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Description
Effective cancer therapy is the most important goal in oncology. The progress in antibody drug conjugates encouraged us to verify if attachment of two different cytotoxins to a targeting protein will lead to a efficient conjugate. The Project stems from our findings that fibroblast growth factor 2 (FGF2) can replace antibody as a targeting protein. FGF2 conjugated with monomethyl auristatin E (MMAE) can destroy cancer cells overexpressing FGF receptor 1. We will produce FGF2 conjugates with two extremely cytotoxic warheads: MMAE and α-amanitin (AMN). Simultaneous application of two drugs is an alternative for conventional combination therapy. MMAE blocks microtubule formation and AMN is RNA polymerase II and III inhibitor. Both warheads should act in concert in a single cancer cell. Further, MMAE can be secreted from cancer cell and via so called bystander effect destroy neighboring cells and AMN shows outstanding activity in cells expressing multi-drug resistant transporters. The most promising dual conjugate will be tested in animal models, including FGFR-dependent xenographs and patient-derived tumor models. The approach is original and addresses trends in anti-cancer drug development: specific delivery, high level of drug loading combined with defined stoichiometry and homogeneiety of conjugate.
The consortium of Oslo University Hospital, University of Wroclaw and Pure Biologics represent a team well-suited for the completion of described tasks and will benefit from their diverse background. Proposed tasks are interconnected and we believe that our laboratories are in many aspects highly complementary, providing better chance of successfully delivering project milestones. The project includes both specialized work packages where we can benefit from a deep experience of a consortium partner, along with more interdisciplinary ones, where tight collaboration will enhance knowledge transfer and all the benefits of our previous collaborations will come into play.
Summary of project results
Effective cancer therapy is the most important goal in oncology. The progress in antibody drug conjugates encouraged us to verify here if attachment of two different cytotoxins to a targeting protein would lead to a more efficient conjugate. The Project stems from our findings that fibroblast growth factor 2 (FGF2) can replace antibody as a targeting protein. The approach is highly original and addresses trends in anti-cancer drug development: specific delivery, high level of drug loading combined with defined stoichiometry and overall homogeneiety of conjugate. Most importantly our proposed targeted strategy will have not only the potential to efficiently kill tumor cells reducing the side effects on healthy cells, but also to limit their ability to develop resistance.
Within the project we have established and optimized methods for obtaining double-warhead conjugates of recombinant FGF2 with two cytotoxic
drugs of two independent modes of action, with a potential to be utilized in targeted treatment for cancers with high FGFR1 expression. In the
first phase of the project we have constructed, produced, purified and thoroughly characterized FGF2 variants specifically designed for chemical
conjugation with cytotoxic drugs via an optimized hydrolysable linker. Subsequently, we have generated a panel of single- and double warhead
conjugates of FGF2 with MMAE and beta-amanitin. Biophysical analysis of obtained proteins and the conjugates revealed their native
conformation and biological competence. The results of a systematic evaluation of cytotoxicity of the generated conjugates using cancer derived
FGFR-positive cell lines allowed to confirm the anticancer potential of the analyzed molecules by showing significant toxicity of selected dualwarhead
FGF2 conjugates towards FGFR-positive cells. Within the last part of the project we have performed in vivo studies of the selected
conjugates in mouse xenograft models for human cancer. After obtaining promising results of the preliminary analysis we are planning to
perform further experiments ensuring a detailed efficacy evaluation.
The vast majority of the scientific data resulting from the project implementation has been published as peer-reviewed publications in best
quality journals, moreover there has been one national patent application submitted. Several master''s and PhD theses have been or will be
written based on results obtained during the project. During the project particular attention was paid to ensure the use and propagation of
results achieved during the project. All purchased and produced construct (vectors), proteins or cell lines were appropriately catalogued,
amplified, aliquoted and frozen for future projects and are freely available for all researchers. New methods and protocols implemented during
the project have become well established and widespread in our laboratories broadening scope of our skills. Experimental results obtained
during the projects will be further used as a basis for new projects development.
Summary of bilateral results
Within the project we developed the fruitful collaboration between Polish and Norwegian groups, providing platform for exchange of practicallaboratory knowledge, experience and ideas, and sharing the results together with experimental methods between project participants.Due to the Polish-Norwegian grant excellent conditions for MS and PhD students were created to develop their scientific skills. Advancedlaboratory techniques applied in both laboratories are available for young scientists. PhD students and scientists involved in the project hadextraordinary opportunities to attend high quality scientific meetings, conferences and trainings to broaden their knowledge and promote theresults obtained within the project. To provide the best cooperation opportunity we organized several visits and meetings, to gather Polish andNorwegian scientists together. All meetings were extremely fruitful and beneficial especially for young PhD students. There were many scientificdiscussions and exchange of new ideas concerning the project.Knowledge exchange and development were accompanied with social benefits. Working in closely collaborating laboratories located in differentcountries also influenced the cognition of different cultures and backgrounds.Both Polish and Norwegian partners are willing to continue collaboration between our laboratories. We are going to further exchange the knowhowbetween laboratories as well as to allow young scientist to learn new technologies during onsite visits.