Education and Training:
1984 B.S. Philadelphia College of Pharmacy and Science (Chemistry)
1986 M.S. University of Kansas (Medicinal Chemistry)
1990 Ph.D. University of Kansas (Medicinal Chemistry)
1990-1992 Postdoctoral Fellow, University of Kansas, Department of Medicinal Chemistry
1992-1994 Pharmacology Research Associate Training (PRAT) Fellow, National Institutes of Health, Laboratory of Cell Biology, National Institute of Mental Health
- Intramembrane Proteases
The Wolfe lab studies intramembrane proteases that play critical roles both in normal biology and in human disease. The last place in the cell to expect hydrolysis is within the hydrophobic environment of the lipid bilayer. Nevertheless, a number of multi-pass membrane proteins appear to carry out this seemingly paradoxical process (Wolfe and Kopan, Science, 2004; Wolfe, Chem Rev, 2009). Such proteases cut within the transmembrane region of their respective substrates, and consistent with this observation, these proteases contain putative catalytic residues located within transmembrane domains.
γ-Secretase and Alzheimer’s Disease
The specific focus of the lab has been on the chemistry and biology of g-secretase. This protease is critical to the pathogenesis of Alzheimer's disease (Esler and Wolfe, Science, 2001) and to cell differentiation during embryonic development. Small organic inhibitors were developed and used as tools to characterize and identify g-secretase (Wolfe et al., J Med Chem, 1998; Wolfe et al., Biochemistry, 1999a). Findings from the lab implicate a multi-pass membrane protein called presenilin as the catalytic component of a larger g-secretase complex (Wolfe et al., Nature, 1999; Esler et al., Nat Cell Biol, 2000). Missense mutations in presenilin cause hereditary Alzheimer's disease, and these mutations specifically affect g-secretase activity.
The lab found that presenilin and a presenilin-associated protein called nicastrin copurify with g-secretase activity from an immobilized inhibitor, evidence that nicastrin is also a member of the protease complex (Esler et al., Proc Natl Acad Sci USA, 2002). Moreover, a g-secretase substrate also copurified, suggesting an initial substrate docking site on the protease complex distinct from the active site. Helical peptides designed to interact with this docking site can potently inhibit g-secretase activity both in cell-free and cell-based assays (Das et al., J Am Chem Soc, 2003; Kornilova et al., Proc Natl Acad Sci USA, 2005). The lab also determined the stoichiometry of the active g-secretase complex, which had been unknown (and, with respect to presenilin, controversial). The four essential components (presenilin, nicastrin, Aph-1 and Pen-2) are each represented only once per complex (Sato et al., J Biol Chem, 2007). Disease-causing mutations in presenilin alter processive proteolysis by g-secretase, leading to increased proportions of long Ab peptides (Quintero-Monzon et al., Biochemistry, 2011; Fernandez et al., J Biol Chem, 2014). Nicastrin functions as a molecular gatekeeper to ensure processing of substrates with short ectodomains (Bolduc et al., PNAS, 2016). Most recent efforts have focused on further understanding substrate recognition and mechanism (Bolduc et al, eLife, 2016; Fernandez, Biochemistry, 2016).
The lab also discovered a nucleotide binding site on the g-secretase complex. Small organic molecules that interact with this site can selectively block g-secretase proteolysis of the amyloid-b precursor protein (APP), critical to the pathogenesis of Alzheimer’s disease, without affecting proteolysis of an alternative substrate, the Notch receptor. Notch signaling, critical in many cell differentiation events, requires proteolysis by g-secretase (De Strooper et al., Nature, 1999), and blocking Notch signaling with g-secretase inhibitors causes severe toxicity in mice. The finding that compounds can selectively block the cleavage of APP without affecting that of Notch (Fraering et al., J Biol Chem, 2005) revived this protease as a therapeutic target. In 2006, Dr. Wolfe cofounded the Laboratory for Experimental Alzheimer Drugs (LEAD) at HMS to advance the development of such selective agents as disease-modifying therapeutics for Alzheimer’s disease (Lu et al, Bioorg Med Chem Lett, 2016; Wei et al, Bioorg Med Chem Lett, 2016; Zhang et al, Bioorg Med Chem Lett, 2016).
