EACR Travel Fellowships are co-sponsored by Worldwide Cancer Research and provide funds up to €3,000 to early-career cancer researchers. For more information on how to apply for Travel Fellowships, you can visit the EACR website.
Home institute: IPATIMUP/I3S and Medical Faculty, University of Porto, Porto, Portugal
Host Institute: Glyco-Oncology, Ovarian Cancer Research, Department of Biomedicine, University Hospital Basel and University of Basel, Basel, Switzerland
Dates of visit: 01 September 2017 – 21 December 2017
Ovarian cancer (OvCa) is the leading cause of cancer death among gynecological malignancies. Peritoneal dissemination is a particular form of metastisation of OvCa, in which cancer cells detach from the primary tumour in the ovary or in agreement to recent proposals in the fallopian tube, disseminate in the peritoneal fluid and implant across the mesothelial layer throughout the entire abdomen, resulting in the formation of peritoneal carcinomatosis.
My PhD thesis is focused on disclosing the role of mucin 16 (MUC16) and Mesothelin (MSLN) and of their interaction in ovarian cancer dissemination. Evidences indicate that the interaction between MUC16 and MSLN, expressed both in cancer cells and in the peritoneal mesothelial lining, plays a crucial role in the adhesion of ovarian cancer cells to the mesothelium covering the entire peritoneal surface. To disclose the role played by these two proteins, I generated ovarian cancer cell lines with downregulation of MUC16 and MSLN expression. Then, we decided to perform functional experiments using in vitro and in vivo models and we observed that the reduction of MUC16 expression decreases the proliferation capacity and increases cell death. On the other hand, downregulation of MSLN expression decreases in vitro cell invasion and in vivo peritoneal dissemination/ascites formation.
During my stay in the lab I was welcomed in the friendliest and most dedicated way. All the lab members helped me in every aspect during my stay.
To expand the data obtained with the downregulation experiments, we decided to generate cell lines with knockout for MUC16 and MSLN using CRISPR/Cas9. To generate these cell lines, we established a collaboration with Dr. Francis Jacob at University Hospital Basel and University of Basel, Switzerland. The group of Dr. Jacob has recently used the CRISPR/Cas9 system in ovarian cancer cells to disrupt glycoprotein/glycolipid integrity. The purpose of the visit was to generate CRISPR/Cas9 mediated MUC16and MSLN knockout cell lines. In order to do this, we applied the paired sgRNA strategy by designing two sgRNAs targeting either MUC16 or MSLN and cloning into pSpCas9 (BB)-2A-GFP vector. Constructs were initially tested for Cas9 activity in transiently transfected HEK293T cells and further applied to ovarian cancer cell lines. We successfully established various knockout cell lines verified on genomic DNA and protein level by genotyping/ DNA Sequencing and Western blot/ FACS/ immunofluorescence, respectively. In order to rescue biological effects of MSLN in vitro and in vivo, we lentivirally transduced ΔMSLN cells for constitutive expression of MSLN.
The whole experience was extremely rewarding as it taught me new and helpful techniques for my future research. This experience also allowed acquaintance with inspiring scientists that shared new ideas and perspectives with me.
During my stay in the lab I was welcomed in the friendliest and most dedicated way. All the lab members helped me in every aspect during my stay. Special thanks goes to Dr. Jacob for sharing his knowledge and for his willingness to discuss the many intricacies of this field, which gave me the opportunity to expand my future research. I enjoyed all the scientific discussions that we had. The results we obtained at the lab are essential to characterise the role of MUC16 and MSLN in the peritoneal dissemination of ovarian cancer. The whole experience was extremely rewarding as it taught me new and helpful techniques for my future research. This experience also allowed acquaintance with inspiring scientists that shared new ideas and perspectives with me.
I am very grateful for the EACR Travel Fellowship that made this experience possible. I am very glad to have had the opportunity to visit an incredible country and work in a high standard laboratory environment.