Novel blood-brain barrier permeable CDNF-derived therapeutic peptides for the protection and regeneration of dopamine neurons

Project Information

THERAPE

Grant agreement ID: 835140

DOI

10.3030/835140

Project closed

EC signature date 11 April 2019

Start date 2 September 2019

End date 3 May 2022

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 202 680,96

EU contribution

€ 202 680,96

Coordinated by

HELSINGIN YLIOPISTO
Finland

Periodic Reporting for period 1 - THERAPE (Novel blood-brain barrier permeable CDNF-derived therapeutic peptides for the protection and regeneration of dopamine neurons)

Reporting period: 2019-09-02 to 2021-09-01

Summary of the context and overall objectives of the project

Parkinson's disease (PD) is the second most common neurodegenerative disease characterized by the progressive loss of midbrain dopamine neurons that leads to the severe movement disorders and several non-motor symptoms. Available PD treatments do not provide long-lasting relief from the symptoms and do not slow down or stop the neurodegeneration.
Cerebral dopamine neurotrophic factor (CDNF) was shown to reverse the PD-associated neurodegeneration in tests on cultures and animals. Despite of these promising results, CDNF doesn’t pass through the blood brain barrier (BBB) and should be delivered to the brain of animals and PD patients through the risky microsurgery. Thus, there is a great need for the CDNF variants that can penetrate through the BBB
avoiding intracranial surgery.
Current project aimed to develop the next generation CDNF-derived peptides able to penetrate the BBB. For that 1) the length of CDNF was be reduced to enable penetration through the BBB, 2) CDNF fragment was modified to improve its stability in the blood and tissues and 3) CDNF fragment was be conjugated to nanoparticles and brain-targeting peptide to increase its stability and ability to penetrate the BBB.
As a result, we characterized the mechanism by which CDNF rescues dopamine neurons and developed new versions of CDNF that enable to deliver CDNF peripherally. This brings us closer to the (non-invasive) symptomatic treatment of PD to help over 10 mln of PD patients.
Please see also our YouTube video introducing the project: https://www.youtube.com/watch?v=W9PCA2JD1Mk&t

