Tissue Microarray and Imaging Core
The Tissue Microarray and Imaging (TMI) Core is dedicated to the construction and evaluation of high quality TMAs for cancer research. The Core performs high-throughput isolation of DNA and RNA from formalin-fixed paraffin-embedded tissues and also provides members with web-based digital pathology services using the Aperio system. These services include slide scanning, image viewing and storage, and quantitative image analysis, which facilitate tissue-based research. Finally, the Core provides access to the Automated Quantitative Analysis [AQUA] system.
Listed below are a few examples of how the core has contributed to various projects.
Geoffrey ShapiroDFCI (Translational Pharmacology and Early Therapeutic Trials Program, Lung Cancer Program): The investigators evaluated the anticancer activity of ganetespib, a novel non-geldanamycin heat shock protein 90 (HSP90) inhibitor, in non-small cell lung cancer (NSCLC) models. Specifically, the activity of ganetespib was compared to that of the geldnamaycin 17- AAG in biochemical assays, cell lines and xenografts, and evaluated in an ERBB2 YVMA-driven mouse lung adenocarcinoma model. Results demonstrated that in genomically-defined NSCLC cell lines, ganetespib caused depletion of receptor tyrosine kinases, extinguishing of downstream signaling, inhibition of proliferation and induction of apoptosis. In mice bearing NCI-H1975 (EGFR L858R/T790M) xenografts, ganetespib was rapidly eliminated from plasma and normal tissues but was maintained in tumor with t1/2 58.3 hours, supporting once-weekly dosing experiments, in which ganetespib produced greater tumor growth inhibition than 17- AAG. However, after a single dose, re-expression of mutant EGFR occurred by 72 hours, correlating with reversal of anti-proliferative and pro-apoptotic effects. Consecutive day dosing resulted in xenograft regressions, accompanied by more sustained pharmacodynamic effects. Ganetespib also demonstrated activity against mouse lung adenocarcinomas driven by oncogenic ERBB2 YVMA. Increased expression of HSP27, consistent with HSP90 inhibition, and reduced expression of ERBB2 was documented in the treated tumors (see figure).
The Core used the Aperio ScanScope System to digitize xenograft tissue slides immunostained for several biomarkers (EGFR, CD31, DiOC7(3), TUNEL, pimonidazole and BrdU, HSP27, Ki67, p-S6) and provided access to Aperio Image Analysis algorithms for quantification of biomarker expression. Shimamura T, Perera SA, Foley KP, Sang J, Rodig SJ, Inoue T, Chen L, Li D, Carretero J, Li YC, Sinha P, Carey CD, Borgman CL, Jimenez JP, Meyerson M, Ying W, Barsoum J, Wong KK, Shapiro GI. Ganetespib (STA-9090), a Non-Geldanamycin HSP90 Inhibitor, has Potent Antitumor Activity in In Vitro and In Vivo Models of Non-Small Cell Lung Cancer. Clin Cancer Res. 2012 Jul 17.
James A. DeCaprioDFCI (Cancer Cell Program, Cancer Genetics Program): Fbw7, a substrate receptor for Cul1-RING-ligase (CRL1), facilitates the ubiquitination and degradation of several proteins, including Cyclin E and c-Myc. In spite of much effort, the mechanisms underlying Fbw7 regulation are mostly unknown. In this study, the investigators demonsrated that Glomulin (Glmn), a protein found mutated in the vascular disorder glomuvenous malformation (GVM), binds directly to the RING domain of Rbx1 and inhibits its E3 ubiquitin ligase activity. Loss of Glmn in a variety of cells, tissues, and GVM lesions results in decreased levels of Fbw7 and increased levels of Cyclin E and c-Myc (see figure). The increased turnover of Fbw7 is dependent on CRL and proteasome activity, indicating that Glmn modulates the E3 activity of CRL1(Fbw7). These data reveal an unexpected functional connection between Glmn and Rbx1 and demonstrate that defective regulation of Fbw7 levels contributes to GVM.
The Core used the Aperio ScanScope System to digitize GVM tissue slides immunostained for biomarkers (Cyclin E, c-Myc, Cul 1) and provided access to Aperio Image Analysis algorithms for quantification of biomarker expression.
Tron AE, Arai T, Duda DM, Kuwabara H, Olszewski J, Fujiwara Y, Bahamon BN, Signoretti S, Schulman BA, DeCaprio JA. The Glomuvenous Malformation Protein Glomulin Binds Rbx1 and Regulates Cullin RING Ligase-Mediated Turnover of Fbw7. Mol Cell. 2012;46(1):67-78.