFR (FFPE) EGFR (Liquid biopsy) KRAS KRAS U.S. regulatory approval in July 2012 U.S. regulatory approval in May 2014 U.S. regulatory approval in July 2013 CE-IVD kit in Europe and other non-u.s. markets CE-IVD kit in Europe In development In development Blood cancer JAK2 In development Lung cancer (NSCLC) EGFR (T790M) In development Various projects Not disclosed Various In development Not disclosed Confidential Oncology/ nononcology Confidential In development 6

7 Transform Biological Samples into Molecular Insights Complete Workflow Bioinformatics QIAsymphony RGQ Analysis Sequencing Interpretation Protein Biomarker Assays Reporting BIOLOGICAL SAMPLE Sample Technologies Assay Technologies VALUABLE MOLECULAR INSIGHTS Extract DNA, RNA and proteins in reliable process Make molecular info visible and available for interpretation 7

8 Our Complete Platforms for Translational Medicine Biobanking & Sample prep NGS and Bioinformatics Genomics/Epigenomics Transcriptomics/miRNomics GeneReader Precision Medicine Biomarker Screening & Discovery Roto-Gene Q PyroMark Q24 NextSeq 500 MiniSeq Pathology Companion Diagnostics Approval Patient Stratification Suzhou Biomarker assay Development & Optimization Biomarker Validation ABI ViiA7 Protein ELISA Nikon ECLIPSE Ni-U ThermoBrite Hybridization oven SpectraMax i3 MSD S 600 Bio-Rad V3 Western Workflow 8

9 Our Expertise: Whole Spectrum Biomarker Development Genomics Epigenomics Transcriptomics mirnomics Whole Genome/Exome Sequencing Targeted sequencing ARMS qpcr (therascreen RGQ PCR Assay) TaqMan qpcr (ipsogen Onco-Hematology PCR Assays) Droplet Digital PCR (ddpcr) Pyrosequencing (therascreen PyroMark Assay) FISH/CISH RNAseq miscript Primer Assays miscript mirna PCR Arrays RNA Interference Proteomics DNA Gene Expression RNA RNAseq RT 2 qpcr Primer Assays RT 2 Profiler PCR Arrays QuantiTect Primer Assays QuantiFast Probe Assays PROTEIN Immunoassay Western blotting Multi-Analyte ELISArray MSD Pathology Promoter Gene DNA Methylation by NGS DNA Methylation by Pyrosequencing (PyroMark CpG Assay) EpiTect Methyl II qpcr System Next-Generation Sequencing PCR-based Pyrosequencing-based Immunoassay Pathology Bioinformatics 9

10 Our Expertise: Biobank Collaboration Others,2654 Colorectal,2399 Specimens Cases Liver,612 Breast,1603 Stomach,1892 lung,1774 Number of Specimens and Cases Tumor classification Other types of specimens Tissue Microarray (Colorectal & Breast cancer) 10

11 Our Expertise: Sample Preparation Automated Workflows Samples collection Isolation Stabilization Disruption Purification Amplification Cells PBMC (Ficoll) RNAlater TissueLyser FFPE kit Tissue (fixed or T cell RNAprotect TissueRuptor exorneasy frozen) B cell Allprotect QIAshredder CTC Blood Single cells PAXgene CfDNA Bone marrow tube REPLI-g Liquid biopsy QIAcard QIAamp QIAsafe DNeasy RNeasy AllPrep mirneasy Qproteome 11

12 Our Complete NGS Sample-to-Insight Solution Cells Tissue Blood FFPE Tumor Actionable Mutations Clinically Relevant Tumor Myeloid Neoplasms Breast Cancer Colorectal Cancer Liver Cancer Lung Cancer Ovarian Cancer Prostate Cancer Gastric Cancer Cancer Predisposition Comprehensive Cancer GeneReader NextSeq 500 Liquid biopsy MiniSeq Sample preparation A variety of samples Targeted enrichment Library Construction WGS WES RNA-seq Targeted sequencing NGS run With any Sequencing Platform Data Analysis Interpretation Open Source pipeline IPA/IVA Brand Analysis 12

