The overall objective of this project is to determine the molecular mechanisms by which Bif-1 activates autophagy and suppresses tumorigenesis. Autophagy, an evolutionarily conserved "self- eating" process, is activated in response to environmental and cellular stresses and plays a fundamental role in maintaining normal cellular homeostasis by delivering unfolded proteins, damaged organelles, and microbial pathogens to lysosomes for degradation. Dysregulation of this cellular self-digestion process can have profound consequences and most likely plays a major role in many types of diseases, including cancer, autoimmune diseases, and neurodegenerative disorders. To date, many autophagy-related (Atg) genes have been identified by independent genetic screens for autophagy-defective mutants in yeast;however, the molecular machinery required for the biogenesis of autophagosomes in mammalian systems has yet to be determined. We have recently discovered that Bif-1 interacts with Beclin 1 through UVRAG and is required for the biogenesis of autophagosomes. Bif-1, also known as SH3GLB1 or endophilin B1, was originally discovered as a Bax-binding protein that contains an amino-terminal N-BAR domain and a carboxy-terminal SH3 domain and demonstrates membrane binding and bending activities. Although the SH3 domain of Bif-1 is sufficient for binding to UVRAG, the N-BAR domain is indispensable for Bif-1 to induce autophagosome formation. Therefore, it is possible that Bif-1 interacts with Beclin 1 through UVRAG at the isolation membrane, or phagophore, during autophagy to regulate vesicle nucleation by inducing membrane curvature through its N- BAR domain.
Specific Aim 1 will test this possibility. While our in vitro studies clearly demonstrated the vital role Bif-1 plays in autophagosome formation, the majority of Bif-1 knockout mice, unlike Atg-deficient mice, developed normally. This phenotypic discrepancy suggests that an unknown factor(s) exists in specific tissues that functionally compensate for the lack of Bif-1 during embryonic and neonatal development. A possible candidate is the Bif-1 homologue endophilin B2. Our preliminary results revealed that endophilin B2 also interacts with Beclin 1 as well as UVRAG and can restore autophagosome formation in Bif-1 deficient cells.
Specific Aim 2 will examine whether endophilin B2 shares a function redundant with Bif-1 in the control of autophagy. Moreover, we found that Bif-1 ablation significantly enhances the development of spontaneous tumors in mice. Since loss of Bif-1 also suppresses Bax/Bak activation and mitochondrial apoptosis, Specific Aim 3 will determine whether Bif-1 induces autophagosome formation and suppresses tumorigenesis independently of its interaction with Bax. We believe that successful implementation of this research will not only gain novel insight into the origin of isolation membranes and the molecular mechanism responsible for autophagic vesicle nucleation and expansion, but will also contribute to the establishment of new strategies for the prevention and treatment of cancer through manipulation of autophagy.

Public Health Relevance

We believe that successful implementation of this research will not only gain novel insight into the origin of isolation membranes and the molecular mechanism responsible for autophagic vesicle nucleation and expansion, but will also contribute to the establishment of new strategies for the prevention and treatment of cancer through manipulation of autophagy.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA129682-05
Application #
8447365
Study Section
Cancer Molecular Pathobiology Study Section (CAMP)
Program Officer
Salnikow, Konstantin
Project Start
2009-04-01
Project End
2015-01-31
Budget Start
2013-02-01
Budget End
2015-01-31
Support Year
5
Fiscal Year
2013
Total Cost
$293,448
Indirect Cost
$104,249
Name
Pennsylvania State University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
129348186
City
Hershey
State
PA
Country
United States
Zip Code
17033
Takahashi, Yoshinori; Young, Megan M; Serfass, Jacob M et al. (2013) Sh3glb1/Bif-1 and mitophagy: acquisition of apoptosis resistance during Myc-driven lymphomagenesis. Autophagy 9:1107-9
Young, Megan M; Kester, Mark; Wang, Hong-Gang (2013) Sphingolipids: regulators of crosstalk between apoptosis and autophagy. J Lipid Res 54:5-19
Young, Megan M; Takahashi, Yoshinori; Khan, Osman et al. (2012) Autophagosomal membrane serves as platform for intracellular death-inducing signaling complex (iDISC)-mediated caspase-8 activation and apoptosis. J Biol Chem 287:12455-68
Watters, Rebecca J; Wang, Hong-Gang; Sung, Shen-Shu et al. (2011) Targeting sphingosine-1-phosphate receptors in cancer. Anticancer Agents Med Chem 11:810-7
Takahashi, Yoshinori; Meyerkord, Cheryl L; Hori, Tsukasa et al. (2011) Bif-1 regulates Atg9 trafficking by mediating the fission of Golgi membranes during autophagy. Autophagy 7:61-73
Coppola, Domenico; Helm, James; Ghayouri, Msoumeh et al. (2011) Down-regulation of Bax-interacting factor 1 in human pancreatic ductal adenocarcinoma. Pancreas 40:433-7
Liao, Aijun; Broeg, Kathleen; Fox, Todd et al. (2011) Therapeutic efficacy of FTY720 in a rat model of NK-cell leukemia. Blood 118:2793-800
Zhang, Yi; Cheng, Yan; Zhang, Li et al. (2011) Inhibition of eEF-2 kinase sensitizes human glioma cells to TRAIL and down-regulates Bcl-xL expression. Biochem Biophys Res Commun 414:129-34
Yang, Jun; Takahashi, Yoshinori; Cheng, Erdong et al. (2010) GSK-3beta promotes cell survival by modulating Bif-1-dependent autophagy and cell death. J Cell Sci 123:861-70
Lee, Jong Woo; Park, Sungman; Takahashi, Yoshinori et al. (2010) The association of AMPK with ULK1 regulates autophagy. PLoS One 5:e15394