Next Generation Sequencing Leads to the Discovery of Novel Associations of miRNA with Myelodysplastic Syndromes
Jose M Gonzalez Berjon, Karla Alvarez, Elissa Hudspeth, Jiango Wen, Federico A Monzon Bordonaba, Randall J Olsen, Youli Zu, Xiabo Zhou, Jeff Chang. The Methodist Hospital/Weil Medical College of Cornell University, Houston, TX; The Methodist Hospital, Houston, TX; Florida Hospital, Orlando, FL; Baylor College of Medicine, Houston, TX
Background: Myelodysplastic Syndromes (MDS) are pre-leukemic disorders with very limited treatment options. Little is known about small regulatory RNAs and how they contribute to pathogenesis progression and transcriptome changes in MDS. After initial steps using next generation sequencing (NGS) technologies, when compared to control, several markedly abnormal levels of miRNA were found. To validate these findings miRNA levels were compared in marrow clot samples from controls, low-grade and high-grade MDS utilizing quantitative real time PCR (qPCR).
Design: This study utilized a total of 35 bone marrow samples. For the discovery set high-throughput next generation sequencing of small RNAs (RNA-seq) on primary bone marrow cells from control, low-grade MDS: refractory anemia (RA) and high-grade MDS: refractory anemia with excess blast (RAEB2) on an Illumina Genome Analyzer IIx were utilized. The validation set consisted of 32 paraffin-embedded bone marrow clots were utilized in the validation cohort (24 MDS cases: 14 low-risk MDS [RARS n=11, RA n=3], 10 high-risk MDS [RAEB n=8, refractory cytopenia with multilineage dysplasia n=3] and 8 negative staging bone marrow from patients with lymphoma and similar age) in which five miRNA (miR) sequences (miR-29a, miR-130, miR-376, miR-155 and miR-451) were measured by qPCR.
Results: Notably miR-451 and miR-29a demonstrated particularly high discrimination between high-risk MDS cases and normal controls (sensitivity [sens]=91%, specificity [Spec]=89% and sens. 83%, spec 80%; respectively). miR-130, miR-376, miR-155 sequences also show high specificity but low sensitivity (Spec: 89%, 100%, 92% and sens: 62%, 63%, 63%, respectively) in MDS (combined low-risk and high-risk) cases when compared to controls.
Conclusions: To our knowledge miR451 and miR29a had not been previously associated with MDS. Our results corroborate that NGS is an important tool to explore the small RNAome for understanding MDS pathogenesis. Further studies are warranted to experimentally substantiate our observations and to develop biomarkers for the diagnosis and treatment of MDS.
Tuesday, March 20, 2012 9:30 AM
Poster Session III # 241, Tuesday Morning