Nanoligomers targeting NF-κB and NLRP3 reduce neuroinflammation and improve cognitive function with aging and tauopathy

Abstract

Neuroinflammation contributes to impaired cognitive function in brain aging and neurodegenerative disorders like Alzheimer’s disease, which is characterized by the aggregation of pathological tau. One major driver of both age- and tau-associated neuroinflammation is the NF-κB and NLRP3 signaling axis. However, current treatments targeting NF-κB or NLRP3 may have adverse/systemic effects, and most have not been clinically translatable. Here, we tested the efficacy of a novel, nucleic acid therapeutic (Nanoligomer) cocktail specifically targeting both NF-κB and NLRP3 in the brain for reducing neuroinflammation and improving cognitive function in old wildtype mice, and in a mouse model of tauopathy. We found that 4 weeks of NF-κB/NLRP3-targeting Nanoligomer treatment strongly reduced neuro-inflammatory cytokine profiles in the brain and improved cognitive-behavioral function in both old and tauopathy mice. These effects of NF-κB/NLRP3-targeting Nanoligomer treatment were associated with reduced glial cell activation in old wildtype mice, less pathology in tauopathy mice, favorable changes in transcriptome signatures of inflammation (reduced) and neuronal health (increased) in both mouse models, and positive systemic effects. Collectively, our results provide a basis for future translational studies targeting NF-κB/NLRP3 in the brain, perhaps using Nanoligomers, to inhibit neuroinflammation and improve cognitive function with aging and neurodegenerative disease.

Brain-Penetrant NF-κB and NLRP3 Targeting Nanoligomers are Therapeutic in Amyotrophic Lateral Sclerosis (ALS) and Alzheimer’s Disease (AD) Human Organoid and Mouse Models

“…We observed similar results in terms of cognitive improvements and neuroinflammation reductions in the rTg4510 AD mouse model with NI112 4-week treatments. 64 Further neuropathological assessment using immunohistochemistry also showed a clear effect of NI112 treatment in the rTg4510 AD mouse model ( Fig. 2J, S3 ).…”

“…40,[53][54][55] Inflammasome inhibition has been intensely investigated as a novel target, as well as a common cure for a range of neurodegenerative and autoimmune diseases, 56,57 and using NF-κB and NLRP3 mRNA targeting and translation-inhibiting Nanoligomer combination (NI112) is a safe, effective, and targeted approach. [58][59][60][61][62][63][64][65] However, even other inflammasome-targeting smallmolecule therapeutics (e.g., MCC950) are limited by: 1) potential off-target effects; [66][67][68] and 2) the fact that they target only NF-κB or NLRP3 alone. NI112, an RNA therapeutic that crosses the blood-brain barrier, addresses problems with pan immunosuppressive therapies (such as steroids), and the cocktail potently inhibits neuroinflammation.…”

“…[58][59][60][61][62][63][64] Thus, whereas clinical trials of generic anti-inflammatory drugs for cognitive decline/AD have been disappointing, [69][70][71] targeting neuroinflammation with NI112 has promising therapeutic potential. We have shown that NI112 reduces pro-inflammatory astrocytes/microglia even in prion-treated mice, 61 agingassociated neuropathology and AD, 64 and EAE mice 63 (models of severe neurodegeneration), and that this is associated with cognitive improvements. Moreover, the high binding specificity (K D ~5 nM 60,62,72 ), minimal off-targeting, 60,62,65 high safety profile, 65 lack of any immunogenic response or accumulation in first pass organs, 65 absence of any observable histological damage to organs even for long (>15-20 weeks) treatments, [60][61][62]65 and facile delivery to the brain to counter neuroinflammation, 58,61,63,65 make Nanoligomers and specifically the NI112 cocktail a promising, safe and targeted approach to mitigate neuropathological inflammation, as well as microglial and astrocytic activation.…”

Large- and Small-Animal Studies of Safety, Pharmacokinetics (PK), and Biodistribution of Inflammasome-Targeting Nanoligomer in the Brain and Other Target Organs

“…We first started with a maximum tolerable dose (MTD) assessment of the NF-κB and NLRP3 inhibitor combination NI112 ( nfkb1 (Sequence: AGTGGTACCGTCTGCTA) and nlrp3 (Sequence: CTTCTACTGCTCACAGG)). Prior studies have shown assessment of NI112 in relevant disease models (MS, AD, Prion/Creutzfeldt-Jacob Disease, neuroinflammation, and autoimmune diseases), 17,18,22,23,25,26 and Nanoligomer selectivity and high-binding affinity (K D 3.17 nM, 19,20 other PNA/RNA and PNA/DNA binding studies showed K D 5-8 nM 27 ) along with rapid biodistribution and clearance. We observed strong safety-toxicity profile in both animal models for longer-term repeat dosing (IP, 150 mg/kg, 3x a week for 12 weeks, Fig.…”

Inflammasome-Inhibiting Nanoligomers are Neuroprotective Against Space-Induced Pathology in Healthy and Diseased 3D Human Motor and Pre-Frontal Cortex Brain Organoids

“…Nanoligomer™ therapeutic platform generates safe, targeted, 34 and highly-specific (K D 3.37 nM 35,36 ) RNA-targeting molecules for facile delivery to targeted organs and both up-and downregulation of desired protein through translational and transcriptional regulation. [37][38][39][40][41][42] Nanoligomers utilize peptide nucleic acids (PNAs) as the sequence-specific binding component that demonstrate strong hybridization and specificity to their RNA (or DNA) sequence targets, 43 and exhibit no known enzymatic cleavage, leading to increased stability in human blood serum and mammalian cellular extracts. 44 Given the low K D , 35,36,43 high binding specificity, minimal off-targeting, 35,36 lack of any immunogenic response or accumulation in first pass organs, 34 absence of any observable histological damage to organs even for long (>15-20 weeks) treatments, [34][35][36]40 and facile delivery to the brain to counter neuroinflammation, 34,[37][38][39][40][41][42] we have demonstrated that Nanoligomers are a safe and targeted approach to screen for therapeutic targets in organoids and rodents for multiple neurodegenerative diseases aboard the International Space Station (ISS).…”