Datum: 20:e September
Tid: Kl. 14.00
Plats: Heilbronnsalen
Opponent: Professor Astrid Gräslund
While most modern pharmaceuticals are small organic molecules, a greater understanding of the molecular and cellular levels of pathogenesis have prompted the development of mechanism-based macromolecular drugs. Gene therapy, the concept of introducing genetic elements in order to treat disease present one promising therapeutic strategy based on such macromolecular agents.
Oligonucleotides (ONs), indispensible tools for gene therapeutic applications, comprise a class of versatile biomolecules capable of modulating the regulation of genes. While unmodified ONs are rapidly degraded, chemi- cal modifications have been developed in order to not only ameliorate poor stability but also enhance both potency and stability. However, the therapeutic potential of ONs and other macromolecules are largely hampered by the impermeability of the plasma membrane to such agents.
Cell-penetrating peptides (CPPs) have gained significant attention as promising deliver vehicles for pharmaceuticals as they possess the ability to translocate across the cell membrane together with cargoes otherwise unable to permeate the lipid bilayer.
In this thesis, a range of potent ON modifications is evaluated for use in the design of therapeutic ON. Additionally, a novel CPP is designed and tested for its capabilies in delivery of ONs. In the first article we study a repertoire of chemically modified anti-miRNAs of different lengths to establish their potency and specificity. All varieties of ONs used in the study target miRNA-21 and are evaluated using a dual luciferase reporter system. In the second article we design a CPP, PepFect 15 (PF15), aiming at combining the desirable properties of improved peptide stability and efficient cellular uptake, with a propensity for endosomal escape, to produce a delivery vector well-suited for delivery of ONs. The performance of this new CPP was evaluated based on its delivery capabilities pertaining to splice-correcting oligonucleotides (SCOs) and anti-miRNAs (antimiRs).