13 Our Wet-Bench Extensively Verified Cancer Panels Type Panel # Genes Size (kb) # Amplicons Specificity Uniformity (0.2x mean) Experimental coverage Tumor Actionable Mutations % 91% 91% Solid tumors Clinically Relevant Tumor % 90% 90% Human BRCA1 & BRCA % 99% 100% Hematologic malignancies Disease-specific Comprehensive Myeloid Neoplasms % 94% 94% Breast Cancer % 91% 91% Colorectal Cancer % 95% 95% Liver Cancer % 96% 96% Lung Cancer % 90% 90% Ovarian Cancer % 96% 96% Prostate Cancer % 94% 94% Gastric Cancer % 93% 93% Cancer Predisposition % 93% 93% Comprehensive Cancer % 92% 92% 14 Catalog Panels (Extensively Validated RUO Kits) Mix-n-Match Panels (Any of 570 gene was wet-bench validated for library performance) Custom Panels (Anywhere in human genome can be designed to amplify) 13

14 Example 1:Mechanism study of Drug resistance QIAGEN GeneRead Lung Cancer Panel is used in this article to detect and compare the ctdna sequence to find the mutations that may cause AZD9291 resistance. The data from above would shed light on the potential mechanisms of resistance to AZD9291 before the availability of resistance biopsy specimens. Reference: Thress KS, et al. Acquired EGFR C797S mutation mediates resistance to AZD9291 in non-small cell lung cancer harboring EGFR T790M. Nature medicine, 2015, 21(6):

15 Example 2:Application on genetic profiling of tumors Focused NGS of samples with a minimum of 8 ng cfdna was performed using the QIAGEN GeneRead Lung Cancer Panel as described by the manufacturer, except input was reduced to as low as 8 ng DNA. Results demonstrate the utility of a simple approach that enabling robust molecular analysis of CTCs and cfdna for genotype-directed therapies in multi-site clinical trials and represent a significant methodological improvement for clinical benefit. Reference: Rothwell DG, et al. Genetic profiling of tumours using both circulating free DNA and circulating tumour cells isolated from the same preserved whole blood sample. Molecular oncology,

16 Human Comprehensive Cancer Panel (V2) Gene list (160 Genes, 0.8Mb,10000X,10G) ABL1 BUB1B DDR2 FGFR2 IDH2 MEN1 PDGFRA SMARCA4 AKT1 CARD11 DICER1 FGFR3 IKZF1 MET PHF6 SMARCB1 AKT2 CBL DNMT3A FH IL6ST MLH1 PIK3CA SMO ALK CBLB ECT2L FLCN IL7R MSH2 PIK3R1 SPOP AMER1 CD79A EGFR FLT3 JAK1 MSH6 PMS2 SRC APC CD79B EP300 FUBP1 JAK2 MTOR PPP2R1A STK11 AR CDC73 EPCAM GATA1 JAK3 MUTYH PRDM1 SUFU ARID1A CDH1 ERBB2 GATA2 KDM6A MYC PRKAR1A TERT ARID2 CDK12 ERBB3 GATA3 KDR MYD88 PTCH1 TNFAIP3 ASXL1 CDK4 ERBB4 GNA11 KIT NF1 PTEN TNFRSF14 ATM CDKN2A ERCC5 GNAQ KLF6 NF2 PTPN11 TP53 ATRX CHEK2 ESR1 GNAS KMT2D NFE2L2 RAC1 TSC1 BAP1 CIC EZH2 GPC3 KRAS NFKBIA RB1 TSC2 BCL6 CREBBP FAM46C GRIN2A MAP2K1 NOTCH1 RET TSHR BCOR CRLF2 FANCA H3F3A MAP2K2 NOTCH2 ROS1 U2AF1 BRAF CSF1R FANCD2 HIST1H3B MAP2K4 NPM1 SDHB VHL BRCA1 CTNNB1 FANCE HNF1A MAP3K1 NRAS SETD2 WT1 BRCA2 CYLD FAS HRAS MAP4K3 PALB2 SF3B1 XPC BRIP1 DAXX FBXO11 HSPH1 MDM2 PAX5 SLC7A8 ZNF2 BTK DDB2 FBXW7 IDH1 MED12 PBRM1 SMAD4 ZRSR2 The Human Comprehensive Cancer GeneRead DNAseq Targeted Panel is a collection of multiplexed PCR primer assays for targeted enrichment of the coding (exonic) regions of the 160 genes that are most commonly mutated in cancers with a recognizable oncogenic consequence. Mutations in these oncogenes and tumor suppressor genes are often relevant for tumor classification, and warrant extensive investigation to enhance the understanding of carcinogenesis. 16

