Personalized Healthcare Update

Transcription

1 Dr. Kai - Oliver Wesche Market Development Manager, Personalized Healthcare QIAGEN Personalized Healthcare Update

2 Pioneering Personalized Medicine through Partnering TOMTOVOK BKM120 Zelboraf QIAGEN partners: and many more...

3 QIAGEN Core Competencies 1 Complex Sample 2 Sample Technology 3 Pure Analyte 4 Assay Technology Target detected 3

4 Your benefit Kits detecting clinical relevant mutations

5 Example: Non-small cell lung carcinoma (NSCLC) Drug efficacy correlates with EGFR mutation status IPASS Study IPASS (IRESSA Pan-ASia Study): randomised, parallel-group study to assess the efficacy, safety and tolerability of gefitinib (novel drug) versus carboplatin/paclitaxel (standard therapy) as first line treatment in a clinically selected patient population from Asia. Gefitinib or Carboplatin Paclitaxel in Pulmonary Adenocarcinoma. TS. Mok, Y-L. Wu, S. Thongprasert, C-H. Yang, D-T. Chu, N, Saijo, et al., NEJM, Sep 2009; 361:

6 Example: metastatic Colorectal carcinoma (mcrc) Drug efficacy correlates with KRAS mutation status Clinical trial demonstrating that patients with a wild type KRAS gene in DNA from colorectal cancer cells, showed significant benefit to treatment with Panitumumab ( Vectibix ). Patients whose tumor contained mutated KRAS gene had no drug response. KRAS mutation positive Panitumumab+BSC Best Supportive Care (BSC) KRAS mutation negative Panitumumab+BSC Best Supportive Care (BSC) "Routine use of KRAS mutational testing in colorectal cancer patients could save the health care system more than $600 million in drug costs alone." Wild-Type KRAS Is Required for Panitumumab Efficacy in Patients With Metastatic Colorectal Cancer. R.G. Amado, M. Wolf, M. Peeters, et al., Journal of Clinical Oncology, Vol 26, (10), 2008: pp

7 therascreen fully validated workflows Patient sample Sample technologies Assay technologies Diagnostic Results Manual: QIAamp (eg. DNA FFPE) PAXgene Tissue PyroMark Q24 Comprehensive & Complex Tasks Classical FFPE Sample Tissue Automated: QS-RGQ RotorGene-Q Selective & Sensitive Tasks

8 therascreen - overview therascreen Reliability Regulatory status (CE-IVD) Workflow solutions Short Turn around time ARMS Scorpions qpcr Highest Sensitivity on the market Selective mutation detection Ease of use One-step procedure Pyrosequencing Comprehensive results in real-time Complex mutation testing with simple data interpretation Small amount of starting material

9 Pyrosequencing workflow DNA purification PCR Sample prep Pyrosequencing ~ 2h ~ 15 min ~ min therascreen Pyro Kits PyroMark Q24 Vacuum Prep Workstation PyroMark Q24 PyroMark Q96ID PyroMark Q96MD Pyrosequencing provides quantitative results in a sequence context in as little as 1 hour after PCR

10 Features of the therascreen Pyro portfolio TAT Regulatory Status Information output Starting material Workflow Solution ~ 3h CE-IVD marked assays Sequencing data Highly fragmented DNA in small amounts (<10ng per reaction) Workflow validation from Sample to Result Robustness +++ Result interpretation one-click result easy interpretation Sensitivity ~ 5%

