Dong-Hua Yang, MD, PhD
Assistant Research Professor
Office Phone: 215-214-3242
1) Develop IHC assay for new biomarkers, in collaboration with Dr. Barbara Burtness
Personalized and molecular-based cancer treatment depends on biomarker discovery. Biomarker research and assay development using immunohistochemistry (IHC) is becoming increasingly important because it drives drug discovery & development, clinical diagnostics, prognostics and treatment decisions.
Immunohistochemistry (IHC), which examines protein localization while preserving tissue morphology, is a standard in situ assay to assess protein expression. This invaluable technique has been applied in many facets of biological science, especially in the field of biomarker research. IHC result depends highly on antibody type, procedure protocol, and interpretation criteria. One of the important considerations for IHC staining is the optimization of staining procedure. At BioSample Repository Facility (BRF), we have been at the forefront of developing novel IHC assays for biomarker research. We optimize each staining procedure to achieve low background, high sensitivity, reliability and reproducibility on all assays we developed. IHC assays that were established and validated include:
Cell growth related markers: Ki67, EGFR/pEGFR, Her2/pHer2, Her4/pHer4, PTEN,
AKT/pAKT, ACK/pACK, MAPK/pMAPK, Met
Apoptosis related markers: p53, survivin, PARP
DNA synthesis and repair related markers: ERCC1, RRM1, XPF
Cell cycle regulation related markers: AuroraA/pAuroraA, NEDD9 (HEF1)
HPV status: p16Top
2) Automated Quantitative Analysis (AQUA) assay development and validation, in collaboration with Dr. Barbara Burtness
AQUA (Automatic Quantitative Analysis) is an automated fluorescence IHC image analysis technology that provides unprecedented information about protein expression in tissue. Its unbiased, quantitative, localized and reproducible analyses provide new insights into protein biomarker characterization and discovery. Since its introduction in 2002, AQUA has been rapidly adopted for basic and clinical applications and is now a well-established powerful tool for biomarker research. AQUA algorithm allows accurate and precise image segmentation such that protein expression can be quantified in specific cellular and subcellular compartments. This is an enhancement of other manual image quantification methods where these expression levels must be identified manually and the expression categorically scored by eye.
Accuracy and reproducibility are particular important for the implementation of AQUA-based application. Although AQUA provides objective linear scores of protein expression level, some of the scores might be obscured by the insensitivity or saturation of an assay. By optimizing each AQUA assay and careful validation, we have applied the above IHC biomarkers for AQUA analysis on head and neck squamous cell carcinoma (HNSCC), renal cell carcinoma (RCC), pancreatic carcinoma, ovarian carcinoma and other tumors. The application of these biomarkers in clinical setting will identify factors that are of diagnostic or prognostic importance and predict benefits from adjuvant chemotherapy.Top