Studies fronn the laboratory ofthe PI have focused on elucidating the components, mechanisms, and roles of bioactive sphingolipids, especially ceramide with its emerging roles In regulating cell growth, differentiation, senescence, and inflammation. However, the study of these pathways has been complicated by their multiplicity, metabolic interactions, sub cellular compartmentalization, and lack of molecular tools. These considerations led us to the overall hypothesis that the specific cellular functions of ceramide are dictated by the sub cellular site of ceramide generation which in turn depends on which of the enzymes of ceramide metabolism regulates its site-specific formation. In this proposal we will focus on the neutral sphingomyelinase (SMase) pathway focusing on an extended family of enzymes identified by the PI and others. Specifically, we will address the following alms: 1) To define cellular mechanisms of regulation of nSMases. We will investigate the specific hypothesis that defined signaling pathways lead to the activation/regulation of n-SMase2 in response to TNF and retinoic acid. We will also study the mechanisms by which oxidative stress regulates the activation of ma-nSMase, which we recently identified as a novel mitochondrial sphingomyelinase. 2) To determine molecular and biochemical mechanisms of activation of nSMases. We will determine the mechanisms by which nSMase2 and ma-nSMase interact with phosphatidylserine and cardiolipin, respectively, as activating lipids, and we will determine the mechanisms of phosphorylation and palmitoylation and their roles in activation and translocation of nSMases. 3) To determine the role of nSMases in generating a specific 'pool'of ceramide with specific downstream functions. We will test the specific hypothesis that nSMase2 and ma-nSMase act on a distinct pool of sphingomyelin that resides In the inner leaflet ofthe plasma membrane and in the outer leaflet ofthe mitochondrion, respectively, leading to the corresponding subset of ceramide, with its own specific functions. Taken together, these results should provide a foundation for the study of molecularly-distinct sphingolipid-mediated pathways of cell regulation. Moreover, molecular studies on nSMase2 and ma-nSMsae would also impact our understanding of the mechanisms of an emerging and novel family of phospholipid phosphodiesterases. These pathways are of direct significance to the biology of stress responses and specifically to cancer biology and inflammation.

Public Health Relevance

The PI has identified a novel family of enzymes that regulate the formation of important lipid (fat) molecules that function in transmitting and modulating stress signals to cells. Understanding these pathways is important for our understanding of lipid-mediated biology, membrane and vesicular trafficking, human leukemia, osteogenesis imperfecta (a bone disease that should shed light on osteoporosis), and cancer therapeutics.

Agency
National Institute of Health (NIH)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37GM043825-25
Application #
8667453
Study Section
No Study Section (in-house review) (NSS)
Program Officer
Chin, Jean
Project Start
Project End
Budget Start
Budget End
Support Year
25
Fiscal Year
2014
Total Cost
Indirect Cost
Name
State University New York Stony Brook
Department
None
Type
Schools of Medicine
DUNS #
City
Stony Brook
State
NY
Country
United States
Zip Code
11794
Rajagopalan, Vinodh; Canals, Daniel; Luberto, Chiara et al. (2015) Critical determinants of mitochondria-associated neutral sphingomyelinase (MA-nSMase) for mitochondrial localization. Biochim Biophys Acta 1850:628-39
Spincemaille, Pieter; Matmati, Nabil; Hannun, Yusuf A et al. (2014) Sphingolipids and mitochondrial function in budding yeast. Biochim Biophys Acta 1840:3131-7
Truman, Jean-Philip; García-Barros, Mónica; Obeid, Lina M et al. (2014) Evolving concepts in cancer therapy through targeting sphingolipid metabolism. Biochim Biophys Acta 1841:1174-88
Airola, Michael V; Tumolo, Jessica M; Snider, Justin et al. (2014) Identification and biochemical characterization of an acid sphingomyelinase-like protein from the bacterial plant pathogen Ralstonia solanacearum that hydrolyzes ATP to AMP but not sphingomyelin to ceramide. PLoS One 9:e105830
Wei, Jianwen; Ferron, Mathieu; Clarke, Christopher J et al. (2014) Bone-specific insulin resistance disrupts whole-body glucose homeostasis via decreased osteocalcin activation. J Clin Invest 124:1-13
Airola, Michael V; Hannun, Yusuf A (2013) Sphingolipid metabolism and neutral sphingomyelinases. Handb Exp Pharmacol :57-76
Barbosa, Antonio Daniel; Osorio, Hugo; Sims, Kellie J et al. (2011) Role for Sit4p-dependent mitochondrial dysfunction in mediating the shortened chronological lifespan and oxidative stress sensitivity of Isc1p-deficient cells. Mol Microbiol 81:515-27
Clarke, Christopher J; Wu, Bill X; Hannun, Yusuf A (2011) The neutral sphingomyelinase family: identifying biochemical connections. Adv Enzyme Regul 51:51-8
Milhas, Delphine; Clarke, Christopher J; Idkowiak-Baldys, Jolanta et al. (2010) Anterograde and retrograde transport of neutral sphingomyelinase-2 between the Golgi and the plasma membrane. Biochim Biophys Acta 1801:1361-74
Milhas, Delphine; Clarke, Christopher J; Hannun, Yusuf A (2010) Sphingomyelin metabolism at the plasma membrane: implications for bioactive sphingolipids. FEBS Lett 584:1887-94

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