The mechanistic target of rapamycin (mTOR), an evolutionarily-conserved protein kinase, coordinates signaling networks to regulate fundamental cellular processes including cell growth and proliferation, cell metabolism, cell survival, immunity, and ageing. Dysregulation of mTOR complex (mTORC) signaling contributes to myriad diseases including diabetes, obesity, cardiovascular disorders, and tumorigenesis. Indeed, clinicians employ mTOR inhibitors for immunosuppression after organ transplantation, for suppression of coronary artery stent restenosis after angioplasty, and for treatment of kidney cancer. Despite the clear physiologic and therapeutic importance of mTOR, fundamental gaps exist in our scientific knowledge of cellular mTOR regulation, especially with regard to the full set of molecular pathways and mechanisms that regulate mTOR activity in response to diverse signals. Exciting work from our laboratory revealed that mTOR phosphorylation plays an important and previously unrecognized role in mTORC1 function. Using phospho specific antibodies and an in vitro kinome screen as innovative tools, we discovered that the non-canonical IKK (IkB kinase)-related kinases TBK1 and IKKe phosphorylate mTOR directly on S2159, which occurs in vitro, in cultured cells, and in vivo. Our preliminary data indicate that in several cell types (i.e. MEFs;HEK293;RAW264.7 macrophages) in response to pathogen-associated inflammatory signals (i.e. bacterial LPS;dsRNA) and a subset of growth factors (i.e. EGF but not insulin), TBK1/IKKe-promotes mTORC1 and mTORC2 signaling. Importantly, TBK1/IKKe-driven mTORC1 signaling requires mTOR S2159 phosphorylation and induces IFNb production, an important early event in the host response to microbial infection. These preliminary data present the exciting hypotheses that TBK1 and IKKe function as novel mTORC1/2 activators and that mTORC1/2 represent novel TBK1 and IKKe substrates.
In Aim 1 we will elucidate biochemical mechanisms by which TBK1/IKKe promotes mTORC1 and mTORC2 signaling;
in Aim 2 we will identify upstream signaling intermediates that mediate the action of TBK1/IKKe on mTORC1 and mTORC2;and in Aim 3 we will understand cellular functions controlled by TBK1/IKKe action on mTORC1 and mTORC2. This research in cultured cells represents an essential first step towards the identification of novel drug targets and the development of rationally-designed therapeutic agents to treat clinical disorders linked to aberrant mTOR and TBK1/IKKe network action, such as inflammatory and autoimmune diseases, diabetes, obesity, and cancer.

Public Health Relevance

The protein kinase mTOR (mechanistic target of rapamycin) responds to diverse environmental stimuli to control essential cellular functions that participate in metabolism, cardiovascular disorders, tumorigenesis, and immune function. Despite central roles played by mTOR in cellular and organismal physiology, and its therapeutic importance, fundamental gaps exist in our knowledge regarding cellular mTOR regulation and function. This project investigates the regulation and function of mTOR complex 1 (mTORC1) and mTORC2 by novel upstream molecules and pathways. This research will lead to the identification of novel drug targets and development of rationally-designed therapeutic agents to treat the myriad clinical disorders associated with aberrant mTOR signaling.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
1R01DK103877-01
Application #
8800805
Study Section
Cellular Aspects of Diabetes and Obesity Study Section (CADO)
Program Officer
Silva, Corinne M
Project Start
2014-09-10
Project End
2018-06-30
Budget Start
2014-09-10
Budget End
2015-06-30
Support Year
1
Fiscal Year
2014
Total Cost
$209,813
Indirect Cost
$74,813
Name
University of Michigan Ann Arbor
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109