Characterization of cellular behavior within hydrogels / Investigation of RNA mis-splicing in Glioblastoma and its manipulation with CRISPR-dCas13 as a future therapeutic strategy

Characterization of cellular behavior within hydrogels

Summary: Although metastases are responsible for the majority of deaths associated with cancer, they are the least understood phase of the disease. Thus, there is an urgent need for reliable models for their study. Our goal is to develop an organotypic hydrogel to study breast cancer metastasis. I will present the rheological characterization of the gels and, as a proof of concept, the study of the behavior of an invasive breast cancer line within these gels.

Investigation of RNA mis-splicing in Glioblastoma and its manipulation with CRISPR-dCas13 as a future therapeutic strategy

Glioblastoma (GB) is the most common and lethal primary brain tumour, whose aetiology remains unclear. To better understand the biological mechanisms underlying GB, we study RNA mis-splicing (AS) and its contribution to GB malignancy.

Bioinformatics analysis of RNA-seq data from the Chinese Glioma Genome Atlas (CGGA) identified hundreds of differentially expressed splicing events. According to previous studies, our analysis reveals a mutually exclusive exon (MXE) in PKM gene, which encodes Pyruvate kinase M1/M2, which is differentially regulated in GB. To validate and further characterize these findings, we employ GB samples from Donostia University Hospital, including blood samples and tumour biopsies from which we isolate Glioma Stem Cells (GSCs). Our final objective is to manipulate RNA splicing as a therapeutic strategy in GB. To this end, we employ RNA-targeting CRISPR-dCas13 system, where in vitro preliminary experiments indicate that this system can be employed to switch PKM1/2 splicing. Further experiments are planned to further explore its therapeutic potential in GB models.