UNIVERSITY OF MASSACHUSETTS
My lab studies the circuitry of cellular signaling pathways and one of our long-term goal is to define which connections are critical in vivo to the progression of human cancer and to use that knowledge to facilitate the development of targeted therapies. To this end, my lab studies the mTOR kinase, a central player in a signaling network that is increasingly viewed as crucial to the initiation and progression of many human cancers. The cellular functions of mTOR are mediated through its interaction with regulatory proteins in two distinct complexes, called mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). As part of mTORC1, mTOR functions in the canonical rapamycin-sensitive cell growth pathway. It does so in part by regulating the S6 kinase and 4E-BP1 proteins, both of which control protein synthesis. Considerably less is known about mTORC2, which we discovered more recently. mTORC2 phosphorylates and activates the Akt kinase, which is the major effector of the PI3K signaling pathway. I subsequently generated mice deficient for mTORC2 activity and obtained genetic evidence confirming the role of mTORC2 in Akt regulation. The PI3K pathway is commonly activated in human cancer and intense efforts in the pharmaceutical industry are underway to develop inhibitors of this pathway. A primary goal of my work is to determine the role of mTORC2 in cancer, particularly cancer drive by elevated PI3K pathway activation. To do this, we use genetically engineered mice and human cancer cell lines to study the critical roles of mTORC2 in tumor initiation and progression. The purpose of this award is to purchase a state-of-the art microscope, cameras, and workstation capable of high-resolution brightfield and epifluorescence microscopy. We study cell and tissue samples derived from our genetically engineered mice as well as human cancer cells in our research. Part of our methodology involves extensive histological analysis to study individual fixed cells and proteins within tissues and cells. These analyses include simple stains such as Hematoxylin, Eosin, Massons trichorme and basic immunohistochemical assays for proliferation and apoptosis, such as TUNEL assay, Caspase Cleavage stain, and Ki67 stains. In addition, we use many specialized immunohistochemical and immunofluorescence assays to study specific proteins that are relevant to our research. Furthermore, we are developing assays to look at live cells, which requires a sophisticated microscopy system.