Current efforts are aimed at further elucidating the mechanism of γ-secretase substrate recognition and its multiple proteolytic functions, understanding how genetic mutations that cause Alzheimer’s disease alter g-secretase structure and function, and developing new chemical probes and therapeutic prototypes targeting this protease complex.
Signal Peptide Peptidase and Rhomboid
The Wolfe lab has also investigated the structure, mechanism, and inhibition of other intramembrane proteases, such as the serine protease Rhomboid (Urban and Wolfe, Proc Natl Acad Sci USA, 2005) and the presenilin homolog signal peptide peptidase (Sato et al., Biochemistry, 2006; Narayanan et al., J Biol Chem, 2007; Sato et al., J Biol Chem, 2008), both of which are highly conserved across evolution and play critical roles in biology. In this way, the lab helped establish common biochemical principles and strategies for designing inhibitors for this family of membrane-embedded enzymes. The current focus is developing new chemical probes to understand the roles of these membrane-embedded proteases in biology, particularly in human parasites such as Plasmodium falciparum that causes malaria, and explore the potential of these proteases as therapeutic targets.
- mRNA Processing
Tau in Neurodegenerative Disease
The lab has also combined chemistry and biology toward the study of another factor critical to the pathogenesis of dementias: the microtubule-associated protein tau. Filaments of tau are a common feature in a variety of different neurodegenerative diseases, including Alzheimer’s disease. Mutations in the gene encoding this protein are associated with dominant, familial forms of frontotemporal dementia, and many of these mutations alter pre-mRNA splicing to increase inclusion of exon 10. The Wolfe lab validated the in vivo existence of a hypothetical stem-loop at the end of exon 10, where many of the dementia-associated mutations occur (Donahue et al., J Biol Chem, 2006). These mutations destabilize this RNA stem-loop structure, allowing more ready access to splicing factors. High-throughput screening has led to identification of small molecules that interact with and stabilize this structure (Donahue et al., J Biomol Screen, 2007), and NMR studies have elucidated how one of these compounds interacts with the tau mRNA stem-loop (Zhang et al., Chem Biol, 2009). The lab has worked to improve the potency and selectivity of these agents (Liu et al., J Med Chem, 2009) as well as antisense oligonucleotides (Peacey et al., Nucleic Acid Res, 2012) and conjugates thereof (Liu et al., Bioorg Chem, 2014) to provide new tools for chemical biology as well as new prototype therapeutics. Study of the 3’-UTR of the tau message has also led to identification of miR-34a as a down-regulator of tau levels (Dickson et al., J Neurochem, 2013)
β-Secretase and Alzheimer’s Disease
In addition, the lab studied alternative mRNA splicing of the b-site APP-cleaving enzyme 1 (BACE1; b-secretase), determining that alternative splice isoforms are catalytically inactive and that shunting BACE1 down these alternative pathways with antisense oligonucleotides effectively lowers Ab production in cells (Mowrer and Wolfe, J Biol Chem, 2008). A G-quadruplex structure in exon 3 of BACE1 mRNA partly regulates alternative splicing (Fisette et al., J Neurochem, 2012). Thus, modulation of BACE1 alternative splicing represents a new strategy for developing therapeutics for Alzheimer’s disease.
Future Projects on mRNA
Potential future projects include (1) compound screening to target a hexanucleotide repeat expansion in the gene c9ORF72, involved in 50% of cases of amyotrophic lateral sclerosis and (2) searching for cryptic riboswitches, structural motifs in mRNA that are potential sites for interaction and modulation by synthetic small molecules.
