1. Giving Clinical Significance To Molecular Targets

2. TriStararoundtheworld Tristar Technology Group 9700 Great Seneca Highway Rockville, MD 20850 U.S.A www.tristargroup.us Rockville, MD Headquarters Hamburg, Germany Main Repository Array Manufacturing Contract Research Catania, Italy Tissue Repository

3. Network of Repositories HematologicalMalignancies Washington DC, USA Headquarters Hamburg, Germany Main Repository Array Manufacturing Contract Research Solid Tumors InflammatoryDiseases Hepatic & RenalDiseases CNS Solid Tumors HematologicalMalignancies ProspectiveCollection Projects Catania, Italy Tissue Repository

4. General Information Cancer 120 Tumor Types Over 2.5 millionsamples Over 60,000 microarrayedsamples Formalin fixed & frozen Detailedclinicalinformation CNS Over 100,000 samples Formalin fix & frozen Clinicalinformation Alzheimer‘sDisease, Parkinson‘sDisease, Stroke, MS, VascularDementia, PrionDisease etc. Inflammatory Disease Over 300,000 samples Over 3,000 microarrayedsamples Formalin fixed Rheumatoid Arthritis, InflammatoryBowelDisease, Psoriasis Arthritis etc.

5. Biological Samples Tissue micro arrays with matched large sections Tissue blocks with clinical database Primary cells, RNA & DNA Matched FFPE & Frozen samples Primary Tumors Primary Tumors with Matched Normal Tissues Primary Tumors with Matched Metastases Nodal & Distant Metasteses Samples with Molecular & Clinical Data

6. EthicalConsiderations Informed Donor Consent IRB Approval Fully Anonymized Compliant with Current International & EU Regulations Blocks That Are in Excess of Diagnostic Sample Only Team of Pathologists & Oncologists for Clinical Data Review

7. Prospective Collection Projects Parameters defined by Customer Protocol Established Timeline & Number of Samples IRB Approval Projects Include Collection of Snap-Frozen Samples with Matched Serum/Plasma, CSF as applicable

8. Quality Control Samples are fixed/frozen within 2 – 10 minutes of Excision OCT embedded sample Snap frozen sample Formalin fixed sample 10% Buffered formalin, 10-12 hrs fixation time Morphology (H&E) & IHC Markers for immunogenicity RNA & DNA Quality (Agilent 2100 Bioanalyzer) RIN can be checked & provided upon request

9. Finding an Attractive Drug Target MolecularEpidemiology Howfrequentisexpression in human cancer? Specificcancersubtypesorbiologicalproperties? -prognosticrelevance What normal tissues do express target? Option 2:Performownstudies Option 1:Reviewtheliterature

10. Currentsituation Typicalearlysteps Latersteps usuallynotdone! FunctionalAnalysis MolecularEpidemiology Druggability

11. Finding an Attractive Drug Target Currentsituation Typicalearlysteps Latersteps usuallynotdone! FunctionalAnalysis MolecularEpidemiology Druggability Large sets of Human tissues with clinical Follow-up required. Difficult to get.

12. Finding an Attractive Drug Target Optimal situation Latersteps Earlysteps FunctionalAnalysis MolecularEpidemiology Druggability Identifyclinically relevant targets beforeincurringsignificantcosts

13. Tissue Micro Array (TMA) Technology A tool for the High-Throughput Analysis of Thousands of Tissue samples Kononen, Sauter et al. Nat Med. 1998 Jul;4(7):844-7

14. Frozen OCT Embedded Morphology Formalin Fixed Paraffin Embedded RNA/protein TissueMicroarrays 2. 14. 200. DNA

15. Tristar: A New Dimension in Human Tissue Analysis

17. Colon 1,500

18. Lung 1,500

19. Prostate 3,000

20. Bladder, Pancreatic, Ovarian etc.Cancer-Specific Analyisis (Prognosis / Progression) 5,000 - 10,000 Datapoints Generated Per Antibody Database Generation Genotype vs Phenotype

21. TriStar Antibody Protocol Development Test Various Normal and Cancer Tissues Test Different Tissue Pre-treatment Conditions: Temperature, Pressure, Enzymatic Antibody Dilution Series for Optimal Background: Signal Ratio

22. TriStar Antibody Protocol Development Controls: Pre-absorption Control (Test for Target Specificity) Isotype Control Antibody Test for Unspecific (Fc-mediated) Binding Omit Primary Antibody Test for Unspecific Staining Induced by the Detection System

23. List of Tissues for Cross-Reactivity Study Adrenal * Lung * Spinal Cord Blood Cells * Lymph Node * Spleen Blood vessels (endothelium) * Ovary * Striated (skeletal) Muscle Bone Marrow * Fallopian Tube (oviduct) * Testis Brain – cerebrum (cortex) * Pancreas * Thymus Brain – cerebellum * Parathyroid * Thyroid Breast (mammary gland) * Peripheral Nerve * Tonsil Eye * Pituitary * Ureter Gastrointestinal Tract2 * Placenta * Urinary Bladder Heart * Prostate * Uterus- body (endometrium) Kidney (glomerulus, tubule) * Salivary Gland * Uterus – cervix Liver * Skin

