Mir-17-92 Micromanaging Lupus Nephritis： Discovery from Rna-Rna Interaction, Immunometabolism, to Preclinical Study

Lupus nephritis, one of the most serious manifestations of systemic lupus erythematosus was seen in up to 60% of cases with SLE. Since SLE resulted from autoimmune dis-regulation, for those with lupus nephritis, immune modulation may be their life-saving straw. Regulatory T cells (Treg) play a crucial role in limiting the aberrant immune responses. T follicular helper (TFH) cell provide crucial growth signals to germinal center B cells supporting antibody production. Suppressive T-follicular regulatory (TFR) cells, sharing phenotypic characteristics with TFH and Foxp3+ Treg cells, regulate TFH. According to our previous study, microRNA miR-17-92 modulates Treg function and miR17-92 CD4 specific knockouts (miR17-92 -/-) ameliorate autoimmune disease. Therefore, we will investigate the role of miR17-92 and single microRNA in modulating TFR and TFH of lupus murine model. We then propose a novel therapeutic strategy of using antisense-oligonucleotides for crucial microRNA of the cluster in attempt to improve control of lupus nephritis. Long noncoding RNAs (lncRNAs) have been linked to gene regulation in various tissues. How miR-17-92 modulates TFR and TFH via lncRNA and epigenetic regulation remains an open question. Firstly, we will exam the transcriptome and Chip-Seq regulated by miR-17-92 in TFR and TFH. We then will investigate how lncRNA control miR-17-92 in modulating TFH development. lncRNA enhanced by Bcl6 will be correlated with epigenetic change in miR-17-92 promoter and enhancer region to identify the lncRNA which can enhance miR-17-92 production. We will also explore the interaction between miR-17-92 and lncRNA in Treg. Secondly, to understand how miR17-92 modulates TFH and TFR, we will take advantage of CD4 specific miR-17-92 conditional knockout mice and Foxp3 specific miR-17-92 knockout mice for TFH induction to identify the crucial microRNA in miR-17-92 cluster that modulate TFH. Moreover, to clarify the metabolic regulation of TFR and TFH by miR-17-92, we will analyze metabolite of TFR and TFH from wild type and miR17-92 -/- mice by mass spectrometry, which will be correlated with epigenetic modification. Finally, according to the identified crucial microRNA, we will apply miR antisense-oligonucleotides in MRL/lpr lupus murine model and then access autoantibodies and cells in their circulation, spleen, and kidney to evaluate treatment response. Moreover, we will study pharmacokinetic properties and toxicity of the antisense-oligonucleotides. In sum, this proposal will be the very first study focusing on how miR-17-92 mediate lncRNA and metabolic modulation in TFR and TFH. After identifying the critical microRNA in miR-17-92, we then will treat lupus murine model with its antisense-oligonucleotides to monitor treatment response and toxicity for future clinical application as a novel therapy. Year 1. To determine the effect and epigenetic modification of miR-17-92 on TFR and TFH cells through interaction with long noncoding RNA (lncRNA). Year 2. To understand how miR17-92 cluster mitigate lupus nephritis by modulating TFH and TFR through metabolic regulation. Year 3. To investigate the effect and safety of antisense-oligonucleotides for murine lupus nephritis

Project ID:PC10901-1042
External Project ID:MOST108-2314-B182-016-MY3