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Nanoparticle Transfer Through Endothelial Barrier (NanoTENDO)

 

 

  

 

Project: Nanoparticle Transfer Through Endothelial Barrier

Acronym: NanoTENDO

Call for proposal: M-ERA.NET Call 2018

Contract number: No. 1.1.1.5/ERANET/19/07

Implementation period: 36 months (1 October 2019 – 30 September 2022)

Project coordinator: University of Lodz

Project partners:

University of Alcala, Faculty of Sciences, Department of Organic and Inorganic Chemistry

Riga Stradiņš University

Latvian Biomedical Research and Study Centre

Leader of Latvian team: Dr. biol., Juris Jansons

Total Costs: 170 000.00 EUR

 

Summary

Many brain-associated diseases (multiple sclerosis, Alzheimer’s disease, stroke, etc.) remain undertreated by effective therapies because therapeutic molecules cannot cross the endothelial barrier, which is an important part of the blood-brain barrier.  Some nanoparticles (NP) ‒ gold NP, dendrimers, dendrons ‒ represent a promising new tool for crossing the endothelial and blood-brain barriers and delivering into the brain the biologically active compounds, as drugs and immunotherapeuticals.

 As we do not know what properties they have to possess to be able to cross the barriers, we cannot meaningfully design them. All the NPs described in literature were chosen empirically after numerous candidates had been screened. Therefore, the main aim of present project is to elucidate the mechanisms by which the most promising NPs: gold NPs, dendrimers, dendrons and their complexes with drugs are transferred through the endothelial barrier. The in vitro experiments will use an endothelial barrier model: primary human aortic endothelial cells (HAEC). Isolation of primary cells encounters many obstacles, among them the variability of the reagents and procedures and access to appropriate ethically sourced donor tissue type, therefore, we decided to use the commercially available sources.

The mechanisms of NP transfer (uptake, cytotoxicity and efflux) and the functional state of the cells (active/passive transport, proliferation, morphology) will be examined on the basis of the size, charge, hydrophobicity, and flexibility of the NP alone and when loaded with drugs or immunotherapeuticals. The same mechanisms will be analyzed under cell stress conditions using oxidative stress and an acidic medium (related to stroke), and in the presence of amyloid-beta peptides (related to AD). The data obtained will: (i) elucidate the mechanisms of NP transfer through the endothelial barrier depending on their nature, key functional groups, and attached drug(s) and/or immunotherapeutical(s), and (ii) identify the functioning of the endothelial barrier during transfer of NPs and their complexes with drugs/immunotherapeuticals(s) under physiological and pathological conditions. The results will provide a basis for systematic screening of NPs for the delivery of active compounds/substances into the brain.

The in vivo experiments will use the murine models of AD and brain ischemia to study the therapeutic potential of NP complexes with drugs, nucleic-acids based immunotherapeuticals.

The objects for study will be gold NPs of 2-4 different types, the newly synthesized dendrimers of 1st-4th generations, and their corresponding dendrons; and complexes of NPs with drugs (laquinimod), endothelial cell targeting proteins (transferrin, wheat germ agglutinin (WGA)) or short-chain nucleic acids (including siRNA, and immunomodulating short double stranded DNAs and single stranded RNAs). The complexes will be labelled with fluorescent probes to follow up the transfer.

The subject for investigation will be the endothelial barrier model (commercially available primary HAEC) and murine models of Alzheimer’s disease and brain ischemia in experiments in vivo. Primary cells are isolated directly from the donor tissue and therefore maintain tissue characteristics that are closer to in vivo conditions.

The tasks are to:

(1) Study the uptake and efflux of NPs through the endothelial barrier and their cytotoxicity using molecular and cell biology techniques;

(2) Estimate the functional state of the endothelial barrier cells in the presence of NPs and their complexes;

(3) Repeat (1) and (2) under conditions of endothelial barrier stress induced by oxidative stress and acidification (related to brain ischemia) and the presence of amyloid-beta peptides (related to Alzheimer’s disease);

(4) Establish murine models of brain ischemia and AD; and

(5) Approve the therapeutic potential of NP complexes of drugs/nucleic acids tested above in in vivo system.

Information published 01.10.2019.

 

Progress of the project:

October 1, 2019 - December 31, 2019

During the reporting period, the Project Consortium Agreement has been agreed and signed. Work is underway on the development of the website, the necessary information is being collected from the consortium partners.

Information published on 31.12.2019.

 

Progress of the project:

1 January 2020 - 31 March 2020

During the reporting period, an application is being prepared for the Ethics Commission of the Food and Veterinary Service for a pilot project permit for the use of animals in experiments planned in the project. Work on the website is continuing.

Information published on 31.03.2020.

 

Progress of the project:

April 1, 2020 - June 30, 2020

During the reporting period, the development of the project website http://nanotendo.pl/, was completed and it is supplemented with information from the consortium partners. An experimental design was developed in consultation with the consortium partners, taking into consideration the situation with the Covid19 pandemic and the available infrastructure. The ethical aspects of the research were analyzed.

Information published on 30.06.2020.

 

Progress of the project:

1 July 2020 - 30 September 2020

During the reporting period, the theoretical basis of the project was supplemented with information from current scientific publications on ischemic stroke and Alzheimer's disease treatment methods, the possibilities for testing in mouse models of selected therapeutic agents in a complex with nanoparticles (dendrimers) have been evaluated.

Information published on 30.09.2020.

 

Progress of the project:

1 October 2020 - 31 December 2020

During the reporting period, experiments on the formation of complexes of dendrimer nanoparticles and therapeutic agents and testing of the stability and other properties of the obtained complexes in in vitro experiments were performed. Based on the results, the siRNA that specifically induces silencing of MAPT genes involved in the development of the disease was selected as a treatment agent for Alzheimer's disease.

Information published on 31.12.2020.

 

Progress of the project:

January 1, 2021 - March 31, 2021

During the reporting period, a strategy has been developed for the labeling of dendrimers and transportable substances with fluorescent markers, which allows to study the distribution of injected drugs in the body and the ability to cross the blood-brain barrier in animal experiments. Due to the situation in Covi19 in Spain and Poland, the schedule of experiments was significantly delayed, thus the project partners agree to extend the project implementation deadline.

Information published on 31.03.2021.



Mājas lapas izstrādi finansēja ERAF 2.1.1.2. aktivitātes projekts Nr. 2010/0196/2DP/2.1.1.2.0/10/APIA/VIAA/004 "Latvijas biomedicīnas pētījumu integrācija Eiropas zinātnes telpā".