1994-98 Assistant Professor of Medicinal Chemistry, University of Tennessee
1998-99 Associate Professor of Medicinal Chemistry, University of Tennessee
1999-2008 Associate Professor of Neurology, Harvard Medical School
1999-2008 Associate Scientist, Center for Neurologic Diseases, Brigham and Women’s Hospital
2008-2016 Professor of Neurology, Harvard Medical School
2008-2016 Scientist, Center for Neurologic Diseases, Brigham and Women’s Hospital
2016-present Mathias P. Mertes Professor of Medicinal Chemistry, University of Kansas
Liu Y, Rodriguez L, Wolfe MS. Template-directed synthesis of a small molecule-antisense conjugate targeting an mRNA structure. Bioorg. Chem. 2014; 54:7-11.
Holmes O, Paturi S, Wolfe MS, Selkoe DJ. Functional analysis and purification of a Pen-2 fusion protein for γ-secretase structural studies. J. Neurochem. 2014; 131(1):94-100
Holmes O, Paturi S, Selkoe DJ, Wolfe MS. Pen-2 is essential for γ-secretase complex stability and trafficking but partially dispensable for endoproteolysis. Biochemistry. 2014; 53(27):4393-406.
Fernandez MA, Klutkowski JA, Freret T, Wolfe MS. Alzheimer Presenilin-1 Mutations Dramatically Reduce Trimming of Long Amyloid β-Peptides (Aβ) by γ-Secretase to Increase 42-to-40-Residue Aβ. J Biol Chem 2014; 289(45):31043-52.
Park HJ, Ran Y, Jung JI, Holmes O, Price AR, Smithson L, Ceballos-Diaz C, Han C, Wolfe MS, Daaka Y, Ryabinin AE, Kim SH, Hauger RL, Golde TE, Felsenstein KM. The stress response neuropeptide CRF increases amyloid-β production by regulating γ-secretase activity. EMBO J 2015; 34(12): 1674-86.
Bolduc DM, Montagna DR, Gu Y, Selkoe DJ, Wolfe MS. Nicastrin functions to sterically hinder γ-secretase-substrate interactions driven by substrate transmembrane domain. Proc Natl Acad Sci USA 2016 113(5):E509-18.
Lu D, Wei HX, Zhang J, Gu Y, Osenkowski P, Ye W, Selkoe DJ, Wolfe MS, Augelli-Szafran CE. Part 1: Notch-sparing γ-secretase inhibitors: The identification of novel naphthyl and benzofuranyl amide analogs. Bioorg Med Chem Lett 2016, 26(9):2129-32.
Wei HX, Lu D, Sun V, Zhang J, Gu Y, Osenkowski P, Ye W, Selkoe DJ, Wolfe MS, Augelli-Szafran CE. Part 2. Notch-sparing γ-secretase inhibitors: The study of novel γ-amino naphthyl alcohols. Bioorg Med Chem Lett 2016, 26(9):2133-37.
Zhang J, Lu D, Wei HX, Gu Y, Selkoe DJ, Wolfe MS, Augelli-Szafran CE. Part 3: Notch-sparing γ-secretase inhibitors: SAR studies of 2-substituted aminopyridopyrimidinones. Bioorg Med Chem Lett 2016, 26(9):2138-41.
Bolduc DM, Montagna DR, Seghers MC, Wolfe MS, Selkoe DJ. The amyloid-beta forming tripeptide cleavage mechanism of γ-secretase. eLife 2016, DOI: http://dx.doi.org/10.7554/eLife.17578
Fernandez MA, Biette K, Dolios G, Seth D, Wang R, Wolfe MS. Transmembrane substrate determinants for γ-secretase processing of APP CTF-β. Biochemistry 2016,55(40):5675-5688.
Selected Awards & Honors
2003 Sato Memorial International Award, Pharmaceutical Society of Japan
2007 Alzheimer Drug Discovery Foundation and Elan Pharmaceuticals, Award for Innovative Research
2007 Annual Alumni Award, University of the Sciences in Philadelphia
2008-11 Zenith Fellows Award, Alzheimer’s Association
2008 MetLife Award for Medical Research, MetLife Foundation
2009 Potamkin Prize for Research in Pick’s, Alzheimer’s and Related Diseases, American Academy of Neurology