© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Imaging Genomic mapping of Edema/Cellular Invasion MRI-Phenotypes in Glioblastoma Multiforme PO Zinn 1, B Mahajan 2, P Sathyan 1, S Singh 1, S Majumdar 1, A Flanders 3, E Huang 4, R Jain 5, D Gutman 6, S Hwang 6, J Kirby 7, J Freyman 7, TCGA Glioma Phenotype Research Group,, F Jolesz 2, RR Colen 2, 1 M.D. Anderson Cancer Center, Houston, TX, USA. 2 Brigham and Women's Hospital, Boston, MA, USA. 3 Thomas Jefferson University Hospital, Philadelphia, PA, USA. 4 National Cancer Institute, Bethesda, MD, USA. 5 Henry Ford, Detroit, MI, USA. 6 Emory University, Atlanta, GA, USA. 7 SAIC-Frederick, Bethesda, MD, USA.

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Disclosure  No Disclosures. R25 CA089017(RRC) P41 RR (FAJ)

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Cellular invasion is one of the major reasons for therapy failure of modern multimodal GBM treatment The discovery of targetable genes responsible for cell spread and invasion can be expected to impact modern therapy and patient survival. Glioblastoma Multiforme

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Imaging genomics Imaging genomics has emerged as a new field which links the specific imaging traits (radiophenotypes) with gene-expression profiles. We present our paper on the first comprehensive imaging genomic analysis using quantitative MRI volumetrics and large scale gene- and micro-RNA expression profiling in GBM. This was first described by our group (Zinn et al PLOSOne 2011) in 2011 where an invasive gene and microRNA was uncovered using the FLAIR volume as an MRI biomarker (radiophenotype).

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Purpose In this presentation, we seek to demonstrate the imaging-genomic mapping of an invasive MRI radiophenotype, linking MR imaging traits with gene- and miRNA expression profiles, in GBM patients to determine genomic correlates of an edema/invasion MRI radiophenotype In order to: - Discover - Discover new genomic targets for GBM treatment (decreasing edema/invasion) - Predict - Predict key genetic events by MRI - Identify - Identify patients who are candidates for the targeted treatment

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 We retrospective identified 78 treatment naïve GBM patients whom had: Pre-treatment MRI neuroimaging from The Cancer Imaging Archive (TCIA) Gene- and miRNA- expression profiles The Cancer Genome Atlas (TCGA) Methods and Materials

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Methods and Materials All Image Analysis was performed on Slicer 3.6 (slicer.org) - The Segmentation module was used to obtain volumes of the peritumoral non-enhancing FLAIR hyperintensity reflecting tumor invasion/edema - T2/FLAIR was registered to the post- contrast T1WI. - Volumetric segmentation was performed in a simple hierarchical model of anatomy, proceeding from peripheral to central. - 3 distinct structures were segmented: edema/invasion enhancing tumor necrosis

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Methods and Materials Tumor Segmentation. 65 year old male patient with a right temporal GBM. Segmentation of tumor edema (blue), enhancement (yellow) and necrosis (red) was performed and edema volume was obtained.

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Methods and Materials Image- Genomic Biostatistics analysis 12, 674 genes(22,267 hybridization probes) and 555 microRNAs (1,510 hybridization probes) were analyzed (Affymetrix/Agilent chip technology) in each patient Quantitative edema volumes for each patient (N=78) were obtained High and low groups corresponding to volumes of high and low peritumoral edema/invasion were analyzed/compared for differential genomic expression profiles Comparative Marker Selection (Broad Institute) Statistical method to identify preferentially upregulated genomic events in one vs. another predefined patient group (high_low) Ingenuity Pathway Analysis (IPA) was used to determine gene ontology

Example of a patient with high edema/tumor infiltration © NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Methods and Materials High FLAIR Radiophenotype

Example of a patient with low edema/tumor infiltration © NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Methods and Materials Low FLAIR Radiophenotype

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Results Genes associated with invasion were seen in the high FLAIR discovery and validation sets.

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Results Top concordant genes across discovery and validation sets

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Results T OP CONCORDANT MICRO RNA S ACROSS DISCOVERY AND VALIDATION SETS Top Cellular function migration/invasion

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Results B IOINFORMATICALLY PREDICTED GENE - MICRO RNA REGULATORY NETWORKS IN HIGH FLAIR SIGNAL GBM S.

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Results Our genes and microRNAs were more predictive of patient survival than the current subclassification schema used in GBM today.

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Conclusion In this presentation, we present the first comprehensive image- genomic analysis using quantitative MRI volumetrics and large-scale gene- and microRNA expression profiling in GBM We identified MRI as a possible screening tool and imaging surrogate for genes and microRNAs involved in GBM cellular migration and invasion Specifically, we identified miR-219 and Periostin as a potential therapeutic target against GBM invasion.

© NlH National Center for Image Guided Therapy, 2012 ASNR 2012 Acknowledgements Helping make cancer history using imaging Thank you! This work is supported R25 CA089017(RRC), P41 RR (FAJ). A collaborative project between BWH and M.D. Anderson.