Freeman Lab

The basic investigation and clinical research activities of the Freeman Laboratory focus on the eukaryotic cell production, reactions and signal transduction properties of oxidizing and free radical inflammatory mediators (e.g., superoxide, hydrogen peroxide, nitric oxide (NO), peroxynitrite, nitrogen dioxide, oxidized/nitrated lipids).  In particular, we are interested in the actions of these species as both redox signaling mediators, as pathogenic agents in inflammatory diseases and in the case of nitro-fatty acids as drug candidates.  Our observations regarding O2 and NO-derived reactive species have lent new insight into mechanisms of redox-dependent cell signaling, post-translational protein modification and have revealed new therapeutic strategies for treating acute inflammation, metabolic syndrome, respiratory disorders and cardiovascular diseases.

Gregory Buchan
Predoctoral Fellow

Veronika Cechova, PhD
Health Sciences Fellow

Marco Fazzari, PhD
Research Assistant Professor

Bruce A. Freeman, PhD
Irwin Fridovich Distinguished Professor and Chair

Nicholas Khoo, PhD
Research Assistant Professor

Lihua Li
Research III

Crystal Uvalle
Research Specialist IV

Dario Vitturi, PhD
Research Assistant Professor

Stacy Gelhaus Wendell, PhD
Assistant Professor

Steven R. Woodcock, PhD
Research Instructor

Gregory Buchan

Bilinovich SM, Davis CM, Morris DL, Ray LA, Prokop JW, Buchan GJ, Leeper TC. 2014, The C-terminal domain of SRA1p has a fold more similar to PRP18 than to an RRM and does not directly bind to the SRA1 RNA STR7 region. J Mol Biol. 2014 Apr 17;426(8):1753-65. doi: 10.1016/j.jmb.2014.01.007. Epub 2014 Jan 29. PMID: 24486611

Marco Fazzari, PhD

M Fazzari, N Khoo, SR Woodcock, DK Jorkasky, L Li, FJ Schopfer and BA Freeman. Nitro-fatty acid pharmacokinetics in the adipose tissue compartment. Journal of Lipid Research 58:375-385, 2017. 
SR Salvatore, DA Vitturi, M Fazzari, DK Jorkasky and FJ Schopfer. Evaluation of 10-nitro oleic acid bio-elimination in rats and humans. Scientific Reports 7: 39900, 2017.
M Fazzari, N Khoo, SR Woodcock, L Li, BA Freeman and FJ Schopfer. Generation and esterification of electrophilic fatty acid nitroalkenes in triacylglycerides. Free Radical Biology & Medicine 87:113-124, 2015. 
DA Vitturi, L Minarrieta, SR Salvatore, EM Postlethwait, M Fazzari, G Ferrer-Sueta, JR Lancaster, BA Freeman and FJ Schopfer. Convergence of biological nitration and nitrosation reactions via symmetrical nitrous anhydride. Nature Chemical Biology 11:504-510, 2015.
M Fazzari, A Trostchansky, F.J Schopfer, SR Salvatore, B Sanchez-Calvo, D Vitturi, R Valderrama, JB Barroso, R Radi, BA Freeman and H Rubbo. Olives and olive oil are sources of electrophilic Fatty Acid nitroalkenes. PLoS One 9:e84884, 2014.

