A Method to Monitor the NAD<sup>+</sup> Metabolome—From Mechanistic to Clinical Applications
Nicotinamide adenine dinucleotide (NAD<sup>+</sup>) and its reduced form (NADH) are coenzymes employed in hundreds of metabolic reactions. NAD<sup>+</sup> also serves as a substrate for enzymes such as sirtuins, poly(ADP-ribose) polymerases (PARPs) and ADP-ribosyl cyclases. G...
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2021-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/19/10598 |
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| author | Maria Pilar Giner Stefan Christen Simona Bartova Mikhail V. Makarov Marie E. Migaud Carles Canto Sofia Moco |
| author_facet | Maria Pilar Giner Stefan Christen Simona Bartova Mikhail V. Makarov Marie E. Migaud Carles Canto Sofia Moco |
| author_sort | Maria Pilar Giner |
| collection | DOAJ |
| description | Nicotinamide adenine dinucleotide (NAD<sup>+</sup>) and its reduced form (NADH) are coenzymes employed in hundreds of metabolic reactions. NAD<sup>+</sup> also serves as a substrate for enzymes such as sirtuins, poly(ADP-ribose) polymerases (PARPs) and ADP-ribosyl cyclases. Given the pivotal role of NAD(H) in health and disease, studying NAD<sup>+</sup> metabolism has become essential to monitor genetic- and/or drug-induced perturbations related to metabolic status and diseases (such as ageing, cancer or obesity), and its possible therapies. Here, we present a strategy based on liquid chromatography-tandem mass spectrometry (LC-MS/MS), for the analysis of the NAD<sup>+</sup> metabolome in biological samples. In this method, hydrophilic interaction chromatography (HILIC) was used to separate a total of 18 metabolites belonging to pathways leading to NAD<sup>+</sup> biosynthesis, including precursors, intermediates and catabolites. As redox cofactors are known for their instability, a sample preparation procedure was developed to handle a variety of biological matrices: cell models, rodent tissues and biofluids, as well as human biofluids (urine, plasma, serum, whole blood). For clinical applications, quantitative LC-MS/MS for a subset of metabolites was demonstrated for the analysis of the human whole blood of nine volunteers. Using this developed workflow, our methodology allows studying NAD<sup>+</sup> biology from mechanistic to clinical applications. |
| first_indexed | 2024-03-10T06:58:31Z |
| format | Article |
| id | doaj.art-0820e414ecc345bd87701d172f06110c |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T06:58:31Z |
| publishDate | 2021-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-0820e414ecc345bd87701d172f06110c2023-11-22T16:12:22ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-09-0122191059810.3390/ijms221910598A Method to Monitor the NAD<sup>+</sup> Metabolome—From Mechanistic to Clinical ApplicationsMaria Pilar Giner0Stefan Christen1Simona Bartova2Mikhail V. Makarov3Marie E. Migaud4Carles Canto5Sofia Moco6Nestle Research, EPFL Innovation Park, H, 1015 Lausanne, SwitzerlandNestle Research, EPFL Innovation Park, H, 1015 Lausanne, SwitzerlandNestle Research, EPFL Innovation Park, H, 1015 Lausanne, SwitzerlandMitchell Cancer Institute, University of South Alabama, 1660 Springhill Avenue, Mobile, AL 36604, USAMitchell Cancer Institute, University of South Alabama, 1660 Springhill Avenue, Mobile, AL 36604, USANestle Research, EPFL Innovation Park, H, 1015 Lausanne, SwitzerlandNestle Research, EPFL Innovation Park, H, 1015 Lausanne, SwitzerlandNicotinamide adenine dinucleotide (NAD<sup>+</sup>) and its reduced form (NADH) are coenzymes employed in hundreds of metabolic reactions. NAD<sup>+</sup> also serves as a substrate for enzymes such as sirtuins, poly(ADP-ribose) polymerases (PARPs) and ADP-ribosyl cyclases. Given the pivotal role of NAD(H) in health and disease, studying NAD<sup>+</sup> metabolism has become essential to monitor genetic- and/or drug-induced perturbations related to metabolic status and diseases (such as ageing, cancer or obesity), and its possible therapies. Here, we present a strategy based on liquid chromatography-tandem mass spectrometry (LC-MS/MS), for the analysis of the NAD<sup>+</sup> metabolome in biological samples. In this method, hydrophilic interaction chromatography (HILIC) was used to separate a total of 18 metabolites belonging to pathways leading to NAD<sup>+</sup> biosynthesis, including precursors, intermediates and catabolites. As redox cofactors are known for their instability, a sample preparation procedure was developed to handle a variety of biological matrices: cell models, rodent tissues and biofluids, as well as human biofluids (urine, plasma, serum, whole blood). For clinical applications, quantitative LC-MS/MS for a subset of metabolites was demonstrated for the analysis of the human whole blood of nine volunteers. Using this developed workflow, our methodology allows studying NAD<sup>+</sup> biology from mechanistic to clinical applications.https://www.mdpi.com/1422-0067/22/19/10598NAD<sup>+</sup>metabolomicsmass spectrometry |
| spellingShingle | Maria Pilar Giner Stefan Christen Simona Bartova Mikhail V. Makarov Marie E. Migaud Carles Canto Sofia Moco A Method to Monitor the NAD<sup>+</sup> Metabolome—From Mechanistic to Clinical Applications International Journal of Molecular Sciences NAD<sup>+</sup> metabolomics mass spectrometry |
| title | A Method to Monitor the NAD<sup>+</sup> Metabolome—From Mechanistic to Clinical Applications |
| title_full | A Method to Monitor the NAD<sup>+</sup> Metabolome—From Mechanistic to Clinical Applications |
| title_fullStr | A Method to Monitor the NAD<sup>+</sup> Metabolome—From Mechanistic to Clinical Applications |
| title_full_unstemmed | A Method to Monitor the NAD<sup>+</sup> Metabolome—From Mechanistic to Clinical Applications |
| title_short | A Method to Monitor the NAD<sup>+</sup> Metabolome—From Mechanistic to Clinical Applications |
| title_sort | method to monitor the nad sup sup metabolome from mechanistic to clinical applications |
| topic | NAD<sup>+</sup> metabolomics mass spectrometry |
| url | https://www.mdpi.com/1422-0067/22/19/10598 |
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