Energy, Longevity, Brain Health Anti-Inflammatory

NAD+ Energy, longevity, brain health anti-inflammatory

Ray Griffiths MSc Clinical Education Associate Webinar Disclosure

Webinars such as this are for educational purposes only and are intended for health care practitioners.

These therapies are not substitutions for standard medical care. Practitioners are solely responsible for the care and treatment provided to their own patients. Rajman, L., Chwalek, K. and Sinclair, D.A., 2018. Therapeutic potential of NAD- boosting molecules: the in vivo evidence. Cell metabolism, 27(3), pp.529-547. Covered in this presentation:

Energy production Sirtuins (SIRTs) our anti-ageing proteins Poly(ADP-ribose) polymerases (PARPs) – DNA repair CD38 – Immune cell surface marker which degrades NAD+ Enzymes - dehydrogenases NAD+ For energy NAD+ a vital part of the energy web of life

Flow of energy largely determines the structure of ecosystems via food webs and taste sensors with appetite and quality controls, that avoid toxins but allow self- medication…

…diet, along with domestication of microbes as gut symbionts, provide or manufacture vitamins, including nicotinamide/niacin. This vitamin has evolved to supply and translocate energy and information via NAD+ and NADH

Williams, A.C., Hill, L.J. and Ramsden, D.B., 2012. Nicotinamide, NAD (P)(H), and methyl-group homeostasis evolved and became a determinant of ageing diseases: hypotheses and lessons from pellagra. Current gerontology and geriatrics research, 2012. Pellagra – in effect an NAD+ deficiency?

Overt niacin or NAD+ deficiency is called pellagra

Symptoms are dementia, depression, dermatitis, diarrhoea, and death

But could subclinical pellagra be driving many chronic Western diseases?

NAD+ exists in two forms – oxidised and reduced Our mitochondria are effectively solar powered

Energy from sunlight is used to remove electrons from water to synthesise plants. We eat the plants, to extract the high energy electrons, and then shuttle them via NADH to the mitochondrial electron transport chain. The high energy electrons are contained in the bond of NAD+ and H+ to form NADH Our mitochondria are effectively solar powered You would think that higher NADH levels are better for health?

…but not so fast! When not carrying ‘energy bricks’ NAD+ has other vital roles to play The mitochondrial NAD+ pool is relatively distinct from that of the rest of the cell, while cytoplasmic NAD+/NADH ratios range between 60 and 700 in a typical eukaryotic cell, mitochondrial NAD+/NADH ratios are maintained at 7 to 8

These ratios decline as we age

Zhu, X.H., Lu, M., Lee, B.Y., Ugurbil, K. and Chen, W., 2015. In vivo NAD assay reveals the intracellular NAD contents and redox state in healthy human brain and their age dependences. Proceedings of the National Academy of Sciences, p.201417921. Stein, L.R. and Imai, S.I., 2012. The dynamic regulation of NAD metabolism in mitochondria. Trends in Endocrinology & Metabolism, 23(9), pp.420-428. NAD + decreases as we age NADH increases as we age

It’s estimated that by 40-50, most people will have half the NAD+ levels of their youth

Zhu, X.H., Lu, M., Lee, B.Y., Ugurbil, K. and Chen, W., 2015. In vivo NAD assay reveals the intracellular NAD contents and redox state in healthy human brain and their age dependences. Proceedings of the National Academy of Sciences, p.201417921. Schultz, M.B. and Sinclair, D.A., 2016. Why NAD+ declines during aging: it’s destroyed. Cell metabolism, 23(6), pp.965-966. A lower NAD+/NADH ratio is strongly implicated in mitochondrial disorders and, age- related disorders including diabetes, obesity, neurodegeneration and cancer

The NAD+/NADH ratio is in equilibrium with the pyruvate/lactate ratio

Srivastava, S., 2016. Emerging therapeutic roles for NAD+ metabolism in mitochondrial and age-related disorders. Clinical and Translational Medicine, 5(1), pp.1-11. Mintun, M.A., Vlassenko, A.G., Rundle, M.M. and Raichle, M.E., 2004. Increased lactate/pyruvate ratio augments blood flow in physiologically activated human brain. Proceedings of the National Academy of Sciences, 101(2), pp.659-664. Parkinson’s patient organic acid results from Great Plains and Genova Same urine sample! Oxidative stress in schizophrenia The importance of the NAD+/NADH Ratio

