Gregg Fields

Matrix metalloproteinase mechanisms and inhibition

The dissolution of the collagen triple-helix has been implicated in diseases where the structural integrity of various components of the body is compromised (for example, arthritis and multiple sclerosis). Collagen also provides a barrier to “compartmentalize” cellular activities; destruction of this barrier plays a role in tumor cell invasion and metastasis. The matrix metalloproteinase (MMP) family has been recognized for their collagenolytic activities, and has thus been the subject of intense research efforts, in order to elucidate their mechanisms of action and allow for rational design of inhibitors. Our laboratory has created and utilized a “mini-library” of collagen-model triple-helical substrates to gain significant insight into the mechanism of collagenolysis, as well as identifying secondary binding sites (exosites) that contribute to collagenolysis.

The work on triple-helical peptidase activity has led our laboratory to consider unique approaches for designing inhibitors targeted for collagenolytic proteases. Rather than focus on small molecules, we have utilized a mini-protein approach to develop agents that interact with multiple sites within the protease, increasing selectivity. The first generation of these inhibitors, triple-helical transition state analogs, have provided high affinity and unique selectivities not observed previously in the MMP field.

Tumor cell interactions with collagen and targeted nanotechnology-based drug delivery systems (nanoDDSs)

Our laboratory has utilized mini-protein approaches to create structurally accurate models of collagen to (a) identify specific regions within collagen that were bound by metastatic melanoma cells, (b) identify the melanoma cell receptors that bound to these regions, (c) evaluate the results of outside-in signal transduction mediated by those receptors, (d) correlate the three-dimensional structural state of collagen with promotion of melanoma activities, and (e) “proteolytically profile” metastatic potential. As a result, triple-helical ligands for the α2β1 and α3β1 integrins and CD44/chondroitin sulfate proteoglycan (CSPG) were developed. These ligands were used in a novel approach for creating targeted nanoDDSs, involving the direct incorporation of peptide amphiphiles (PAs) into liposomal vesicles. Stable PA nanoDDSs allowed for selective delivery to CD44-positive melanoma cells. Overall, the progression of melanoma was found to be partially dictated by the structural state of collagen.

In the course of the studies described above, glycosylation was found to strongly modulate melanoma cell activities, as adhesion and spreading were dramatically decreased due to the presence of glycosylated hydroxylysine residues within the collagen triple-helix. Overall, this (a) was the first demonstration of the prophylactic effects of glycosylation on tumor cell interaction with the basement membrane, (b) provided a rare example of an apparent unfavorable interaction between carbohydrates, and (c) suggested that sugars may mask “cryptic sites” accessible to tumor cells containing cell-surface or secreted glycosidase activities.

Dr. Gregg B. Fields received his B.S. and Ph.D. degrees in chemistry from the University of Florida and Florida State University, respectively. He was a Postdoctoral Scholar with Ken A. Dill at the University of California San Francisco. Dr. Fields joined the faculty at the University of Minnesota in 1991 as an assistant professor. He was promoted to associate professor with tenure in 1995 and then achieved the rank of full professor of chemistry & biochemistry at Florida Atlantic University in 1997. In 2008, Dr. Fields became a Robert A. Welch Foundation Distinguished University Chair in Chemistry in the Department of Biochemistry at The University of Texas Health Science Center at San Antonio. Dr. Fields recently relocated to the Torrey Pines Institute for Molecular Studies, where he is a Full Member, Director of Research, and Distinguished Chair of Metalloproteinase and Multiple Sclerosis Research. Dr. Fields' research interests are in the use of chemical approaches to better understand how protein three-dimensional structures influence cellular and enzymatic behaviors. Early studies by Dr. Fields included the systematic examination of mild methodologies for solid-phase synthesis of small proteins. Chemical approaches were used to develop “mini-protein” models for the study of cellular recognition processes, which in turn allowed for the mapping of protein domains involved in tumor cell binding and signal transduction. Mini-protein models have subsequently been utilized to dissect the mechanisms of collagen catabolism, and in the process have provided new avenues for protease inhibitor design. Dr. Fields has authored or coauthored more than 200 scientific publications and presented over 100 invited lectures. Dr. Fields is currently President of the American Peptide Society (2009-2011).

