Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - MMBio (Molecular Tools for Nucleic Acid Manipulation for Biological Intervention)

Teaser

The action of genes is responsible for all organismal processes of life. If mutations in these genes arise, the processes may go wrong and lead to diseases. To treat such genetic malfunctions, it would be desirable to manipulate the deficient genes directly, but that requires...

Summary

The action of genes is responsible for all organismal processes of life. If mutations in these genes arise, the processes may go wrong and lead to diseases. To treat such genetic malfunctions, it would be desirable to manipulate the deficient genes directly, but that requires novel reagents that can be administered without causing harm and are selective and efficient. Such novel reagents would have profound impact on disease prevention and cure and benefit society as they enable new approaches to disease treatment.

The objective of MMBio is to provide the ability to ultimately cure cellular disease phenotypes by employing molecular, chemical and biological approaches to interfere gene expression precisely and selectively. MMBio will focus on unconventional new therapeutic approaches, namely “genetic” drugs that are typically based on oligonucleotides (ONs) or the action of genome-editing nuclease enzymes. Genetic drugs have the advantage that their design can be programmed to act at a specific target much more predictably than conceivable for small molecule drugs. The challenges of this ambitious scientific objective include the synthesis of drug conjugates (e.g. therapeutic antisense oligonucleotides), drug delivery reagents and artificial chemical and protein-based nucleases (e.g. the CRISPR/Cas system, a novel genetic pair of scissors with huge potential for genome editing), a quantitative mechanistic understanding of their action, efficient and selective delivery into the cell and consideration of the effects on disease development.

To address this broad challenge MMBio brings together representatives of classically separate disciplines ranging from organic synthesis, mechanistic chemistry and nanochemistry to microengineering and experimental medicine. MMBio builds on a successful FP7 network that focused on fundamental aspects of phosphate transfer and recognition, and has now been extended to become a consortium covering the development pipeline of experimental drug conjugates from the reaction flask into cells. Many of the experimental approaches are rooted in Chemical Biology, a supradisciplinary field based on fundamental molecular research that employs selective intervention in biological systems by chemical means. But MMBio attempts to reach beyond this field, through consideration of the druggability of the constructs made and aiming, as far as possible, at the integration of in vitro, cell-based and in vivo studies. The network assembles experts in all the disciplines required across the entire R&D pipeline for the development of new gene therapeutics, including small and large private sector companies, academics focusing on fundamental and applied aspects and many opportunities for collaboration. The six industrial stakeholders in MMBio are ready to commercialize output of our network research.

Work performed

The MMBio consortium consists of 12 European partner groups (8 academic groups, 1 medical school, 1 national research institute and 2 companies). 15 ESRs, representing 13 different nationalities (9 female and 6 male), have been recruited and started their PhDs between January 2017 and June 2018. The consortium has already run several network meetings and training workshops that covered scientific topics (state of the art, quantitative analysis of complex bioorganic systems, why drugs fails, drug development, CRISPr, nucleic acid chemistry and the use of nucleic acids for therapy) as well as ‘soft skills’: innovation & entrepreneurship, intellectual property (run by an EPO patent examiner), ethics, science communication and project management. All ESRs were given a tour of the RISE (Research Institutes of Sweden ) and a tour of the Gotheburg Astrazeneca site, from the chemistry labs to in-vivo testing. In addition to these workshops and company visits, each ESR is required to exchange ideas with each other, enhance their research project and learn new research techniques through secondments to industry and to other academic laboratories within the MMBio network. In order to recruit the next generation of scientists, we participated in the Cambridge Science Festival and our both was visited by around 100 future scientists below the age of 12.

To date, results from the ongoing projects include:

• The study of peptide dendrimers as nucleic acid drug delivery reagents
doi: 10.1016/j.ejpb.2018.09.002.

• Novel artificial nucleases, e.g.
• synthetic metal-free RNA cleavers
• protein-nucleic acid-based artificial nucleases, ‘PNAzymes’

• The directed evolution of proteins that transfer phosphate groups (i.e. kinases and nucleases) at ultrahigh-throughput (starting from libraries of > 10e8 members)

• High-throughput cell-based assays for drug testing using only nanoliters of reagent

Final results

The first half of the project has focused on the molecular engineering of chemical and biochemical systems that have never been made before. This is in itself interesting as the functional tests with these systems will tell us about their behaviour and yield structure-function relationships. We are now starting to find out whether our designs (or our combinatorial approaches) were successful – and, if the efficiency of these systems is sufficient, we are in a position to probe their value in an applied biotechnological or even pre-therapeutic context.

Website & more info

More info: http://www.bioc.cam.ac.uk/hollfelder/Research/mmbio.