Projecten per jaar
The use of gene expression profiling (GEP) in cancer management is rising, as GEP can be used for disease classification and diagnosis, tailoring treatment to underlying genetic determinants of pharmacological response, monitoring of therapy response, and prognosis. However, the reliability of GEP heavily depends on the input of RNA in sufficient quantity and quality. This highlights the need for standard procedures to ensure best practices for RNA extraction from often small tumor biopsies with variable tissue handling. We optimized an RNA extraction protocol from fresh-frozen (FF) core needle biopsies (CNB) from breast cancer patients and from formalin-fixed paraffin-embedded (FFPE) tissue when FF CNB did not yield sufficient RNA. Methods to avoid ribonucleases andto homogenize or to deparaffinize tissues and the impact of tissue composition on RNA extraction were studied. Additionally, RNA’s compatibility with the nanoString nCounter® technology was studied. This technology platform enables GEP using small RNA fragments. After optimization of the protocol, RNA of high quality and sufficient quantity was obtained from FF CNB in 92% of samples. For the remaining 8% of cases, FFPE material prepared by the pathology department was used for RNA extraction. Both resulting RNA end products are compatible with the nanoString nCounter® technology.
|Nummer van het tijdschrift||5|
|Status||Published - mei 2021|
VingerafdrukDuik in de onderzoeksthema's van 'Overcoming the Challenges of High Quality RNA Extraction from Core Needle Biopsy'. Samen vormen ze een unieke vingerafdruk.
SRP48: Strategic Research Programme: Cancer Cell Targeting in Myeloma and Melanoma (MyMe)
Vanderkerken, K., Thielemans, K., Vanderkerken, K. & Breckpot, K.
1/11/17 → 31/10/24
IWTOO25: Baekelandmandaat: In situ mRNA vaccinatie: het gevecht naar de tumor brengen
Breckpot, K., Thielemans, K. & Locy, H.
1/01/18 → 30/11/22