Location, Location, Location: Compartmentalization of NAD+ Synthesis and Functions in Mammalian Cells
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The numerous biological roles of NAD+ are organized and coordinated via its compartmentalization within cells. The spatial and temporal partitioning of this intermediary metabolite is intrinsic to understanding the impact of NAD+ on cellular signaling and metabolism. We review evidence supporting the compartmentalization of steady-state NAD+ levels in cells, as well as how the modulation of NAD+ synthesis dynamically regulates signaling by controlling subcellular NAD+ concentrations. We further discuss potential benefits to the cell of compartmentalizing NAD+, and methods for measuring subcellular NAD+ levels.
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NAD+ is highly compartmentalized in the cytoplasm, mitochondria and nucleus, which are its main subcellular pools.
“NAD+ is highly compartmentalized in the cytoplasm, mitochondria and nucleus, which represent its main subcellular pools (Supplementary Box 1).”
From NAD+ metabolism and its roles in cellular processes during ageing (2021) · cites this paper as [2]
The enzymes involved in NAD+ biosynthesis or degradation are highly compartmentalized.
“These pools are regulated independently of each other, and consistent with this, the enzymes involved in the biosynthesis or degradation of NAD+ are highly compartmentalized as well2–4.”
From NAD+ metabolism and its roles in cellular processes during ageing (2021) · cites this paper as [2]
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NAD+ metabolism and its roles in cellular processes during ageing
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Molecular mechanisms of ischemia and glutamate excitotoxicity
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ADP-ribosylation from molecular mechanisms to therapeutic implications
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Principles and functions of metabolic compartmentalization
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PARPs in lipid metabolism and related diseases
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Self-citationNAD metabolism: Role in senescence regulation and aging
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“Synthesis of NAD in the body occurs through both extracellular and intracellular pathways (Cambronne & Kraus, 2020; Cantó et al., 2015; Liu, Su, et al., 2018).”
BackgroundCarboranes as unique pharmacophores in antitumor medicinal chemistry
Chen, Y., Du, F., Tang, L., Xu, J., Zhao, Y., Wu, X., Li, M., Shen, J., Wen, Q., Cho, C. H., et al. (2022) · Molecular Therapy — Oncolytics · doi:10.1016/j.omto.2022.01.005 · Cited by 124 · see citing works
Crosstalk between metabolism and epigenetics during macrophage polarization
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“This compartmentalization appears crucial for the immune response to foreign invaders but poses significant challenges for investigation as techniques to measure sirtuin targets and compartment-specific NAD + levels are limited and not yet widely accessible [156, 173].”
The autophagy–NAD axis in longevity and disease
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Emerging role of PARP‐1 and PARthanatos in ischemic stroke
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“Interestingly, NAD+ pools in these subcellular compartments have non- redundant functions (Cambronne & Kraus, 2020).”
BackgroundAxonal energy metabolism, and the effects in aging and neurodegenerative diseases
Yang, S., Park, J. H., & Lu, H.-C. (2023) · Molecular Neurodegeneration · doi:10.1186/s13024-023-00634-3 · Cited by 94 · see citing works
Tryptophan metabolism and disposition in cancer biology and immunotherapy
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PARPs and ADP-ribosylation: Deciphering the complexity with molecular tools
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The therapeutic perspective of NAD+ precursors in age-related diseases
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Subcellular NAD+ pools are interconnected and buffered by mitochondrial NAD+
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The Drama of Wallerian Degeneration: The Cast, Crew, and Script
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Age-Dependent Decline of NAD+—Universal Truth or Confounded Consensus?
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“We know from animal studies that mitochondrial, nuclear and cytoplasmic NAD+ levels are all regulated individually [100].”
BackgroundImpact of NAD+ metabolism on ovarian aging
Liang, J., Huang, F., Song, Z., Tang, R., Zhang, P., & Chen, R. (2023) · Immunity & Ageing · doi:10.1186/s12979-023-00398-w · Cited by 56 · see citing works
NAD+ metabolism-based immunoregulation and therapeutic potential
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“Thus, PARPs and sirtuins in the same subcellular compartmentation have counterbalancing roles in the regulation of NAD+ homeostasis [55].”
BackgroundPu, Q., Guo, X., Hu, J.-J., Li, A.-L., Li, G., & Li, X. (2022) · Biomedicine & Pharmacotherapy · doi:10.1016/j.biopha.2022.112659 · Cited by 51 · see citing works
Poljšak, B., Kovač, V., Špalj, S., & Milisav, I. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms24032959 · Cited by 58 · see citing works
“NAD+ is also phosphorylated by NAD+ kinase to generate NADP+, which in turn is reduced by dehydrogenases to generate NADPH [12].”
