Diagnostic leukapharesis (DLA) as tool to maximize the number of circulating tumour cells
Circulating tumour cells (CTCs) are acknowledged as important tumour biomarkers. However, their rarity (typically 1-10 in 1L of blood) and the small volume of blood that can be analysed from cancer patients (typically 10-20ml) impose severe limitations to assess their informative value. Our group has introduced diagnostic leukapheresis (DLA) as a powerful approach to enrich circulating tumour cells (CTCs) from litres of blood. This approach has now been adopted by several international groups and it has been instrumental in the activities of CTC-Trap and CANCER-ID European consortia. We believe that the higher number of CTCs that can be collected by this minimally invasive method provide unprecedented opportunities for CTC-based research and clinical applications in the sense of a true liquid biopsy. Our group is committed to further develop the technique and prove the clinical relevance of DLA-CTCs.
CANCER-ID is an European consortium that aims to validate technologies for Circulating Tumour Cells (CTCs), circulating tumour DNA (ctDNA), circulating free micro RNAs (cfmiRNAs) and exosome-bound miRNA as blood-based biomarkers to determine the absence/presence of drug targets and/or assess response to treatment in non-small cell lung carcinomas (NSCLC, large indication) and treatment refractory HER2-positive metastatic breast cancer (Her2RMBC, small indication).
In the project our group leads task 1.3 aiming at enumerating CTCs in diagnostic leukaphaeresis (DLA) products, and task 1.4 aiming at comparing isolation methods of single CTCs.
This project is funded by the Innovative Medicines Initiative Joint Undertaking (IMI JU) [Grant Agreement #115749].
The clonal evolution of systemic mCRPC cancer during therapy monitored by analysis of individual circulating tumour cells
In this project we aim to evaluate the potential of circulating tumour cells to follow the genetic evolution of castration-resistant prostate cancer (mCRPC) during systemic therapy. We aim to explore the utility of CTCs as source of molecular biomarkers of therapeutic resistance. For that we analyse the genetic makeup of multiple single circulating tumour cells isolated from blood samples collected at different moments of treatment and investigate changes in the clonal architecture of the CTC population. The objective is to identify molecular traces able to predict sensitivity and/or resistance to specific drugs.
This project is being conducted in cooperation with the group of Professor Johann de Bono at the Institute of Cancer Research (ICR, London, UK), and the group of Dr. Nicolò Manaresi at Menarini Silicon Biosystems spa (Bologna, Italy).
New tools for the enrichment of pancreatic circulating tumour cells from blood and DLA samples
Diagnostic leukapheresis (DLA) is a powerful approach to enrich circulating tumor cells (CTCs) from litres of blood. Nevertheless, due to the excess of white blood cells (WBCs) in DLA products, isolation of CTCs remains very challenging. Different technologies are available for the enrichment of circulating tumour cells (CTCs) from blood samples while their performance to enrich CTCs from DLAs is barely explored. In this project we aim to find strategies to better enrich CTCs from DLA products and to maximize the volume of DLA product that can be processed for CTC-enrichment. Working closely with different companies we test the performance of new technologies and participate actively in the development of workflows adapted to the specificities of DLA products.
This project is in part funded by the Brigitte und Dr. Konstanze Wegener Foundation [Project #01).
The epigenetic signature of circulating tumour cells
In this project we want to analyse the epigenetic signature of single circulating tumour cells. More concretely, we aim to analyse DNA methylation at the promoter of genes with the potential to contribute to the metastatic potential of CTCs. We want to explore the potential epigenetic heterogeneity of CTCs as tumour biomarker. For this project we have been using our in-house developed multiplexed single-cell agarose-embedded bisulphite sequencing (multiplexed-scAEBS) to analyse DNA methylation at single-CpG resolution from multiple genes from individual cells.