Session: Innovative Technologies

Transcription

1 Session: Innovative Technologies Abstract Title: Tissue Microarray Technology: New opportunity for Tissue Banks to contribute to Cancer Research and Diagnostics. Pasquale De Blasio Founding President of ESBB Member of Marble Arch Int. Biobanking Working Group President of ISENET-USA LLC

2 Tissue Microarray Technology Pathology archives PPFT Tissue Blocks & Slides Tissue Microarrayer Combines tens to hundreds of paraffinembedded tissue specimens into a single paraffin block. TMA Block

3 Tissue Microarray Technology It is possible to slice up to 200 consecutive sections of 4-5μm from each TMA block, mounted on a microscope slide The TMA slide can be processed like ordinary tissue sections with a wide range of techniques: Histo staining, Immuno histo Immuno fluorescent. FISH

4 Advantages to use Pathology Archives widest collection of clinical tissues available with the entire clinical heterogeneity range Tissue samples are related to clinical records and have follow-up and outcome information Are under the responsibility of pathologists that have access to all the information and are bound by professional secrecy It is now possible to perform many types of molecular analysis on this type of tissues.

5 TMAs in Biomarker Development In Vitro Screening Methods Clinical Trials Epidemiology Based Studies Multi-tumor Systems Micro-Arrays Other Methods Tissue Microarrays Validation In Real Patient Samples TMAs advantages: the speed of molecular analyses is increased by more than 100-fold. precious tissues are not destroyed large number of molecular targets can be analysed from consecutive TMA sections (Cost reduction). Compliments of S. Hewitt, NCI TARP Laboratory

6 First scientific pubblications on TMA

7 From Tissue To Arrays Develop Hypothesis Identify Tissue Prepare Tissue Array Tissue Section Arrays Perform Experiment Analyze Data Publish Repeat Compliments of S. Hewitt, NCI TARP Laboratory

8 Quality of Tissue is everything Tissue Quality Histology Proteins Nucleic Acids Diagnostic Reporting Clinical Annotation Storage Of Biomaterial Compliments of S. Hewitt, NCI TARP Laboratory

9 Challenges in the Construction of a TMA Identifying Adequate/Appropriate Material Negotiating MTA/IRB Approval Specimens Present Special Challenges Biopsy & Small Specimens Are Hard To Array Prior Use of Tissue for diagnostic Diversity of Specimen Sources Regional Differences In Diagnosis & Treatment Differences In Specimen Handling Informatics TRACEABILITY Managing Expectations Pathologist Vs Scientist

10 Clinical Database Integrated Tissue MicroArray Pathology Platform workflow Tissue Microarrayer Platform Quantitative Digital Pathology Digital Scanners Fish Fluorescence Histo Immuno

11 Summary FFPETs donor blocks Pick cell or tissue cores from donor blocks Place cell/tissue cores in specific vessels Spot cell/tissue cores in recipient paraffin blocks DNA and RNA extraction Harvest and re-suspend cells in agarose Embed cells in paraffin donor blocks Immunohisitochemical/ immunofluorescemnce analysis

12 Summary Tissue microarrays (TMAs) have become a mainstay in preclinical and translational research, especially for the development of biomarker assays for characterization of disease. They allow for analysis of extremely small amounts of tissue, thus preserving valuable tissue blocks. At the same time, they dramatically increase the efficiency and cost effectiveness of performing tissue-based studies by enabling the examination of 10s to 100s of different patient samples on the same slide.

13 Summary Platform For High-Throughput Pathology A block of Samples from hundreds of Tissue Blocks Multiple Samples (3-4) from Paraffin Embedded or Frozen Tissue blocks Arranged in an organized way Immuno Histo Fluorescence Fish Compliments of S. Hewitt, NCI TARP Laboratory

14 Summary Platform For High-Throughput Proteomics A Protein Array with Retained Histomorphology Cell Type Localization Cell Type Quantification Subcellular Localization Immuno Histo Fluorescence Fish Compliments of S. Hewitt, NCI TARP Laboratory

15 Conclusion ,888 2, SINGLE SLIDES TMA H&E IHC IHC (75 cases, dupl) 3, , , , , MATERIALS H&E IHC INDIRECT COSTS IHC on TMA LABOR Total cost per unit Cost per patient Cost per 75 patients Compliments from G. Cattoretti, UNIMIB- Italy

16 Summary Brest, Gult1, IHC, Membrane (A, I) Connectivity Brest, PgR (DAB), Pos Rat (N, A) Melanoma KI-67 + MART1 (DAB, AEC) Proliferation

17 Conclusion Courtesy of: Giorgio Cattoretti Associate Professor, Universitá degli Studi Milano-Bicocca Director, Division of Surgical Pathology, Cytology, Medical Genetics and Nephropathology Azienda Ospedaliera San Gerardo, Monza This material may be copyrighted. Do not Compliments diffuse without from authorization. G. Cattoretti, UNIMIB- Italy

18 Tuesday 21st October Training Course on Tissue Microarrays and Quantitative Digital Pathology. Topics covered include: planning a biomarker research project using TMAs, diagnostic use of TMAs quantitative digital pathology analysis. Includes a hands-on session to make your own TMA and analyse your slides (bring your own blocks and slides).

19 Acknowledgments Maurizio Andrea Aikaterini Tatiana Davide Monica Simona Alberto Alessandria Valeria