MicroRNA profiling of patient plasma for clinical trials using bioinformatics and biostatistical approaches

• Published in: OncoTargets and therapy Vol. 9; pp. 5931 - 5941
• Main Authors: Markowitz, Joseph, Abrams, Zachary, Jacob, Naduparambil K, Zhang, Xiaoli, Hassani, John N, Latchana, Nicholas, Wei, Lai, Regan, Kelly E, Brooks, Taylor R, Uppati, Sarvani R, Levine, Kala M, Bekaii-Saab, Tanios, Kendra, Kari L, Lesinski, Gregory B, Howard, J Harrison, Olencki, Thomas, Payne, Philip R, Carson, 3rd, William E
• Format: Journal Article
• Language: English
• Published: New Zealand Dove Medical Press Limited 01-01-2016; Taylor & Francis Ltd; Dove Medical Press
• Subjects: Analysis; Bioinformatics; Biomarkers; Biometry; Blood; Blood plasma; Cancer therapies; Cell cycle; Clinical trials; Computational biology; Cyclin-dependent kinases; Disease; DNA microarrays; Health aspects; Informatics; Innovations; Kinases; Medical prognosis; Melanoma; Metastasis; MicroRNA; MicroRNAs; Oncology; Original Research; Patients; Phlebotomy; Principal components analysis; Skin cancer; United States > US; Ohio; melanoma; miRNA; profiling; rank-based statistic
• ISSN: 1178-6930; 1178-6930
• DOI: 10.2147/OTT.S106288

MicroRNAs (miRNAs) are short noncoding RNAs that function to repress translation of mRNA transcripts and contribute to the development of cancer. We hypothesized that miRNA array-based technologies work best for miRNA profiling of patient-derived plasma samples when the techniques and patient populations are precisely defined.
Plasma samples were obtained from five sources: melanoma clinical trial of interferon and bortezomib (12), purchased normal donor plasma samples (four), gastrointestinal tumor bank (nine), melanoma tumor bank (ten), or aged-matched normal donors (eight) for the tumor bank samples. Plasma samples were purified for miRNAs and quantified using NanoString
arrays or by the company Exiqon. Standard biostatistical array approaches were utilized for data analysis and compared to a rank-based analytical approach.
With the prospectively collected samples, fewer plasma samples demonstrated visible hemolysis due to increased attention to eliminating factors, such as increased pressure during phlebotomy, small gauge needles, and multiple punctures. Cancer patients enrolled in a melanoma clinical study exhibited the clearest pattern of miRNA expression as compared to normal donors in both the rank-based analytical method and standard biostatistical array approaches. For the patients from the tumor banks, fewer miRNAs (<5) were found to be differentially expressed and the false positive rate was relatively high.
In order to obtain consistent results for NanoString miRNA arrays, it is imperative that patient cohorts have similar clinical characteristics with a uniform sample preparation procedure. A clinical workflow has been optimized to collect patient samples to study plasma miRNAs.