Adenosome-Based Nucleic Acid Delivery to Prostate Cancer Cells

T. W.A.N. Mooijenkind

Research output: Types of ThesisMaster's ThesisAcademic

Abstract

Introduction: The incidence of prostate cancer (PCa) in the Netherlands has been increasing since 1989, with roughly 13.500 new PCa diagnoses in 2019, making it the most common cancer type among men in the Netherlands (1). Treatment depends on the stage of the cancer and risk profile of the patient; therefore, patients with late-stage, metastasised PCa no longer receive treatments with curative intent; therapy is instead given to increase lifespan and quality of life (2). Furthermore, only 41% of patients with PCa and distant metastases are alive after 5 years (1), so new and effective treatment options are essential to decrease cancer related mortality. Extracellular vesicles (EVs) have been studied more extensively in the past years due to their beneficial characteristics compared to existing delivery systems. EVs are naturally occurring, non-immunogenic, and can pass biological barriers, such as the blood brain barrier (3-6). This report aims to analyse the potential of adenosomes to deliver functionally active small RNAs to PCa cells.
Methods: Adenosome and EV uptake by PCa cells was assessed by adding adenosomes carrying GFP to DU145 PCa cells on a 384-well plate. The plates were imaged using the Opera Phenix® High-Content Screening System. Furthermore, a lentiviral vector was constructed carrying a short hairpin RNA against sheGFP to assess the delivery of functionally active genesilencing agents. However, the validation of the delivery of gene-silencing agents could not be performed due to time constraints.
Results: Adenosome uptake is very efficient, reaching efficiencies up to 96%, compared to 91% for EV uptake. Furthermore, the delivery and processing of GFP was promising, with roughly 16% of cells that took up adenosomes having a GFP intensity above the threshold value. The sheGFP vector was constructed but no further experiments were performed with the vector due to time constraints.
Discussion: Adenosomes show great potential as a delivery system for therapeutic nucleic acids (TNAs). This study aimed to assess the efficacy of adenosomes to deliver functionally active small RNAs to PCa cells, and found that adenosome uptake efficiency is comparable to (or higher than) EV uptake efficiency. Moreover, the delivery and processing of functional cargo in the form of genetic information was effective as 16% of cells that took up adenosomes carrying GFP, started expressing GFP above the threshold intensity. Overall, adenosomes show enormous potential to become the new delivery system for TNAs.
Original languageEnglish
Supervisors/Advisors
  • Martens, Elena, Supervisor
  • van Royen, Martin, Supervisor
  • de Vrij, Jeroen, Supervisor
Publication statusPublished - 2023
Externally publishedYes

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