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

2019 - MSCA-IF Information day organized – 30 people attended - WP2
09.2019 – Posts about the projects on social media (Facebook, LinkedIn, ca 50 researchers and ca 200 general public persons reached) - WP2
09.2019 - Meetings with all (co-)supervisors and administration. Roles and research and publication plans agreed - WP1
17.09.2019 – Career Development Plan agreed with the supervisor - WP1
30.09.2019 – In vivo course completed and certificate obtained - WP1
1.10.2019-30.10.2019 - Training with dopamine neuron cultures and artificial blood-brain-barrier (BBB) in vitro model to assess the activity and BBB permeability of CDNF and its peptides - WP3
1.11.2019-4.02.2020 - Functionalization of 61 aa CDNF fragment with PEG (further referred as C-CDNF-PEG) to increase the stability of CDNF fragment. Testing of CDNF fragments in dopamine neuron cultures, 6-OHDA model. Testing is completed. 26 aa, 33 aa, 37 aa, 61 aa CDNF and C-CDNF-PEG are biologically active in this test model - WP3 and WP4
19.02.2020 - Public lecture at the Institute of Biotechnology, Helsinki, Title: "Novel blood-brain barrier permeable CDNF-derived THERApeutic PEptides for the protection and regeneration of dopamine neurons (THERA-PE)", ca 30 attendees - WP2
25.02.2020 - Isotope safety course completed - WP1
26.02.2020-1.05.2020 Optimization of indirect ELISA for the detection of CDNF fragments shorter than 61 aa and C-CDNF-PEG - WP4
22.04.2020 – ZOOM-enabled lecture at the University of Tartu, Estonia; Title: "Regulatory aspects of nanomedicines", ca 10 attendees - WP2
1.05.2020 - 15.06.2020 It was not possible to conduct experimental work at the University of Helsinki due to COVID-19 situation. Together with Mart Saarma we planned and started to draft the review article for the journal “Frontiers in Neuroscience”.
15.06.2020 – 20.09.2020 – Practical work resumed. Testing of CDNF fragments in artificial blood-brain-barrier (BBB) in vitro model. Testing is completed. The data show that the efficiency of penetration of CDNF peptides depends on their length (as hypothesised in the beginning) but also on the number of helices (new finding). In this model 49 aa CDNF is the best fragment. We plan to use it for in vivo studies – WP3
1.07.2020 – 1.09.2020. Supervision of two students, Bcs student Beatrice Marlene-Metsaorg and Msc student Sven Sakson – WP1
1.09.2020 – 1.11.2020. Collaboration with Prof. Tambet Teesalu on brain-homing peptide. Original plan was to screen for the new brain-homing peptide during the secondment in Prof. Laakkonen's lab. However, since we are in the middle of pandemics, the visits to the other labs are complicated. Thus, we decided to go on with the existing brain-homing peptide (unpublished data by Prof. Tambet Teesalu). I observed exciting results: 49 aa CDNF-brain-homing peptide fusion is 1) active in dopamine neurons and 2) significantly increases the penetration of 49 aa CDNF-homing peptide (compared to 49 aa CDNF without fusion) in artificial BBB in vitro model. - WP5
11.2020 - Two ERC grant writing courses completed: from Yellow Research and at the University of Helsinki. - WP1
9.12.2020 - ZOOM-enabled lecture "Can CDNF prevent neurodegeneration in Parkinson's disease" together with Prof. Mart Saarma for Estonian Parkinson's Disease Society (in Estonian and Russian, https://www.youtube.com/watch?v=74uzQcNBFJY&t=272s). - W2
11.2020-04.2021 - Writing of review article for the journal “Frontiers in Neuroscience”.
21.04.2021: ZOOM-enabled lecture at the University of Tartu; Title: "Regulatory aspects of nanomedicines", ca 6 attendees - WP2
23.04.2021: Article "Neurotrophic Factors in Parkinson's Disease : Clinical Trials, Open Challenges and Nanoparticle-Mediated Delivery to the Brain" for the journal “Frontiers in Neuroscience” was accepted for publication.
1.05.2021: Drafting of article 2 (potential of CDNF fragments to rescue dopamine neurons) started
9.08.2021: YouTube video with supervisors Prof. Saarma and Prof. Voutilainen: "Neurotrophic factor CDNF as a drug for Parkinson's disease", https://www.youtube.com/watch?v=W9PCA2JD1Mk
14.08.2021: Presentation with Prof. Saarma "Life and death of the dopamine neurons in Parkinson's disease" at the event organized by the Estonian Parkinson's Disease society.
20.09.2021 - 01.2022: Supervision of student Hend Missonen (Finnish school of technicians) on tests with dopamine neurons.
22.09.2021: Presentation at Researcher's Night Event, ca 20 attendees.
1.10.2021-21.12.21 Effects of the inhibitors of unfolded protein response sensors on ability of CDNF fragments and mutants to rescue dopamine neurons.
2.01.2022-30.04.2022 Studies of full-size CDNF mutants: whether their mechanism of action is similar to CDNF fragments (MST assays, competition assays, circular dichroism, Alpha Fold predictions of structure)
13.04.2022: ZOOM-enabled lecture at the University of Tartu; Title: "Regulatory aspects of nanomedicines", ca 10 attendees - WP2
26.04.2022: Presentation at 3rd International Conference of Nanomaterials, ca 50 attendees.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

The most important results of the project are the following:
We reduced the length of CDNF to enable penetration through the BBB and demonstrated that short CDNF fragments penetrated BBB better compared to the full-size CDNF in vitro and in vivo.
For the first time, we demonstrated the mechanism of action of CDNF fragments to rescue dopamine neurons in different cell death paradigms.
We showed that conjugation of CDNF fragments to brain-targeting peptide significantly increased the passage of CDNF fragments through the BBB.

This brings us closer to the (non-invasive) symptomatic treatment of PD to help over 10 mln of PD patients.

Key steps of my MSCA-IF projects with illustrations