AUSTRALIAN TWO DOCTORAL (PHD) CENTRE FOR...
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AUSTRALIAN CENTRE FOR MICROSCOPY & MICROANALYSIS A/Prof. Filip Braet’s research covers nanobiology, structural cell biology and cancer biology, culminating in developing targeted drug-delivery approaches for colorectal cancer (CRC), including CRC hepatic metastasis. His research uses combinatorial labeling approaches and correlative light & electron microscopy (CLEM) to determine the precise subcellular target(s) of these anti-cancer compounds at the supramolecular level. By drawing on these structural insights and mechanistic understanding, A/Prof. Braet and his collaborators are designing and assessing new therapeutic strategies for treatment of cancer, such as hydrogel-based carriers for sustained, local application of anti-cancer drugs and metal-based nanoparticles, such as iron–gold complexes, that show selective activity towards cancer cells. PROJECT 1 Investigation of the cellular distribution of metal-based anti- cancer compounds by a correlative morphomics approach Cells exposed to anti-cancer drugs exhibit varied cellular responses, such as structural reorganisation, complexation and aggregation of molecules and/or proteins or relocation and/or activation of subcellular compartments. In order to obtain an in-depth view of anti- cancer drug induced cell responses, systematic cellular analysis of the cells’ architecture and functional machinery is a prerequisite. This can be achieved by combining CLEM with combinatorial labeling thereby permitting the direct correlation of dynamic subcellular events and macromolecules in 4-D with the ‘undisturbed’ nano-environment in 3-D on the same cell. The aim of the first part of this project is to devise methodology and standardised protocols for quantifying (sub)cellular responses under varying experimental conditions in real-time (i.e., X, Y, Z & t). Besides devising high-throughput live-cell imaging protocols, the student will apply rapid-fixation and sample-relocation approaches for subsequent electron microscopy. In the second part, this methodology will be applied directly to clinically relevant human CRC cell lines exposed to different metal-based anti-cancer drugs. The work will lead to (i) a standardised CLEM model for drug screening purposes and (ii) better understanding of the action of metal-based anti-cancer drugs. PROJECT 2 The role of membrane rafts in anti- cancer drug treatment Membrane rafts are key structural and molecular regulator for various natural compounds and/or drugs in many biomedical settings, including cancer. Our team recently developed established CRC Caco-2 cell clones that differentially express the membrane-associated raft marker GM1. In preliminary studies, we have demonstrated that this differential The Australian Centre for Microscopy & Microanalysis (ACMM) is the largest and most comprehensive facility of its type in Australia, and it offers an exciting variety of research programs, spanning disciplines from archaeology to biology, from chemistry to engineering, and from medicine to physics. TWO DOCTORAL (PHD) SCHOLARSHIP OPPORTUNITIES MEMBRANE AND CANCER CELL BIOLOGY (MCCB) Colon carcinoma cells stained for actin (green), membrane rafts (red) and DNA (blue). Endothelial cells labelled for actin (red) and nuclei (blue).
Transcript of AUSTRALIAN TWO DOCTORAL (PHD) CENTRE FOR...
AUSTRALIAN CENTRE FOR MICROSCOPY & MICROANALYSIS
A/Prof. Filip Braet’s research covers nanobiology, structural cell biology and cancer biology, culminating in developing targeted drug-delivery approaches for colorectal cancer (CRC), including CRC hepatic metastasis. His research uses combinatorial labeling approaches and correlative light & electron microscopy (CLEM) to determine the precise subcellular target(s) of these anti-cancer compounds at the supramolecular level. By drawing on these structural insights and mechanistic understanding,
A/Prof. Braet and his collaborators are designing and assessing new therapeutic strategies for treatment of cancer, such as hydrogel-based carriers for sustained, local application of anti-cancer drugs and metal-based nanoparticles, such as iron–gold complexes, that show selective activity towards cancer cells.
PROJECT 1Investigation of the cellular distribution of metal-based anti-cancer compounds by a correlative morphomics approachCells exposed to anti-cancer drugs exhibit varied cellular responses, such as structural reorganisation, complexation and aggregation of molecules and/or proteins or relocation and/or activation of subcellular compartments. In order to obtain an in-depth view of anti-cancer drug induced cell responses, systematic cellular analysis of the cells’ architecture and functional
machinery is a prerequisite. This can be achieved by combining CLEM with combinatorial labeling thereby permitting the direct correlation of dynamic subcellular events and macromolecules in 4-D with the ‘undisturbed’ nano-environment in 3-D on the same cell.
The aim of the first part of this project is to devise methodology and standardised protocols for quantifying (sub)cellular responses under varying experimental conditions in real-time (i.e., X, Y, Z & t). Besides devising high-throughput live-cell imaging protocols, the student will apply rapid-fixation and sample-relocation approaches for subsequent electron microscopy. In the second part, this methodology will be applied directly to clinically relevant human CRC cell lines exposed to different metal-based anti-cancer drugs. The work will lead to (i) a standardised CLEM model for drug screening purposes and (ii) better understanding of the action of metal-based anti-cancer drugs.
PROJECT 2The role of membrane rafts in anti-cancer drug treatmentMembrane rafts are key structural and molecular regulator for various natural compounds and/or drugs in many biomedical settings, including cancer. Our team recently developed established CRC Caco-2 cell clones that differentially express the membrane-associated raft marker GM1. In preliminary studies, we have demonstrated that this differential
The Australian Centre for Microscopy & Microanalysis (ACMM) is the largest and most comprehensive facility of its type in Australia, and it offers an exciting variety of research programs, spanning disciplines from archaeology to biology, from chemistry to engineering, and from medicine to physics.
TWO DOCTORAL (PHD) SCHOLARSHIP OPPORTUNITIESMEMBRANE AND CANCER CELL BIOLOGY (MCCB)
Colon carcinoma cells stained for actin (green), membrane rafts (red) and DNA (blue).
Endothelial cells labelled for actin (red) and nuclei (blue).
AUSTRALIAN CENTRE FOR MICROSCOPY & MICROANALYSIS
expression of the ganglioside GM1 resulted in a different cellular response to well-described anti-cancer drugs, such as cisplatin and doxorubicin. Therefore, these Caco-2
GM1- and
Caco-2GM1+
cell clones a unique cell-culture model for drug screening studies in which the uptake, transport and resistance to anti-cancer drugs rely on membrane rafts.
The overall aim of this project is to assess the involvement of membrane rafts in anti-cancer drug efficacy in further detail. The differential responses of the human CRC cell lines will be assessed by CLEM, by different viability and cytotoxicity assays, by cell-sorting analysis, by protein blotting, and by drug-accumulation
studies combined with transepithelial electrical resistance measurements. A central aspect of the work will include labeling studies with live-cell probes and fine-structure immunolabeling for electron microscopy. This project will result in a better understanding about the key-regulatory actions that membrane rafts have on existing and new anti-cancer drugs, particularly addressing whether membrane rafts or their associated molecules (i) facilitate or inhibit the uptake of drugs; and/or (ii) initiate a cascade of intracellular (cell-death) pathways; and/or (iii), constitute a platform for the regulation of multi-drug-resistant proteins.
CONTACT
A/Prof. Filip Braet Australian Centre for Microscopy & MicroanalysisMadsen Building F09The University of Sydney T +61 2 9351 7619F +61 2 9351 7682E [email protected]/acmm
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Correlative live cell and scanning electron microscopy (CLEM) observations on colorectal cancer cells exposed to anti-actin drugs.
