Apoptosis is the orchestrated process by which cells self-destruct in response to specific internal and external signals. Programmed cell death is regulated by a complex network of checks and balances governed by the """"""""BCL-2 family"""""""" of proteins. Faulty regulation of apoptotic pathways is a seminal event in the pathogenesis of a variety of hematologic and other diseases. BCL-2 is a survival protein, and its overproduction facilitates pathologic cell survival death. The specific molecular choreography that triggers mitochondrial apoptosis remains unknown. Bid's structure is defined by 8 alpha-helices, one of which is a critical death effector helix. We hypothesize that specific alpha-helical domains within BID engage in distinct interactions that contribute to its overall function. The goal of this research is to fuse chemical and biological techniques to elucidate the mechanism of BID-induced apoptosis and develop molecules to manipulate apoptosis in vivo. Thus, the specific aims of this proposal are: 1) Synthesize and characterize Stabilized Alpha Helices of BID (SAHBs), 2) Define the role(s) of BID's alpha-helical domains in triggering mitochondrial apoptosis, and 3) Utilize SAHBs to validate and manipulate BID activities in cultured cells and mouse models. A preliminary panel of SAHBs modeled after BID's death effector helix has been synthesized. Compared to unmodified peptide, two of these SAHBs demonstrate: (a) more than 5-fold increase in alpha-helicity; (b) up to 3.5-fold enhancement in protease resistance; (c) up to 25-fold increased potency in mitochondrial cytochrome c release, a key step in apoptosis; and (d) apoptosis induction in cultured Jurkat cells. SAHBs have the dual potential to serve as biological tools to dissect apoptotic pathways, and as prototype therapeutics for diseases characterized by aberrant cellular hypo- or hyperplasia. A diverse team of internationally recognized mentors, collaborators, and advisors with expertise in the fields of synthetic chemistry, structural biology, apoptosis biology, and clinical hematology/oncology will provide an idea training environment for Dr. Walensky's development as an independent investigator at the interface of these notable fields.
| Danial, Nika N; Walensky, Loren D; Zhang, Chen-Yu et al. (2008) Dual role of proapoptotic BAD in insulin secretion and beta cell survival. Nat Med 14:144-53 |
| Pitter, Kenneth; Bernal, Federico; Labelle, James et al. (2008) Dissection of the BCL-2 family signaling network with stabilized alpha-helices of BCL-2 domains. Methods Enzymol 446:387-408 |
| Bird, Gregory H; Bernal, Federico; Pitter, Kenneth et al. (2008) Synthesis and biophysical characterization of stabilized alpha-helices of BCL-2 domains. Methods Enzymol 446:369-86 |
| Verdine, Gregory L; Walensky, Loren D (2007) The challenge of drugging undruggable targets in cancer: lessons learned from targeting BCL-2 family members. Clin Cancer Res 13:7264-70 |
| Bernal, Federico; Tyler, Andrew F; Korsmeyer, Stanley J et al. (2007) Reactivation of the p53 tumor suppressor pathway by a stapled p53 peptide. J Am Chem Soc 129:2456-7 |
| Walensky, L D (2006) BCL-2 in the crosshairs: tipping the balance of life and death. Cell Death Differ 13:1339-50 |
| Walensky, Loren D; Pitter, Kenneth; Morash, Joel et al. (2006) A stapled BID BH3 helix directly binds and activates BAX. Mol Cell 24:199-210 |
| Walensky, Loren D; Kung, Andrew L; Escher, Iris et al. (2004) Activation of apoptosis in vivo by a hydrocarbon-stapled BH3 helix. Science 305:1466-70 |