17 Human Comprehensive Cancer Panel (V3) Gene list (275 Genes, 1.7 Mb, 10000X, 20G) ABL1 BCL2L1 CDK12 DOT1L FBXW7 HIST1H3B KMT2C MYC PIK3CA RHEB STK11 ACVR1B BCL6 CDK4 EED FGF4 HNF1A KMT2D MYCL PIK3R1 RHOA SUFU AKT1 BCOR CDK6 EGFR FGF6 HOXB13 KRAS MYCN PIK3R2 RIT1 SUZ12 AKT2 BCORL1 CDKN2A EGLN1 FGFR1 HRAS LRP1B MYD88 PIM1 RNF43 TAL1 AKT3 BCR CDKN2B EP300 FGFR2 HSP90AA1 MAP2K1 NF1 PLCG1 ROS1 TCF3 ALK BIRC3 CDKN2C EPAS1 FGFR3 ID3 MAP2K2 NF2 PMS1 RUNX1 TERT AMER1 BLM CEBPA EPHA3 FGFR4 IDH1 MAP2K4 NFE2L2 PMS2 SDHB TET2 APC BRAF CHEK1 EPHA5 FH IDH2 MAP3K1 NFKBIA POLD1 SETBP1 TGFBR2 AR BRCA1 CHEK2 ERBB2 FLCN IGF1R MAP3K14 NKX2-1 POLE SETD2 TNFAIP3 ARAF BRCA2 CIC ERBB3 FLT3 IKZF1 MAPK1 NOTCH1 PPM1D SF3B1 TNFRSF14 ARID1A BRIP1 CREBBP ERBB4 FLT4 IKZF3 MCL1 NOTCH2 PPP2R1A SMAD2 TP53 ARID1B BTK CRLF2 ERG FOXL2 IL7R MDM2 NOTCH3 PRDM1 SMAD4 TRAF3 ARID2 CALR CSF1R ESR1 FUBP1 INHBA MDM4 NPM1 PRKAR1A SMARCA4 TSC1 ASXL1 CARD11 CSF3R ETV6 GALNT12 IRF4 MED12 NRAS PRKDC SMARCB1 TSC2 ATM CBL CTCF EXO1 GATA1 JAK1 MEF2B NSD1 PRSS1 SMC1A TSHR ATR CBLB CTNNA1 EZH2 GATA2 JAK2 MEN1 NTRK1 PTCH1 SMC3 U2AF1 ATRX CBLC CTNNB1 FAM175A GATA3 JAK3 MET NTRK2 PTEN SMO U2AF2 AURKA CCND1 CUX1 FAM46C GEN1 KAT6A MITF NTRK3 PTPN11 SOCS1 VHL AURKB CCND3 CXCR4 FANCA GNA11 KDM5C MLH1 PAK3 RAC1 SOX2 WHSC1 AURKC CCNE1 CYLD FANCC GNAQ KDM6A MPL PALB2 RAD21 SOX9 WT1 AXIN1 CD274 DAXX FANCD2 GNAS KDR MRE11A PAX5 RAD50 SPOP XPO1 AXIN2 CD79A DDR2 FANCE GREM1 KEAP1 MSH2 PBRM1 RAD51 SRC XRCC2 B2M CD79B DICER1 FANCF GRIN2A KIT MSH6 PDGFRA RAF1 SRSF2 XRCC3 BAP1 CDC73 DNM2 FANCG H3F3A KMT2A MTOR PDGFRB RB1 STAG2 ZNF217 17

