The aim of this study was to investigate the microRNA expression pattern in neutrophils from rheumatoid arthritis patients and its contribution to their pathogenic profile and to analyze the effect of specific autoantibodies or inflammatory components in the regulation of microRNAs in rheumatoid arthritis neutrophils and its modulation by biological therapies. Neutrophils were isolated from paired peripheral blood and synovial fluid samples of 40 patients with rheumatoid arthritis and from peripheral blood of 40 healthy donors. A microRNA array was performed using nCounter technology. Neutrophils from healthy donors were treated in vitro with antibodies to citrullinated protein antigens isolated from rheumatoid arthritis patients and tumor necrosis factor-α or interleukin-6. A number of cytokines and chemokines were analyzed. In vitro treatments of rheumatoid arthritis-neutrophils with tocilizumab or infliximab were carried out. Transfections with pre-microRNAs and DICER downregulation were further performed. Rheumatoid arthritis-neutrophils showed a global downregulation of microRNAs and genes involved their biogenesis, alongside with an upregulation of various potential mRNA targets related to migration and inflammation. Decreased levels of microRNAs and DICER correlated with autoimmunity, inflammation and disease activity. Citrullinated protein antigens and tumor necrosis factor-α decreased the expression of numerous microRNAs and their biogenesis-related genes, increasing their potential mRNA targets. Infliximab reversed those effects. Transfections with pre-miRNAs-223, -126 and -148a specifically modulated genes regulating inflammation, survival and migration. DICER depletion influenced the neutrophils inflammatory profile. Taking together rheumatoid arthritis neutrophils exhibit a global low abundance of microRNAs induced by autoantibodies and inflammatory markers, which might contribute to their pathogenic activation. microRNA biogenesis is significantly impaired in rheumatoid arthritis-neutrophils and further associated with a greater downregulation of microRNAs mainly related to migration and inflammation in synovial neutrophils. Finally, anti-tumor necrosis factor-α and anti-interleukin-6 receptor treatments can modulate microRNA levels in the neutrophils, minimizing their inflammatory profile.