BackgroundInfluentialEmerging strategies, applications and challenges of targeting NAD+ in the clinic
Zhang, J., Wang, H., Lautrup, S., Nilsen, H. L., Treebak, J. T., Watne, L. O., Selbæk, G., Wu, L. E., Omland, T., Pirinen, E., et al. (2025) · Nature Aging · doi:10.1038/s43587-025-00947-6 · Cited by 58 · see citing works
“Additionally, NAD + turnover rates vary across tissues and physiological or pathological states, making steady-state measurements insufficient for capturing these dynamic differences 197 .”
Sirtuins: The NAD + -Dependent Multifaceted Modulators of Inflammation
Santos, L., Benítez-Rosendo, A., Bresque, M., Camacho-Pereira, J., Calliari, A. J., & Escande, C. (2023) · Antioxidants and Redox Signaling · doi:10.1089/ars.2023.0295 · Cited by 46 · see citing works
“Reported NAD + concentrations in cells vary between studies and methodologies, but are generally in the range of 50–120 l M , making this a very abundant molecule in cells (Cambronne and Kraus, 2020).”
SLC25A51 decouples the mitochondrial NAD+/NADH ratio to control proliferation of AML cells
Lu, M., Busquets, J., Impedovo, V., Wilson, C. N., Chan, H., Chang, Y.-T., Matsui, W. H., Tiziani, S., & Cambronne, X. A. (2024) · Cell Metabolism · doi:10.1016/j.cmet.2024.01.013 · Cited by 45 · see citing works
Self-citationBalancing NAD+ deficits with nicotinamide riboside: therapeutic possibilities and limitations
Cercillieux, A., Ciarlo, E., & Cantó, C. (2022) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-022-04499-5 · Cited by 41 · see citing works
Coordinated metabolic transitions and gene expression by NAD+ during adipogenesis
Sánchez-Ramírez, E., Ung, T. P. L., del Carmen, A. A., del Toro-Ríos, X., Fajardo-Orduña, G. R., Noriega, L. G., Cortés-Morales, V. A., Tovar, A. R., Montesinos, J. J., Orozco-Solís, R., et al. (2022) · The Journal of Cell Biology · doi:10.1083/jcb.202111137 · Cited by 41 · see citing works
Local and dynamic regulation of neuronal glycolysis in vivo
Wolfe, A. D., Koberstein, J. N., Smith, C. B., Stewart, M. L., Gonzalez, I. J., Hammarlund, M., Hyman, A. A., Stork, P. A., Goodman, R. H., & Colón‐Ramos, D. A. (2024) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2314699121 · Cited by 36 · see citing works
“There is considerable evidence that different regions within a neuron have distinct metabolic capabilities depending on their differing and often dynamic energy requirements (29–31).”
BackgroundZeng, H., Xu, J., Wang, X., Li, S., & Han, Z. (2022) · Ecotoxicology and Environmental Safety · doi:10.1016/j.ecoenv.2022.113441 · Cited by 39 · see citing works
Current Uncertainties and Future Challenges Regarding NAD+ Boosting Strategies
Poljšak, B., Kovač, V., & Milisav, I. (2022) · Antioxidants · doi:10.3390/antiox11091637 · Cited by 39 · see citing works
Exploring the role of parthanatos in CNS injury: Molecular insights and therapeutic approaches
Zhang, J., Hu, X., Geng, Y., Xiang, L., Wu, Y., Li, Y., Yang, L., & Zhou, K. (2024) · Journal of Advanced Research · doi:10.1016/j.jare.2024.04.031 · Cited by 37 · see citing works
“Nicotinamide adenine dinucleotide (NAD + ) is a coenzyme for redox reactions and is involved in multiple catabolic pathways, including glycolysis, glutaminolysis, and fatty acid oxidation, thus making it central to energy metabolism [39–41].”
Chen, Y., Yang, L., Lu, Y., Liu, N., Ma, W., Fan, H., Hu, Q., Han, X., Gan, W., & Li, D. (2022) · Cancer Letters · doi:10.1016/j.canlet.2022.215689 · Cited by 34 · see citing works
Metabolic Fuel for Epigenetic: Nuclear Production Meets Local Consumption
Boon, R. (2021) · Frontiers in Genetics · doi:10.3389/fgene.2021.768996 · Cited by 31 · see citing works
“These pathways converge in the generation of nicotinic acid mononucleotide (NaMN) or nicotinamide mononucleotide (NMN) that are then condensed with AMP through the activity of compartmentalized NMN (NMNAT) enzymes (Cambronne and Kraus, 2020).”
