Tracking nanoparticles: from cell culture to in vivo delivery

Project Overview

The aim of this ARC Laureate Fellowship is to define the fundamental mechanisms by which a nanoparticle travels from the bloodstream to its site of action inside a cell in vivo. This involves the use of advanced microscopy, novel nanoparticle delivery systems, and unique in vivo systems to dissect how a nanoparticle reaches target cells in a living organism and then reaches its site of action within those cells.

The pathway taken by a protein or nanoparticle to reach its site of action inside a cell involves two distinct processes, as indicated by these key questions:

• How does a naturally occurring protein, or an artificial nanoparticle carrying a fluorescent cargo protein in the bloodstream, cross the cellular barrier formed by the endothelial cells of the vasculature (transcellular transport)?

• How does the protein or nanoparticle become internalised by target cells (e.g. cancer cells) and then reach the cytosol or nucleus of that cell, overcoming the barrier of the endosomal or lysosomal membrane (endocytosis and endosomal escape)?

Objectives

• Quantitatively map trafficking of proteins and nanoparticles from the bloodstream to tissues by 4D light microscopy and correlative 3D electron microscopy using the zebrafish as a model system

• Genetically manipulate and quantitatively assess endocytosis in vivo across a range of cell types using superresolution light microscopy and electron microscopy

• Develop systems to map trafficking of nanoparticles to mammalian xenografts in zebrafish embryos