NMN Transporter Slc12a8 and the future of NMN clinical trials with Dr Alessia Grozio
The need for NMN Clinical trials
In a recent post, we discussed a key barrier in our knowledge of NMN supplementation in humans - namely the very limited clinical trial data.
However, this barrier is soon to be removed, thanks to high profile research + business collaborations which a directly confronting this lack of information.
Dr Alessia Grozio is Chief Scientist at Elevant and a researcher at the Buck Institute, ‘insanely’ interested in all things NMN and longevity.
With their execution and success, we will soon have a clear understanding of the benefits of NMN supplementation in human subjects (well beyond simpler experimental results, such as how NMN improved our Founder Nick’s biological age by 3.5 years).
Collaborations on NMN Clinical Research
One such collaboration we recently highlighted was the the work between aging research leader the Buck Institute, Elevant, and its parent company Seneque.
Together they have developed a healthy pipeline of NMN clinical trials in humans, which will focus on muscle recovery after exercise, potential benefits to skin health and much more (details in our interview).
Today, we host Elevant’s Chief Scientist and Buck Institute Researcher Dr. Alessia Grozio (PhD) for an exclusive interview on the future clinical trial outlook for NMN.
Interview with NMN researcher and expert Dr Alessia Grozio
Longevity Blog contacted Dr. Grozio with a number of important questions on the topic of NMN supplementation in humans.
Seneque’s mission is to increase healthspan by 20% in the next decade.
One key topic area we addressed was Dr Grozio’s co-discovery of a transporter that delivers NMN directly into cells called slc12a8.
This discovery was the first time that a pathway for NMN to enter cells directly had been discovered. A key development which well renowned NMN expert Dr. David Sinclair was very quick to highlight in Nature Metabolism (read it here) .
This discovery has not been without controversy, as NAD biologist and researcher Dr. Charles Brenner was quick to criticise the study. This has led to a bit of a schism in the research community on whether or not slc12a8 does what Dr Grozio says it does.
So we wanted to hear it from the expert directly, asking Dr. Grozio to help us better understand the significance of slc12a8, particularly around how NMN enters the cell (which is critical for it to fulfil its NAD boosting function).
As always, we also ask her about her personal longevity strategy.
Without further delay - onwards!
This interview was conducted in written format over email correspondence during May 2021
Longevity Blog (LB): Dr Grozio, thanks for joining us here on Longevity Blog.
To kick us off - you work diligently at both a research organisation and a commercial venture. Surely its hard work! Tell us - what drives you? What brings you into work each day?
Alessia Grozio (AG): Thanks for inviting me. The passion for my work and an “insane” curiosity for anything related to aging/longevity and NAD+ biology that brings me to work every day.
After earning my PhD in biochemistry, I started working on the enzymes (sirtuins, NAMPT, CD38/CD73 ecto-enzymes) involved in NAD+ metabolism.
NAD biology is quite complex. This is an area Dr. Grozio finds quite fascinating. We do too!
In a previous post with Dr Grozio’s college, Dr. Eric Verdin, we discuss this topics in further detail.
I have always been fascinated by the pivotal role that NAD+ plays in maintaining homeostasis in our body and the plethora of cellular processes in which it is involved.
Therefore, investigating the effects of NMN (nicotinamide mononucleotide), a critical endogenous compound for NAD+ biosynthesis, in different physio-pathological conditions where NAD+ levels are impaired, was a natural prosecution of my scientific interests.
More than “hard”, I would say that my job, as for many other scientists, is never boring!
What is NMN-C
LB: Thank you for sharing those personal insights. One of the reasons we’re speaking with you today is to help introduce ‘Elevant’, and your efforts to make ‘NMN-C’ available to a broad audience.
Please introduce us to NMN-C. How does this formulation differ from other forms of NMN that our readership may already be taking?
“What makes our NMN-C special is the extremely high level of safety and development that has gone into it. ”
AG: What makes our NMN-C special is the extremely high level of safety and development that has gone into it. We think what is most important is providing consumers with NMN-based products that have the highest levels of safety and quality possible.
NMN-C is exclusively available through Elevant in their “Prime” product (pictured)
What’s critical is, once NMN has been synthetized, to characterize the impurities, filter them as much as possible to reduce them to the minimum, and make sure that the few impurities remaining are safe.