BackgroundInfluentialDwivedi, S. K., Arachchige, D. L., Waters, M. G. J., Jaeger, S., Mahmoud, M. E., Olowolagba, A. M., Tucker, D. R., Geborkoff, M. R., Werner, T., Luck, R. L., et al. (2023) · Sensors and Actuators B Chemical · doi:10.1016/j.snb.2023.135073 · Cited by 30 · see citing works
A metabolic signature for NADSYN1-dependent congenital NAD deficiency disorder
Szot, J. O., Cuny, H., Martin, E. M. M. A., Sheng, D. Z., Iyer, K. R., Portelli, S., Nguyen, V., Gereis, J. M., Alankarage, D., Chitayat, D. A., et al. (2024) · Journal of Clinical Investigation · doi:10.1172/jci174824 · Cited by 34 · see citing works
Uncovering the Invisible: Mono-ADP-ribosylation Moved into the Spotlight
Hopp, A.-K., & Hottiger, M. O. (2021) · Cells · doi:10.3390/cells10030680 · Cited by 30 · see citing works
“The salvage pathway depends on the expression level of nicotinamide phosphoribosyltransferase (NAMPT) and the nicotinamide mononucleotide adenylyl transferases (NMNATs), NMNAT1, 2 and 3 [42,43].”
BackgroundNew cases that expand the genotypic and phenotypic spectrum of Congenital NAD Deficiency Disorder
Szot, J. O., Slavotinek, A. M., Chong, K., Brandau, O., Nezarati, M. M., Cueto-González, A. M., Patel, M. S., Devine, W. P., Rego, S. M., Acyinena, A. P., et al. (2021) · Human Mutation · doi:10.1002/humu.24211 · Cited by 34 · see citing works
“Owing to an inability to import NAD, mammalian cells synthesize NAD de novo from L‐tryptophan via the kynurenine pathway, or from the conversion of its dietary precursors nicotinamide, nicotinic acid, and nicotinamide riboside via the Preiss‐Handler and salvage pathways (Cambronne & Kraus, 2020).”
BackgroundOdoh, C. K., Guo, X., Arnone, J. T., Wang, X., & Zhao, Z. K. (2022) · Biogerontology · doi:10.1007/s10522-022-09958-x · Cited by 29 · see citing works
“The NAD molecule along with the biosynthesis and degradation enzymes are compartmentalized in the cell cytoplasm and also regulated within other cellular organelles (mitochondria and nucleus) (Cambronne and Kraus 2020; Verdin 2015).”
BackgroundOrgan Protection by Caloric Restriction Depends on Activation of the De Novo NAD+ Synthesis Pathway
Späth, M. R., Hoyer-Allo, K. J. R., Seufert, L., Höhne, M., Lucas, C., Bock, T., Isermann, L., Brodesser, S., Lackmann, J., Kiefer, K., et al. (2023) · Journal of the American Society of Nephrology · doi:10.1681/asn.0000000000000087 · Cited by 28 · see citing works
Channeling Nicotinamide Phosphoribosyltransferase (NAMPT) to Address Life and Death
Reddy, V. G., Krider, I. S., Alves, E. T. M., Courey, J. M., Laham, M. S., & Thatcher, G. R. J. (2024) · Journal of Medicinal Chemistry · doi:10.1021/acs.jmedchem.3c02112 · Cited by 27 · see citing works
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The evolution of small molecule enzyme activators
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“…and, especially, microbiota-derived catabolites that reach extraintestinal tissues on delayed timelines set by distinctive pharmacokinetics [76]. γ -VLs from flavan-3-ols align with AhR-linked outputs alongside AMPK/PPAR tuning, consistent with modest-affinity ligands gaining leverage via…”
- Authors
- Cambronne, X. A., & Kraus, W. L.
- Affiliations
- The University of Texas at Austin (US)
- The University of Texas Southwestern Medical Center (US)
- Corresponding author
- Cambronne, Xiaolu A.
- Kraus, W. Lee
- Venue
- Trends in Biochemical Sciences
- Publisher
- Elsevier BV
- Published
- 2020-10
- Volume
- 45
- Issue
- 10
- Pages
- 858-873
- ISSN
- 0968-0004 (ISSN-L)
- PMC
- PMC7502477
- OpenAlex
- W3037844642
- Cited by
- 137
- References
- 130
- Citations by year
- 2026: 25 · 2025: 23 · 2024: 26 · 2023: 25 · 2022: 21 · 2021: 15 · 2020: 2
- FWCI
- 1.95
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Trends in Biochemical Sciences (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health
- Topics
- Health Sciences › Medicine › Geriatrics and Gerontology › Sirtuins and Resveratrol in Medicine
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Calcium signaling and nucleotide metabolism
- Keywords
- NAD+, compartmentalization, NAD+ synthesis, cellular metabolism, signal transduction, subcellular compartmentalization
- MeSH terms
- Animals, Cell Compartmentation, NAD, Subcellular Fractions, Signal Transduction
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