Project #7

Discovery and characterization of cell biological barriers to therapeutic RNA delivery

RNA based therapeutics, e.g., siRNA, antisense, mRNA- and CRISPR-based drugs, are a novel class of drugs that that are potential ideal precision drugs for oncology given their highly specific and targeted nature. However, cytosolic delivery remains the major obstacle to macromolecular RNA drug development.

In my (Anders Wittrup) lab we have developed several new techniques both for detection and quantification of RNA delivery. To further advance our possibilities to visualize the processes involved in cytosolic RNA delivery we will in this project leverage newly developed imaging strategies including event triggered microscopy automation and a novel strategy to extract ultra-high resolution information from lower resolution images, We will exploit these advances to characterize the cell biology and biophysics of lipid nanoparticle (LNP) mediated RNA delivery. The ultimate aim is to bring targeted nucleic acid drugs to the clinic for cancer therapeutics and regenerative medicine.

Improved understanding of the molecular mechanisms involved in LNP delivery and endosomal escape of large RNA cargos will enable rational design of new delivery vehicles. The lessons from this work will be applicable to multiple cancer types for which there is currently a need for novel therapeutic strategies. Enhanced delivery of RNA cargos in general will also be highly beneficial for any efforts to reprogram cells or tissues for regenerative purposes. Understanding current RNA delivery inefficiency and coming up with strategies to overcome this barrier is the focus of this project.

Main Principal Investigator

• Anders Wittrup

Co-supervisor

• Pontus Nordenfelt