Periodic Reporting for period 1 - ENDOSCAPE (ENDOSCAPE, a clinically applicable non-viral gene delivery technology)
Reporting period: 2019-01-01 to 2020-06-30
The ENDOSCAPE technology aims to create a non-viral gene delivery technology comprising a scaffold that carries all required components, the EEE, a targeting ligand, and the effector gene. Proof of concept of the ENDOSCAPE technology will have a major impact on the therapeutic opportunities for current and future biopharmaceuticals with intracellular sites of action. Thus, overall objectives of the ENDOSCAPE project are to solve the longstanding problem of cytosolic and nucleic delivery of gene therapeutics, to minimize treatment risks by circumventing virusmediated gene transfer, to enhance the efficacy of targeted gene therapeutic treatment in patients, to reduce the costs of gene therapy and make it available for a broad patient base, and to be compatible with personalized gene therapies.
Another objective is the functional characterization of the ENDOSCAPE prototypes in cell cultures and mouse models, and to investigate their molecular mode of action. Here we validated genes for hemophilia therapy and suicide genes for cancer treatment. In addition, we tested the expression of target receptors on different cell lines for several candidate ligands that have been recombinantly expressed and purified, or purchased. Currently, we are investigating the escape of nucleic acids from endosomes in living cells by confocal and light sheet microscopy using fluorescently labeled dendrimers modified with EEEs. We demonstrated efficient delivery of effector DNA in cell culture with low toxicity. The use of plant natural products such as EEEs entails the risk of heterogeneous source material and lack of availability. Therefore, an independent production process is vitally important. We established various plant culture formats and evaluated different plant growth systems and culturing conditions.
We further conducted an expression analysis towards gene discovery in EEE producing plants with the intention to manipulate the biosynthesis of EEEs for higher yields. To date we are already able to extract and purify target EEEs from the biomass of greenhouse grown plants.
In addition, we started an early health economic evaluation by defining clinical example applications and planning analyses based on literature review. We are further collecting information on the expected cost of the ENDOSCAPE production process. Our project is regularly communicated, e.g. by a public website, a project video and a LinkedIn account.