Abstract 3973: Comparison of different highly sensitive miRNA array platforms for the investigation of circulating cell-free microRNAs in blood

Abstract

Abstract Introduction: Circulating free (cf) microRNAs hold great promises in early detection and prognosis of human diseases such as cancer. However due to the very low amount of cf miRNA in plasma and serum, highly informative miRNAs are lost. Thus, there is an urgent need for highly efficient, sensitive, cost efficient technologies to measure low abundant miRNA in a quantitative manner. Previously, we have successfully measured and validated miRNA profiles of early onset breast cancer cases, healthy controls and breast cancer patients who had developed breast cancer metastasis during follow-up by applying TaqMan Low density arrays. We established and validated miRNA signatures for the early detection of early stage breast cancer, and prognosis and early detection of breast cancer metastasis. However more sensitive microarray platforms with highly dynamic ranges could even improve identified diagnostic signatures especially in respect of early detection of diseases and relapse. Here, we compared the sensitivity of the previously applied array platform TaqMan Low density array (TLDA) with Toray innovative 3D-Gene microarray technology by measuring plasma cf miRNA profiles of 40 individuals (10 healthy controls, 10 early stage breast cancer, 20 metastatic breast cancer patients). Experimental Procedures: Plasma samples were processed within 2 h after blood collection. EDTA blood was centrifuged for 15 min at 1200 g and the supernatant centrifuged again at 12.000 g for 10 min in order to remove any cell debris. Cf RNA was isolated from plasma. Cf miRNA profiles of plasma samples of 40 individuals (10 healthy controls, 10 early stage breast cancer, 20 metastatic breast cancer patients) were investigated A: via TaqMan low density arrays (TLDA) and B: via 3D-Gene microarray technology. Expression profiles between both array types and between cases, early stage breast cancer and metastatic breast cancer patients were compared. In addition selected miRNAs were measured with single TaqMan assays. These results were compared with the array based results. Conclusion: The result will be of importance for all scientists who intend to develop and improve diagnostic miRNA panels. Based on our results recommendations for a microarray platform for the investigation of cf miRNA will be given. Citation Format: Melanie Maierthaler, Eric Mennesson, Katarina Cuk, Dharanija Madhavan, Manuela Zucknick, Isabelle Fixe, Alexandra Foucher, Markus Wallwiener, Andreas Schneeweiss, Nadia Normand, Barbara Burwinkel. Comparison of different highly sensitive miRNA array platforms for the investigation of circulating cell-free microRNAs in blood. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3973. doi:10.1158/1538-7445.AM2015-3973