Human Cancer Genetics Programme

Molecular Cytogenetics Unit

Head of Unit:  Sandra Rodríguez
Research highlights
Optimising CRISPR-Cas9 to model cancer aberrations in primary cells

In vitro modelling of complex tumour-associated chromosome translocations at native loci is feasible with CRISPR. However, the generation of translocations must be optimised, especially for mimicking events in human primary cells. We have optimised our CRISPR protocol to efficiently obtain those cells, thereby enabling the rescue of translocation+ populations of human primary cells, including induced pluripotent stem (iPS) cells and mesenchymal stem cells (MSCs). These models can surely help us to understand the molecular mechanisms underlying the initiation of human cancers, and can also be used for high-throughput drug screening, toxicological testing and biomarker identification.

From the patient’s chromosome translocations to their functional effects

We have worked on the oncogenic role of the translocation t(8;21)(q22;q22)/RUNX1-RUNX1T1, which occurs in 4% of acute myeloid leukaemia patients. We deciphered a new function for the activation of MAPK8, observed in t(8;21)+ cells, which is responsible for the stabilisation of SP1. Our data show the essential role of SP1 in t(8;21)+ cell maintenance through the regulation of key genes, such as CDKN1A. These results provide new evidence for the inclusion of pharmacological approaches leading to degradation of SP1 in the treatment of these patients.

Technological and translational activities

We provide state-of-the-art molecular cytogenetics and genome editing services. The Unit makes available various techniques to the CNIO Research Groups; these techniques provide more sensitive and accurate tools to analyse cancer cells, such as RNA-FISH, chromosome stability studies based on a combined array CGH-FISH approach, or the use of CRISPR libraries to perform high-throughput functional analysis. For gene editing experiments, we have set up a specific FISH analysis to detect genomic integration sites of small constructs including LV particles. In 2016, we carried out over 1,000 assays for experimental and clinically-oriented projects.