In our NMN-C the level of impurities is very low, complying with the pharmaceutical grade guidelines.
Our product is then tested at high dosage, pre-clinically and in human, to confirm it’s safe to use.
Is NMN Safe for Humans?
LB: There was a recent study specifically looking into the ‘safe’ level of NMN supplementation, could you share some information on this work?
“NMN-C at doses up to 1500 mg/kg/d appears to be safe”
AG: Our NMN-C acquired self-affirmed GRAS (Generally Recognized as Safe) status in accordance with US FDA regulatory guidelines, performed by an independent panel of toxicology and nutrition experts.
Moreover, we have recently published the first OECD 408 toxicology study performed on NMN. This study analyses the No Observable Adverse Effect Level (NOAEL) of our NMN-C and demonstrated that repeated oral administration of NMN-C at doses up to 1500 mg/kg/d appears to be safe and did not promote adverse effects in rats up to 90 days.
Diagram showing the role of Slc12a8 and how it enables NMN to enter the cell. Source: Nature Metabolism
Does NMN enter the cell intact?
LB: You made some waves in the research community with your published results on a NMN ‘transporter’ slc12a8.
This was a particularly exciting discovery, as the ‘doorway’ for NMN to enter the cell was previously unknown.
Could you briefly explain why this discovery is significant for the prospects of NMN supplementation?
AG: The identification of a novel NMN transporter, Slc12a8, further advances our knowledge regarding NAD+ metabolism by providing a new mechanistic insight through which the absorption and distribution of NMN is regulated to fuel the NAD+ biosynthesis in different tissue.
In the manuscript published in 2019 in Nature Metabolism, we employed several biochemical and in vivo (testing done in a whole organism) analyses to demonstrate that the Slc12a8 gene encodes a selective NMN transporter that facilitates the direct uptake of NMN into cells and intestinal tissue of mice.
We found that Slc12a8 is regulated by NAD+, specifically, Slc12a8 expression is increased when intracellular NAD+ levels are declined, as a compensatory mechanism, in order to maintain the cellular NAD+ homeostasis.
The Debate Around Slc12a8
LB: Proponents of alternative NAD precursors (NAD biologist Dr. Charles Brenner amongst them) such a Nicotinamide Riboside have contested that slc12a8 is an ‘intestinal cation-calcium transporter’ and not an NMN transporter.
The NMN proponents predicted that NMN would still work as NMN in cells that don't have NR kinase but we and others showed that when NR kinase is knocked out, NMN can only raise NAD+ by acting as nicotinamide. That's because NMN works by conversion to NR and needs NR kinase /14
— Charles Brenner, PhD (@CharlesMBrenner) January 13, 2020
Surely the ‘devil in the details’, but what is your reply to such claims?
How can we trust that NMN is making it ‘into the cell’ intact, rather than only through the Nicotinamide Riboside Kinase pathway as NR?
AG: Slc12a8 is an orphan member of the SLC12 family of the electroneutral cation–chloride co-transporters, this means that so far, its function as cation–chloride co-transporter has not been demonstrated.
Indeed, Slc12a8 shares low level of similarity with its family members for what concerns amino acid sequence and predicted membrane topology.
As described in our Nature Metabolism paper, we have conducted isotopic tracing experiments using side by side isotopic NMN and nicotinamide riboside in Slc12a8-OE NIH3T3 cells (the cell line overexpressing Slc12a8 protein) and primary hepatocytes from Slc12a8 knockout mice (mice completely lacking the Slc12a8 protein).
Both experimental conditions demonstrate direct NMN uptake without any previous conversion of NMN to nicotinamide riboside outside of cells.
LB: Thanks for answering that very important question.
NMN as a Topical Cream for Skin Health
LB: Moving on from transporters, and onto Elevant’s impressive research efforts, you are engaging with some very significant topic areas - namely beauty and sports performance, which are very significant consumer markets.
Could you share with us the future of NMN based ‘creams’ for topical use? Is this a future development area for beauty products? And do we know if NMN can enter skin cells through a topical? (We are very interested in measuring & preserving aesthetic age)
AG: Yes, previous research has shown that NAD+ appears to play an enormous potential role in skin health.
Research has shown that NAD+ energizes aging skin cells to function more like young cells, counters transepidermal water loss and improves the moisture content of the outer layer of the skin, along with several other actions that support healthy skin.