In the 1960s, psychiatrists Abraham Hoffer and Humphry Osmond gave as much as 17g of niacin to schizophrenic patients with impressive results

Recent research has pointed to a dramatic drop in the NAD+/NADH Ratio during a patient’s first episode of schizophrenia. This fall in the ratio points to very high levels of oxidative stress within the central nervous system of the patient

It is likely that Hoffer and Osmond were effectively increasing the NAD+/NADH ratio to support schizophrenic patients

Carter, S., 2019. Origins of Orthomolecular Medicine. Integrative Medicine: A Clinician's Journal, 18(3), p.76. Kim, S.Y., Cohen, B.M., Chen, X., Lukas, S.E., Shinn, A.K., Yuksel, A.C., Li, T., Du, F. and Öngür, D., 2017. Redox dysregulation in schizophrenia revealed by in vivo NAD+/NADH measurement. Schizophrenia bulletin, 43(1), pp.197-204. NAD+ and caffeine to support motor neurons

Green tea also supports Nmnat2

Zwilling, M., Theiss, C. and Matschke, V., 2020. Caffeine and NAD+ improve motor neural integrity of dissociated wobbler cells in vitro. Antioxidants, 9(6), p.46 Lau, C., 2008. Functional characterisation of three human nicotinamide mononucleotide adenylyltransferases. Human PD patient stem cell-derived dopaminergic neurons exhibit reduced NAD+ levels

Fulleylove-Krause, B., Sison, S. and Ebert, A., 2020. Nicotinamide mononucleotide treatment increases NAD+ levels in an iPSC Model of Parkinsons Disease. bioRxiv. Parkinson’s disease is often linked with mitochondrial complex I dysfunction

Complex I dysfunction is translated to an increased production of reactive oxygen species and a decreased energy supply

NNAD+/NADH

Tapias, V., McCoy, J.L. and Greenamyre, J.T., 2019. Phenothiazine normalizes the NADH/NAD+ ratio, maintains mitochondrial integrity and protects the nigrostriatal dopamine system in a chronic rotenone model of Parkinson's disease. Redox biology, 24, p.101164. Marella, M., Seo, B.B., Yagi, T. and Matsuno-Yagi, A., 2009. Parkinson’s disease and mitochondrial complex I: a perspective on the Ndi1 therapy. Journal of bioenergetics and biomembranes, 41(6), pp.493-497. NAD+ proteoxicity and AGEs

Lowered NAD+ availability can lead to generation of methylglyoxal from glycolytic intermediates

…which in turn can damage proteins (AGEs), promote generation of reactive oxygen species and induce mitochondrial dysfunction Methylglyoxal = AGEs Low NAD+ blocks glycolysis with catastrophic results Hipkiss, A.R., 2010. NAD+ and metabolic regulation of age-related proteoxicity: A possible role for methylglyoxal?. Experimental gerontology, 45(6), pp.395-399. So is obesity really an NAD+ deficiency?

Is it a desperate attempt to shunt excess energy into adipose tissue to regenerate vital NAD+?

French, S.W., 2016. Chronic alcohol binging injures the liver and other organs by reducing NAD+ levels required for sirtuin's deacetylase activity. Experimental and molecular pathology, 100(2), pp.303-306. Pau D’arco helps regenerate NAD+ from NADH

β-lapachone from the herb Pau D’arco is a metabolite that modulates cellular NAD+ by conversion of NADH to NAD+ via the enzymatic action of NADH: quinone oxidoreductase 1 (NQO1)

https://thegutclub.org/enduracell-sulforaphane/

Kim, H.J., Cao, W., Oh, G.S., Lee, S., Shen, A., Khadka, D., Lee, S.B., Sharma, S., Kim, S.Y., Choe, S.K. and Kwak, T.H., 2019. Augmentation of cellular NAD+ by NQO1 enzymatic action improves age-related hearing impairment. Aging cell, 18(5), p.e13016. Niacin Increases NAD+ Significantly – supports mitochondrial function

NAD+ levels can be rescued by a potent NAD+ booster niacin. Niacin remarkably restores muscle and systemic NAD+ and provides metabolic and functional benefits for patients with mitochondrial myopathy

Up to 1g a day of niacin was used for 4 months

Pirinen, E., Auranen, M., Khan, N.A., Brilhante, V., Urho, N., Pessia, A., Hakkarainen, A., Kuula, J., Heinonen, U., Schmidt, M.S. and Haimilahti, K., 2020. Niacin cures systemic NAD+ deficiency and improves muscle performance in adult-onset mitochondrial myopathy. Cell metabolism, 31(6), pp.1078-1090. NAD+ can be increased by exercise, calorie restriction and nutritional precursors

Nutritional Ketosis Increases NAD+/NADH Ratio in the Human Brain Nutritional ketosis – blood levels 0.5 to 3 mg/dL

Xin, L., Ipek, Ö., Beaumont, M., Shevlyakova, M., Christinat, N., Masoodi, M., Greenberg, N., Gruetter, R. and Cuenoud, B., 2018. Nutritional ketosis increases NAD+/NADH ratio in healthy human brain: an in vivo study by 31P-MRS. Frontiers in nutrition, 5, p.62. Poljsak, B. and Milisav, I., 2016. NAD+ as the link between oxidative stress, inflammation, caloric restriction, exercise, DNA repair, longevity, and health span. Rejuvenation research, 19(5), pp.406-413. Four ways to generate NAD +

Plus don’t forget calorie restriction, exercise, healthy diet and not too much alcohol!

Stein, L.R. and Imai, S.I., 2012. The dynamic regulation of NAD metabolism in mitochondria. Trends in Endocrinology & Metabolism, 23(9), pp.420-428. Supplements to generate NAD+

Nicotinamide Riboside Nicotinamide Dietary tryptophan Nicotinamide mononucleotide Niacin (extended release niacin may be preferable to avoid flushing) Some sources warn of the risk of liver toxicity when supplementing niacin/nicotinamide in excess of 3000mg per day

Stein, L.R. and Imai, S.I., 2012. The dynamic regulation of NAD metabolism in mitochondria. Trends in Endocrinology & Metabolism, 23(9), pp.420-428. Connell, N.J., Houtkooper, R.H. and Schrauwen, P., 2019. NAD+ metabolism as a target for metabolic health: have we found the silver bullet?. Diabetologia, 62(6), pp.888-899. Habibe, M.N. and Kellar, J.Z., 2020. Niacin Toxicity. StatPearls [Internet]. The Sirtuins (SIRT 1-7) NAD+ dependent anti-ageing proteins

The sirtuin family were named after their similarity to a gene in the yeast Saccharomyces Cerevisiae, named silent information regulator 2 (Sir2)

Sirtuins mediate the effects of calorie restriction on the extension of lifespan

Dali-Youcef, Nassim, et al. "Sirtuins: the'magnificent seven', function, metabolism and longevity." Annals of medicine 39.5 (2007): 335-345. Sirtuins are anti-ageing proteins

They ‘deacetylate’ proteins

Therefore excess acetyl-CoA is incredibly ageing!

Warning: blood levels of acetate can be 20 fold higher after alcohol ingestion!

Pownall, H.J., Rosales, C., Gillard, B.K. and Gotto, A.M., 2015. Alcohol: a nutrient with multiple salutary effects. Nutrients, 7(3), pp.1992-2000. NAD+ is required for SIRT 1 activation

NAM; nicotinamide NMN; nicotinamide mononucleotide NMNAT NAMPT; Nicotinamide phosphoribosyltransferase NMNAT; nicotinamide mononucleotide Adenylyltransferase

http://www.cell.com/cms/attachment/2007954463/2030564437/gr1.jpg NAD+ activation of SIRT 1 helps protect against multiple diseases and extend lifespan NAD+

Duntas, L.H., 2011. Resveratrol and its impact on aging and thyroid function. Journal of endocrinological investigation, 34(10), pp.788-792. NAD+ boosters promote healthy ageing

https://www.foundmyfitness.com/topics/sirtuin SIRT 1 is an activator of PGC-1α – a protein required for mitochondrial biogenesis

Ray Griffiths. Mitochondria in Health and Disease – Singing Dragon, 2019 Resveratrol Improves Mitochondrial Function and Protects against Metabolic Disease by Activating SIRT 1 and PGC-1α

Lagouge, M., Argmann, C., Gerhart-Hines, Z., Meziane, H., Lerin, C., Daussin, F., Messadeq, N., Milne, J., Lambert, P., Elliott, P. and Geny, B., 2006. Resveratrol improves mitochondrial function and protects against metabolic disease by activating SIRT1 and PGC-1α. Cell, 127(6), pp.1109-1122. NAD+ supports vitamin D receptor activity

SIRT 1 potentiates 1,25-dihydroxyvitamin D3 signaling via vitamin D receptor deacetylation

Sabir, M.S., Khan, Z., Hu, C., Galligan, M.A., Dussik, C.M., Mallick, S., Stone, A.D., Batie, S.F., Jacobs, E.T., Whitfield, G.K. and Haussler, M.R., 2017. SIRT1 enzymatically potentiates 1, 25-dihydroxyvitamin D3 signaling via vitamin D receptor deacetylation. The Journal of steroid biochemistry and molecular biology, 172, pp.117-129. Browning of Adipose Tissue and Sirtuin Involvement

The browning of white adipose tissue is the process during which the enhancement of mitochondrial biogenesis occurs converting white adipocytes to metabolically active beige adipocytes. Sirtuin 1 (SIRT 1) plays an essential role

Energy expenditure by fat utilisation and oxidation predominately occurs in brown/beige adipose tissue

Favero, G., Krajčíková, K., Bonomini, F., Rodella, L.F., Tomečková, V. and Rezzani, R., 2018. Browning of adipose tissue and sirtuin involvement. In Adipose Tissue. IntechOpen. Browning of Adipose Tissue – Mitochondria are ‘the brown’

University of Pennsylvania School of Medicine Antagonistic crosstalk between NF-κB and SIRT 1 in the regulation of inflammation and metabolic disorders

SIRT 1 stimulates oxidative energy, inhibits NF-κB signaling and so suppresses inflammation.

On the other hand, NF-κB signaling down- regulates SIRT 1 activity, particularly through the increased production of reactive oxygen species.

Kauppinen, A., Suuronen, T., Ojala, J., Kaarniranta, K. and Salminen, A., 2013. Antagonistic crosstalk between NF-κB and SIRT1 in the regulation of inflammation and metabolic disorders. Cellular signalling, 25(10), pp.1939-1948. Tristetraprolin (TTP) is a destabiliser of pro-inflammatory cytokine mRNAs. TTP is dependent on SIRT 1 and NAD+

Elevated NAD+ concentrations activate SIRT 1, which enhance tristetraprolin activity by deacetylation and accelerate the resolution of inflammation

Park, J.M., Lee, T.H. and Kang, T.H., 2018. Roles of tristetraprolin in tumorigenesis. International journal of molecular sciences, 19(11), p.3384. Rappl, P., Brüne, B. and Schmid, T., 2021. Role of Tristetraprolin in the Resolution of Inflammation. Biology, 10(1), p.66. SIRT 1 for Brain Integrity

SIRT 1 Is essential for normal cognitive function and synaptic plasticity

SIRT 1 is involved in processes that maintain brain integrity, such as chromatin remodelling, DNA repair, cell survival, and neurogenesis

SIRT 1 is expressed in neurons of the hippocampus, a key structure in learning and memory

Michán, S., Li, Y., Chou, M.M.H., Parrella, E., Ge, H., Long, J.M., Allard, J.S., Lewis, K., Miller, M., Xu, W. and Mervis, R.F., 2010. SIRT1 is essential for normal cognitive function and synaptic plasticity. Journal of Neuroscience, 30(29), pp.9695-9707. Sirtuins as antivirals

Sirtuins are now known to act as antiviral factors, modifying host metabolism to help defend against many viruses, including RNA viruses

Viruses upregulate glycolysis and lipid synthesis to support their proliferation

Herpes simplex virus depletes NAD+… as does obesity

Koyuncu E, Budayeva HG, Miteva Y V., Ricci DP, Silhavy TJ, Shenk T, et al. Sirtuins are evolutionarily conserved viral restriction factors. MBio 2014;5:2249–63. Non-alcoholic fatty liver disease – Benefits of NAD+ and sirtuins

CD38 – to be explained in the next section…

Bock, K.W., 2020. Modulation of aryl hydrocarbon receptor (AHR) and the NAD+-consuming enzyme CD38: Searches of therapeutic options for nonalcoholic fatty liver disease (NAFLD). Biochemical pharmacology, 175, p.113905. SIRT 1 controls many levels of T cell activation, proliferation and tolerance

Chen, X., Lu, Y., Zhang, Z., Wang, J., Yang, H. and Liu, G., 2015. Intercellular interplay between Sirt1 signalling and cell metabolism in immune cell biology. Immunology, 145(4), pp.455-467. SIRT 3 supports beta oxidation, MnSOD, TCA cycle, and the ETC

LCAD; long-chain acyl-CoA dehydrogenase

IDH2; isocitrate dehydrogenase 2

GDH; glutamate dehydrogenase 1

AceCS; Acetyl-CoA synthetase

Li, Xiaoling, and Nevzat Kazgan. "Mammalian sirtuins and energy metabolism." International journal of biological sciences 7.5 (2011): 575. The electron transport chain

SIRT 3 deacetylates and activates mitochondrial respiratory chain complexes, including complex I, complex II and ATP synthase

Mice lacking SIRT 3 consume

10% less O2 and produce up to 50% less ATP than normal

Newman, John C., Wenjuan He, and Eric Verdin. "Mitochondrial protein acylation and intermediary metabolism: regulation by sirtuins and implications for metabolic disease." Journal of Biological Chemistry 287.51 (2012): 42436-42443. Lipid Metabolism

SIRT 3 allows the metabolism of fatty acids in the liver and also the use of ketone bodies during fasting

Hor, J.H., Santosa, M.M., Lim, V.J.W., Ho, B.X., Taylor, A., Khong, Z.J., Ravits, J., Fan, Y., Liou, Y.C., Soh, B.S. and Ng, S.Y., 2020. ALS motor neurons exhibit hallmark metabolic defects that are rescued by SIRT3 activation. Cell Death & Differentiation, pp.1-19. NAD+ CD38 - T cell surface marker with a link to inflammation and an NAD+ catabolic enzyme (NADase) CD38 depletes NAD+

Camacho-Pereira, J., Tarragó, M.G., Chini, C.C., Nin, V., Escande, C., Warner, G.M., Puranik, A.S., Schoon, R.A., Reid, J.M., Galina, A. and Chini, E.N., 2016. CD38 dictates age-related NAD decline and mitochondrial dysfunction through an SIRT3-dependent mechanism. Cell metabolism, 23(6), pp.1127-1139. CD38

The T cell surface marker and multifunctional enzyme CD38 appears to provide a link between inflammation, age and disease-related decline in tissue homeostasis and, therefore, represents a critical target for therapeutic intervention

CD38 is expressed predominately on immune cells in response to stimulation by cytokines, endotoxins, and interferon

CD38 is the main nicotinamide dinucleotide (NAD+) catabolic enzyme (NADase)

CD38 expression in macrophages mediate “inflammaging.” The senescence associated senescent phenotype (SASP) appears to drive the expression of CD38 in macrophages. During inflammaging, CD38+ macrophages accumulate in aging tissues and potentiate the decline of precursors necessary for NAD+ synthesis.

