Abstract: The actin cytoskeleton shapes cells and also organizes internal membranous compartments. In particular, it interacts with membranes for intracellular transport of material in mammalian cells, yeast or plant cells. Tubular membrane intermediates, pulled along microtubule tracks, are formed during this process, and destabilize into vesicles. While the role of actin in tubule scission is still debated, literature also provides examples of actin-mediated stabilization of membranous structures.
To directly address this apparent contradiction, we mimic the geometry of tubular intermediates with preformed membrane tubes. The growth of an actin sleeve at the tube surface is monitored spatio-temporally. Depending on network cohesiveness, actin is able to entirely stabilize, or locally maintain membrane tubes under pulling. Indeed, on a single tube, thicker portions correlate with the presence of actin. Such structures relax over several minutes, and may provide enough time and curvature geometries for other proteins to act on tube stability.
In parallel, we developed a new platform to study the local mechanics of membrane tubes using atomic force microscopy (AFM). On a single coverslip we quickly generate millions of substrate-bound membrane tubes, out of which dozens can be imaged by AFM in a single experiment. In addition, we mimic in vivo actin polymerization on tubes, and use AFM to assess the induced changes in mechanical properties

Abstract: Kanako has extensive experience in gynecological diseases including endometriosis and ovarian/endometrial cancer, uterine biology and reproductive toxicology. Furthermore, she has a good track record for leadership of research teams, which are necessary to successfully carry out the proposed study. Kanako, also has a broad background in cellular, molecular, genetic and epigenetic techniques to understand etiology and mechanisms of the diseases using animal models, primary human tissues and cells, and data sets from the public sources. Also expertise in the targetable signaling mechanisms for developing therapeutic strategies. For example, we have published our findings in Oncogene 2015 and Biol Reprod 2016 and 2018. In those studies, we have found not only the signaling mechanisms of the disease but also therapeutic target(s) by small molecule(s) in the signaling pathway. Recently, the team and Union Therapeutics A/S in Denmark have agreed to further develop niclosamide for clinical use for treatment of endometriosis through a licensing agreement. The first clinical trial has been planned and pre-clinical pharmacokinetics and toxicokinetics studies for future phase II and III trials are currently ongoing. Thus, we have been successfully expanding potential therapeutic drugs in a pre-clinical setting to clinical translational research. In addition, laying the groundwork for the research by developing necessary animal and cell culture models. In the past, we have developed a new mouse model to examine and understand etiology of endometrial cancer. Together these studies have been well received, and as a result, in 2014 Kanako was named the recipient of New Investigator Award from The Society for the Study of Reproduction, and has successfully collaborated with many topnotch researchers and produced several peer-reviewed publications from each project