RALA-mediated dual delivery of pMIR-143 and gemcitabine triphosphate as a novel nanotherapy for the treatment of pancreatic cancer

Research output: Contribution to conferencePoster

Abstract

Introduction: Pancreatic cancer has a low incidence compared to other types of solid tumours, however, it is an extremely lethal disease ranking among the top 5 most frequent causes of cancer related death in the UK1. Our group has previously shown that the RALA peptide can encapsulate negatively charged moieties and achieve efficient intracellular delivery 2-3. This project is designed to develop and characterise two novel RALA-based nanoparticles (NPs) that encapsulate a miR143-encoded plasmid and the active form of the first-line chemotherapeutic agent in pancreatic cancer, gemcitabine-triphosphate.

Experimental: The size and charge of the designed NPs were measured using DLS and the stability measured under different conditions. Consequently, pancreatic cancer cell lines (PANC-1 and MIA-PaCa2) were treated with RALA/pEGFP-N1 NPs to evaluate the cellular uptake efficiency and cell viability, via flow cytometry and MTS cytotoxicity assay 48 h post-transfection. qRT-PCR and western blot analysis were conducted on RALA/pCMV-MIR-143 treated cells, to evaluate the expression levels of miR-143 and K-RAS as a target gene respectively.

Results: NPs were formed with a diameter ≤ 150 nm and a charge of 20-40 mV. Cells treated with RALA/pEGFP-N1 and RALA/FITC-12-UTP expressed the highest fluorescence at N:P 10 and w:w 10 respectively in all cell lines with no discernible toxicity ˂10%. miR-143 showed increased levels in PANC-1 after 48 h of transfection while no change was recorded in MIA-PaCa2. As a result, the K-RAS target protein levels were decreased in PANC-1 after the transfection with RALA-pCMV-MIR-143 NPs.

Discussion and Conclusions: The designed NPs were formed in the optimal size and charge for cellular uptake with successful delivery of an intact therapeutic cargo which was translated by up-regulation of miR-143 after the transfection. Further studies will be conducted to assess the functionality and the therapeutic effect of the delivery system both in vitro and in vivo.
Original languageEnglish
Publication statusPublished - 20 Sep 2018
EventInternational Conference On Nanomedicine And Nanobiotechnology – ICONAN 2018. - Europe, Rome, Italy
Duration: 26 Sep 201828 Sep 2018
https://premc.org/conferences/iconan2018/

Conference

ConferenceInternational Conference On Nanomedicine And Nanobiotechnology – ICONAN 2018.
Abbreviated titleICONAN
CountryItaly
CityRome
Period26/09/201828/09/2018
Internet address

Fingerprint Dive into the research topics of 'RALA-mediated dual delivery of pMIR-143 and gemcitabine triphosphate as a novel nanotherapy for the treatment of pancreatic cancer'. Together they form a unique fingerprint.

  • Cite this

    Daoud, A., Donnelly, R., & McCarthy, H. (2018). RALA-mediated dual delivery of pMIR-143 and gemcitabine triphosphate as a novel nanotherapy for the treatment of pancreatic cancer. Poster session presented at International Conference On Nanomedicine And Nanobiotechnology – ICONAN 2018., Rome, Italy.