Education

• 1976-1979    A.A., Chemistry, Broward Community College, Davie, FL 33314
• 1979-1982    B.S., Department of Chemistry, University of Florida, Gainesville, FL 32611
• 1983-1988    Ph.D., Department of Chemistry, Florida State University (FSU), Tallahassee, FL 32306

Positions

• 5/82-9/82     Quality Control Analytical Chemist, Key Pharmaceuticals, Incorporated, North Miami Beach, FL 33161
• 5/83-4/84     Teaching Assistant in Biochemistry and Organic Chemistry, Department of Chemistry, FSU
• 5/84-11/88    Research Assistant, Department of Chemistry, FSU
• 12/88-1/91    Postdoctoral Scholar with Professor Ken A. Dill, Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94143
• 3/90-1/91      Visiting Scientist, Peptide Synthesis Research and Development, Applied Biosystems, Incorporated, Foster City, CA 94404
• 2/91-5/95      Assistant Professor, Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN 55455
• 2/91-5/95      Assistant Professor, Biomedical Engineering Center, University of Minnesota
• 2/93-5/95     Assistant Professor (Joint Appointment), Department of Biochemistry, University of Minnesota
• 5/95-12/97    Associate Professor, Department of Laboratory Medicine and Pathology, University of Minnesota
• 5/95-12/97    Associate Professor, Biomedical Engineering Center, University of Minnesota, Minneapolis
• 5/95-12/97    Associate Professor (Joint Appointment), Department of Biochemistry, University of Minnesota
• 12/97-7/08    Professor, Department of Chemistry and Biochemistry, Florida Atlantic University (FAU), Boca Raton, FL 33431-0991
• 10/99-7/08    Professor, Department of Biomedical Science, FAU
• 8/00-7/08     Chair, Department of Chemistry and Biochemistry, FAU
• 7/05-present Adjunct Professor, Advanced Technology, The Scripps Research Institute/Scripps Florida, Jupiter, FL 33458
• 3/06-7/08      Member, H. Lee Moffitt Comprehensive Cancer Center & Research Institute, Tampa, FL 33612
• 10/06-12/09  Member, Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, Miami, FL 33136
• 1/08-12/10    Member, Cancer Therapy & Research Center, UTHSCSA
• 8/08-12/10    Professor, Department of Biochemistry, UTHSCSA
• 6/09-12/10    Adjunct Professor, Department of Chemistry, The University of Texas at San Antonio, San Antonio, TX 78249
• 1/11-present  Full Member and Director of Research, Torrey Pines Institute for Molecular Studies (TPIMS), Port St. Lucie, FL 34987

Professional Affiliations

• Member, American Chemical Society, 1986-2009; European Peptide Society, 1996-2001; American Society for Cell Biology, 1998-2005; Protein Society, 2001-2005; American Society for Biochemistry and Molecular Biology, 2001-present; Society for Melanoma Research, 2004-2008.
• American Peptide Society Charter Member, 1990-present.
• Association of Biomolecular Resource Facilities (ABRF) Research Committee on Peptide Synthesis and Mass Spectrometry Member, 1991-1996.
• Facility Director, ABRF, 1992-present.
• Co-Chair, ABRF Research Committee on Peptide Synthesis and Mass Spectrometry, 1992.
• Chair, ABRF Research Committee on Peptide Synthesis and Mass Spectrometry, 1992-1994.
• Chairman, American Peptide Society Publications Committee, 1993-1995.
• Member, American Peptide Society Publications Committee, 1995-1997.
• New York Academy of Sciences Active Member, 1994-1997.
• Full Member, University of Minnesota Comprehensive Cancer Research Center, 1996-2003.
• American Peptide Society Council, 1997-2003.
• President-Elect, American Peptide Society, 2007-2009.
• President, American Peptide Society, 2009-2011.

