Abstract

Multiple Sclerosis (MS) is a debilitating autoimmune disease that impacts millions of patients worldwide, disproportionately impacting women (4:1), and often presenting at highly productive stages of life. This disease affects the spinal cord and brain, and is characterized by severe neuroinflammation, demyelination, and subsequent neuronal damage, resulting in symptoms like loss of mobility. While untargeted and pan-immunosuppressive therapies have proven to be disease-modifying and manage (or prolong the time between) symptoms in many patients, a significant fraction are unable to achieve remission. Recent work has suggested more targeted neuroinflammation mitigation through selective inflammasome-inhibition can offer relief to patients, while preserving key components of immune function. Here we show a screening of potential therapeutic targets using inflammasome-inhibiting Nanoligomers (NF-{kappa}B1, TNFR1, TNF-, IL-6), that meet or far-exceed commercially available small-molecule counterparts like Ruxolitinib, MCC950, and Deucravacitinib. Using the human brain organoid model, top Nanoligomer combinations (NF-{kappa}B1+TNFR1: NI111, and NF-{kappa}B1+NLRP3: NI112) were shown to significantly reduce neuroinflammation, without any observable negative impact on organoid function. Further testing of these top Nanoligomer combinations in an aggressive Experimental Autoimmune Encephalomyelitis (EAE) mouse model for MS using intraperitoneal (IP) injections showed that NF-{kappa}B1and NLRP3 targeting Nanoligomer combination NI112 rescues mice without observable loss of mobility or disability, minimal inflammation in brain and spinal cord histology, and minimal to no immune cell infiltration of the spinal cord and no demyelination, similar to or at par with mice that received no EAE injections (negative control). Mice receiving NI111 (NF-{kappa}B1+TNFR1) also showed reduced neuroinflammation compared to saline (sham) treated EAE mice and at par/similar to other inflammasome-inhibiting small molecule treatments, although it was significantly higher than NI112 leading to subsequent worsening clinical outcomes. Furthermore, treatment with an oral formulation of NI112 at lower doses showed a significant reduction in EAE severity, albeit with higher variance owing to administration and formulation/fill-and-finish variability. Overall, these results point to the potential of further development and testing these inflammasome targeting Nanoliogmers as an effective neuroinflammation treatment for multiple neurodegenerative diseases, and potentially benefit several patients suffering from such debilitating autoimmune diseases like MS.