Preclinical evaluation of Targeted IL-1β Knockdown via CD44-Immunoliposomes: A Nano-therapy against the Inflammatory Microenvironment

Abstract

Chronic inflammation, characterized by the infiltration of macrophages and the heightened release of pro-inflammatory cytokines, is the underlying cause of the pathogenesis of many critical diseases. Therapeutic interventions for controlling inflammation via gene knockdown of inflammatory mediators have emerged as a promising approach for regulating uncontrolled inflammation. This study explores the potential of siIL-1{beta}-anti-CD44-Liposomes (SIL) as a potent anti-inflammatory therapy against pro-inflammatory RAW264.7 macrophages via gene specific knockdown of IL-1{beta} mRNA through RNAi, and the subsequent down-regulation of the pro-inflammatory cytokine loop. The designed SIL exhibited a uniform size of 131.1 {+/-} 0.5 nm with a quasi-spherical morphology and sustained release of siIL-1{beta} within 24 hours. The reduction in pro-inflammatory cytokines like IL-1{beta}, TNF-, and IL-6 and inflammatory enzymes iNOS and COX-2; and the simultaneous increase in the anti-inflammatory cytokine IL-4, is indicative of the formulations therapeutic efficacy in reducing inflammation at a cellular level. The effects of SIL on the Macrophage-T cell crosstalk also uncovers the liposomes efficacy in reducing cytokine-mediated T cell effector functions. The nuanced effects of siIL-1{beta}-anti-CD44-Liposomes on in-vivo model of chronic inflammation underscore their potential for precise therapeutic interventions in inflammatory conditions, with multifaceted anti-inflammatory effects on tissue levels and cytokine levels. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=147 SRC="FIGDIR/small/683015v1_ufig1.gif" ALT="Figure 1"> View larger version (61K): org.highwire.dtl.DTLVardef@f4d0f9org.highwire.dtl.DTLVardef@c3c24aorg.highwire.dtl.DTLVardef@8c115org.highwire.dtl.DTLVardef@bc863d_HPS_FORMAT_FIGEXP M_FIG Schematic representation of the study: CD44 receptors are elevated in LPS activated macrophages, which can be targeted by using anti-CD44 Liposomes, for RNA therapy. IL-1 knockdown via siRNA leads to lower inflammatory nature of macrophages, compromising its the antigen presentation and T-cell activation. This lowers the cytokine storm in inflammatory milieu and lower tissue damage can be achieved. Created with BioRender.com. C_FIG