11 therascreen Pyro Kits Specification Assay Design Mutations KRAS Prescription of Erbitux & Vectibix in mcrc patients, only when KRAS wt is present in the cells EGFR Detection of mutations & deletions in E18 E21. Important for Iressa and Tarceva in mnsclc first line therapy BRAF Mutations in the BRAF occur mainly in Melanoma (Zelboraf (Vemurafinib)) NRAS Mutations in NRAS codon 12, 13 & 61 metastatic colorectal cancer (mcrc) & melanoma tumors 2 PCR + 2 Sequencing rxn Plug-in 4 PCR + 5 Sequencing rxn Plug-in 2 PCR + 2 Sequencing rxn Reverse assay 2 PCR + 2 Sequencing rxn G12A G12D G12S G12V G12R G12C G13D Q61H Q61E Q61L G719S G719A G719C S768I T790M L858R L861Q L861R Deletions (20 most common ones) V600E G464V G469A V600M G464E G469V V600A G466V G469E V600G G466E G12D G12S G12C G12V G12A G12R G13D G13R G13V G13C G13A G13S Q61L Q61P Q61E Q61R Q61K Q61H MGMT Involved in repairing alkylated DNA. Inactivation of the MGMT-gene leads to tumor formation (Neuro-Onkology) 1 PCR + 1 Sequencing rxn 48 rxn Kit! 4 CpG sites UGT1A1 Only commercial assay on the market that can detect var *6 & var *28 in the UGT1A1 gene 2 PCR + 2 Sequencing rxn *6 = Gly Arg *28 = 6 7 TA repeats

12 therascreen KIT/PDGFRA screening Kit GIST screening kit Coming 2012/13 Detection of KIT exon 9 insertion Gleevec dose 400mg// 800mg PDGFR exon 18 point and complex mutations Gleevec resistance The kit will contain 2 sequencing and 2 PCR primers ( 2 rxn per patient) wt PDGFR B1: CCAGAKACATCATGCATGATTCGAACTA G: 99% T: 1% E S T G C A G A G T A C T A T C A T G % mt PDGFR G>T A2: CCAGAKACATCATGCATGATTCGAACTA 5 G: 60% T: 40% E S T G C A G A G T A C T A T C A T G

13 therascreen RGQ PCR workflow DNA purification DNA Assessment Mutation Testing Result Interpretation optional ~ 140main ~ 2,5 h ~ 15 min QIAamp DNA FFPE Tissue Kit (56404) therascreen (RGQ) PCR Kits Rotor-Gene Q Analysis Software The PCR procedure takes less than 3h.

14 Features of the therascreen PCR portfolio therascreen RGQ PCR Kits Lower Limit of Detection <1% Analytical Sensitivity 100% Amplicon Size ~100bp Procedure time (ex DNA ext n ) <3h Complexity Result Output Low Objective

15 therascreen PCR/RGQ Kits Specification Assay Design Mutations PI3K The PI3K Mutation Test Kit is intended to detect 4 mutations in codons 9 and 20 of the PIK3CA gene for research use. Incl. a control assay and 3 mutation assays H1047R, E542K, E545D, E545K, *1 KRAS The therascreen KRAS RGQ PCR Kit is intended to detect 7 mutations in codons 12 and 13 of the KRAS gene. 2step procedure: 1)DNA assessment 2)mutation detect. 12ALA,12ASP,12ARG,12CYS,12SER, 12VAL,13ASP EGFR The therascreen EGFR RGQ PCR Kit enables detection of 29 somatic mutations in the EGFR gene. 2step procedure: 1)DNA assessment 2)mutation detect. 19 deletions in exon 19* ;T790M; L858R; L861Q; G719X ; S768I; 3 insertions in exon 20 * Detects the presence of any of 19 deletions but does not distinguish between them. Detects the presence of G719S, G719A, or G719C but does not distinguish between them. Detects the presence of any of 3 insertions but does not distinguish between them BRAF Research kit for detection of V600 mutations in the BRAF gene. 2step procedure: 1)DNA assessment 2)mutation detect. V600E common (GAG)* V600E complex (GAA) and V600D (GAT) V600K (AAG) * Down to 1.27% sensitivity. Detects the presence of the two but does not distinguish between them. Detection of this mutation is highly dependent on sufficient copies of V600K present. At 300 copies of V600K, mutant template with a C T of 34.7 cycles is typically observed *1: E545K and E545D are detected in a single reaction thus this kit will be able to detect both mutations but not discriminate between them.