The use of diagnostic FFPE material in cancer epidemiology research
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1 The use of diagnostic FFPE material in cancer epidemiology research Neil O Callaghan Genetic Epidemiology Laboratory Department of Pathology The University of Melbourne
2 Overview Who we are What we do Why FFPE (Formalin Fixed Paraffin Embedded) Challenges of using FFPE Future uses of FFPE Gain Support for collections in the future
3 PEDIGREE Cancer Study Pathology Epidemiology, DNA, Informatics and Genetics: a Research Enabling Enterprise (
4 PEDIGREE Cancer Study Endeavours to generate evidence so that cancer genomics can be used to prevent cancer through: Leading and contributing to, the discovery of new genomic and molecular advances that help identify people at genetic risk of cancer Using epidemiological and epigenetic data to find the environmental and lifestyle exposures relevant to people at genetic risk of cancer Develop novel statistic and high performance computing methods Nurturing and building the expert multi-disciplinary research team Curating and building all aspects of the resource Growing national and international collaborators
5 PEDIGREE The Bio-repository Bio- Specimen Processing Sample Storage Tissue Culture Resource for Research Processing Blood Saliva Histology 1 Million+ Frozen Blood Fractions Saliva FFPE Cases (slides) Guthrie Cards (duplicates in 2 locations) Cell Lines Active
6 MCCS Colon Prostate Breast Kidney Gastric ABC Study
7 ABC Australian Breakthrough Cancer Study / blood samples / saliva samples Future New Incidents of Cancer
8 Study Cycle Study Development Funding - Australia and International(NIH) Ethics - Hrec - Site Specific Ethics Participants - Joining a Study - Cancer Registries - Family Cancer Clinics Recruitment (up to 1 year) - Informed Consent - Questionnaire - Lifestyle - Family History - Clinical History Blood /Saliva Collection (Base Line) ± Tissue Collection Pedigree ± Pathology Review Molecular Testing Data / Publication Translational Pathology Outcomes Extension Follow Up (FUP) 5 years
9 Study Cycle Follow Up Study Modification Funding - Australia and International(NIH) Ethics - Hrec - Site Specific Ethics Participants - Targeted Family Members Re- Recruitment (up to 1 year) - New Informed Consent - Questionnaire - Lifestyle - Family History - Clinical History Blood /Saliva Collection (FUP) New Incident Tissue Collection - Cancer Registry Searches Death Registry Searches Pedigree Updates Pathology Review Molecular Testing Data / Publication Translational Pathology Outcomes Extension Follow Up (FUP2)
10 Blood and FFPE Blood Saliva Normal Tissue FFPE
11 Genetic and Epigenetic Testing Epigenetic Factors Methylation Genetic Factors New Genetic Variants IHC Screening Tool Prognostic Markers
12 Translational Pathology - Outcomes FFPE Tumour Classification Prognosis Risk Measurements Treatment Options Somatic Tumour Testing Germline Testing Blood/Saliva Predictive Testing Risk Measurements Prevalence Penetrance Carriers Non Carriers Intensive Screening Less Intensive Screening Relative Surveillance Other Cancers
13 Translational Pathology Molecular Subtypes of Breast Cancer Luminal A 40% Luminal B 20% Her2 Enriched 15-20% Basal 15-20% Basal Like Triple negative (BRCA1 80%) 70%
14 Translational Pathology Expression-based classification of CRCs from 4,151 patients CMS1 MSI immune Hypermutated, MSI-High and strong immune CMS2 canonical Epithelial, marked WNT and MYC signalling CMS3 Metabolic Epithelial, metabolic dysregulation CMS4 Mesenchymal TGF-β activation, stromal invasion, angiogenesis
15 Translational Pathology MSI Immune Colorectal Cancer 1)MMR Deficient 2) Immune Rich Present Absent Treatment- Check Point Inhibitor Therapy Germline Testing MSI High Pembrolizumab (PDL1) Carrier PDL-1
16 Collection Challenges Collection Issues Availability Quality Storage Cost
17 Availability Retention times Long cohorts Pedigrees Remote archiving Storing and not accessing Workload Amalgamated / closed laboratories / Custodians? Amount of tissue remaining in blocks Resistance to access High price to deter Consented / ethics Collection Challenges
18 Collection Issues Quality DNA Antigenicity Storage time Storage Conditions Transport Temperature Sunlight Exposure UV Oxidation air Hydrolysis Endogenous water Exogenous water e.g. 4 o C Fridge Tissue processing time Inadequate Prolonged Fixation time / type 1 Fresh cut sections from Block 2 Sections stored 4 to -20 O C
19 Collection Challenges Storage Bundling Dark Air tight 4 o C Lab Storage Limited space Cost
20 Collection Challenges Cost Cost of collection is rising Impact on existing funding Budgets Funding decreasing / harder Cost recovery
21 The Future of FFPE Testing?? Independent Repository New diagnostic tests Molecular technologies Cheaper Better Molecular Testing / IHC Research tests = Diagnostic tests = Revenue for laboratories IHC to better define areas of interest Tumour heterogeneity Collecting more Normal and other pre-cancers lesions Predictive tests for patients and families Clinicians driving change
22 Future Normal Testing Normal Colon from MMR Deficient Carriers MSH2 MSH6
23 Future - Using IHC for Macro/Micro-Dissection Laser Capture Micro-Dissection Ducts or Crypts
24 The Victorian Cancer Registry Cancer prevalence statistics
25 Concluding Focus on FFPE Tumour New gene variants Normal Feedback on our requesting What can we give back to keep your labs invested Our participants want their tissues included in our research. Thank you for your contributions to our studies Researchers need your support
26 Acknowledgements Our Participants Funding bodies All laboratories that have provided tissue for our studies Copyright The University of Melbourne 2011
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