ABSTRACT

First Person is a series of interviews with the first authors of a selection of papers published in Journal of Cell Science, helping early-career researchers promote themselves alongside their papers. Ameer Elaimy is first author on ‘Real-time imaging of integrin β4 dynamics using a reporter cell line generated by Crispr/Cas9 genome editing’, published in JCS. Ameer is a MD/PhD student at University of Massachusetts Medical School in the lab of Arthur M. Mercurio, where he works on identifying mechanisms that contribute to tumorigenesis and therapy resistance in breast cancer.

Ameer Elaimy (left) and Art Mercurio (right) celebrating following Ameer's successful PhD defense.

How would you explain the main findings of your paper in lay terms?

The β4 integrin plays a critical role in cell migration and thereby contributes to mammary biology and breast cancer progression. Work from our group and others has suggested that migrating carcinoma cells rapidly upregulate β4 expression. However, these observations were based primarily on immunofluorescence microscopy of fixed cells using antibodies and not real-time imaging in live cells. To more rigorously address the role of β4 in cell migration, our study pioneered the use of Crispr/Cas9 genome editing to tag endogenous β4 and visualize, in real-time, β4 dynamics in carcinoma migration. This tool has numerous potential applications in the study of β4 in mammary development as well as in breast tumor initiation and metastasis.

Were there any specific challenges associated with this project? If so, how did you overcome them?

Using Crispr/Cas9 technology to knock-in fluorescent tags to endogenous genes comes with several challenges. We initially selected the location of the endogenous β4 gene to insert the tag and designed sgRNAs targeting that specific region, which are efficient in cutting to achieve homologous donor recombination. In addition, we engineered the corresponding donor plasmid so that Cas9 does not cut the endogenous β4 locus. Finally, we performed several experiments establishing that the cytoplasmic β4 tag does not alter its function or cell physiology. We overcame these challenges in genome editing and cell biology by assembling a uniquely qualified group of scientists to troubleshoot issues as they arose.

When doing the research, did you have a particular result or ‘eureka’ moment that has stuck with you?

Indeed, we did. We were very excited about the live-cell movies we obtained that showed that YAP-transformed comma-d1 cells rapidly upregulate β4 in response to a scratch-wound. This is because this information is consistent with our prior work and it substantiated the potential applications of the tool we developed, which can be used to monitor β4 in developmental and tumorigenic processes in real-time.

Why did you choose Journal of Cell Science for your paper?

JCS has an outstanding reputation for publishing impactful papers in all aspects of cell biology. Given the numerous applications of our Tools and Resources article in cell and cancer biology, we felt that our work was most appropriate for the JCS readership.

Have you had any significant mentors who have helped you beyond supervision in the lab? How was their guidance special?

My PhD mentor, Art Mercurio, has provided me with outstanding mentorship not only during my PhD years, but also throughout my entire career at UMASS Medical School in the MD/PhD program. He worked with me to develop this challenging project and provided the necessary support and motivation when things became difficult. His unique combination of knowledge and patience is unmatched and I am grateful for his mentorship.

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?

I initially pursued research in clinical oncology during my undergraduate years at Cancer Care Northwest and the University of Washington School of Medicine. This sparked an interest in the molecular basis of cancer, and I spent two years after college working in a laboratory in the Department of Radiation Oncology at the University of Michigan Medical School. This experience taught me the importance of translational research and how it can be applied to improvements in patient care. I subsequently matriculated into the MD/PhD program at UMASS Medical School, which is an outstanding institution to pursue translational cancer research.

Live-cell image of migrating integrin β4 reporter cells. Example images at 6 h and 12 hours are shown. Arrows indicate an example of a integrin β4-positive cell migrating more rapidly than neighboring cells.

Live-cell image of migrating integrin β4 reporter cells. Example images at 6 h and 12 hours are shown. Arrows indicate an example of a integrin β4-positive cell migrating more rapidly than neighboring cells.

What's next for you?

I am planning to pursue a physician-scientist residency in radiation oncology following my MD/PhD training. My ultimate goal is to translate laboratory findings into improvements in the care of cancer patients.

Ameer Elaimy's contact details: UMASS Medical School, 55 Lake Ave N, Worcester, MA 01655 E-mail: ameer.elaimy@umassmed.edu

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