Development of a new polymericDNA delivery system
Author
Santos Llinas, Miquel
Term
4. term
Education
Publication year
2021
Submitted on
2021-06-03
Pages
113
Abstract
Gene therapy aims to treat diseases with a genetic origin by delivering healthy DNA into patients’ cells. A major challenge is getting DNA into human cells safely and efficiently. This thesis presents a new, non‑viral DNA delivery approach that avoids using viruses: a tiny “nanocarrier” built from the amphiphilic polymer PVP‑OD (which has both water‑loving and water‑repelling parts) combined with a short, positively charged peptide (a cationic peptide). We characterized the nanocarrier’s size, shape, and safety, and tested how well it can trigger expression of an introduced gene in human cells (transfection). The nanocarrier shows promising characteristics for gene delivery, but high transfection levels have not yet been achieved, indicating that further optimization is needed. Because traditional flat, two‑dimensional cell culture tests (2D transfection experiments) have technical limitations, we also developed a COMSOL computer model to support the future design of a microfluidic chip—a small device with tiny channels—so this DNA delivery system can be tested under conditions that better mimic the human body.
Genterapi sigter mod at behandle sygdomme med genetisk oprindelse ved at levere sundt DNA til patienters celler. En stor udfordring er at få DNA sikkert og effektivt ind i menneskeceller. I denne afhandling præsenteres en ny, ikke‑viral metode til DNA‑levering, som ikke bruger virus: en lille “nanotransportør” bygget af den amphifile polymer PVP‑OD (med både vandelskende og vandafvisende dele) kombineret med et kort, positivt ladet peptid (et kationisk peptid). Vi karakteriserede nanotransportørens størrelse, form og sikkerhed og testede, hvor godt den kan få et indført gen til at blive udtrykt i menneskeceller (transfektion). Nanotransportøren viser lovende egenskaber for genlevering, men høje transfektionsniveauer er endnu ikke opnået, hvilket peger på behov for yderligere optimering. Fordi traditionelle, flade todimensionelle cellekulturforsøg (2D‑transfektion) har tekniske begrænsninger, udviklede vi også en COMSOL‑computermodel til at støtte den fremtidige udvikling af en mikrofluidik‑chip—en lille enhed med bittesmå kanaler—så systemet kan testes under forhold, der bedre ligner menneskekroppen.
[This apstract has been rewritten with the help of AI based on the project's original abstract]