Bruce A. Freeman, PhD

Vitturi DA, L Minarrieta, SR Salvatore, EM Postlethwait, M Fazzari, G Ferrer-Sueta, JR Lancaster, BA Freeman* and FJ Schopfer*.       Convergence of biological nitration and nitrosation via symmetrical nitrous anhydride. Nature Chem Biol  doi: 10.1038/nchembio.1814, 2015. *Corresponding authors
Wendell SG, F Golin-Bisello, S Wenzel, RW Sobol, F Holguin and BA Freeman. 15-hydroxy-prostaglandin dehydrogenase generation of electrophilic lipid signaling mediators from hydroxy Ω-3 fatty acids. J Biol Chem 290:5868-5880, 2015.
Woodcock SR, SR Salvatore, G Bonacci, FJ Schopfer and BA Freeman. Biomimetic nitration of conjugated linoleic acid: Formation, characterization and synthesis of naturally occurring conjugated nitrodienes. J Org Chem 79:25-33, 2014.
Charles RL, O Rudyk, O Prysyazhna, A Kamyina, J Yang, C Morisseau, BD Hammock, BA Freeman and P Eaton. Protection from hypertension in mice by the Mediterranean diet is mediated by nitro fatty acid inhibition of soluble epoxide hydrolase.       Proc Nat Acad Sci 111:8167-8172, 2014.
Rudolph V, TK Rudolph, FJ Schopfer, G Bonacci, SR Woodcock, MP Cole, PRS Baker, R Ramani and BA Freeman.  Endogenous generation and protective effects of nitro-fatty acids in a murine model of focal cardiac ischaemia and reperfusion.  Cardiovascular Research 85:155-166, 2010.
Groeger AL, C Cipollina, MP Cole, SR Woodcock, G Bonacci, TK Rudolph, V Rudolph, BA Freeman and FJ Schopfer.  Cyclooxygenase-2 generates anti-inflammatory mediators from omega-3 fatty acids.  Nature Chemical Biology 6:433-441, 2010.
Batthyany C, FJ Schopfer, PR Baker, R Duran, LM Baker, Y Huang, C Cervenansky, BP Branchaud and BA Freeman. Reversible post-translational modification of proteins by nitrated fatty acids in vivo. J Biol Chem 281: 20450-20463, 2006.
Schopfer F, Y Lin, P Baker, T Cui, M Garcia-Barrio, J Zhang, K Chen, Y Chen and BA Freeman.       Nitrolinoleic acid – an endogenous PPARγ ligand. Proc Nat Acad Sci 102:2340-2345, 2005.
Eiserich JP, S Baldus, M-L Brennan, W Ma, C Zhang, A Tousson, L Castro, AJ Lusis, CR White and BA Freeman. Myeloperoxidase: A leukocyte-derived vascular ·NO oxidase. Science 296:2391-2394, 2002.
Eiserich JP*, M Hristova, CE Cross, AD Jones, BA Freeman, B Halliwell and A van der Vliet*. Formation of nitric oxide-derived inflammatory oxidants by myeloperoxidase in neutrophils. Nature 391:393-397, 1998. *PhD students.
Rubbo H, R Radi, M Trujillo, R Telleri, B Kalyanaraman, S Barnes, M Kirk and BA Freeman. Nitric oxide regulation of superoxide and peroxynitrite-dependent lipid peroxidation: Formation of novel nitrogen-containing oxidized lipid derivatives. J Biol Chem 265:26066-26075, 1994.
Radi R, JS Beckman and BA Freeman. Peroxynitrite oxidation of sulfhydryls: The cytotoxic potential of endothelial-derived superoxide and nitric oxide. J Biol Chem 266:4244-4250, 1991.
Beckman JS, TW Beckman, J Chen, PA Marshall and BA Freeman. Apparent hydroxyl radical production by peroxynitrite: Implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci 87:1620-1624, 1990.
Rosen GR and BA Freeman. Detection of superoxide in endothelial cells. Proc Natl Acad Sci 81:7269-7273, 1984.

Complete List of Published Work in MyBibliography - ~225 research articles, 65 reviews, H index 91