24. TriStar Multi-Tumor TissueMicroarray 10-50 Samples Each Of 120 Human Cancers Skin: Squamous Cell Carcinoma, Basal Cell Carcinoma, Merkel Cell Carcinoma. Uterine Corpus: Endometrioid Adenocarcinoma, Serous. Parathyroid Gland: Adenoma, Carcinoma. Mammary Gland: Intraductal Carcinoma, Lobular Carcinoma In Situ, Invasive Ductal Carcinoma, Invasiv Lobular Carcinoma, Mucinous Carcinoma, Papillary Carcinoma, Tubular Carcinoma. Kidney: Clear Cell Type, Papillary Type, Chromophobe Cell Type. Urinary Bladder: Non-Invasive Papillary Tumor (Pta), Transitional Cell Carcinoma, Squamous Cell Carcinoma, Adenocarcinoma, Small Cell Carcinoma. Salivary Glands: Mixed Tumor, Adenolymphoma, Adenoma, Mucoepidermoid Carcinoma, Acinic Cell Carcinoma, Adenocarcinoma, Adenoid Cystic Carcinoma. Esophagus: Squamous Cell Carcinoma, Adenocarcinoma. Stomach: Adenocarcinoma Diffuse Type, : Adenocarcinoma Intestinal Type. Adrenal Gland: Adrenal Cortical Adenoma, Adrenal Cortical Carcinoma, Pheochromocytoma. Pancreas: Adenocarcinoma, Adenoma. Mediastinum: Thymoma. Small Intestine: Adenocarcinoma, Carcinoid. Large Intestine: Adenoma, Adenocarcinoma. Appendix: Adenocarcinoma, Carcinoid. Anal: Small Cell Carcinoma. Prostate: Prostatic Adenocarcinoma Untreated, Hormone Refractory Adenocarcinoma Adenocarcinoma, Clear Cell Adenocarcinoma, Atypical Hyperplasia. Cervix: Squamous Cell Carcinoma, Adenocarcinoma. Vagina: Squamous Cell Carcinoma, Adenocarcinoma. Vulva: Squamous Cell Carcinoma. Thyroid Gland: Follicular Carcinoma, Papillary Carcinoma, Anaplastic Carcinoma, Medullary Carcinoma, Adenoma. Lung: Squamous Cell Carcinoma, Adenocarcinoma, Undifferentiated Large Cell Carcinoma, Small Cell Carcinoma, Carcinoid. Testis: Seminoma, Teratoma, Embryonal Carcinoma, Choriocarcinoma, Yolk-Sac-Tumor, Teratocarcinoma. Ovary: Serous Carcinoma, Mucinous Carcinoma, Endometrioid Carcinoma, Brenner Tumor, Germ Cell Tumors. Liver: Hepatocellular Carcinoma, Cholangiocarcinoma. Fibrohistiocytic: Fibrosarcoma, Benign Histiocytoma, Dermatofibrosarcoma Protuberans, Atypical Fibroxanthoma, Malignant Fibrous Hiostiocytoma Lipomatous: Lipoma, Lioposarcoma. Smooth Muscle: Leiomyoma, Leiomyosarcoma, Leiomyoblastoma. Skletal Muscle: Rhabdomyoma, Rhabdomyosarcoma. Blood And Lymph Vessels: Angioma, Epitheloid Hemangioma, Hemangioendothelioma, Angiosarcoma, Kaposi Sarcoma. Perivascular: Glomus Tumor, Hemangiopericytoma. Synovial: Benign Giant Cell Tumor Of Tendon Sheath, Synovial Sarcoma. Mesothelial: Solitary Fibrous Tumor Of Pleura And Peritoneum, Adenomatoidtumor, Malignes Mesothelioma. Neural: Neurofibroma, Neurinoma. Granular Cell Tumor, Malignant Peripheral Nerve Sheath Tumor. Clear Cell Sarcoma. Paraganglioma, Ganglioneuroma. Pnet: Ganglioneuroblastoma, Neuroblastoma, Neuoepithelioma, Extraskelettal Ewings-Sarcoma. Malignant Mesenchymoma. Alveolar Soft Part Sarcoma. Epitheloid Sarcoma. Osseous: Osteoidosteoma, Osteoblastoma, Osteosarcoma. Chondrous: Chondroblastom, Chondrom, Chondrosarcoma, Chordomas. Ewing Sarcoma. Giant Cell Tumor Of The Bone. Brain: Astrocytoma, Glioblastoma Multiforme, Oligodendroglioma, Ependymoma, Medulloblastoma, Medulloepithelioma, Craniopharyngeoma, Esthesioneuroblastoma, Retinoblastoma. Nevus Naevocellularis, Malignant Melanoma, Gastrointestinal Stromatumor, Endometrioid Stromal Sarcoma, Mixed Malignent Mesodermal Tumor, Aml, Cml, Cll, Immunocytic Lymphoma, Plasmocytoma, Centrocytic Lymphoma, Centroblastic Centrocytic Lymphoma, Centroblastic Lymphoma, Immunoblastic Lymphoma, Burkitt Lymphoma, T-Cell Lymphoma Low Grade, T-Cell Lymphoma High Grade, M Hodgkin Lymphocytic Depletion, M Hodgkin Mixed Cell Type, M Hodgkin Nodular Sclerosing etc. In which cancers is the target expressed?

