QIAGEN's Growing Immuno-Oncology Testing Portfolio
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1 QIAGEN's Growing Immuno-Oncology Testing Portfolio QIAGEN Your Partner in Translational Medicine
2 Current Biomarkers of Immuno-Oncology Focus: Immuno-Oncology Testing Portfolio Tumor mutation burden (TMB) by NGS PD-L1 protein level by IHC Microsatellite Instability (MSI) by PCR Mismatch repair deficiency (dmmr) by IHC Overview: QIAGEN Translational Medicine QIAGEN Your Partner in Translational Medicine
3 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. Biomarkers Companion Diagnostics Clinical Testing Pathway to Biomarkers Pathway to Translational Science Pathway to Precision Medicine 3
4 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 Suzho u Biomarker assay Development & Optimization Biomarker Validation ABI ViiA7 Protein ELISA Nikon ECLIPSE Ni-U ThermoBrite Hybridization oven QIAGEN Your Partner in Translational Medicine SpectraMax i3 MSD S 600 Bio-Rad V3 Western Workflow 4
5 Our One-Stop Shop Business Model CDx Development and Marketing Biomarker Products Nucleic acid DNA & RNA Protein Diagnostic assay development Clinical applications Service Platform Patient stratifications for Clinical Studies Commercial testing Bioassay Development Clinical Testing/ Central Lab Bioinformatics GMP-IVD Manufacturing 5
6 Current Biomarkers of Immuno-Oncology Focus: Immuno-Oncology Testing Portfolio Tumor mutation burden (TMB) by NGS PD-L1 protein level by IHC Microsatellite Instability (MSI) by PCR Mismatch repair deficiency (dmmr) by IHC Overview: QIAGEN Translational Medicine QIAGEN Your Partner in Translational Medicine
7 Immunotherapy: Possible Cure for Cancer Immune checkpoint inhibitors, which unleash a patient s own T cells to kill tumors, are revolutionizing cancer treatment. Schumacher TN et al. Biomarkers in cancer immunotherapy. Cancer Cell. 2015;27(1):12-4. Chen et al, Immunity. 2013;39(1):
8 Growing Immuno-Oncology Testing Portfolio Genomics & Epigenomics Transcriptomics Proteomics Whole Genome/Exome Sequencing Targeted TMB by Sequencing NGS ARMS qpcr (therascreen RGQ PCR Assay) TaqMan MSI qpcr by (ipsogen PCR Onco-Hematology PCR Assays) Droplet Digital PCR (ddpcr) Pyrosequencing (therascreen PyroMark Assay) FISH/CISH >>> IRP panel by qpcr array or NGS RNA-seq miscript Primer Assays miscript mirna PCR Arrays >>> RNA Interference DNA Promoter Gene Expression Gene RNA RNA-seq RT 2 qpcr Primer Assays RT 2 Profiler PCR Arrays QuantiTect Primer Assays QuantiFast Probe Assays DNA Methylation by NGS DNA Methylation by Pyrosequencing (PyroMark CpG Assay) EpiTect Methyl II qpcr System PROTEIN Immunoassay Western PD-L1 by blotting IHC Multi-Analyte ELISArray dmmr by IHC MSD Pathology >>> Bioinformatics 8
9 Current Biomarkers of Immuno-Oncology Focus: Immuno-Oncology Testing Portfolio Tumor mutation burden (TMB) by NGS PD-L1 protein level by IHC Microsatellite Instability (MSI) by PCR Mismatch repair deficiency (dmmr) by IHC Overview: QIAGEN Translational Medicine QIAGEN Your Partner in Translational Medicine
10 TMB as Biomarker for Response to 1 st -line Opdivo Retrospective study of CheckMate 026 shows TMB as a better biomarker in NSCLC patient stratification other than PD-L1 IHC Impact of Tumor Mutation Burden on the Efficacy of First-Line Nivolumab in Stage IV or Recurrent Non-Small Cell Lung Cancer: An Exploratory Analysis of CheckMate 026. AACR Title, Location, Date 10
11 TMB as Biomarker for Response to 1 st line Tecentriq No significant enrichment by PD-L1 expression status on infiltrating immune cells (ICs) metastatic urothelial cancer Tumor mutation load (TMB) was associate with the response (> 16 mutation/mb) Balar, et al. Lancet Title, Location, Date 11
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14 TMB Testing - Assay Flowchart (1) 14
15 TMB Testing - Assay Flowchart (2) 15
16 TMB Testing:Tumor tissue and Plasma ctdna Clinical researches demonstrated TMB is a validated, quantitative genomic biomarker associated with response to immunotherapy High TMB levels is a promising indicator for response to immunotherapy in Lung cancer (BMS CHECKMATE-026 Newest Retrospective Analysis) TMB was examined by WGS/WES on tumor tissues with challenges: Tumor tissue/biopsy was not easy to obtain for WGS/WES testing Cost of WGS/WES was high with deep sequencing in demand At QIAGEN, TMB Testing by targeted DNA-seq NGS method on Tumor tissue (FFPE) or Plasma ctdna sample is developed and kept upgrading Ultra-Deep sequencing (coverage >10000X) by panel can provide more accurate TMB results in FFPE and ctdna samples. QIAseq V3 Panel is empowered with molecular barcode technology to deliver most precise quantitative measure of TMB 16
