POS0406\u2005miRNAs DEREGULATED IN RESPONSE TO IL17A INHIBITORS IN PSORIATIC ARTHRITIS REGULATE GENE PRODUCTS IN Rho-GTPase PATHWAYS

Abstract

Using transcriptomic data at initiation of therapy, we recently identified differentially expressed genes (DEGs) that separated IL-17Ai response from non-response1. Integration of cell-type-specific DEGs with protein-protein interactions (PPIs) and further comprehensive pathway enrichment analysis revealed Rho GTPase signaling pathway exhibited a strong signal specific to IL-17Ai response and particularly the genes, RAC1 and ROCKs.To characterize microRNA (miR) profiles among IL-17Ai responders and non-responders, as it relates to RHO GTPase pathway.We interrogated 20 psoriatic arthritis (PsA) patients initiating IL-17Ai. Patients achieving at least low disease activity according to the Disease Activity Index for PsA (DAPSA) at three months were classified as responders. There were seven responders (35%) and thirteen non-responders (65%) in the IL-17Ai group, with biologic treatment naïve (bio-naïve) and previously-exposed (bio-exposed) patients exhibiting a 50% (4/8) and a 25% (3/12) response rate, respectively. For the miR analysis, CD4 positive T cells were isolated from peripheral blood mononuclear cells using DynabeadsTM CD4 beads (ThermoFisher). Total RNA was extracted from the CD4+ T cells using Lexogen’s Split RNA Extraction Kit (D-Mark Biosciences). Libraries were prepared from 200ng total RNA with the NEXTFLEX Small RNA-Seq Kit v3 with UDIs (Bioo Scientific) and sequenced on the Illumina NovaSeq 6000. Raw sequencing fastq data assessed the quality using FastQC. The miRDeep2 was used to trim the adapter, align and quantify human mature miRs from miRbase (Release 22). The abundance of miRs was converted to read counts per million, normalized and limma R package was used to identify pre- and post-differentially expressed miRs.We obtained 2,889 miRs. After removing miRs with low reads in >90% of samples, 1902 high quality miRs remained for further analysis. Using mirDIP v4.1 we identified gene targets for differential miRs, and focused on recently identified DEGs related to RHO GTPase pathway. The miRs on the left of the figure 1 are those deregulated in pre-treatment, and the miRs on the right of the figure 1 show the post-treatment deregulated miRs. hsa-miR-3691-5p and hsa-miR-3161 represent the miRs that were deregulated in both conditions. The red highlighted nodes represent the most connected miRs and genes; thus, representing miRs that are the most RHO-pathway centric regulators (hsa-miR-495-3p, 16-5p, 129-5p, 520h, 520g-3p), and genes representing the most strongly regulated RHO-pathway gene products (ROCK1, RHOQ, PFN2, TAOK1, DYNC1L12, MAPRE1, PAFAH1B1, ARHGAP5, MAPK1, CALM1, DIAPH2, PKN2, ITSN1).Pre- and post-treatment differential miRs related to IL-17Ai response regulate multiple genes from RHO GTPase pathway.[1]Rahmati S, O’Rielly DD, Li Q, Codner D, Dohey A, Jenkins K, Jurisica I, Gladman DD, Chandran V, Rahman P. Rho-GTPase pathways may differentiate treatment response to TNF-alpha and IL-17A inhibitors in psoriatic arthritis. Sci Rep. 2020 Dec 10;10(1):21703.Figure 1.Proton Rahman Speakers bureau: AbbVie, Amgen, BMS, Celgene, Eli Lily, Janssen, Merck, Novartis, Pfizer, UCB, Consultant of: AbbVie, Amgen, BMS, Celgene, Eli Lily, Janssen, Merck, Novartis, Pfizer, UCB, Grant/research support from: Janssen, Novartis, Quan Li: None declared, Dianne Codner: None declared, Darren O’Rielly: None declared, Amanda Dohey: None declared, Kari Jenkins: None declared, Dafna D Gladman Speakers bureau: AbbVie, Amgen, BMS, Eli Lily, Galapagos, Gilead, Janssen, Novartis, Pfizer, UCB, Consultant of: AbbVie, Amgen, BMS, Eli Lily, Galapagos, Gilead, Janssen, Novartis, Pfizer, UCB, Grant/research support from: AbbVie, Amgen, Eli Lily, Janssen, Novartis, Pfizer, UCB, Vinod Chandran Speakers bureau: AbbVie, Amgen, BMS, Eli Lily, Janssen, Novartis, Pfizer, UCB, Paid instructor for: AbbVie, Amgen, BMS, Eli Lily, Janssen, Novartis, Pfizer, UCB, Grant/research support from: AbbVie, Amgen, Eli Lily, Employee of: Spousal Employment Eli Lilly, Igor Jurisica: None declared