Monique van Eijndhoven

 

Research Technician

Exosomes Research Group

www.exosomes.nl

VU University Medical Center (VUmc)

Cancer Center Amsterdam (CCA)

Department of Pathology

 

 

E-mail:

m.vaneijndhoven@vumc.nl

 

 

Telephone:

+31 (0)20 4441776

Fax:

+31 (0)20 4442964

 

 

Address:

VUmc, CCA, room CCA3.04

De Boelelaan 1117

1081HV Amsterdam

The Netherlands

Project number 3.1

Dr. D.M.Pegtel, VU University Medical Center, Amsterdam

Prof. Dr. G.W. Jenster, Department of Urology, Erasmus MC, Rotterdam

Project goals:

Determination of the molecular content of EVs derived from biofluids

Analysis of the genetic content of EV for cancer biomarker discovery

Circulating EVs contain genetic information that reflect tumor cells raising possibilities for non-invasive tumor status diagnosis and monitoring. Previous studies show that NGS can identify subtle changes in ncRNA expression, modification and processing all of which can have a role in cancer development and may thus be considered for biomarker studies. These studies however were performed with suboptimal sample preparations (ultracentrifugation, precipitation) resulting in the co-isolation of contaminating non-EV non-cancer derived RNAs. To overcome this we have recently turned to size-exclusion chromatography, a method that separates urine and plasma EVs from protein complexes and lipid particles. This approach yields high recovery, purity and quality of EV RNA, which is essential for obtaining robust NGS data from patient samples.

Tasks:

·

Optimize current protocols and develop novel methods to separate and purify EVs from frozen urine and serum/plasma repositories and fresh samples

·

Develop a combined small and long RNA sequencing protocol optimized for EV RNA

·

Molecular characterization of EV RNA and validation of diagnostic and prognostic profiles

·

Develop and test gene expression profile detection assays

Figure 1: Schematic of size exclusion chromatography. EVs are collected in fractions 9-10, protein/HDL in fractions 20-21.

Figure 2: smallRNAs are differently distributed when comparing circulating EVs with protein/HDL

Collaboration:

Ibiomics Peter Beyerlein, Berlin, Germany

BitBrains Gjalt van Rutten, Amstelveen, The Netherlands

Cyto-Barr Jaap M. Middeldorp, Zuidhorn, The Netherlands

ExosomeDX Johan Skog, New York, USA