18 Example 3 : Clinically-relevant mutation identification Sequencing libraries were prepared from DNA samples (40 ng input) by targeted enrichment of exonic regions using the QIAGEN GeneRead Human Comprehensive Cancer Panel. The sensitivity and specificity of variant identification achieved with this technique are potentially clinically relevant, and its optimization could provide a true liquid biopsy for molecular characterization of pancreatic cancer. Reference: Tepper CG, et al. Utilization of next-generation sequencing to identify clinically-relevant mutations in cell-free circulating tumor DNA from patients with advanced pancreatic cancer. Cancer Research, 2015, 75(15 Supplement):

19 Example 4 : molecular barcode used in panel The researchers used the QIAGEN-V3 panel to accurately detect somatic mutations with allele fractions as low as 0.1% at 10000X depth in coding regions using our enrichment protocol and variant caller. Reference: Xu et al..detecting very low allele fraction variants using targeted DNA sequencing and a novel molecular barcodeaware variant caller. BMC Genomics (2017) 18:5 DOI /s

20 Advantages of GeneRead DNAseq Targeted Panels(V2) Benefits of GeneRead Targeted Panels: Adapted to all NGS platforms Average coverage can reach 5000X-10000X Well-suited for FFPE and ctdna samples Use as little as 40 ng starting DNA Quick library construction (3 hours) Total analysis in 2 weeks Clinically and Biologically relevant content GeneRead DNAseq Targeted Panels focus on specific mutations, exons, and genes that are most relevant to a particular disease. The content of these panels was selected from the College of American Pathologists (CAP) guidelines, NCCN guidelines, late-stage clinical trials, The Cancer Genome Atlas (TCGA), and Ingenuity Knowledge Base. 20

21 Advantages of QIAseq Targeted Panels(V3) Benefits of QIAseq Targeted DNA Panels: Digital sequencing enabled by molecular barcodes to remove PCR duplicates Complete solution streamlines the workflow Compatibility with low-quality DNA enables efficient sequencing of FFPE and cfdna samples Minimal DNA input to preserve precious samples(20ng) Optimized buffers and conditions to achieve high coverage of GC-rich regions Principle workflow 21

22 Our Expertise: Biomarker and Patient Stratification with MDx MDx Nucleic acidbased Protein-based Pathology Genomic alterations Mutations; indels o therascreen RGQ PCR kit o therascreen PyroMark sequencing o qbiomarker DNA Mutation PCR Arrays and Assays Amplifications o qbiomarker Copy Number PCR Arrays and Assays Gene expression dysregulation Epigenetics MicroRNA profiling Conventional Western blotting ELISA Electrochemiluminescencebased MSD assay Immunohistochemistry (IHC) Immunofluorescence (IF) Fluorescence In situ hybridization (FISH) Chromogenic In situ hybridization (CISH) 22

23 Our Solutions for Gene Mutation Testing Mutation testing therascreen RGQ Kit ARMS Scorpions qpcr therascreen Pyro Kit Pyrosequencing Highest sensitivity on the market Selective mutation detection Ease of use One-step procedure Quantification of alleles Detection of unknown variants Fast delivery of direct and unambiguous data 23