25. TriStar Normal Human TissueMicroarray 76 tissue types, 532 cell types, 8 donors each Mesenchymal tissues: aorta/intima, aorta/media, heart (left ventricle), sceletal muscle, sceletal muscle/tongue, myometrium, appendix (muscular wall), esophagus (muscular wall), stomach (muscular wall), ileum (muscular wall), colon descendens (muscular wall), kidney pelvis (muscular wall), urinary bladder (muscular wall), penis (glans/corpus spongiosum), ovary (stroma), fat tissue (white), Surfaces: skin (surface), skin (hairs, sebaceous glands), lip (epithelium), oral cavity, tonsil (surface epithelium), anal canal (skin), anal canal (transition epithelium), exocervix, esophagus, kidney pelvis, urinary bladder, amnion/chorion, stomach (antrum), stomach (fundus and corpus), small intestine, duodenum, small intestine, ileum, appendix, colon descendens, rectum, gallbladder, bronchus, paranasal sinus. Solid organs: lymph node, spleen, thymus, tonsil, liver, pancreas, parotid gland, submandibullary gland, sublingual gland, lip (small salivary gland), duodenum (Brunner gland), kidney cortex, kidney medulla, prostate, seminal vesicle, epididymis, testis, lung (parenchyma), lung (bronchial glands), breast, endocervix, endometrium (proliferation), endometrium (secretion), fallopian tube, endometrium (early decidua), ovary (stroma), ovary (corpus luteum), ovary (follicular cyst), placenta (first trimenon), placenta (mature), adrenal gland, parathyroid gland, thyroid, cerebellum, cerebrum, pituitary gland (posterior lobe), pituitary gland (anterior lobe) In which normal tissues is the target expressed?

26. Validation CaseStudy EstrogenReceptor (ESR1) Gene Amplification Target Identified using aCGH Isitreallytrue? Howfrequent? Histo-pathologicalsubtypes? Isitclinically relevant? Commercial value? ER: Target forhormonetherapy in breastcancer

27. Validation Platform TriStarBreastCancerPrognosis Array pTstage pNstage Number of nodesexamined Number of positive nodes Tumor diameter BRE grade Polymorphy Tubulusformation Mitoses 2,200 Breast Cancers with 5 yr.follow-up information Survivalmonths (99%) Survivaltumorspecific (40%) Sometherapyinformation (40%) Molecularparameters: FISH: HER2, EGFR, MDM2, CCND1, MYC IHC: ER, PR, p53, Cytokeratins, EGFR, HER2, CD117, others

28. BreastCancerPrognosis TMA Analysis ESR1 amplification (FISH) ESR1 Amplification* in 358/1739 (21%) of Breast Cancers Holst, Simon et al, Nat Gen (39), 655-660, 2007

29. Does ESR1 Amplification cause ER Overexpression? ??? ER IHC datafromourdatabase Isthe ESR1 AmplificationFunctionallyRelevant ?

30. ER expression level (Allred score) ESR1 Amplification Are Linked To ER Protein Overexpression

31. ESR1 Amplificationarelinked to EarlyStage and Low Grade BreastCancers p=0.7295 p<0.0001 Holst, Simon et al, Nat Gen (39), 655-660, 2007

32. 1.0 ESR1 amplification (n=43) 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 0 20 40 60 80 100 ESR1 amplification and anti ER treatment 175 Patients Treated With Tamoxifen No Adjuvant Therapy ER IHC positive (n=109) Surviving ER IHC negative(n=23) p<0.0001 months surv Holst, Simon et al, Nat Gen (39), 655-660, 2007 ESR1 amplificationpredictsresponse to tamoxifen

33. ESR1 Validation StudyusingPrognosis TMA: Timeline Affymetrix experiment, ESR1 amp identification March 2006 Low density TMA validation April 14th, 2006 High density TMA studies April-May, 2006 Patent filed July 15th, 2006 Paper accepted, Holst et al, Nature Genetics February 2nd, 2007 Start to Finish: 11 months

34. Tristar: A New Dimension in Human Tissue Analysis

35. Products & Services Product Groups Archived Human Tissue Repository High-Density Tissue Micro Arrays (>60,000 donor samples) Large sections, Blocks, RNA, DNA & Primary Cells Cancer Stem Cells Arrays Clinical Data – Prognosis, Progression, Treatment & Response Molecular Database Services Compound Screening using Cancer Stem Cells Antibody Protocol Development (IHC) & Characterization FISH analysis Large-Scale Multi-Tumor & Normal Tissue Array Analysis Large-Scale Analysis of Prognosis Cross-Reactivity Screening in Normal Tissue (GLP)

36. Thanks! Tristar Technology Group 9700 Great Seneca Highway Rockville, MD 20850 U.S.A www.tristargroup.us Rockville, MD Headquarters Hamburg, Germany Main Repository Array Manufacturing Contract Research Catania, Italy Tissue Repository