micrornas developing new tools for diagnostics Join forces with IMGM Laboratories to make your mirna project a success Dr. Carola Wagner IMGM Laboratories GmbH Martinsried, Germany qpcr 2009 Symposium March 10 th, 2009 Agenda IMGM - your outsourcing partner (in brief) Our service portfolio (in brief) mirna in diagnostics workflow: microarray-based mirna profiling Summary 10.03.2009 IMGM Laboratories 2009 Slide 2
IMGM Laboratories your outsourcing partner 10.03.2009 IMGM Laboratories 2009 Slide 3 IMGM your outsourcing partner some basic facts IMGM Laboratories was founded d in 2001 We are located on the biotech campus Martinsried / Munich We operate as service provider offering advanced genomic services Our customers come from academia, pharma and biotech a flexible and reliable partnership (e.g. small and dedicated team) 10.03.2009 IMGM Laboratories 2009 Slide 4
IMGM your outsourcing partner some basic facts strategic partnership with the Center for human genetics and laboratory medicine Dr. Klein & Dr. Rost full accreditation according to DIN EN ISO/IEC 17025 full accreditation for expression analysis based on whole genome microarrays and quantitative real-time PCR Agilent Certified Service Provider 10.03.2009 IMGM Laboratories 2009 Slide 5 IMGM Laboratories Service Portfolio 10.03.2009 IMGM Laboratories 2009 Slide 6
Service Portfolio 10.03.2009 IMGM Laboratories 2009 Slide 7 Service Portfolio RNA services RNA services: Microarrays (Agilent platform, multiple options) Real-Time PCR (AB 7900HT, TaqMan arrays, 96- and 384-well blocks, gene expression studies) microrna profiling RNA isolation RNA quality control (Agilent Bioanalyzer Nanodrop) 10.03.2009 IMGM Laboratories 2009 Slide 8
micrornas and their potential in diagnostics 10.03.2009 IMGM Laboratories 2009 Slide 9 mirnas in diagnostics mirnas / mirna profiles: association to ~70 diseases (HMDD, human microrna disease database, last update 2008-10-17) active players in human oncogenesis e.g. mir-21 onco-mirna for breast cancer (Qian et al. (2008) Breast Cancer Res Treat; c.f. Sassen et al. (2008) Virchows Arch 452:1-10 10 classify human cancers e.g. Lu et al. (2005) Nature 435:834-838 Volinia et al. (2006) Proc Natl Acad Sci USA 103:2257-2261 10.03.2009 IMGM Laboratories 2009 Slide 10
mirnas in diagnostics mirnas / mirna profiles associated with: disease prognosis: mir-221/222 cluster associated with poor prognostic in thyroid papillary carcinomas, pancreatic adenocarcinoma & glioblastoma (Lesage et al. (2007) Cell Cycle 6) Low let-7(a) expression associated with shorter survival and poor prognosis for lung cancer patients t (Takamizawa et al. (2004)Cancer Res 64:3753-3756; Yanaihara et al. (2006) Cancer Cell 9:189-198) prognosis of therapy outcome and response high expression of mir-21 associated with poor survival and poor therapeutic outcome of colon adenocarcinoma (Schetter et al. (2008) JAMA 299(4):425-436) prediction of chemotherapeutic response in patients with breast cancer according to mirna signature (Salter et al. 2008) PLoSOne 3(4):e1908) 10.03.2009 IMGM Laboratories 2009 Slide 11 mirnas in diagnostics Actual examples of mirna diagnostic tests: Chronic pancreatitis vs. pancreatic cancer (Asuragen) Prediction of colon cancer (launch early 2009, Exiqon A/S) Differentiation of mesothelioma from other cancers (mirview meso, Rosetta Genomics) Identification of squamous-type lung cancers (mirview squam, Rosetta Genomics) Identification of tissue-of-origin of metastatic tumors (mirview mets, Rosetta Genomics) 10.03.2009 IMGM Laboratories 2009 Slide 12