24 Our Current MDx Portfolio therascreen qpcr Assays Biomarker Product Line Disease Platform RUO CE FDA CDx EGFR therascreen NSCLC RGQ X X X GILOTRIF (BI) IRESSA (AZ) KRAS therascreen mcrc RGQ X X X Erbitux (Lilly/BMS) KRAS therascreen NSCLC RGQ X X 201X Abemaciclib (Lilly) melanoma; BRAF therascreen Glioma; mcrc RGQ X X IDH1/2 therascreen Glioma RGQ X X ALK therascreen NSCLC RGQ X 2015 T315I therascreen CML RGQ X HSP110 therascreen CRC RGQ AKT therascreen Breast Cancer RGQ 2018 Bayer component PI3K therascreen therascreen Pyrosequencing Assays Biomarker Product Line Disease Platform RUO CE FDA CDx EGFR therascreen NSCLC Pyro X X KRAS therascreen mcrc Pyro X X melanoma; BRAF therascreen Glioma; mcrc Pyro X X melanoma; NRAS therascreen mcrc Pyro X X KIT therascreen GIST Pyro X PDGFRA therascreen GIST Pyro X UGT1A1 therascreen mcrc Pyro X X MGMT therascreen Glioma Pyro X X RAS Extension therascreen Pyro X 24

25 Liquid biopsy: monitoring cancer-genetics in the blood 25

26 QIAGEN and AstraZeneca Co-developed the first-ever liquid biopsy-based CDx for IRESSA QIAGEN s circulating tumor DNA test is now CE-IVD marked to assess EGFR mutation status in non-small cell lung cancer (NSCLC) patients based on plasma samples therascreen EGFR RGQ Plasma PCR kit helps physicians to identify patients who could benefit from treatment with IRESSA when tumor tissue sample is not evaluable QIAGEN pioneering the use of liquid biopsy-based companion diagnostics as a lessinvasive option to complement surgical biopsies for genomic profiling of cancers 26

27 Sensitive somatic mutation detection for liquid biopsy QIAamp Circulating Nucleic Acid extraction. The PCR-based, targeted pre-amplification solution enriches the target regions of interest in ctdna by 4000-fold, therefore dramatically increasing the effective template input for downstream mutation analysis. qbiomarker somatic mutation assays can be used to detect, quantify, and monitor potentially 1201 mutations in 126 genes from the ctdna sample. 27

