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Principal Investigator: Prem Khovabutr Premsrirut
Organization: MIRIMUS, INC.
Fiscal Year: 2024
Award: $350,000
Funding agency: National Institute of General Medical Sciences
Abstract
RNA interference (RNAi) – a natural biological mechanism by which eukaryotic cells control gene expression –
can be used to target and silence with high specificity virtually any expressed gene, making it a potentially
powerful therapeutic approach. Importantly, RNAi mechanisms utilize DNA/RNA sequences to specify its target,
enabling fine-tuned silencing of a single gene belonging to a family of structurally similar proteins that typically
share small molecule binding pockets. For these reasons, RNAi-based therapeutics have the potential to provide
effective on-target benefits while avoiding off-target toxicities that are commonly seen with promiscuous protease
and kinase small molecule inhibitors. Despite their therapeutic potential, the most challenging aspect of RNAi-
based drug development involves achieving effective delivery to the cells or tissues of interest, while avoiding
systemic off-target effects. Indeed, only a handful of RNAi-based drugs have received FDA approval to treat liver
diseases since this is the most readily accessible tissue for this class of drugs. More recently, alternative delivery
mechanisms, such as ligand-coupled oligonucleotides, lipid nanoparticles, viral vectors, and extracellular
vesicles (EVs) show great promise with the potential to penetrate tissues and have cellular uptake where they
can engage the endogenous RNAi machinery to induce gene silencing. However, a systematic method for
evaluating these delivery modalities in vivo has yet to be implemented. To overcome this hurdle, we generated
a murine model – the Optimus model, that contains a dual fluorescence reporter cassette where one reporter
harbors unique RNAi responsive elements. Using this Optimus model, we aim to generate an effective pipeline
to readily test an array of new RNAi-based therapeutic biologics generated by coupling our optimized siRNAs or
artificial miRNA scaffolds to established delivery vehicles, beginning with three unique modalities: 1) peptide-
based platform developed by SRI International for specific delivery to lung adenocarcinoma cells; 2) aptamer-
based modality developed by Aptamer group for the silencing in activated hepatic stellate cells under liver fibrotic
conditions; and 3) recombinant adeno-associated viruses (rAAV) we previously generated carrying ubiquitous
and tissue-specific RNAi expression cassettes. Importantly, these delivery platforms have already demonstrated
successful delivery of oligonucleotide payloads, both in vitro and preliminary in vivo. Our collaboration enables
us to further test these new therapeutic biologics in live animals and assess their biodistribution, the features
that promote cellular uptake and enable trafficking of RNAi payload for functional engagement, and the systemic
off-target effects at very early stages of pre-clinical development. This research will define a new paradigm to
accelerate the creation of novel RNAi-based (and potentially oligonucleotide-based) drugs and readily evaluate
their therapeutic potential in live animals.
Terms: <Acceleration><Adenocarcinoma Cell><Algorithms><Animals><Binding><Bioavailability><Biodistribution><Biological><Biological Agent><Biological Availability><Biological Products><Biological Response Modifier Therapy><Biological Therapy><Body Tissues><Cell Body><Cell Line><Cell Transplantation><CellLine><Cells><Chemical Injury><Clinical><Collaborations><Combined Modality Therapy><Coupled><Coupling><DNA><DNA cassette><Deoxyribonucleic Acid><Dose><Drug Therapy><Drugs><Elements><Engineering><Enzyme Gene><Enzymes><Esteroproteases><Eukaryotic Cell><Evaluation><Event><Family><Feasibility Studies><Fluorescence><Future><Gene Expression><Gene Inactivation><Gene Silencing><Gene Targeting><Genes><Hepatic Disorder><Hepatic Stellate Cell><In Vitro><In vivo analysis><International><Intravenous><Ito Cell><Kinases><Ligands><Liver Fibrosis><Liver diseases><Lung Adenocarcinoma><Malignant Glandular Cell><Measures><Medication><Methods><Micro RNA><MicroRNAs><Modality><Modeling><Molecular Interaction><Monitor><Multimodal Therapy><Multimodal Treatment><Oligo><Oligonucleotides><Penetration><Peptidases><Peptide Hydrolases><Peptides><Pharmaceutical Preparations><Pharmacotherapy><Phase><Phosphotransferase Gene><Phosphotransferases><Physiologic Availability><Post-Transcriptional Gene Silencing><Posttranscriptional Gene Silencing><Process><Protease Gene><Proteases><Protein Family><Protein Inhibition><Proteinases><Proteins><Proteolytic Enzymes><RNA Interference><RNA Interference Therapy><RNA Sequences><RNA Silencing><RNA interference therapeutics><RNA interference-based therapy><RNAi><RNAi therapeutics><RNAi therapy><RNAi-based therapeutics><RNAi-based therapy><Recombinant adeno-associated virus><Recombinant adeno-associated virus (rAAV)><Reporter><Reporter Genes><Research><SBIR><Schedule><Screening procedure><Sequence-Specific Posttranscriptional Gene Silencing><Serotyping><Short interfering RNA><Small Business Innovation Research><Small Business Innovation Research Grant><Small Interfering RNA><Specific qualifier value><Specificity><Specified><Spinal Column><Spine><Strains Cell Lines><System><Technology><Testing><Therapeutic><Therapeutic Uses><Tissues><Toxic effect><Toxicities><Transphosphorylases><Validation><Vertebral column><Viral Interference><Viral Vector><Whole Organism><Work><aptamer><backbone><biologic><biological therapeutic><biological treatment><biologically based therapeutics><biologics><biopharmaceutical><biotherapeutic agent><biotherapeutics><biotherapy><cellular transplant><chemical trauma><combination therapy><combined modality treatment><combined treatment><cultured cell line><delivery vector><delivery vehicle><dosage><drug development><drug treatment><drug/agent><enhancer cassette><experiment><experimental research><experimental study><experiments><expression cassette><extracellular vesicles><fibrotic liver><gene cassette><genetic cassette><hepatic disease><hepatic fibrosis><hepatopathy><human disease><improved><in vivo><in vivo Model><in vivo evaluation><in vivo testing><inhibit protein><inhibit proteins><innovate><innovation><innovative><integration cassette><interest><interfering virus><lipid based nanoparticle><lipid nanoparticle><liver disorder><lung cancer cell><miRNA><miRNAs><mouse model><multi-modal therapy><multi-modal treatment><murine model><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation><next generation therapeutics><novel><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><nucleic acid therapy><nucleic acid-based therapeutics><oligonucleotide delivery><oligos><particle><pre-clinical development><preclinical development><promoter cassette><protein inhibitions><rAAV><recombinant AAV><reporter cassette><resistance cassette><scaffold><scaffolding><screening tools><selectable cassette><selection cassette><siRNA><small molecular inhibitor><small molecule><small molecule inhibitor><stop cassette><subcutaneous><subdermal><synthetic lethal interaction><therapeutic nucleic acids><tool><trafficking><transcription cassette><transcriptional cassette><transcriptional silencing><transgene cassette><uptake><validations><virtual>