• Young Investigator Award, Matrix Metalloproteinase Conference, 1989.
• Young Investigator Award, 12th American Peptide Symposium, 1991.
• McKnight-Land Grant Professorship, University of Minnesota, 1993-1995.
• National Institutes of Health Research Career Development Award, 1994-1999.
• Invited Lecturer, Carlsberg Laboratory, Valby, Denmark, 1994.
• Invited Guest Editor, Peptide Science: Comprehensive Reports & Reviews, 1994-1996.
• Invited Editor, Methods In Enzymology, 1995-1998.
• Profiled in Today's Life Sciences 9(1), January 1997, “Tumours and techniques,” p. 36.
• Association of Biomolecular Resource Facilities Excellence Award in Peptide Synthesis Research, 1997.
• Selected as one of “2000 Outstanding Scientists of the 21st Century,” 2000.
• Researcher of the Year Award (Full Professor), FAU, 2000-2001.
• Profiled in Boca Raton Magazine 23(3), May/June 2003, “8 Florida Scientists Who Are Saving Your Life: Breakthrough Medicine - Stopping The Spread Of Cancer,” pg. 98-100.
• Invited Editor, Methods In Molecular Biology, 2003-2007.
• Profiled in Time Magazine 163(23), June 7, 2004, “The Secrets Of Their Success,” pg. 107-110.
• Distinguished Visiting Professor, Imperial College London, 2004.
• Faculty of 1000 Biology, Chemical Biology Faculty, Protein Chemistry & Proteomics section, 2004-present.
• Profiled in NanoBiotech News 3(36), September 21, 2005, “Florida Atlantic University, Moffitt get $1M to develop liposome-based drugs,” p. 7.
• Researcher of the Year Award (Full Professor), FAU, 2005-2006.
• Profiled in Chemical Biology & Drug Design 67(2), February 2006, “Who’s Who In Chemical Biology & Drug Design,” p. 187.
• Keynote Speaker, PepCon-2008, Shenzhen, China, 2008.
• Roche U.S.-Sponsored Keynote Speaker, Chemistry & Biology of Peptides 2008, University of Nottingham, England.
• BIT Life Sciences Lifetime Membership Award, 2008.
• Robert A. Welch Foundation Distinguished University Chair in Chemistry, UTHSCSA, 2008-2010.
• Texas Higher Education Science and Technology Acquisition and Retention (STAR) Award, 2008.
• Texas Higher Education Science and Technology Acquisition and Retention (STAR) Plus Award, 2008.
• Keynote Speaker, PepCon-2009, Seoul, South Korea, 2009.
• Commencement Speaker, UTHSCSA, School of Graduate Sciences, 2009.
• Keynote Speaker, 2nd World Cancer Congress, Beijing, China, 2009.
• Plenary Lecturer, 2nd Modern Solid Phase Synthesis & Its Applications Symposium, Brisbane, Australia, 2009.
• Distinguished Chair of Metalloproteinase and Multiple Sclerosis Research, TPIMS, 2010-present.

Selected Publications (10 of 143)

1. Diane Baronas-Lowell, Janelle L. Lauer-Fields, Jeffrey A. Borgia, Gian Franco Sferrazza, Mohammad Al-Ghoul, Dmitriy Minond, and Gregg B. Fields. Differential Modulation of Human Melanoma Cell Metalloproteinase Expression by α₂β₁ Integrin and CD44 Triple-Helical Ligands Derived from Type IV Collagen. J. Biol. Chem. 279, 43503-43513 (2004).
2. Dmitriy Minond, Janelle L. Lauer-Fields, Mare Cudic, Christopher M. Overall, Duanqing Pei, Keith Brew, Robert Visse, Hideaki Nagase, and Gregg B. Fields. The Roles of Substrate Thermal Stability and P₂ and P₁’ Subsite Identity on Matrix Metalloproteinase Triple-Helical Peptidase Activity and Collagen Specificity. J. Biol. Chem. 281, 38302-38313 (2006).
3. Dmitriy Minond, Janelle L. Lauer-Fields, Mare Cudic, Christopher M. Overall, Duanqing Pei, Keith Brew, Marcia L. Moss, and Gregg B. Fields. Differentiation of Secreted and Membrane-Type Matrix Metalloproteinase Activities Based on Substitutions and Interruptions of Triple-Helical Sequences. Biochemistry 46, 3724-3733 (2007).
4. Evonne M. Rezler, David R. Khan, Janelle L. Lauer-Fields, Mare Cudic, Diane Baronas-Lowell, and Gregg B. Fields. Targeted drug delivery utilizing protein-like molecular architecture. J. Am. Chem. Soc. 129, 4961-4972 (2007).
   
