Marta Valenti is an EACR Travel Fellowship recipient who returned from the DREAM Lab Group at Aarhus University, Denmark in December 2022.
The EACR has joined forces with Worldwide Cancer Research to provide Travel Fellowships of up to €3,000 to enable early-career cancer researchers to gain new skills through a short-term visit to a lab or research group in another country.
You can read about other Travel Fellowship awardees and their experiences here.
Name: Marta Valenti
Job title: PhD Student
Home institute: Cancer Biotherapeutics Group, National Institute of Cellular Biotechnology, Dublin City University, Dublin, Ireland
Host institute: DREAM Lab Group, Department of Biomedicine, Aarhus University, Denmark
Dates of visit: 15 August – 02 December 2022
Research: Skin cancer, the most prevalent cancer in Ireland, showed in a clinical database (cBioPortal) a particularly high mutation frequency (17.34%) of a receptor belonging to the HER-family, known as HER4. The aim of my research is to investigate whether mutations in HER4 could predict response to an HER-family targeting drug called neratinib (NER), clinically used for HER2-amplified breast cancer. By using computational analysis, we identified HER4 mutations that may have a cancer-causing role in skin cancer. We then develop CRISPR/Cas9 genome-edited skin cancer cell lines harbouring these HER4 mutations to investigate their role in relation to NER response.
Why did you decide to apply for an EACR Travel Fellowship?
I applied for the EACR Travel Fellowship to learn a technique called “CRISPR/Cas9” essential for the completion of my PhD project. The Travel Fellowship provided an excellent chance to support my trip to Denmark and learn from experts who have the ultimate knowledge regarding optimal experimental set-up and troubleshooting.
Why did you choose this host lab?
Dr. Yonglun Luo is an established expert in the area of CRISPR/Cas9 (https://orcid.org/0000-0002-0007-7759), publishing most recently on optimising gRNA efficiency in CRISPR/Cas9 [PMID: 35637227]. Dr. Yonglun Luo has been collaborating with my supervisor Dr. Denis M. Collins as part of the Science Foundation Ireland project ACORN (20-SPP-3684) focusing on HER-targeting TKIs.
Can you shortly summarise the research you did and what you learned on your visit?
The main objective of my training in Dr. Yonglun Luo’s lab was to insert site-specific HER4 mutations in skin cancer (melanoma) cell line models by using CRISPR/Cas9. Given the limited time of the research trip, the work plan focused on two melanoma cell lines (WM115, SKMEL24). We performed preliminary experiments to check the efficiency of the CRISPR/Cas9-machinery delivery, and we designed and validated PCR primers and optimised a Sanger sequencing approach to assess the CRISPR/Cas9 efficiency of targeting.
We then single-cell sorted via FACS the pooled cell population subjected to CRISPR/Cas9 to perform a single-cell cloning assay. We monitored the single-cell for colony formation, and we then screened the single-cell colonies by PCR, and Sanger sequencing to isolate pure selection-free HER4-CRISPR/Cas9-modified skin cancer cell lines. The next stage of the project will consist in the in vitro functional characterisation of the isolated HER4-CRISPR/Cas9 knock-ins to evaluate their role as biomarkers of response to an HER-family targeting drug called neratinib (NER), currently used to treat HER2-amplified breast cancer.
“I will also provide organised training about the CRISPR/Cas9 technique to Ph.D. and postdoctoral students to ensure the knowledge is retained in my home institute”
Was the host institution very different from your own?
The Skou Building of the Biomedicine Department is a new and innovative research centre. It has a small canteen inside, which provides fresh food daily, and several coffee machines where you can get free hot beverages to either recharge yourself after a long experiment or have some quality time with your colleagues. I also really enjoyed having breakfast every Friday morning with my lab group; they called it “Friday bread”, and having beers or board games on Friday afternoon in the canteen during what they called “Friday bar”. A few times, I also joined and enjoyed the morning stretching exercises organised weekly by a group of people within the Skou Building; it was a very relaxing moment before the working routine.
What were you able to do that you could not have achieved in your home lab?
Thanks to the EACR Travel Fellowship, I had access to facilities and very expensive devices used to perform CRISPR/Cas9, a technique that we do not do in my home lab. Moreover, I had the opportunity to work in person and learn from experts in the CRISPR/Cas9 field like Dr. Yonglun Luo.
What was a personal highlight on your trip?
A personal highlight of my trip was building lovely friendships and strong relationships with colleagues that surely enriched my personal life beyond my career and PhD. It can seem scary to travel alone to a different country even for such a short period, however, meeting new people and building new friendships is very thrilling.
Did you take part in any interesting local or cultural activities?
During my stay in Aarhus, I really enjoyed visiting around and taking part in different local and cultural activities. Aarhus is a small but very cute city full of traditional and cultural festivals in the summer period, and different social activities all year. For instance, I really enjoyed feeding the deer in the Marselisborg Deer Park. Around Denmark, I was very impressed by Skagen, a small town at the north end of Denmark where the Baltic Sea and the North Sea meet.
Did you have a personal mentor or anyone who particularly helped you?
I would really like to thank Dr. Younglun Luo for helping and supporting me during my stay in his DREAM lab. I would also like to thank Trine Skov Petersen and Yan Zhou for all their support, and all the people of the DREAM lab group (too many to mention singularly) – I think everyone helped and supported me at least one time during these months in Dr. Younglun Luo’s lab. I am really glad I found such a friendly and supportive work environment, I am sure I will miss my Danish lab family!
Have you brought back any specific knowledge or technique that has benefited your home lab?
One of the reasons I joined Dr. Luo’s lab at Aarhus University in Denmark is because we do not perform the CRISPR/Cas9 technique in my home lab. Therefore, I will present the work carried out and the CRISPR/Cas9 technique learned in Denmark to my home group, and other groups in my department, and I will also provide organised training about the CRISPR/Cas9 technique to Ph.D. and postdoctoral students to ensure the knowledge is retained in my home institute.
Does your lab plan to do any future collaboration or publication with the host lab?
I think one of the keys of this trip was to build a strong collaboration between my home lab and the host lab. I am sure there will be future collaborations between Dr. Denis M. Collins and Dr. Younglun Luo. My work in Dr. Luo’s lab has already resulted in the submission of an abstract for the AACR 2023 annual meeting.
How has this visit been beneficial to your research and/or your career?
Learning the CRISPR/Cas9 technique provided me with a critical tool to properly interrogate the impact of HER4 mutations in cell line models without the over-expression of the proteins of interest, a common problem with techniques like transient or viral transfection. The opportunity to learn in person this technique from Dr. Luo was the best way to ensure I learned the technique correctly and as completely as possible.
Is there anything else you’d like to mention?
I would really like to thank my supervisor Dr. Denis M. Collins and Dr. Yonglun Luo for giving me this amazing opportunity. Dr. Denis M. Collins and Dr. Yonglun Luo were always able to guide/mentor me during these months in Denmark. I would also like to thank my funding bodies, Science Foundation Ireland (SFI) and PUMA Biotechnology, for financing this research project, and the EACR Travel Fellowship for supporting this trip to Denmark. Last, but not least, I would like to thank all the people and colleagues that made these months in Denmark remarkable.
If you are interested in applying for the Travel Fellowship scheme, please click here for more information: EACR Travel Fellowships.