17 Current Biomarkers of Immuno-Oncology Focus: Immuno-Oncology Testing Portfolio Tumor mutation burden (TMB) by NGS PD-L1 protein level by IHC Microsatellite Instability (MSI) by PCR Mismatch repair deficiency (dmmr) by IHC Overview: QIAGEN Translational Medicine QIAGEN Your Partner in Translational Medicine
18 PD-L1 Expression in Lung Squamous Cell Carcinoma Rabbit Anti-PD-L1/CD274 Monoclonal Antibody :Clone SP142 (200X) (400X) Result: Positive staining, 3+ Tumor and/or infiltrating immune cells 0 ( 1%), 1 ( 1% but <5%), 2 ( 5 but <10%), 3 ( 10%)
19 PD-L1 Expression in Lung adenocarcinoma Rabbit Anti-PD-L1/CD274 Monoclonal Antibody :Clone SP142 (200X) (400X) Result: Positive staining, 2+ Tumor and/or infiltrating immune cells 0 ( 1%), 1 ( 1% but <5%), 2 ( 5 but <10%), 3 ( 10%) Title, Location, Date 19
20 PD-L1 Expression in Lung adenocarcinoma Rabbit Anti-PD-L1/CD274 Monoclonal Antibody :Clone SP142 (200X) (400X) Result: Positive staining, 1+ Tumor and/or infiltrating immune cells 0 ( 1%), 1 ( 1% but <5%), 2 ( 5 but <10%), 3 ( 10%) Title, Location, Date 20
21 Inside Study: PD-L1 Expression in Cancer Patients Mesothelioma Gastric Carcinoma Lung Carcinoma Esophageal Carcinoma 21
22 Current Biomarkers of Immuno-Oncology Focus: Immuno-Oncology Testing Portfolio Tumor mutation burden (TMB) by NGS PD-L1 protein level by IHC Microsatellite Instability (MSI) by PCR Mismatch repair deficiency (dmmr) by IHC Overview: QIAGEN Translational Medicine QIAGEN Your Partner in Translational Medicine
23 MSI-H or dmmr are the 2 nd GOLDEN key for Opdivo 23
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27 Our Results of MSI Analysis System Normal Sample MSI-H positive control sample Microsatellite instability assays using the MSI Analysis System. Matching normal sample (top panel) and MSIpositive test sample (bottom panel) were analyzed using the MSI Analysis System. Two nanograms of genomic DNA was amplified and analyzed using an ABI PRISM 3100 Genetic Analyzer with POP-4 polymer and a 36cm capillary, and the allelic patterns of the normal and test samples are shown. The presence of new alleles in the test sample (indicated by arrows) that were not present in the normal sample indicates MSI. PCR product sizes and respective fluorescent dyes for loci contained in the MSI Analysis System are provided in Table 1. Title, Location, Date 27
28 MSI Analysis Results Interpretation The parameters used for the diagnosis of MSI using the loci recommended by the NCI. MSI-High (MSI-H) requires shifting of two of the five NCI recommended loci ( 30 to 40% of loci tested if more than five loci are analyzed). MSI-Low (MSI-L) is identified by shifting of one of the five NCI-recommended loci ( 30 to 40% of loci tested if more than five loci are analyzed). microsatellite stable (MSS) requires that none of the loci analyzed be shifted. References: 1. Berg KD et al. Detection of microsatellite instability by fluorescence multiplex polymerase chain reaction. J Mol Diagn 2000, 2(1): Boland Cr et al. A National Cancer Institute workshop on microsatellite instability for cancer detection and familial predisposition: development of international criteria for the determination of microsatellite instability in colorectal cancer. Cancer Res 1998, 58: Stadler ZK. Diagnosis and management of DNA mismatch repair-deficient colorectal cancer. Hematol Oncol Clin North Am 2015, 29(1): Title, Location, Date 28
29 MSI analysis CRC patient MSI-H CRC patient (B210) showed a MSI-H genotype followed MSI analysis IHC: PD-L1 MSI-H Penta C B210C NR-21 BAT-26 BAT-25 NR-24 MONO-27 Penta D 400x 200x Penta C B210B NR-21 NR-24 BAT-26 BAT-25 MONO-27 Penta D Promega kit JOE labeled MSI loci (NR-21, BAT-25, MONO-27) FL labeled MSI loci (BAT-26, Penta D) TMR labeled MSI loci (NR-24, Penta C) 29
30 MSI analysis CRC patient MSI-H CRC patient (B274) showed a MSI-H genotype followed MSI analysis IHC: PD-L1 MSI-H MONO-27 B274C Penta C NR-21 NR-24 Penta D BAT-26 BAT x 200x B274B MONO-27 Penta C NR-21 BAT-26 BAT-25 NR-24 Penta D Promega kit JOE labeled MSI loci (NR-21, BAT-25, MONO-27) FL labeled MSI loci (BAT-26, Penta D) TMR labeled MSI loci (NR-24, Penta C) 30
31 Current Biomarkers of Immuno-Oncology Focus: Immuno-Oncology Testing Portfolio Tumor mutation burden (TMB) by NGS PD-L1 protein level by IHC Microsatellite Instability (MSI) by PCR Mismatch repair deficiency (dmmr) by IHC Overview: QIAGEN Translational Medicine QIAGEN Your Partner in Translational Medicine
32 Contact US Nick Zhang, Ph.D. CEO Address: Suite 901, B5 Building, 218 Xinghu St. Suzhou Industrial Park, China ZIP: Business Contact Wei Zhang (Wesley), Ph.D., Director of Business Development Phone: Mobile: Technical Contact Congmao Wang, Ph.D., Manager of NGS & Bioinformatics 32
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