Nitric oxide (NO), which is synthesized in the body via enzymatic oxidation of L-arginine, is critical to numerous physiological functions but also can contribute to the severity of diseases such as cancer or pathophysiological conditions such as stroke. This diversity in the responses to NO biosynthesis is a reflection of the diverse chemistry of NO. For instance NO can alter the function of enzymes by binding to metal centers. This type of interaction could result in outcomes as disparate as control of blood pressure or death of an invading bacterium. NO can also be readily converted to higher nitrogen oxides such as N₂O₃, NO₂ or ONOOH, which have discrete chemical and biological properties. The ultimate result will depend upon numerous factors, particularly the location and concentration of NO produced. Therefore, site-specific modulation of NO concentration offers intriguing therapeutic possibilities for an expanding list of diseases.

The overall goal of my research is to provide pharmacological therapies for diseases such as cancer, heart failure and stroke. Accomplishing this goal requires both the development of compounds to deliver NO and other nitrogen oxides and a full understanding of the biological consequences of nitrogen oxide production by these compounds. To this end, we are currently engaged in the investigation of the reactivity of nitrogen oxides at the molecular, biomolecular, systemic and whole organism levels.

Our current projects are designed to answer questions of potential medical importance through a multi-disciplinary approach, including analytical, synthetic, inorganic and biochemical techniques. The project categories include:

1. development and utilization of analytical techniques for detection and measurement of NO and other nitrogen oxides as well as the resultant chemistry of these species

2. synthesis of potential donors of NO and other nitrogen oxides

3. chemical characterization of these compounds (spectroscopic features, kinetics, mechanisms and profiles of nitrogen oxide release, etc.)

4. biological characterization of these compounds (assay of effects on biological compounds, mechanisms and pathways, in vitro determination of potential for therapeutic utility, etc.)

5. identification of potential targets, such as enzymes, for treatment of disease through exposure to nitrogen oxide donors

Katori T., Donzelli S., Miranda K.M., Thomas D.D., Tocchetti C.G., Lee M.J., Mancardi D., Kass D.A., Wink D.A. and Paolocci N. "Peroxynitrite and myocardial contractility: in vivo versus in vitro effects". Free Radic. Biol. Med., 15, 1601-1618 (2006)
Donzelli S., Espey M.G., Thomas D.D., Mancardi D., Tocchetti C.G., Ridnour L.A., Paolocci N., King S.B., Miranda K.M., Lazzarino G., Fukuto J.M. and Wink D.A., "Discriminating formation of HNO from other reactive nitrogen oxide species". Free Radic. Biol. Med., 40, 1056-1066 (2006)
Dutton A.S., Suhrada C.P., Miranda K.M., Wink D.A., Fukuto J.M. and Houk K.N., Mechanism of pH dependent decomposition of monoalkylamine diazeniumdiolates to form HNO and NO, deduced from Density Functional Theory and CBS-QB3 calculations". Inorg. Chem., 45, 2448-2456 (2006)
Miranda K.M., "The chemistry of nitroxyl (HNO) and implications in biology". Coord. Chem. Rev., 249, 433-455 (2005)
Miranda K.M., Dutton A.S., Ridnour L.A., Foreman C.A., Ford E., Paolocci N., Katori T., Mancardi D., Thomas D.D., Espey M.G., Houk K.N., Fukuto J.M. and Wink D.A., "Mechanism of aerobic decomposition of Angeli’s salt (sodium trioxodinitrate) at physiological pH". J. Am. Chem. Soc., 127, 722-31 (2005)
Miranda K.M., Nagasawa H.T. and Toscano J.P., "Donors of HNO". Curr. Top. Med. Chem., 5, 649-664 (2005)
Miranda K.M., Katori T., Torres de Holding C.L., Thomas L., Ridnour L.A., McLendon W.J., Cologna S., Dutton A.S., Champion H.C., Mancardi D., Tocchetti C.G., Saavedra J., Keefer L.K., Houk K.N., Fukuto J.M., Kass, D.A., Paolocci N. and Wink D.A., "Comparison of the NO and HNO donating properties of diazeniumdiaolates. Primary amine adducts release HNO in vivo". J. Med. Chem., 48, 8220-8228 (2005)
Miranda K.M., Ridnour L., Espey M., Katori T., Tocchetti C.G., Thomas D., Mancardi D., Ferlito M., Donzelli S., Fukuto J., Paolocci N. and Wink D.A., "The chemical biology of NO and HNO: an inorganic perspective". Prog. Inorg. Chem., 54, 349-384 (2005)
K.M. Miranda, N. Paolocci, T. Katori, D.D. Thomas, M.D. Bartberger, M.G. Espey, D.A. Kass, M. Feelisch, J.M. Fukuto and D.A. Wink, "A biochemical rationale for the orthogonal behavior of nitroxyl and nitric oxide in the cardiovascular system", Proc. Natl. Acad. Sci. USA, 100, 9196-9201 (2003)
D.A. Wink, K.M. Miranda, T. Katori, D. Mancardi, D. Thomas, L. Ridnour, M.G. Espey, M. Feelisch, C. Colton, J. Fukuto, D.A. Kass, N. Paolocci, "The orthogonal properties of the redox siblings nitroxyl (HNO) and nitric oxide (NO) in the cardiovascular system: a novel redox paradigm", Am. J. Physiol. Heart Circ. Physiol., 285, 2264-2276 (2003)
N. Paolocci, T. Katori, H.C. Champion, M.E. St. John, K.M. Miranda, J.M. Fukuto, D.A. Wink, D.A. Kass, "Positive inotropic and lusitropic effects of HNO/NO^- in failing hearts: independence from β-adrenergic signaling", Proc. Natl. Acad. Sci. USA 100, 5537-5542 (2003). A “From the cover” article.
K.M. Miranda, R.W. Nims, D.D. Thomas, M.G. Espey, D. Citrin, M.D. Bartberger, N. Paolocci, J.M. Fukuto, M. Feelisch and D.A. Wink, "Comparison of the reactivity of nitric oxide and nitroxyl with heme proteins. A chemical discussion of the differential biological effects of these redox related products of NOS", J. Inorg. Biochem. 93, 52-60 (2003)
D.D. Thomas, K.M. Miranda, M.G. Espey, D. Citrin, D. Jourd’heuil, N. Paolocci, S. Hewett, C.A. Colton, M. Grisham, M. Feelisch and D.A. Wink, "Guide for the use of nitric oxide (NO) donors as probes of the chemistry of NO and related redox species in biological systems", Methods Enzymol. 359, 84-105 (2003)
M.D. Bartberger, W. Liu, E. Ford, K.M. Miranda, C. Switzer, J.M. Fukuto, P.J. Farmer, D.A. Wink and K.N. Houk, "The reduction potential of nitric oxide (NO) and its importance to NO biochemistry", Proc. Natl. Acad. Sci. USA 99, 10958-10963 (2002)
K.M. Miranda, M.G. Espey and D.A. Wink, "A rapid, simple spectrophotometric method for detection of nitrate and nitrite", Nitric Oxide, 5, 62-71 (2001). Cover feature.