A. Personal Statement 

I study the formation and signaling of bioactive lipids in cell and animal models while translating these basic science discoveries to clinical collaborations. Bioactive lipids signal through G protein coupled receptor (GPCR) pathways as well as non-GPCR mediated pathways and are responsible for many cellular functions including the regulation of inflammatory responses and cellular proliferation. The overall scope of my work focuses on understanding key events that regulate inflammation and proliferation in lung and airway disease as well as the role of dehydrogenase enzymes relevant to the formation of bioactive, electrophilic fatty acids. Recent work on the role of a particular dehydrogenase, 15-hydroxyprostaglandin dehydrogenase, has led to the discovery that it is involved in omega-3 fatty acid metabolism, thus leading to the formation of omega-3-derived bioactive lipids. Other studies focus on the formation and signaling actions of nitro-fatty acids (NO2-FA). A recent pilot study demonstrated that oral supplementation of 15N-labeled nitrate and nitrite in conjunction with conjugated linoleic acid resulted in the formation of the anti-inflammatory nitro-fatty acid, nitro-conjugated linoleic acid (NO2-cLA), in healthy humans. This pilot study along with other basic research studies in cell and animal models related to anti-inflammatory signaling of bioactive lipids provide the background and expertise necessary to move this project into the obese asthmatic phenotype. In addition to my research pursuits, I am the Director of Metabolomics for the Biomedical Mass Spectrometry Center at the University of Pittsburgh and I have extensive experience and expertise in the development of liquid chromatography mass spectrometry (LC-MS) methods for the analysis of fatty acids, exogenous and endogenous biomarkers of disease, and drug metabolites. I have published several reviews and book chapters on metabolite identification by LC-MS and fatty acids in disease. In this proposed study I will be responsible for the LC-MS analysis of steroids. 

B. Positions and Honors 

Positions and Employment

2005 - 2005 Scientist, UPM Pharmaceuticals, Baltimore, MD 

2010 - 2011 Research Associate, University of Pennsylvania, Depart. of Pharmacology, Philadelphia, PA

2011 - Research Assistant Professor, University of Pittsburgh, Department of Pharmacology & Chemical Biology, Pittsburgh, PA 

2015 - Director of Metabolomics, University of Pittsburgh, Biomedical Mass Spectrometry Center, Pittsburgh, PA 

Other Experience and Professional Memberships 

2001- American Chemical Society 

2005- National Postdoctoral Association 

2006- American Society for Mass Spectrometry 

2008-2011 National Postdoctoral Association, Board of Directors Member 

2008-2012 American Association for the Advancement of Science 

2008-2011 American Association for Cancer Research 

2009-2010 Chair, National Postdoctoral Association, Board of Directors 

2011- American Thoracic Society 

2011 Treasurer, National Postdoctoral Association, Board of Directors 

2013- Society for Free Radical Biology and Medicine 

2013- Member of Faculty Senate Student Affairs Committee 

2014- American Society for Biochemistry and Molecular Biology 

Honors 

1999 American Chemical Society Outstanding Achievement Award in Chemistry 

1999 American Institute of Chemists Foundation Student Award for Outstanding Biochemistry Major 

1999 The James M. Murray Memorial Prize in Chemistry 

2007 Graduate Research Conference, First Place Poster Award, UMBC 

2007 NPA Alfred P. Sloan Foundation Postdoctoral Leadership Mentoring Travel Award 

2013 AB Sciex Young Investigator Award 

2013 NHLBI NIH Loan Repayment Program 

C. Contribution to Science

16 research articles, 3 reviews, H index 15 

1. My graduate thesis work focused on the development of novel analytical purification techniques for modified DNA/RNA primers and secondary structures. These structures are often synthesized with modified bases to test polymerase activity or to determine how modifications alter nucleic acid secondary structure. The subtlety of these modifications, such as N2-methyl- or O6-methyl-guanine, could not be distinguished and purified from a sequence without modification using conventional technology (electrophoresis or ion chromatography). I developed an ion-pairing reversed phase chromatography method to separate and purify identical primers differing only by a methyl group modification. This analytical method development was superior to all other chromatographic methods and insured that the identity of DNA and RNA primers being used in biochemical analyses was accurate and the purity was > 99%.

a. Gelhaus SL, LaCourse WR, et al. Rapid purification of RNA secondary structures. Nucleic Acids Res. 2003 Nov 1;31(21):e135. PubMed PMID: 14576335; PubMed Central PMCID: PMC275492. 

b. Gelhaus SL, LaCourse WR. Separation of modified 2'-deoxyoligonucleotides using ion-pairing reversed-phase HPLC. J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Jun 25;820(2):157-63. PubMed PMID: 15899369) 

c. Perrino FW, Blans P, Harvey S, Gelhaus SL, McGrath C, et al. The N2-ethylguanine and the O6-ethyl- and O6-methylguanine lesions in DNA: contrasting responses from the "bypass" DNA polymerase eta and the replicative DNA polymerase alpha. Chem Res Toxicol. 2003 Dec;16(12):1616-23. PubMed PMID: 14680376. 

d. Perrino FW, Harvey S, Blans P, Gelhaus SL, LaCourse WR, et al. Polymerization past the N2-isopropylguanine and the N6-isopropyladenine DNA lesions with the translesion synthesis DNA polymerases eta and iota and the replicative DNA polymerase alpha. Chem Res Toxicol. 2005 Sep;18(9):1451-61. PubMed PMID: 16167838)