Nicholas Khoo, PhD

Fazzari M, NK Khoo, SR Woodcock, L Li, BA Freeman and FJ Schopfer.  Generation and esterification of electrophilic fatty acid nitroalkenes in triacylglycerides.  Free Radic Biol Med 87:113-124, 2015.
Chartoumpekis DV, DL Palliyaguru, N Wakabayashi, NK Khoo, G Schoiswohl, RM O'Doherty and TW Kensler. Notch intracellular domain overexpression in adipocytes confers lipodystrophy in mice. Molecular Metabolism 4:543-550, 2015.
Kelley EE, J Baust, G Bonacci, F Golin-Bisello, JE Devlin, CM St Croix, SC Watkins, S Gor, N Cantu-Medellin, ER Weidert, JC Frisbee, MT Gladwin, HC Champion, BA Freeman and NK Khoo. Fatty acid nitroalkenes ameliorate glucose intolerance and pulmonary hypertension in high-fat diet-induced obesity. Cardiovasc Res 101:352-363, 2014.
Khoo NK, L  Mo, S Zharikov, C Kamga-Pride, K Quesnelle, F Golin-Bisello, L Li, Y Wang and S Shiva. Nitrite augments glucose uptake in adipocytes through the protein kinase A-dependent stimulation of mitochondrial fusion. Free Radic Biol Med 70:45-53, 2014.
Khoo NK, S Hebbar, W Zhao, SA Moore, FE Domann and ME Robbins. Differential activation of catalase expression and activity by PPAR agonists: Implications for astrocyte protection in anti-glioma therapy. Redox Biol 1:70-79, 2013. 
Khoo NKH, N Cantu-Medellin, JE Devlin, CM St Croix, SC Watkins, AM Fleming, HC Champion, RP Mason, BA Freeman and EE Kelley.  Obesity-induced tissue free radical generation:  An in vivo immuno-spin trapping study.  Free Radical Biol Med 52:2312-2319, 2012.
Khoo NKH, V Rudolph, MP Cole, F Golin-Bisello, SR Woodcock, C Batthyany and BA Freeman.  Activation of vascular endothelial nitric oxide synthase and heme oxygenase-1 by electrophilic nitro-fatty acids.  Free Radical Biology and Medicine 48:230-239, 2010.
Khoo NKH, CR White, L Pozzo-Miller, F Zhou, C Constance, T Inoue, RP Patel and DA Parks.  Dietary flavonoid quercetin stimulates vasorelaxation in aortic vessels.  Free Radical Biology and Medicine 49:339-447, 2010.

Dario Vitturi, PhD

Vitturi DA, CS Chen, SR Woodcock, SR Salvatore, G Bonacci, JR Koenitzer, NA Stewart, N Wakabayashi, TW Kensler, BA Freeman and FJ Schopfer.  Modulation of nitro-fatty acid signaling:  Prostaglandin reductase-1 is a nitroalkene reductase. J Biol Chem 288:25626-25637, 2013.
Salvatore SR, DA Vitturi, PR Baker, G Bonacci, JR Koenitzer, SR Woodcock, BA Freeman and FJ Schopfer.  Characterization and quantification of endogenous fatty acid nitroalkene metabolites in human urine.  J Lipid Res 54:1998-2009, 2013.
Vitturi DA, CW Sun, VM Harper, B Thrash-Williams, N Cantu-Medellin, BK Chacko, N Peng, Y Dai, JM Wyss, T Townes and RP Patel.  Antioxidant functions for the hemoglobin ß93 cysteine residue in erythrocytes and in the vascular compartment in vivo.  Free Radic Biol Med 55:119-129, 2013.
Rodriguez C, DA Vitturi, J He, M Vandromme, A Brandon, A Hutchings, LW Rue, JD Kerby and RP Patel.  Sodium nitrite therapy attenuates the hypertensive effects of HBOC-201 via nitrite reduction.  Biochem J 422:423-432, 2009.
Vitturi DA, X Teng, JC Toledo, S Matalon, JR Lancaster and RP Patel.  Regulation of nitrite transport in red blood cells by hemoglobin oxygen fractional saturation.  Am J Physiol Heart Circ Physiol 296:H1394-H1407, 2009.
Isbell TS, CW Sun, LC Wu, X Teng, DA Vitturi, BG Branch, CG Kevil, N Peng, JM Wyss, N Ambalavanan, L Schwiebert, J Ren, KM Pawlik, MB Renfrow, RP Patel and TM Townes.  SNO-hemoglobin is not essential for red blood cell-dependent hypoxic vasodilation.  Nat Med 14:773-777, 2008.
Möller MN, Q Li, DA Vitturi, JM Robinson, JR Lancaster and A Denicola.  Membrane "lens" effect:  Focusing the formation of reactive nitrogen oxides from the *NO/02 reaction.  Chem Res Toxicol 20:709-714, 2007.

