Engineering the immunosuppressive microenvironment in mesothelioma with targeted LipidNanoParticles encapsulating self-amplifying RNA

Informazioni relative al progetto

OncoNanoR

ID dell’accordo di sovvenzione: 101090312

DOI

10.3030/101090312

Progetto concluso il 23 Aprile 2024

Data della firma CE 19 Settembre 2022

Data di avvio 1 Gennaio 2023

Data di completamento 30 Giugno 2025

Finanziato da

• Widening participation and spreading excellence

Costo totale

Nessun dato

Contributo UE

€ 215 773,20

Coordinato da

UNIVERSIDADE DE COIMBRA
Portugal

Periodic Reporting for period 1 - OncoNanoR (Engineering the immunosuppressive microenvironment in mesothelioma with targeted LipidNanoParticles encapsulating self-amplifying RNA)

Periodo di rendicontazione: 2023-01-01 al 2025-06-30

Sintesi del contesto e degli obiettivi generali del progetto

Malignant pleural mesothelioma (MPM) is a highly aggressive disease and standard chemotherapy treatment achieves an overall survival of 12 months. Anti–programmed cell death 1 protein immunotherapy has a low response rate and does not improve the patient's survival due to the immunosuppressive tumor microenvironment. The EU-funded OncoNanoR project aimed to develop innovative immunotherapy for the treatment of MPM, involving activation of the innate and adaptive tumor microenvironment. The general goal was to achieve an intracellular delivery of potentially therapeutic RNA molecules using intravenous injection of MPM-targeted lipid nanoparticles. The specific objective for this period was to obtain and characterize an saRNA-LNP system with appropriate features for biomedical administration; with the ability to target MPM cells.

Lavoro eseguito dall’inizio del progetto fino alla fine del periodo coperto dalla relazione e principali risultati finora ottenuti

In particular, during the last period, we were able to design, formulate under reproducible conditions, and characterize lipid nanoparticles (LNPs) containing mRNA. In order to standardize the procedures, three methods were used: NanoAssemblr® Ignite microfluidic platform from Precision Nanosystems Inc.’s (PNI, Canada), vortex mixing, and pipette mixing. Different ionizable lipids, RNA working concentration, total volume, volume rate ratios (aqueous to ethanol phase) and phase addition order, N/P ratios, mixing times, and speed were tested when possible; and the resulting LNPs were analyzed to determine the amount of encapsulated RNA, the amount of cholesterol and total lipids, particle size, polydispersity index (PdI), and zeta potential. Two different buffers were used for particle dilution and conservation, and possible changes in particle size and PdI were analyzed for several weeks.

Progressi oltre lo stato dell’arte e potenziale impatto previsto (incluso l’impatto socioeconomico e le implicazioni sociali più ampie del progetto fino ad ora)

Similar and positive results were achieved by the three methods used for particle production. In general, particle size was between 80-160 nm, polydispersity index presented values under 0,15; and zeta potential was always close to neutrality, as expected due to the presence of pegylated molecules on particle surface. Encapsulation efficiency was over 70% and up to 87% in some cases; and the particles produced presented high stability over time when stored at 4 0C, in terms of size, PdI, and encapsulated RNA.
Partial results of the project were included in a poster presentation at the 6th ASPIC International Congress 2024 (Faro, Portugal), and we also participated in a collaborative review article (in preparation), which includes aspects related to the generation of RNA-LNPs systems to fight cancer.
Conclusions:
mRNA-LNP systems with relevant features for biomedical applications were efficiently generated using three different methods. The systems proved to be stable over time when stored at 4 0C, and the mRNA was encapsulated inside LNPs at high percentages. Having an extensive standardization phase allowed us to select the best conditions for subsequent production and functionalization steps.
Group URL: https://cibb.uc.pt/en/research-group/tumor-microenvironment-and-targeted-therapies

General scheme of the project