28 Liquid Biopsy: Mutations in cfdna of Lung Cancer Gene COSMIC ID Nucleotide Amino Acid Gene COSMIC ID Nucleotide Amino Acid COSMIC Nucleotide Amino Acid Gene Change Change Change Change ID Change Change AKT c.49g>a p.e17k KRAS 516 c.34g>t p.g12c TP c.818g>a p.r273h BRAF 451 c.1397g>t p.g466v KRAS 521 c.35g>a p.g12d TP c.818g>t p.r273l BRAF 460 c.1406g>c p.g469a KRAS 522 c.35g>c p.g12a TP c.844c>t p.r282w BRAF 470 c.1789c>g p.l597v KRAS 520 c.35g>t p.g12v TP c.856g>a p.e286k BRAF 476 c.1799t>a p.v600e KRAS 527 c.37g>t p.g13c TP c.892g>t p.e298* CTNNB c.110c>g p.s37c KRAS 532 c.38g>a p.g13d AKT CN* CN* CTNNB c.110c>t p.s37f NRAS 580 c.181c>a p.q61k BRAF CN* CN* EGFR c.2125g>a p.e709k PIK3CA 763 c.1633g>a p.e545k CTNNB CN* CN* EGFR c.2126a>c p.e709a PIK3CA 775 c.3140a>g p.h1047r EGFR CN* CN* EGFR 6252 c.2155g>a p.g719s STK c.1062c>g p.f354l ERBB CN* CN* EGFR 6253 c.2155g>t p.g719c STK c.109c>t p.q37* HRAS CN* CN* EGFR 6239 c.2156g>c p.g719a TP c.461g>t p.g154v KRAS CN* CN* EGFR 6223 c.2235_2249del15 p.e746_a750del TP c.469g>t p.v157f NRAS CN* CN* EGFR 6225 c.2236_2250del15 p.e746_a750del TP c.473g>t p.r158l PIK3CA CN* CN* EGFR c.2237_2251del15 p.e746_t751>a TP c.488a>g p.y163c STK CN* CN* EGFR c.2237_2255>t p.e746_s752>v TP c.517g>t p.v173l TP CN* CN* EGFR 6218 c.2239_2247del9 p.l747_e749del TP c.524g>a p.r175h SMPC PC** PC EGFR c.2239_2248>c p.l747_a750>p TP c.527g>a p.c176y SMPC PC PC EGFR c.2239_2251>c p.l747_t751>p TP c.527g>t p.c176f EGFR 6254 c.2239_2253del15 p.l747_t751del TP c.536a>g p.h179r EGFR 6255 c.2239_2256del18 p.l747_s752del TP c.574c>t p.q192* EGFR c.2240_2254del15 p.l747_t751del TP c.578a>g p.h193r EGFR c.2240_2257del18 p.l747_p753>s TP c.614a>g p.y205c EGFR 6241 c.2303g>t p.s768i TP c.637c>t p.r213* EGFR c.2307_2308ins p.v769_d770ins TP c.641a>g p.h214r EGFR 6240 c.2369c>t p.t790m TP c.659a>g p.y220c EGFR c.2497t>g p.l833v TP c.701a>g p.y234c EGFR 6224 c.2573t>g p.l858r TP c.711g>t p.m237i EGFR 6213 c.2582t>a p.l861q TP c.713g>t p.c238f ERBB2 682 c.2322_2323ins p.m774_a775ins TP c.725g>t p.c242f ERBB c.2325_2326ins p.a775_g776ins TP c.733g>t p.g245c HRAS 498 c.182a>t p.q61l TP c.734g>t p.g245v KRAS 552 c.182a>g p.q61r TP c.742c>t p.r248w KRAS 553 c.182a>t p.q61l TP c.743g>a p.r248q KRAS 554 c.183a>c p.q61h TP c.743g>t p.r248l KRAS 555 c.183a>t p.q61h TP c.745a>t p.r249w KRAS 512 c.34_35gg>tt p.g12f TP c.746g>t p.r249m KRAS 517 c.34g>a p.g12s TP c.747g>t p.r249s KRAS 518 c.34g>c p.g12r TP c.817c>t p.r273c 28

29 Complete list of 126 genes in qbiomarker TM Mutation PCR Assays # Gene Name Mutations # Gene Name Mutations # Gene Name Mutations # Gene Name Mutations 1 ABL EZH MLH PTPRS 1 2 ADAMTS FAM123B 0 66 MLST PTPRT 2 3 ADAMTS FBXW MMP RB1 7 4 AKT FGFR MPL RET 16 5 AKT FGFR MSH RHEB 1 6 ALK FGFR MTOR RHOA 1 7 APC FLT MYD RICTOR 1 8 ARID1A 4 40 FOXL NF RPS6KB1 1 9 ASXL GATA NF RUNX ATM 6 42 GATA NFE2L SF3B ATRX 3 43 GNA NOS SLC17A AXIN GNAQ 6 76 NOTCH SMAD BCOR 1 45 GNAS 6 77 NOTCH SMARCB BRAF GRIN2A 1 78 NPM SMO 1 15 BRCA HNF1A 7 79 NRAS SOCS CARD HRAS NTRK SRC 1 17 CBL 9 49 IDH PAX SRSF CDC IDH PDGFRA STK CDH IL7R 1 83 PDGFRB TET CDKN2A IRS PDK TMEM132B 1 21 CEBPA IRS PDK TNFAIP CHRM IRS PIK3CA TP CREBBP 7 55 JAK PIK3R TRRAP 1 24 CRLF JAK PIK3R TSC CSF1R 4 57 KIT PPP2R1A TSC CTNNB KRAS PRDM TSHR CYLD 1 59 LRRN PTCH U2AF DNMT3A 2 60 MAP2K PTEN VHL EGFR MAPKAP PTPN WT EIF4EBP MEK PTPN XPO EP MEN PTPRB 1 32 ERBB MET PTPRD 1 Total

