Research

Endocytosis and the Endocytic Cytoskeleton

While we have a general interest in the process of endocytosis, we place a specific emphasis on Clathrin-Mediated Endocytosis (CME). A key question about this process is whether it is a "monolithic" event with highly stereotypic behavior or a variable process with multiple functional variations. One obvious way to test this hypothesis is to study the internalization of various receptors across different cells and physiological conditions. However, tools capable of measuring each individual receptor are not widely available. To tackle this issue, we developed a general strategy to study receptor internalisation (). Currently, we are exploring the use of synthetic protein binders generated with BindCraft and RFdiffusion to further refine our technology.

We hypothesize that the actin cytoskeleton is a key factor providing robustness or variability to endocytosis. However, many questions remain regarding how the cytoskeleton surrounding endocytic structures is formed, organized, and regulated. We detailed many of these questions in our recent review (), where we proposed the term “endocytic cytoskeleton.” Beyond its importance in endocytosis, the endocytic cytoskeleton serves as an excellent system for understanding general actin biology, as the formation of these actin networks has well-defined start and end points. We currently have several projects in the lab aimed at tackling these fundamental questions.

Reticular Adhesions

We have recently discovered that flat clathrin lattices are required for the formation of reticular adhesions (). We also found that the extracellular matrix protein fibronectin inhibits reticular adhesions via activation of its receptor (integrin α5β1). Moreover, we showed that the onset of cell migration is accompanied by the disappearance of these structures. Reticular adhesions were only recently discovered (Lock, NCB 2018) and they differ from typical focal adhesions for their higher stability, lack of direct connection to the actin cytoskeleton and absence of classical adhesion markers (paxillin/talins/FAK and etc). Results from our lab and from others suggests that reticular adhesions are markers of non-migrating cells. One could say that reticular adhesions are counter-migratory structures. We have a series of projects in the lab trying to understand the details of how these structures are formed, regulated and their role in cell migration during development and cancer.

Others

It would be disingenuous to say that we don’t dabble on other projects. We are curious and creative people and we like to try new things and develop new methods. We are not afraid to follow our noses into directions that are not the core of our science. One of these directions include the use of synthetic protein binders.  Recently, we have developed a tool to facilitate the generation of such binders ()

Our philosophy: Be kind

In my lab, we work as a family. I enforce respect, cooperation and kindness towards lab members and everyone around us (). I actively participate in the projects by doing experiments, data analysis and regular discussions on results. I encourage cross-project collaborations, a relaxed atmosphere, and a healthy work-life balance. I know that relaxed and engaged people produce better science.

Typical techniques we use (or have used) in the lab

CRISPR/Cas9, microscopy (confocal, live cell imaging, super-resolution microscopy), computational image analysis, cell migration and invasion assays, protein biochemistry, FACS, in vitro reconstitutions, Crystallography, CryoEM, and Bioinformatics.