Stacy Gelhaus Wendell, PhD

Delmastro-Greenwood M, BA Freeman and SG Wendell.  Redox-dependent anti-inflammatory signaling actions of unsaturated fatty acids.  Annual Review of Physiology 76:79-105, 2014.
Woodcock S, G Bonacci, SL Gelhaus and F Schopfer.  Nitrated fatty acids:  Synthesis and measurement.  Free Radical Biology and Medicine 59:14-26, 2013.
Fajt ML, SL Gelhaus, B Freeman, CE Uvalle, JB Trudeau, F Holguin and SE Wenzel.  Prostaglandin D2 pathway upregulation:  Relation to asthma severity, control, and Th2 inflammation.  J Allergy and Clin Immunol 131:1504-1512, 2013.
Gelhaus SL, O Gilad, TM Penning and IA Blair.  Multidrug resistance protein (MRP) 4 attenuates benzo[a]pyrene-mediated DNA-adduct formation in human bronchoalveolar H358 cells.  Toxicology Letters 209:58-66, 2012.
Gelhaus SL, RG Harvey, TM Penning and IA Blair.  Regulation of benzo[a]pyrene-mediated DNA- and glutathione-adduct formation by 2,3,7,8-tetrachlorodibenzo-p-dioxin in human lung cells.  Chemical Research in Toxicology 24:89-98, 2011.
Bhat S, SL Gelhaus, C Mesaros, A Vachani and IA Blair.  A new liquid chromatography-mass spectrometry method for analysis of urinary 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL).  Rapid Communications in Mass Spectrometry 25:115-121, 2011.
Basu S, C Mesaros, SL Gelhaus and IA Blair.  Stable isotope labeling by essential nutrients in cell culture (SILEC) for preparation of labeled coenzyme A and its thioesters.  Analytical Chemistry 83:1363-1369, 2011.
Park J-H, SL Gelhaus, S Vendantam, AL Oliva, A Batra, IA Blair, AB Troxel, J Field and TM Penning.  The pattern of p53 mutations caused by PAH o-quinones is driven by 8-oxo-dGuo formation while the spectrum of mutations is determined by biological selection for dominance.  Chemical Research in Toxicology 21:1039-1049, 2008.

Steven R. Woodcock, PhD

Woodcock SR, SR Salvatore, G Bonacci, BA Freeman and FJ Schopfer.  Biomimetic nitration of conjugated linoleic acid:  Characterization and synthesis of naturally-occurring conjugated dienes.  J Org Chem 79:25-33, 2014.
Vitturi DA, CS Chen, SR Woodcock, N Stewart, N Wakabayashi, SR Salvatore, G Bonacci, JR Koenitzer, TW Kensler, BA Freeman and FJ Schopfer.  Modulation of nitro-fatty acid signaling:  Prostaglandin reductase-1 is a nitroalkene reductase.  J Bio Chem 288:25626-25637, 2013.
Woodcock SR, G Bonacci, SL Gelhaus and FJ Schopfer.  Nitrated fatty acids:  Synthesis and measurement.  Free Rad Biol Med 59:14-26, 2013.
Bonacci G, PR Baker, SR Salvatore, D Shores, NK Khoo, JR Koenitzer, DA Vitturi, SR Woodcock, F Golin-Bisello, MP Cole, S Watkins, C St. Croix, CI Batthyany, BA Freeman and FJ Schopfer.  Conjugated linoleic acid is a preferential substrate for fatty acid nitration.  J Biol Chem 287:44071-44082, 2012.
Rudolph V, FJ Schopfer, NK Khoo, TK Rudolph, MP Cole, SR Woodcock, G Bonacci, AL Groeger, F Golin-Bisello, CS Chen, PR Baker and BA Freeman.  Nitro-fatty acid metabolome:  Saturation, desaturation, beta-oxidation, and protein adduction.  J Biol Chem 284:1461-1473, 2009.
Woodcock SR, AJV Marwitz, P Bruno and BP Branchaud.  Synthesis of nitrolipids:  All four possible diastereomers of nitro-oleic acids:  (E)- and (Z)-, 9- and 10-Nitro-octadec-9-enoic acids.  Org Lett 8:3931-3833, 2006.