We have conducted clinical trials that have directly shown the beneficial effects of NMN on the skin.
The trials demonstrated improved skin moisturization, elasticity and brightness, and a reduction in skin roughness and visibility of wrinkles. The results are to be published soon and we will share them with you.
NMN for Exercise Recovery
Longevity Blog is very interested in how anti-aging compounds might improve athletic performance and the benefits of exercise.
Another longevity supplement “Urolithin A”, which is derived from pomegranates, also has benefits on improving physical recovery, through the mitophagy pathway. Read more!
LB: An ongoing clinical trial is now recruiting participants, which will analyse the benefits of NMN for ‘physical capacity and recovery’.
Looking forward, do you see a potential future role for NMN in human performance (i.e. sport)? If so, do you have any insights on how it might improve an athlete’s ability to compete in their sport of choice?
AG: We have preliminary results showing NMN is efficient on physical recovery, at any age. We will keep you informed, prior to the publication of our results.
Upcoming NMN Clinical Trials
LB: Beyond these two clinical trial areas, Seneque are also ‘preparing’ several more clinical trials covering many areas with NMN-C: impact on NAD+ levels, NMN vs NR, sleep, cognitive function, immunity - each is a significant undertaking (self-titled ‘the world largest pipeline’).
What timeline do you expect for the results on these projects? They will each go quite a long ways toward demonstrating the efficacy of NMN supplementation.
Seneque has a very impressive pipeline of NMN based clinical trials at various stages of maturity. Find the latest on their website.
AG: Two more studies started this month (May 2021), a tolerance and a pharmacokinetic study with 20 participants each and dosage of 400mg per day. Preliminary results to be announced next July (2022).
The first results of further trials will be published Q3 2021 and several publications will occur until July 2022. You can see our extensive clinical research pipeline here.
Some additional confidential studies are being conducted at the moment, so pending the timing and results of those we hope to start communicating about them at the end of this year.
We are confident it won’t be long until we can demonstrate the efficacy of NMN supplementation in several clinical trials.
We aim to be able to share results sometime between Q3 and Q4 2021.
Research Business Partnerships for Longevity Research
“We are confident it won’t be long until we can demonstrate the efficacy of NMN supplementation in several clinical trials”
LB: Some elements of these clinical trial efforts will occur in partnership with the Buck Institute, namely the pre-clinical studies and basic research efforts.
Could you describe the merits/value of private longevity technology companies collaborating with leading aging research institutions? What are the challenges for you personally, ‘wearing two hats, so to speak (as you’re also a scientist at the Buck Institute)?
AG: The main interest in collaborating with leading aging research institutions is to work and share knowledge with scientists coming from various backgrounds and expertise, foster new ideas and drive our research to constantly innovate.
This allows diverse new research leads and areas of study, all undertaken in line with scientific principles.
The powerful and high-end equipment and infrastructure at the Buck is of course a huge asset.
Finally, as you know, time is of the essence. These partnerships allow invaluable gains of time.
NMN-C is Manufactured to a High Standard
LB: Elevant is now manufacturing NMN-C in Europe, which differentiates from other suppliers of NMN, who rely on manufacturing in China. This undoubtedly introduces additional costs, but likely as a trade-off for many benefits.
Could you comment on what ‘Made in Europe’ means for consumers?
AG: We strictly control every element of our proprietary manufacturing process, which we undertake at cGMP-certified facilities in France. Consumers receive what we believe are pharmaceutical grade and the highest levels of quality and safety available in an NAD+ booster today.
Dr Grozio’s Longevity Strategy
LB: Dr Grozio, changing gears, as we close off the interview - could you share with our audience one or two items from your personal longevity strategy?
AG: I am Italian, therefore I am following a Mediterranean diet, rich in vegetables, legumes, fish and extra virgin olive oil.
Moreover, I am taking our NMN-C every morning to boost my physical and mental energy needed to go throughout the entire day.
LB: Dr. Grozio, thank you such much for taking the time to answer our equations today!
Want more NMN Focused Content? We got it!
FDA & TGA DISCLAIMER
This information is intended for educational purposes only and is not meant to substitute for medical care or to prescribe treatment for any specific health condition. These blog posts are not intended to diagnose, treat, cure or prevent any disease, and only may become actionable through consultation with a medical professional.
NMN Nicotinamide Mononucleotide Before and After
NMN Before and After with Biological Age
In mid 2020, I set out on a self-experiment with NMN. The purpose of this experiment was three-fold
First - my primary intention was to lay out some suggestions for how you could structure your own self-experiments, and not just for NMN
Second - I of course was quite curious what my own results would be, and wanted to have a well-controlled self-experiment to answer the question: should I continue to invest in NMN as part of my longevity supplement stack?
Third - to raise awareness of NMN, biological age testing and encourage more of you to join in on being a biohacker!
Designing a self-experiment with NMN in mid 2020 was a unique challenge, as at the time, there was no direct way to test intracellular NAD levels.
That has however changed, and in upcoming posts, I am going to show you how to test your NAD levels directly. If that sounds interesting, be sure to subscribe below!
Takeaway: Over 6 months, a combination of 500mg NMN, 500mg TMG and 500mg resveratrol improved my biological age by 3.5 years. Read on to learn more!
Use the code ‘longevityblog’ to save 10%.
Looking for an NMN supplier? I recommend DoNotAge, who:
offer a third party tested product
have a bulk supply option (100g)
offer further savings when you subscribe to regular shipments
plus I can get you a discount: Use the code ‘longevityblog’ to save 10%!
Self-Experimenting With NMN
It is not easy to run a self-experiment. To review, all self-experiments are:
Difficult to control
Often hard to measure directly
Prone to bias and placebo effect
Even when well-controlled, don’t necessarily provide clear insights
Let’s take a minute to address each of these, how this particular self-experiment addressed each over the course of the 6-months of NMN supplementation.
Control
Over the course of my 6-month experiment, I made several efforts to control the experiment:
My exercise ‘Load’ as per my Garmin Fenix 6 Pro, which I use to track all of my workouts. At top, near the start, at bottom, near the end of the 6 month period.
I did not change my exercise load or volume
I did not change my diet
I did not start taking any other supplements
I did not make any other major lifestyle changes
It is of course not possible to completely control your life! However this does not mean you should not try.
Of these items, I would suggest that consistency in diet and supplementation are of paramount importance.
You can find many “NMN before and after” testimonials on YouTube, for example, but each of these has many problems with control.
The most common issue is making too many changes, mostly around supplementation protocol, during the ‘before’ and ‘after’ period.
Your chief goal here is to decide: is taking NMN worth my hard earned cash?
You can’t make that call if you’re also introducing many other longevity supplements at the same time! You’d be surprised how often this is the case! Be patient and be consistent.
Measurement
In the absence of a direct NAD test, the decided measurements before & after were set according to the 2x clinical NMN trials which were ongoing at the time, as well as adding biological age:
Lipid profile
Blood pressure
Biological age
Epigenetic age (Saliva Test via Chronomics)
Phenotypic age (Blood Test)
Bias & Placebo
I put serious thought into how to conduct a double-blind experiment, but ultimately elected not to.
Given the objective measurements we could draw from the above, and the ‘n of one’ experimental design, this was not practical to implement.
It also reduced the likelihood that everyday folks like yourself would be able to repeat the experiment effectively.
In the effort to minimise placebo effect, I made it my mindful intention to not overhype my personal attitude toward the experiment.
To be quite honest - I adopted quite a sceptical attitude.
Personally, I was unconvinced that NMN would make much of a difference in my day-to-day experience, as there was relatively little science in human subjects & most of these ‘hot trends’ turn out to be over-hyped!
NMN
NMN Self-experimentation Protocol
The supplement protocol for this self-experiment was designed to match the widely used combination of NMN, Resveratrol and TMG:
NMN at 250mg/day (first two months), 500mg (following four months)
For risk management, 1:1 dosage of Tri-Methyl Glycine (TMG)
Resveratrol at 500mg/day, taken with a high fat meal
Note: this was not a change for me, I’d been taking it for 2+ years, and therefore did not interfere with the need for supplemental control
NMN Before & After: Our Hypothesis
This experiment was undertaken to test the following hypothesis (all self-experiments should have one!):
NMN supplementation will boost intracellular NAD+ levels
Boosting NAD+ levels with NMN will measurably improve select characteristics of youthfulness
Combining boosted NAD+ with a sirtuin activator (resveratrol) will enhance DNA repair
Combined, these effects are hypothesised to be likely to:
change blood lipid profile (as per clinical trial designs)
change blood pressure (as per clinical trial designs)
slow or reverse biological age (Longevity Blog hypothesis)
NMN Before & After: Results
The self-experiment was run from 1 August 2020 for 6 months until 31 January 2021.
What follows are the results, including both hard data points and anecdotal observations.
Blood Lipid Profile:
Blood lipids refer to the commonly assessed HDL, LDL and Total Cholesterol measurements. These are also joined by Triglycerides to form the ‘Blood Lipid Panel’ of tests.
As you can see in the provided image, there was no significant change in any of these parameters over the course of my NMN self-experiment.
Blood Pressure
Blood pressure measurements at the start of the experiment were ~125/85.
At the end of the experiment, 126/84.
There was no significant change in blood pressure from the NMN self-experiment.
Biological Age: Phenotypic Age
Phenotypic age is calculated using 9 blood based biomarkers, to calculate a biological age.
This is based on work led by Yale researcher and longevity thought-leader Dr. Morgan Levine (whom we hope to interview on the blog in the future!).
We’ve previously reviewed this test, including providing information on how you can use it yourself here (its free!).
Blood markers in this test include:
White Blood Cell Count (WBC)
Red Cell Distribution Width (RDW)
Mean Corpuscular Volume (MCV)
Fasting Blood Glucose
Lymphocyte %
Creatinine
Albumin
Alkaline Phosphatase (ALP)
C-reactive protein (CRP)
If you’d like to learn more about any of these blood markers, checkout Lab Tests Online.
Pictured: My Phenotypic Age (Biological Age measure #1) over time.
At the start of the self-experiment, my Phenotypic Age was 28 (Chronological Age 34).
At the end of the self-experiment, my Phenotypic Age was 25 (Chronological Age 35).
Given that 6 months passed, this equates to a total change of -3.5 years of Biological Age.
Did I reverse my biological age with NMN?
Now - the key question is, was this change significant? Was it due to the NMN based protocol?
This comes back around to one of my initial opening points about self-experiments: “Even when well-controlled, they don’t necessarily provide clear insights”
For our analysis, we should extrapolate from the baseline measurements a bit further.
Over the preceding 3 years, my Phenotypic Age was an average of 25.7 years old. It was as high as 28 and as low as 23.
More importantly, we need to consider the difference between biological age and chronological age.
My average difference between biological age and chronological age over the preceding 3 years was -7.7 years. This was as high a -10 years and as low as -5 years.
At the start of the NMN self-experiment, this difference was -7 years (below the average), whereas at the end, it was -10 years (at the previous extreme).
There is a degree of subjective interpretation which must occur here, as the data volume does not lend itself to statistical analysis.
Based on the data and trends, there is sufficient evidence to suggest that this NMN based protocol reversed my biological age.
It is however is not conclusive evidence, and as a scientist, I choose to approach that statement conservatively.
There is a bit more to explore however, to let’s continue.
Biological Age: Epigenetic Age
This one was a real bummer. My follow-up test with Chronomics failed their quality control step, meaning the biological age result was not valid.
This was a crushing blow, as it was the chief data point, in my opinion.
However, I have re-tested it, but it was not in-line within the 6-month period self-experimentation period.
Due to my choice to begin another 6-month self-experiment immediately following this one, the next biological age result will not be valid for this experiment.
I’ve since dramatically increased my exercise routine (I’m now training for triathlons), made major dietary changes and increased my NMN supplementation to 1000mg/day in a separate 6-month self-experiment aimed a direct feedback from Chronomics on how to reverse my biological age.
I will update this post with those results, when they are returned, however they will not be well-controlled for NMN experimental purposes.
I know - major bummer! 😔
Such is self-experimenting life!
NMN Before and After: Anecdotal Observations
SF-36 Questionnaire
My results from the SF-36 questionnaire before & after did show a slight change in a few key areas.
While I did not experience any changes in general health, physical health problems or limitation of activities, I did have interesting results in the energy & emotions section.
Excerpt from my SF-36 questionnaire, before and after the NMN self-experiment.
At the start of the self-experiment, I think I was feeling many of the things that folks in their mid-30s start to feel. Notably less energy than in their 20s, less upbeat and more likely to feel worn out at the end of a long day.
Over the course of the self-experiment, this changed in notable ways, in a sustained way that continues to today.
You can see for yourself the meaningful parts of the questionnaire that changed for me in the following image.
I have to add - it is completely possible that these results are placebo based. I was taking NMN, most folks who supplement with NMN boast ‘increased energy levels’.
But of course, since NMN is hypothesised to impact intracellular energy, this is what we expect to happen.
When it comes to ‘energy levels’, NMN is a prime placebo influence candidate.
Nicotinamide Mononucleotide Before and After Conclusions
To summarise, I was able to run a well-controlled self-experiment with NMN supplementation at 500mg/day.
I recorded before and after blood tests, blood pressure and detailed subjective experience related data.
Initial results do indicate improvement in my Biological Age (as per the Phenotypic Age) by approximately 3.5 years.
Subjective data (qualitative data) support this quantitative data, and therefore the conclusion is: NMN left me both feeling younger and perhaps biologically younger.
However, this cannot be placebo controlled, and warrants further testing. In particular, direct testing of NAD levels is now available, and that is what I will be testing next!
Also, we have not been able to test the much more robust and accurate epigenetic age via Chronomics, which was unable to be measured. Ultimately, this was the most powerful data point, and though disappointing - we will revisit it.
Coming soon, I will review the results from testing intracellular NAD levels directly, before and after 1000mg of NMN.
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FDA & TGA DISCLAIMER
This information is intended for educational purposes only and is not meant to substitute for medical care or to prescribe treatment for any specific health condition. These blog posts are not intended to diagnose, treat, cure or prevent any disease, and only may become actionable through consultation with a medical professional.
NMN-C, Elevant and the future of NAD boosters: An Interview with Dr Eric Verdin
NAD Precursor NMN-C - A New NMN Supplement
As we have discussed in recent posts on NAD boosters, clinical trial data for NMN in humans remains quite limited.
In fact, up until recently, it wasn’t even well established that NMN was safe to use as a supplement. Hence, why we’ve had to discuss this issue and how minimise any risks in involved should you choose to self-experiment with NMN.
One key outstanding question has been - is NMN safe to take as a supplement?
IS NMN SAFE?
Excitingly, this question has now been answered, thanks to innovative work by the company ‘Elevant’ (in collaboration with two other parties, more on this shortly), who recently published results from a toxicology study for NMN supplementation (read it here).
On the back of this successful study, Elevant is now offering this a novel form of NMN, matching that which was used in the research effort - the product is called ‘Prime’, and the specific NMN formulation is called NMN-C.
What is NMN-C?
NMN-C is differentiated by its quality and extremely high-level of safety where the processes for characterising and removing as many potential impurities has been standardised and follows pharmaceutical grade guidelines.
Elevant’s NMN-C product, “Prime”
There are not any changes to the molecular structure, rather NMN-C is the outcome of this highly rigorous processing approach.
This outcome of this method for producing NMN has then been tested at high dosages (i.e. the study we just mentioned) and also in pre-clinical testing in humans.
This has made NMN-C the only form of NMN to be categorised by the US Food and Drug Administration as “Generally Recognised as Safe” - so called GRAS status.
Elevant, Seneque and the Buck Institute
These recent efforts to create and study NMN-C are the fruits of an exciting trifecta of collaboration between Elevant, its parent company Seneque and the Buck Institute.
They have an exciting pipeline of future NMN related studies in humans, including skin health, muscle recovery and more. We’ll explore these topics in an upcoming post with Elevant’s Chief Scientist.
Together, these companies boast some very grand visions, which should get any longevity enthusiast (that’s you) quite excited.
The Buck Institute for Research on Aging is a global leader in the study of aging, and increasingly, interventions in age-related disease.
Seneque, the parent company of Elevant, is a Swiss life sciences company, founded by French entrepreneur Guillaume Bermond, who now leads the company on its mission to “increase healthspan by 20% in the next decade”.
Rounding out the three-way collaboration is the Buck Institute for Research on Aging, who are global leaders in the study of aging, whose missions is equally grand - “to end the threat of age-related disease for this and future generations”.
Interview with Longevity Pioneer Dr Eric Verdin
Playing a key cross-over role between these three entities, is Dr Eric Verdin, CEO and President of the Buck Institute, and more recently, Chair of the Scientific Advisory Board at both Seneque and Elevant.
Longevity Blog reached out to Dr. Verdin to discuss this new role as at Seneque, and in order to better gauge his view on the promise of NMN supplementation in humans.
We discuss the use of NMN in human subjects, the role of inflammation and CD38 in NAD biology, the future of NAD boosters and even his personal longevity strategy.
This interview was conducted in written format over email correspondence during May 2021
Does NMN actually work? Is NMN effective?
Longevity Blog (LB): Clinical trial data for NMN supplementation in humans is at a very early stage, and we’ve yet to see conclusive results. Yet, the new venture Seneque will deliver NMN based products. What gives you confidence in the efficacy of NMN supplementation in humans?
Dr. Eric Verdin (EV): That’s a great question. I think that one should know that not everything from mouse or from animal model experimentation will actually translate into humans, and so there is always some degree of uncertainty in terms of what will happen.
Most NMN research has been performed in rodents, what will this mean for humans? Image credit
However, there is now growing evidence that NMN supplementation in animal models really has profound effects in the biology of every organism on which we have tested it. Based on this, one can be reasonably optimistic that we’re going to be seeing the same thing in humans.
One key thing that we will have to consider is the dose at which we give the NMN. Right now, NMN is typically commercially available at lower concentrations than what we’ve given to animals, and eventually we will have to account for some of the potential differences.
“NMN supplementation in animal models really has profound effects in the biology of every organism on which we have tested it”
CD38 Inhibitors for NAD Boosting?
LB: One of the more recent discoveries related to NMN supplementation, is a need to pay attention to the role of the CD38 enzyme. The Buck Institute recently published a study on the role of inflammation and macrophages in CD38 regulation. Could you comment on the potential role of CD38 ‘inhibitors’ alongside NMN supplementation? What compounds might prove useful in this role?
EV: Yes. What this paper actually showed is that, as we age, the activity of this enzyme called CD38 increases. CD38 is an NAD hydrolase, and so it is able to take NAD and to cleave it into its byproducts – ADP-ribose and nicotinamide – and we think that it is one of the major pathways that leads the progressive degradation of NAD that occurs during aging.
Longevity Blog did a deep dive on CD38’s role in NAD biology with Nuchido Founder Dr. Nichola Conlon - be sure to check it out!
So, on this basis, one hypothesis would be that to restore or maintain NAD levels in aging, we have to inhibit CD38, but you also have to replenish depleted NAD levels.
This is where NMN could come in. One thing to also consider is that CD38 also cleaves NMN, so this makes it even more of an important potential synergy between providing inhibitors of CD38 and providing NMN.
I think this is one way to move forward into the future – to have specific CD38 inhibitors and also to provide NMN to restore NAD levels.
“one hypothesis would be that to restore or maintain NAD levels in aging, we have to inhibit CD38”
Chronic Inflammation: CD38 and Senescent Cells
LB: As a follow-up, given the role of chronic inflammation and senescent cell activity (SASP) to create pressure on NAD levels in the cell, does this suggest that senolytic compounds (e.g. fisetin, quercetin, etc) may be a natural partner in boosting cellular NAD levels in addition to NMN?
EV: That’s a very good question. In the paper you discussed just before, which we published last December in Nature Metabolism, we identified the mechanism for the progressive increase in CD38 during aging, and one of the mechanisms was the progressive accumulation of senescent cells.
As a part of the ageing process, CD38 and Senescent cell activity both increase, lowering NAD+ levels. Image credit: Buck Institute
The senescent cells secrete a series of pro-inflammatory cytokines, the so-called SASP, which means ‘senescence associated secretory phenotype’.
What we showed is that the SASP is one of the mechanisms by which CD38 increases during aging.
Based on this, one could predict that if we were able to eliminate a senescent cell, we would eliminate the SASP and then we would eliminate the induction of the CD38 expression.
One thing to remember is that the SASP is not the only mechanism that was responsible for the induction of CD38. There were other substrates including bacterial products such as LPS (lipopolysaccharide), and other series’ of factors that we will also have to control.
So I believe that fighting the induction of CD38 is going to be harder than inhibiting CD38 expression or providing NMN as an exogenous substrate.
“if we were able to eliminate a senescent cell, we would eliminate the SASP and then we would eliminate the induction of the CD38”
NAD Boosting Supplements: More Options on the Horizon?
