Novel bioengineering platform to produce natural RNAi agents

PROJECT SUMMARY Noncoding RNAsncRNAshave revolutionized biomedical research as master regulators of all known biological processes through the modulation of target gene expressionFurthermorencRNAs are attractive candidates for the treatment of human diseasesHowevercurrent ncRNA research and development is limited to the use of chemically engineered orsynthesized ncRNA agentse gmiRNAs and siRNAsthat are decorated with various and extensive artificial modificationsThis is in accordance with the termination of a number of siRNA miRNA oligonucleotide drugs from further clinical investigationsdue to the induction of adverse immune responses in patientsAlthough the exact causes of immunogenicity remain undisclosedartificial modifications have been well documented to provoke immunogenicity as recognized by toll like receptorsThereforewe have made large efforts to develop novel bioengineering techniques for the production of natural ncRNA agentsOur efforts have led to the identification of stable tRNA pre miRNA hybrid carriers for large scale expression of recombinant miRNAs and siRNAs in a common strain of EcoliOur studies have demonstrated that bioengineered miRNA siRNA agentsBERAscarry no or just a few natural posttranscriptionally modified nucleosides and exhibit favorable stability within human cellsFurthermoreBERAs are biologically active in modulating target gene expression and cancer cellular processese gcell proliferation and chemosensitivityetcin vitroand suppress tumor progression in animal models in vivoGiven these exciting preliminary findingswe hypothesize that the tRNA pre miRNA based technology may be refined and standardized to achieve a more consistenthigher yield production of fully humanized natural RNAi agentsTo test this hypothesiswe will optimize pre miRNA to improve expression yieldandgtof total RNAsand incorporate human tRNAs to offer fully humanized ncRNA carriersAimIn additionwe will develop a standardized pipeline to produce a collectionof ready to use BERAsand critically compare the efficacy and safety of model BERAs with synthetic miRNA mimicsAimThis project will establish a novel ncRNA bioengineering platform toward high yield production of fully humanized RNAi agents and offer BERAs as a new class of natural ncRNA agents for research and development PROJECT NARRATIVE The proposed research is to establish a novel ncRNA bioengineering platform to achieve a consistenthigher yield production of natural and fully humanized ncRNA moleculesand to employ this technology to produce a panel of ready to use biologic miRNA siRNA agentsThis technology is distinguished from existing technologies pursued by all other companies that produce and study chemicallyengineered synthetized miRNA siRNA molecules bearing various and extensive artificial modificationsResults from this project are anticipated to open a new avenue for the production of natural ncRNA agents for research and development