Possibilities for you to join the field of mirna research!!! 10.03.2009 IMGM Laboratories 2009 Slide 13 mirna profiling the general workflow Sample Size IMGM /Customer Agilent Bioanalyzer Catalog/Custom Arrays Project Consulting Sample Preparation Quality Control Agilent DNA Microarrays Functional analysis mirna mimic or inhibition Hit Validation with qpcr Bioinformatic Data Analysis Customer AB7900HT/TLDAs e.g. mrna microarrays (GEx) 10.03.2009 IMGM Laboratories 2009 Deregulated Genes Slide 14
mirna profiling sample preparation Sample types: Cells Tissues Blood and PBMCs FFPE e.g. Li et al. (2007) BMC Biotechnology 7:36 Xi et al. (2007) RNA 13:1668-1674 Siebolts et al. (2009) J Clinc Path 62(1):84-88 Body fluids e.g. serum, urine, salvia, amniotic fluid, pleural fluid e.g. Chen et al. (2008) Cell Research 18:997-1006 Gilad et al. (2008) PLoS One 3 (9):3148 Hunter et al. (2008) PLoS One 3 (11):e3694 10.03.2009 IMGM Laboratories 2009 Slide 15 mirna profiling (mi)rna quality control & quantification www.agilent.com/chem/labonchip 10.03.2009 IMGM Laboratories 2009 Slide 16
Tissot (2008) Agilent Application Note, Publication Number 5989-7878EN; www.agilent.com/chem/laponchip 10.03.2009 IMGM Laboratories 2009 Slide 17 mirna profiling e.g. Agilent Microarray Workflow Total RNA incl. small RNAs (100ng) Labeled RNAs direct RNA 3 end Labeling with Cy3-pCp hybridization on microarrays mirna profile 10.03.2009 IMGM Laboratories 2009 Slide 18
mirna profiling Data analysis Technical Note 15 (3), www.ambion.com 10.03.2009 Yanaihara et al. (2006) Cancer Cell 9(3):189-198 IMGM Laboratories 2009 Slide 19 mirna profiling Sample treated log2 data Data analysis Collaborative work with SIRION BIOTECH: Thirion et al. (2009) qpcr 2009 RNAi: microrna sirna applications SSample_untreated l d log2 data 10.03.2009 IMGM Laboratories 2009 Slide 20
mirna profiling Hit validation by qpcr linear Fold Change 6 4 * ** * ** 2-2 2 0 2 calibrator mir 103 calibrator mir 27a calibrator mir 23b + mir_103 4 Collaborative work with SIRION BIOTECH: Thirion et al. (2009) qpcr 2009 RNAi: 66 microrna sirna applications 8 10 * p<0.05 ** p<0.01 10.03.2009 IMGM Laboratories 2009 Slide 21 mirna profiling e.g. functional analysis by mrna expression profiling mirna overexpression mirna inhibition mrna profiling = gene expression analysis www.thco.com.tw 10.03.2009 IMGM Laboratories 2009 Slide 22
Summary Single mirnas / mirna profiles in relation to diagnostics: Well established mirna technologies Increased number of publications with promising results First diagnostic tests t launched mirna research for you powered by IMGM laboratories: Advanced genomic services with one focus on RNA services and mirna services State-of-the-art technology to provide top-quality data Full accreditation especially for microarray- and qpcr-based expression analysis Sound bioinformatics to extract meaningful results Expert knowledge in molecular genetics and medical diagnostics 10.03.2009 IMGM Laboratories 2009 Slide 23 Thank you very much for your attention! We are open for business The IMGM Laboratories Team D. Wiegand, M. Hirt, S. Brauer, S. Kotschote, H-G. Klein, R. Oehlmann, C. Wagner, M. Sohns www.imgm.com 10.03.2009 IMGM Laboratories 2009 Slide 24