30 Electrochemiluminescence-based assay (Meso Scale Discovery,MSD) 30

31 Protein Assay Platform: MSD Description Akt Signaling Akt Signaling (Total Protein) Akt Signaling II Apoptosis EGFR Family ERK-STAT3 Cascade Insulin Signaling Insulin Signaling (Total Protein) MAP Kinase MAP Kinase (Total Protein) Phospho-STAT Panel Intracellular Signaling Panels Analytes pakt (Ser 473) pp70s6k (Thr 421/ Ser 424) pgsk-3β (Ser 9) Akt p70s6k GSK-3β pakt (Ser 473) pp70s6k (Thr 389) pgsk-3β (Ser 9) ps6rp (Ser 240/244) p53 (total) pp53 (Ser 15) cleaved PARP cleaved Caspase-3 pegfr perbb2 pigf-1r pmek1/2 (Ser 217/221) perk-1/2 (Thr/Tyr: 202/204; 185/187) pstat3 (Tyr 705) pigf-1r pir pirs-1 IGF-1R IR IRS-1 pjnk (Thr 183/ Tyr 185) pp38 (Thr 180/ Tyr 182) perk-1/2 (Thr/Tyr: 202/204; 185/187) JNK p38 ERK-1/2 pstat3 pstat4 pstat5a,b Marker Phospho/Total Panels Phospho/Total Akt Ser 473 Phospho/Total BAD Ser 112 Cleaved/Total Caspase-3 p20 / p17 Phospho/Total EGFR Tyr 1173 Phospho/Total ErbB2 Tyr 1248 Activation Site Phospho/Total ERK Thr 202 / Tyr 204, Thr 185 / Tyr 187 Phospho/Total GSK-3β Ser 9 Phospho/Total HSP27 Ser 15 Phospho/Total HSP27 Ser 78 Phospho/Total HSP27 Ser 82 Phospho/Total JNK Thr 183 / Tyr 185 Phospho/Total c-kit Tyr 721 Phospho/Total MEK1/2 Ser 217 / 221 Phospho/Total Met Tyr 1349 Phospho/Total mtor Ser 2448 Phospho/Total p38 Thr 180 / Tyr 182 Phospho/Total p53 Ser 15 Phospho/Total p70s6k Thr 421 / Ser 424 Phospho/Total Rb Ser 608 Phospho/Total Rb Ser 780 Phospho/Total S6RP Ser 240 / 244 Phospho/Total STAT5a/b Tyr

32 Protein Assay Platform: MSD Inflammation Assays 32

33 Pathology Platform Biomarker: c-met Drug target Biomarker Test Detection assay Material needed Platform Kit/antibody etc References c-met c-met Expression of total c-met/phospho-c-met Immunohistochemistry (IHC) FFPE tissue Molecular Pathology CONFIRM anti-total c-met Rabbit Monoclonal Primary Antibody (Ventana) Anti-Met (c-met) (phospho Y1349) antibody (Abcam) 1. Santoro, A. et al. The Lancet. Oncology; 14, (2013). 2. Klotz, M. et al. Biomarkers ; 17, , (2012). Drug target Biomarker Test Detection assay Material needed Platform Kit/antibody etc References c-met c-met Amplification of c-met Fluorescent in situ hybridization (FISH) FFPE tissue Molecular Pathology Vysis MET SpectrumRed FISH Probe Kit (Abbott Molecular) CEP 7 SpectrumGreen Probe (Abbott Molecular) 1. Ou S-HI et al., Journal of Thoracic Oncology; 6, (2011). 2. F. Cappuzzo et al., Journal of Clinical Oncology; 27, (2009). 3. J. Bean et al., PNAS; 104, (2007) 33