5. Janelle L. Lauer-Fields, Keith Brew, John K. Whitehead, Shunzi Li, Robert P. Hammer, and Gregg B. Fields. Triple-helical transition-state analogs: A new class of selective matrix metalloproteinase inhibitors. J. Am. Chem. Soc. 129, 10408-10417 (2007).
   
6. Janelle L. Lauer-Fields, John K. Whitehead, Shunzi Li, Robert P. Hammer, Keith Brew, and Gregg B. Fields. Selective modulation of matrix metalloproteinase 9 (MMP-9) functions via exosite inhibition. J. Biol. Chem. 283, 20087-20095 (2008).
7. Janelle L. Lauer-Fields, Dmitriy Minond, Peter S. Chase, Pierre E. Baillargeon, S. Adrian Saldanha, Roma Stawikowska, Peter Hodder, and Gregg B. Fields. High throughput screening of potentially selective MMP-13 exosite inhibitors utilizing a triple-helical FRET substrate. Bioorg. Med. Chem. 17, 990-1005 (2009).
8. Janelle L. Lauer-Fields, Michael J. Chalmers, Scott A. Busby, Dmitriy Minond, Patrick R. Griffin, and Gregg B. Fields. Identification of Specific Hemopexin-like Domain Residues That Facilitate Matrix Metalloproteinase Collagenolytic Activity. J. Biol. Chem. 284, 24017-24024 (2009).
   
9. Gregg B. Fields. Synthesis and biological applications of collagen-model triple-helical peptides. Org. Biomol. Chem. 8, inside cover + 1237-1258 (2010).
10. Jianxi Xiao, Janelle L. Lauer, Gregg B. Fields, and Jean Baum. Local conformation and dynamics of isoleucine in the collagenase cleavage site provides recognition signal for matrix metalloproteinases. J. Biol. Chem. 285, 34181-34190 (2010).

• Gregg B. Fields. Mild Solid-Phase Synthesis of Aligned, Branched Triple-Helical Peptides. Patent issued November 19, 1996, United States Patent #5,576,419. Licensed for application as an anti-cancer therapeutic by BioStratum, Inc., July 25, 1994.
• Gregg B. Fields. Mild Solid-Phase Synthesis of Aligned, Branched Triple-Helical Peptides. Patent issued March 10, 1998, United States Patent #5,726,243.
• Gregg B. Fields, James B. McCarthy, and Leo T. Furcht. Polypeptides With Type I Collagen Activity. Patent issued March 24, 1998, United States Patent #5,731,409.
• Hideaki Nagase and Gregg B. Fields. Discriminatory Substrates For MMP Hydrolysis. Patent issued June 23, 1998, United States Patent #5,770,691. Licensed by Bachem Biosciences, July 8, 1997, and Peptides International, December 11, 1998. Marketed by Peptide Institute, Inc. (Japan)/Peptides International, Inc. (U.S.A.) as product #3168-v, 1995-present, and Bachem Biosciences as product #M-2110, 1995-present.
• Gregg B. Fields, and Daniel L. Mooradian. Solid-Phase Method Attaching a Biomolecule to a Substrate Surface With a Photoreactive Crosslinking Agent. Patent issued December 29, 1998, United States Patent #5,853,744.
• Matthew Tirrell and Gregg B. Fields. Self-Assembling Amphiphiles for Construction of Peptide Secondary Structures. International Publication #WO98/07752, February 26, 1998. Patent issued August 1, 2000, United States Patent #6,096,863.
• Gregg B. Fields and Robert Hammer. Selective Inhibition of Matrix Metalloproteinases. Application filed November 21, 2006, U.S. Patent Application No. 11/562,295.