A novel multi-functional nanocomposites against Parkinson’s disease for the protection and regeneration of dopamine neurons with anti alpha-Synuclein aggregation properties.

Project Information

NEUROCURE

Grant agreement ID: 101027379

DOI

10.3030/101027379

Project closed

EC signature date 19 April 2021

Start date 1 September 2021

End date 16 September 2023

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 190 680,96

EU contribution

€ 190 680,96

Coordinated by

HELSINGIN YLIOPISTO
Finland

Periodic Reporting for period 1 - NEUROCURE (A novel multi-functional nanocomposites against Parkinson’s disease for the protection and regeneration of dopamine neurons with anti alpha-Synuclein aggregation properties.)

Reporting period: 2021-09-01 to 2023-08-31

Summary of the context and overall objectives of the project

Parkinson’s disease (PD) is the second most common neurodegenerative disorder that has affected over 10 million people around the world. PD is characterized by progressive degeneration of nigrostriatal dopamine neurons in the substantia nigra. PD progression leads to severe movement disorders and non-motor symptoms. Currently, PD treatment strategies are mostly based on dopamine replacement medicines that temporarily alleviate the symptoms. However, none of the available treatment schemes has shown an ability to slow down the progression of neurodegeneration. Thus, there is an urgent need for novel disease-modifying PD therapies.

Preclinical studies have shown that neurotrophic factors have ability to support neuronal functions and reverse neurodegeneration. Therefore, NTFs could not only protect remaining dopamine neurons but also stimulate their regeneration and capacity to make up for already lost cells. One of the most potent NTFs for PD therapy so far is the cerebral dopamine neurotrophic factor (CDNF). However, despite great potential, CDNF does not pass through the blood-brain barrier (BBB) and requires complicated stereotactic brain surgery to administer. Therefore, there is a great need for the development of efficient systemic CDNF delivery methods to avoid intracranial surgery.

The current project aimed to develop new and effective methods for CDNF delivery to the brain by utilizing nanotechnology and molecular biology. Specifically, novel nanoparticles functionalized with brain-targeting (homing) peptides and loaded with CDNF were developed and the ability to rescue dopamine neurons was tested. In addition, the ability to penetrate through the BBB was analyzed. Another delivery approach was based on a direct fusion of CDNF with brain-targeting peptides. These novel approaches in CDNF delivery to the brain bring us closer to the non-invasive treatment of PD to help over 10 million people worldwide.

Please also see our Youtube video introducing the project: https://youtu.be/6BDo1jcgxBc

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

09.2021 – Posts about the projects on social media (Facebook, ca 25 researchers and ca 5 general public persons reached) - WP2
09.2021 - Meetings with all (co-)supervisors and administration. Roles and research and publication plans agreed - WP1
23.09.2021 – Career Development Plan agreed with primary supervisor - WP1
12.10.2021-5.11.2021 - Training with dopamine neuron cultures to assess the activity of CDNF - WP3
9.11.2021- 4.02.2020 – Comparing activity of CDNF fragment – homing fusion peptides with full length CDNF in dopamine neuron cultures, 6-OHDA model. 6-OHDA model did not show activity of CDNF in the control sample. I change the model to MPTP model – WP3
14.02.2022 – Started synthesizing nanoparticles (liposomes) for use as vehicle for CDNF delivery across the blood brain barrier (BBB) to the brain – WP3
14.03.2022– In vivo course completed, and certificate obtained - WP1
24.03.2022 - Testing toxicity of nanoparticles in neuronal culture. Results proved that synthesized nanoparticles are nontoxic and biocompatible – WP3
04.04.2022 – Started synthesis of homing peptide functionalized nanoparticles – WP3
05.04.2022 – Started testing homing peptide functionalized nanoparticles in neuronal culture – WP4
12.04.2022 – Started synthesis of CDNF fragment loaded nanoparticles – WP3
19.04.2022 – Started testing activity of CDNF loaded nanoparticles in dopamine culture – WP4
03.05.2022 – Started in vitro quantification of loaded CDNF in the nanoparticles using ELISA assay – WP3
10.05.2022 - Isotope safety course completed – WP1
12-15.06.2022 - Presented at the Sigrid Jusélius Symposium “Neuronal Plasticity in Brain Disorders and Their Treatment”, ca 100 attendees – WP2
20.06.2022 – Quantification of homing peptide conjugation to nanoparticles – WP3
18.07.2022 – Characterization of synthesized nanoparticles – measurement of the hydrodynamic size – WP3
24.08.2022 - Continued testing activity of CDNF fragment - homing peptide fusion peptides in dopamine culture. MPTP model – WP3
19.10.2022 - Started in vivo blood brain barrier (BBB) permeability experiment of CDNF-homing peptide fusion peptides and CDNF loaded nanoparticles – WP4
11-12.10.2022 – Completed “Physiologically based pharmacokinetic modeling (PBPK) course” – WP1
09.01.2023 – Started testing activity of fusion peptides in neuronal culture using another neurotoxicity model (thapsigargin) – WP3
17.03-31.08.2023 - Supervising research assistant Dmitriy Zhilin – WP1
28.05-02.06.2023 – Presented at Neurotrophic Mechanisms in Health and Disease Gordon Research Conference “Neurotrophic Factor Functions in Development, Plasticity, Regeneration and Disease”, ca 120 attendees - WP2
19.07.2023 - Started working on a review article with Prof. Saarma an Prof. Teesalu. for the International Journal of Nanomedicine – WP2
28.07.2023 – Nanoparticle characterization: quantification lipid content in liposomal nanoparticles – WP3
22-24.08.2010 – Visited the University of Tartu to give a talk at the Institute of Biomedicine and Translational Medicine (ca 20 attendees) and to discuss results and future collaboration with Prof. Teesalu.
16.09.2023: YouTube video with supervisors Prof. Saarma and Prof. Teesalu: "Targeted delivery of neurotrophic factors to the brain against Parkinson’s disease", https://youtu.be/6BDo1jcgxBc

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:
CDNF-homing fusion peptides are biologically active, penetrate through the BBB, and rescue the neurons in vitro. We used newly discovered and characterized in our lab CDNF fragment, which is significantly shorter than the full-length protein but is biologically active and has an advantage in the BBB permeability.
We successfully synthesized and characterized brain-homing peptide functionalized nanoparticles with high CDNF loading capacity.
Animal experiments are ongoing to support in vitro results.
The results of this project open new approaches in the brain-targeted delivery of neurotrophic factors to support and regenerate neurons in Parkinson's disease.
These new delivery methods are called upon to overcome BBB permeability limitations and avoid risky and complicated stereotactic brain surgery.

Youtube video with acknowledgment to promote the project