PII-015 - UTILITY OF MIROMICS AND INTEGRATIVE MULTI-OMICS SYSTEMS ANALYSIS FOR IDENTIFICATION OF MIRNAS AS POTENTIAL PHARMACODYNAMIC BIOMARKERS OF IFNβ-1A BIOLOGICS

M. Mehanna¹, L. Chekka¹, D. Samarth¹, E. Decker², E. Mohamed¹, Y. Guo¹, S. Schrieber¹, J. Florian¹, Y. Wang³, D. Strauss¹, P. Hyland¹; ¹U.S. Food and Drug Administration, ²Capitol Technology University, ³U.S. Food and Drug Administration, Silver Spring, United States.

Background: MiRomics may identify pharmacodynamic (PD) biomarkers to support biosimilar development and approval. Interferon beta-1a (IFNβ-1a) biologics have limited well-characterized PD biomarkers and complex mechanisms of action. Here we evaluated the utility of miRomics for identifying IFNβ-1a PD biomarkers and explored miRNA-protein target relationships by integrating proteomic data from the same samples.

Methods: In a single dose randomized clinical study, we sequenced plasma miRNAs in healthy subjects treated with IFNβ-1a (30ug, n=11), pegIFNβ-1a (125ug, n=11) or product specific placebo (n=6 each). MiRNAs were measured at 10 timepoints over 6 days in IFNβ-1a group, and at 12 timepoints over 13 days in pegIFNβ-1a and placebo groups, including baseline. Linear-mixed effect models regressing normalized count changes from baseline with treatment*time interaction was used to identify miRNA signals (false discovery rate (FDR)-corrected P< 0.1). MiRNAs with a significantly different response from placebo were prioritized and predicted targets were identified in proteomic signals (FDR-corrected P< 0.1) using miRDB. Upstream regulatory analysis of predicted targets was conducted using Ingenuity Pathway Analysis tool.

Results: MiR-223-3p, miR-21-5p, and miR-150-5p were identified and prioritized as potential PD candidates for both products. Of the proteomic signals, 14, 27, and 32 IFNβ-1a impacted proteins are targets of miR-223-3p, miR-21-5p, and miR-150-5p, respectively. And 22, 35, and 52 pegIFNβ-1a impacted proteins are targets of hsa-miR-223-3p, hsa-miR-21-5p, and hsa-miR-150-5p, respectively. Mir-223-3p regulates STAT1 and MX1, top protein PD biomarkers for both products. Predicted protein targets for each miRNA were shown to be involved in previously identified master regulatory networks for both products, including immune signaling (STAT3 and immunoglobulin), inflammation (TNF and IL6), and IFNB1 signaling.

Conclusion: Using MiRomics, we identified three miRNAs as potential IFNβ-1a PD biomarker candidates for further investigation to support biosimilar development programs. Our integrative miRNA-protein expression approach supports these miRNAs as candidates by showing links to miRNA targets that are top protein PD biomarkers and to previously identified regulatory networks downstream of both products.