34 IHC: c-met Expression in Liver cancer (patient) IHC Score ( 200) ( 400) IHC (using CONFIRM anti-total c-met (SP44) antibody) analysis in liver cancer. 34

35 MET no amplification in Liver Cancer MET no amplification in Q-HCC-02 PDX model. MET copy number=2.095 by Affymetrix Genome-Wide Human SNP Array 6.0 MET/CCP7 FISH Probe Kit, CytoTest Inc. Original magnification, 1000X 35

36 Correlation of c-met Overexprssion and Amplification MET overexpression by IHC (400X) MET amplification by FISH (1000X) IHC (anti-total c-met antibody) and FISH analysis in Q-HCC-01 patient-derived xenograft tumors. 36

37 Patient Stratification Biomarker for Immunotherapy-I Drug target Biomarker Analyte Detection assay Material needed Platform Kit/antibody etc PD-1 and PD-L1 PD-L1 Expression of total PD-L1 Immunohistochemistry (IHC) FFPE tissue Molecular Pathology Rabbit Anti-Human PD-L1 /CD274 Monoclonal Antibody References 1. Herbst RS et al. Predictive correlates of response to the anti-pd-l1 antibody MPDL3280A in cancer patients. Nature. 2014; 515(7528): Tumeh PC et al. PD-1 blockade induces responses by inhibiting adaptive immune resistance. Nature. 2014; 515(7528):

38 PD-L1 IHC Assay Validation Placenta (Positive control ) Stomach (negative control) PD-L1 ( 400) ( 400) Immunohistochemical analysis using SP142 antibody in normal placenta and stomach tissues( 400). 38

39 PD-L1 Expression in Esophagus Cancer (patient) Esophagus cancer (200X) Esophagus cancer (400X) IHC (using Rabbit Anti-Human PD-L1/CD274 Monoclonal Antibody (Clone SP142)) analysis in esophagus cancer. 39

40 PD-L1 Expression in Gastric Cancer (patient) Gastric cancer (200X) Gastric cancer (400X) IHC (using Rabbit Anti-Human PD-L1/CD274 Monoclonal Antibody (Clone SP142)) analysis in gastric cancer. 40

41 PD-L1 Expression in Lung squamous cell carcinoma Lung squamous cell carcinoma (200X) Lung squamous cell carcinoma (400X) IHC (using Rabbit Anti-Human PD-L1/CD274 Monoclonal Antibody (Clone SP142)) analysis in Lung squamous cell carcinoma. 41

42 Biomarker Solutions Across the Entire Drug R&D Continuum Basic research & Preclinical Clinical FDA Approval & Precision Medicine Biomarker Discovery Biomarker analysis for: Clinical samples In vitro cell-based assays In vivo animal studies Patient Stratification & response monitoring Patient tumors Liquid biopsy Histopathology Genomics Transcriptomics Proteomics Companion Diagnostics (CDx) Approval Personalized HealthCare 42

43 Our Unique Advantages as Your Best Partner Integrated workflow solution From biomarker discovery and companion diagnostics (CDx) development, to clinical testing for personalized medicine. Access to high quality samples QIAGEN Suzhou has access to clinical samples from hospitals, internal and external biobanks with great sample management system. QIAGEN global technology & expertise Work with reliable and high-quality technologies and platforms with global presence focusing on sample preps, biomarkers and CDx as one-stop-shop solution provider. High quality, low cost and timely With the capability of unique high-through put platforms, Global standard quality data can be generated timely. 43

44 Contact Us QIAGEN (Suzhou) Translational Medicine Co.,Ltd Nick Zhang, Ph.D. CEO Address: Suite 901, 5B Building, 218 Xinghu St. Suzhou Industrial Park, China Business Contact Wei Zhang (Wesley), Ph.D., Director of Business Development Phone: Mobile: Technical Contact Ais Zhu, Ph.D., Senior Manager of Central Lab Phone: