ABSTRACT
First Person is a series of interviews with the first authors of a selection of papers published in Journal of Cell Science, helping researchers promote themselves alongside their papers. Anu Prakash is first author on ‘ Centrosome amplification promotes cell invasion via cell–cell contact disruption and Rap-1 activation’, published in JCS. Anu conducted the research described in this article while a PhD student in Dr Emer Bourke's lab at University of Galway, Ireland. She is currently looking for a job in the pharmaceutical industry. Her research interests are focused on clinical research and drug discovery to develop novel treatments for human diseases.
Anu Prakash
How would you explain the main findings of your paper in lay terms?
The centrosome (a tiny cell component) directs normal cell shape and accurate division of one cell into two. We studied how a cellular process called centrosome amplification (CA), which involves cells having too many centrosomes, can lead to cancer.
In non-cancerous breast cells, inducing CA made them more mobile and invasive. We discovered that a protein called Rap-1 became activated during CA, contributing to these cancer-like behaviors. CA also disrupted how the cells stuck together and affected the expression of proteins involved in cell connections. At the same time, CA caused these cells to attach more to surrounding tissue and increased the production of enzymes that break down tissue, such as matrix metalloproteinases (MMPs), aiding in cancer cell spread.
Using a chicken egg in vivo model, we found that cells with CA invaded into surrounding tissue and additionally generated cancer characteristics like dilation of blood vessels and abnormal cell crowding (hyperplasia). Blocking Rap-1 minimised these effects. By contrast, inhibiting CA in breast cancer cells reduced their ability to spread.
In summary, our study shows that CA can turn normal cells into cancer-like ones by altering their behavior and interaction with tissues, without them needing additional pro-tumorigenic changes.
Were there any specific challenges associated with this project? If so, how did you overcome them?
A big challenge for this project was setting up a new in vivo assay, the chicken egg model. We realised that the work would be improved by validation in vivo, and this model was an excellent way to do that (within a reasonable timeframe and, importantly, budget). It was something new both for myself and my PI. I had to acquire and master many of the new techniques needed, from learning to implant cancer cells into the egg, to learning how to remove the tumour and prepare it for pathology. Learning to analyse the results from the assay also added to the challenges, and allowed me to master new methods of 3D imaging and analysis. I was able to get technical advice from a colleague who we discovered was also using this assay. I also used papers and online videos (material from the Journal of Visual Experiments was helpful) to help plan and master techniques. YouTube videos, which explain and teach many techniques and analysis methods, are a very much underappreciated scientific resource for young researchers.
When doing the research, did you have a particular result or ‘eureka’ moment that has stuck with you?
My most memorable ‘eureka’ moment was when using our new in vivo model, I found that centrosome amplification all on its own was able to promote early tumorigenic changes, including angiogenesis. This was a completely surprising and entirely novel finding that had not been reported before. It played a crucial role in facilitating new collaborations (nationally and internationally). This was so exciting, not only as a new discovery, but also as a validation of all my hard work setting up the new model. It came during a tough time in my research, and it reminded me I really could be a researcher that makes exciting discoveries that others would be interested in, which really motivated me.
Why did you choose Journal of Cell Science for your paper?
We chose JCS because the papers it publishes are very well regarded, technically sound and impactful. Other key factors in our choice were the fact that it is a not-for-profit journal, its great reputation for supporting the scientific community, and its support and use of bioRxiv and Review Commons.
Fluorescent microscopy image of MCF10A cells. The cells show normal (≤2 per cell) and amplified centrosomes or CA (≥3 per cell) post treatment with the CDK1 inhibitor RO3306. Immunofluorescence stained for microtubules (α-tubulin, red) and centrosomes (γ-tubulin, green), and counterstained with DAPI (DNA, blue).
Fluorescent microscopy image of MCF10A cells. The cells show normal (≤2 per cell) and amplified centrosomes or CA (≥3 per cell) post treatment with the CDK1 inhibitor RO3306. Immunofluorescence stained for microtubules (α-tubulin, red) and centrosomes (γ-tubulin, green), and counterstained with DAPI (DNA, blue).
Have you had any significant mentors who have helped you beyond supervision in the lab? How was their guidance special?
I am fortunate to have Dr Emer Bourke as my mentor, whose unwavering support for innovative ideas and emphasis on methodical problem-solving has greatly influenced my approach to research. Dr Bourke was also able to give me excellent career advice, which led me to follow my interests into clinical trials.
Dr Bourke focused on helping me build my own professional network, allowing me to coordinate and work with our collaborators (Dr James Brown, University of Limerick, and Dr Ann Hopkins, Royal College of Surgeons in Ireland, Dr Susana Godinho, Queen Mary University, UK) and helped me to discuss our work as the leader of our team, which allowed me to learn how to manage a project with multiple partners, and showed me how, when we work together, we can tackle big problems.
What motivated you to pursue a career in science, and what have been the most interesting moments on the path that led you to where you are now?
My sister was my first biology teacher; she introduced me to the subject of genetics, explaining how our physical traits and characteristics are shaped by the transmission of genes from our parents. One particular narrative that deeply intrigued me was about Dolly the Sheep, ultimately inspiring me to pursue both undergraduate and postgraduate degrees in genetics. However, the turning point in my career happened during my internship at the Adyar Cancer Institute in India. I witnessed the challenges faced by cancer patients, both physically and emotionally. This experience motivated me to obtain a PhD in cancer biology and use my knowledge to improve patient health. My passion for clinical research continues to drive my career forward.
Who are your role models in science? Why?
My role models have been my main supervisor, Emer, but also our collaborators and scientists I have worked with. I took elements from each of them, learning from their different strengths, and seeing how they helped me and others.
Emer, my supervisor, inspired me with her professionalism and collaborative spirit. She not only taught me scientific research, but also promoted my professional development. As a woman, I see how she has overcome barriers to become a scientist that I have modeled myself after. I hold a deep admiration for women in science, and always draw inspiration from researchers and scientists who work hard to contribute to science and the betterment of humanity.
What's next for you?
I left academia to work on clinical trials, which I found very interesting, and which allowed me to gain new specific skills. However, research kept calling to me and I followed this with a post-doctoral experience from the National Institutes of Health (NIH).
Tell us something interesting about yourself that wouldn't be on your CV
I'm an ardent pet lover and support pet animal rescue organizations that save innocent lives from abusive environments.
E-mail: [email protected]