It is thought that CD38, as either an ecto-enzyme or as an intracellular enzyme, promotes metabolic collapse in pathogens by degrading NAD+ and its precursors

Hogan, K.A., Chini, C. and Chini, E.N., 2019. The multi-faceted ecto-enzyme CD38: roles in immunomodulation, cancer, aging, and metabolic diseases. Frontiers in immunology, 10, p.1187. NAD+ depletion by CD38 in lupus patients decreases T cell cytotoxicity and leaves individuals prone to infection

Katsuyama, E., Suarez-Fueyo, A., Bradley, S.J., Mizui, M., Marin, A.V., Mulki, L., Krishfield, S., Malavasi, F., Yoon, J., Sui, S.J.H. and Kyttaris, V.C., 2020. The CD38/NAD/SIRTUIN1/EZH2 axis mitigates cytotoxic CD8 T cell function and identifies patients with SLE prone to infections. Cell reports, 30(1), pp.112-123. The flavonoids luteolinidin, luteolin, quercetin, and apigenin are all inhibitors of CD38 and help to raise NAD+

Food sources: (from Phenol Explorer - phenol-explorer.eu) Luteolinidin – Sorghum Luteolin - Sage, artichoke, extra virgin olive oil, thyme, oregano Quercetin – Black elderberry, beer, capers, cloves, oregano, dark chocolate Apigenin – Sage, oregano, marjoram, rosemary, onion, pistachio, beer (hops)

Rajman, L., Chwalek, K. and Sinclair, D.A., 2018. Therapeutic potential of NAD- boosting molecules: the in vivo evidence. Cell metabolism, 27(3), pp.529-547. NAD+ PARP - poly(ADP-ribose) polymerase

DNA protection – overactivation can lead to catastrophic NAD+ and energy loss followed by cell death PARP is activated by DNA damage

PARP is activated by DNA damage and catalyze the NAD-dependent polymerization of a chain of (poly (ADP-ribose), which signals repair of DNA. PARP leads to intranuclear and intracellular NAD+ consumption and NAD+ collapse

Hogan, K.A., Chini, C. and Chini, E.N., 2019. The multi-faceted ecto-enzyme CD38: roles in immunomodulation, cancer, aging, and metabolic diseases. Frontiers in immunology, 10, p.1187. Ke, Y., Wang, C., Zhang, J., Zhong, X., Wang, R., Zeng, X. and Ba, X., 2019. The Role of PARPs in Inflammation—And Metabolic—Related Diseases: Molecular Mechanisms and Beyond. Cells, 8(9), p.1047. Alpha lipoic acid is a PARP inhibitor

Alpha lipoic acid is able to significantly reduce 8-hydroxy-2-deoxyguanosine (marker for oxidative stress) in laboratory animals

Alpha lipoic acid protects against DNA damage and so decreases the activity of PARP

Chen, J. and Li, Q., 2020. Lipoic acid decreases the expression of poly ADP-ribose polymerase and inhibits apoptosis in diabetic rats. Diabetes, metabolic syndrome and obesity: targets and therapy, 13, p.1725. The flavonoids myricetin, tricetin, quercetin, fisetin, rutin, and naringin are all inhibitors of PARP and help to raise NAD+

Food sources: (from Phenol Explorer - phenol-explorer.eu)

Myricetin – Onion and beer (hops) Tricetin – Cereal, ginkgo, nuts, pulses, tea Quercetin – Black elderberry, beer, capers, cloves, oregano, dark chocolate Fisetin – Strawberry, apple, mango, kiwi, grapes, tomato, onion Rutin – Buckwheat, tea, asparagus, olives, capers, raspberry, buckwheat Naringin – Grapefruit, rosemary, beer Overview of the health conditions impacted by low NAD+ levels

Rajman, L., Chwalek, K. and Sinclair, D.A., 2018. Therapeutic potential of NAD-boosting molecules: the in vivo evidence. Cell metabolism, 27(3), pp.529-547. Overview of NAD+ consuming processes

Chini, C.C., Tarragó, M.G. and Chini, E.N., 2017. NAD and the aging process: Role in life, death and everything in between. Molecular and cellular endocrinology, 455, pp.62-74. Supplements

Intervention Function Dosing

Niacin (and sustained release) NAD+ precursor 500 mg

Nicotinamide NAD+ precursor 500 mg

Nicotinamide Riboside NAD+ precursor 500 mg

Nicotinamide Mononucleotide NAD+ precursor 500 mg

Resveratrol Promotes SIRT 1 activity 100 mg

Alpha Lipoic Acid PARP/DNA damage prevention 200 mg

Thank You Learn more at www.pure-encapsulations.co.uk/education Q&A