Novel Quantitative Image-Analysis Based Scoring Technique for In-Situ Assessment of mRNA in Archival Tumor Tissues: Strong Correlation between Manual and Automated Schemes
Jeffery C Hanson, Timothy R Holzer, Angie D Fulford, Robert J Konrad, Aejaz Nasir. Eli Lilly & Co., Indianapolis; Laboratory for Exp Medicine, Indianapolis, IN
Background: We developed a semi-quantitative scoring technique for manual in-situ mRNA quantification of oncology biomarkers in archival tumor tissues. We also developed an image analysis-based quantitative algorithm as an automated in-situ mRNA evaluation tool. The goal was comparative evaluation of these scoring techniques for novel oncology biomarkers.
Design: Archival sections from 6 non-small-cell lung carcinomas (NSCLCA) were stained for BIRC5 (survivin) mRNA. The in-situ mRNA staining was manually scored by an experienced pathologist as 0, 1+/2+ (<50%/≥50% area of the cell with non-confluent fine, red granules) or 3+/4+ (with partial/total confluence of granules). The sum of weighted mRNA scores (staining scores X respective %ages) was reported as a semi-quantitative (sq-) m-RNA score. Whole slide images were captured (Aperio Scanscope XT) and analyzed by Definiens' EII software. Each tumor section was scored using the manual criteria above and a quantitative (q-) m-RNA score was derived. Both scores (range 0-400) were correlated (Pearson test; GraphPad, Prism).
Results: Manual sq-RNA scores on 6 NSCLCA cases ranged 0-162. The automated algorithm analyzed tens of thousands of cells/section. The positive cells ranged from 2.5% to 75%, and the q-RNA score ranged from 2-120. Key challenges with the automated technique were delineation of various tissue/tumor components and tissue artifacts. The image analysis-based q-RNA scores correlated welll with the manual scores (Pearson's correlation coefficient of 0.967; p=0.002).
Conclusions: Our quantitative image analysis-based in-situ mRNA scoring on archival tumor tissues shows high-correlation with the manual techniqe. Evaluation on larger cohorts of independent test sets of archival tissues will further substantiate its practical utility in oncology clinical trials and practice.
Monday, March 19, 2012 1:00 PM
Poster Session II # 308, Monday Afternoon