2. I discovered that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) paradoxically downregulates B[a]P-diol epoxide (BPDE)-deoxyguanosine adduct levels in lung cancer cell lines. B[a]P is implicated in the promotion of environmentally-induced lung carcinogenesis. Metabolism by CYP1A/1B1 leads to the formation of the mutagenic metabolite BPDE. As a postdoc I discovered that TCDD also induced a glutathione S-transferase isoform leading to increased formation of BPDE-glutathione adduct. The BPDE-GSH adduct was exported by the multi-drug resistance transporter MRP4 and this phenomenon could be inhibited with probenecid. This was my primary project during my postdoctoral work at Penn for which I received an F32 NRSA fellowship. This work resulted in a total of 5 publications.

a. Gelhaus SL, et al. Regulation of benzo[a]pyrene-mediated DNA- and glutathione-adduct formation by 2,3,7,8-tetrachlorodibenzo-p-dioxin in human lung cells. Chem Res Toxicol. 2011 Jan 14;24(1):89-98. PubMed PMID: 21028851; PubMed Central PMCID: PMC3021323 

b. Gelhaus SL, et al. Multidrug resistance protein (MRP) 4 attenuates benzo[a]pyrene-mediated DNA-adduct formation in human bronchoalveolar H358 cells. Toxicol Lett. 2012 Feb 25;209(1):58-66. PubMed PMID: 22155354; PubMed Central PMCID: PMC3256298 

c. Jiang H, Gelhaus SL, et al. Metabolism of benzo[a]pyrene in human bronchoalveolar H358 cells using liquid chromatography-mass spectrometry. Chem Res Toxicol. 2007 Sep;20(9):1331-41. PubMed PMID: 17702526; PubMed Central PMCID: PMC2423818 

d. Park JH, Gelhaus SL, et al. 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. Chem Res Toxicol. 2008 May;21(5):1039-49. PubMed PMID: 18489080; PubMed Central PMCID: PMC2671329). 

3. I contributed to the proof of concept that arachidonic acid (AA) metabolites previously thought to be inactive are conferred with biological activity. 15-hydroxyprostaglandin dehydrogenase (15PGDH) is primarily known for its “inactivation” of prostaglandin E2 (PGE2). However, the metabolite of PGE2, 15-ketoPGE2 is an electrophile that contains an α,β-unsaturated ketone moiety. This moiety confers 15-ketoPGE2 and other arachidonic acid-derived metabolites with anti-inflammatory and anti-proliferative properties. Cell culture studies with AA-derived 15-oxoETE demonstrated that this electrophilic fatty acid was able to induce the Nrf2-mediated antioxidant response pathway while simultaneously inhibiting NF-κB-mediated cytokine expression. These results are consistent with emerging results in the literature showing that electrophilic fatty acids signal via Michael addition adduct formation with reactive nucleophiles in transcriptional regulatory proteins and pro-inflammatory enzymes. This work started at the end of my postdoc and I have continued it in my junior faculty position.

a. Delmastro-Greenwood M, Freeman BA, Wendell SG. Redox-dependent anti-inflammatory signaling actions of unsaturated fatty acids. Annu Rev Physiol. 2014;76:79-105. PubMed PMID: 24161076; PubMed Central PMCID: PMC4030715 and 

b. Snyder NW, Golin-Bisello F, Gao Y, Blair IA, Freeman BA, Wendell, SG. 15-Oxoeicosatetraenoic acid is a 15-hydroxyprostaglandin dehydrogenase-derived electrophilic mediator of inflammatory signaling pathways. Chem Biol Interact. 2014 Nov 4;PubMed PMID: 25450232; NIHMS640480; PMCID: PMC4414684 In Process 

4. I have identified novel bioactive lipid mediators derived from 15-hydroxyprostaglandin dehydrogenase metabolism. I have demonstrated that HDoHEs [hydroxylated metabolites of docosahexaenoic acid (DHA)] are substrates of 15PGDH and contribute to the overall beneficial effects of omega-3 fatty acids. Metabolism of DHA-derived HDoHEs by 15PGDH leads to the formation of electrophilic fatty acids that are conferred with anti-inflammatory signaling properties. These metabolites inhibited NF-κB-mediated cytokine expression in primary alveolar macrophages. I am the primary driver of this work in my faculty role. 

a. Wendell SG, Golin-Bisello F, Wenzel S, Sobol RW, Holguin F, et al. 15-hydroxyprostaglandin dehydrogenase generation of electrophilic lipid signaling mediators from hydroxy ω-3 Fatty acids. J Biol Chem. 2015 Feb 27;290(9):5868-80. PubMed PMID: 25586183; PubMed Central PMCID: PMC4342494 

b. Wendell SG, Baffi C, Holguin F. Fatty acids, inflammation, and asthma. J Allergy Clin Immunol. 2014 May;133(5):1255-64. PubMed PMID: 24613565; PubMed Central PMCID: 4417548.

5. I have demonstrated that the dietary constituents, nitrate, nitrite, and conjugated linoleic acid are responsible for the formation of nitro-conjugated linoleic acid (NO2-cLA) the primary endogenous nitro-fatty acid electrophile that promotes anti-inflammatory signaling by forming Michael adducts with nucleophilic amino acids (primarily cysteine) in key reactive transcription factors and enzymes such as Nrf2, NF-κB, PPARγ, heat shock factor, and soluble epoxide hydrolase, to name a few. 

a. Delmastro-Greenwood, Meghan, Hughan, KS, Vitturi, DA, Salvatore, SR, Grimes, G, Potti, G, Shiva, S, Schopfer, FJ, Gladwin, MT, Freeman, BA, and SG Wendell. Nitrite and nitrate-dependent generation of anti-inflammatory fatty acid nitroalkenes. FRBM. 2015. DOI 10.1016/j.freeradbiomed.2015.07.149. PubMed PMID: 26385079; NIHMS723667; PMCID: PMC4684780 In Progress. 

View a complete list of published work in MyBibliography