LB: With a forward looking approach on NAD boosting, NAD precursors and their role in alleviating some of the pressures of aging, do you believe we are likely to see a wider variety of NAD boosting supplements emerge over the next 5-10 years?
If so, what form might these (strategies) take? The recent discoveries around the reduced form of NMN (NMNH) is one such example.
EV: This is a very good point, and one I think is exciting. Clearly, what the data shows is the interconnectedness between multiple processes is something that we had actually not fully appreciated.
One is NAD degradation, that’s the role of CD38. The role of senescence. The role of what we call the PAMPs (pathogen associated molecular phenotypes) such as LPS and others.
So, if you think about fighting aging and its manifestations, I would predict that in the future, we will likely have a combination of these approaches to really maintain NAD levels, to eliminate senescent cells, and to restore NAD levels, using a variety of precursors.
“I can predict, based on what has been published and what we know, that there will be other [NAD boosting] molecules.”
Right now, the field is focusing on NR (nicotinamide riboside) and NMN (mononucleotide nicotinamide). Both of those have shown remarkable activities, which is why there is so much excitement in this field.
But I can predict, based on what has been published and what we know, that there will be other molecules.
One thing that is really important to realize is that to be truly demonstrated as efficacious, I think it will be important for these molecules – whatever they are, whether they are CD38 inhibitors or NAD precursors – we need to conduct clinical trials to make sure that whatever we’re giving to patients is not only given at the right dose, but is safe and actually has the intended safety profile and the intended benefit.
I think this is the way that truly evidence-based medicine has been progressing over the last 100 years and I hope that our field of aging research will abide by this principle and demonstrate the efficacy of what we’re providing to patients.
Wearing Two Hats: Bridging the Gap Between Ageing Research and Deploying Interventions
LB: Dr. Verdin, could you comment on the importance of translational roles, such as your role as Chair of the Seneque Scientific Advisory Board, in bridging the gap between ageing research and making relevant treatments available to the general public?
While keeping a foot in both the ‘research’ and ‘commercial’ camps can be difficult to navigate, surely they are likely to become more common as this field matures?
EV: I am very excited by the ongoing opportunity and ability to not only conduct basic research and make discoveries – that’s been my whole life’s work.
But as I’m getting older I have been increasingly interested in taking things one step further and really pushing the envelope to make these discoveries closer to translation in humans.
This is certainly the approach we’re taking at the Buck Institute. We’re starting companies, we’re collaborating with established pharmaceutical companies, we’re collaborating with biotech companies such as Seneque.
I very much view my role not only as Director of a basic research institute, but as an advocate for pushing these discoveries into the clinics. And I really think the science is there in terms of the promise.
“I hope that in 20 years from now, we’ll look back at the Buck and say that this was an institute that really took the bull by the horns and really tried to change the way we age”
The deep questioning and the hard work start now. Which is to say really bring this into humans. We know from data in the pharma industry that it is much harder to translate into humans, especially given the diversity of the human population both in terms of genetics and lifestyle and so on.
One of the reasons we work closely with Seneque is the value they place on proper clinical research. They have a significant program of clinical trials in play which will drive the understanding of the effects of NMN in humans for all of us.
Seneque’s mission is to increase healthspan by 20% in the next decade.
In the future we will hopefully see that in some way the Buck successfully redefined what the basic research institute is doing.
In this case we are aggressively building a translational infrastructure – by collaborating with pharma and biotech and also by starting our own company.
I hope that in 20 years from now, we’ll look back at the Buck and say that this was an institute that really took the bull by the horns and really tried to change the way we age, not only in animal models but in humans. I think this is a little different from what most places do, but I find it very exciting.
“As I’m getting older I have been increasingly interested in taking things one step further and really pushing the envelope to make these discoveries closer to translation in humans.”
Dr. Verdin’s Personal Approach to Longevity. Does Dr. Eric Verdin take NMN? What supplements does Dr Verdin Take?
LB: Dr Verdin, changing gears, as we close off the interview - could you share with our audience one or two of your personal longevity strategies?
EV: I have to be careful here, because these are not ‘recommendations’, this is strictly what I do as a person. I believe in lifestyle effect being very critical in determining longevity, so I really focus on five different aspects.
Nutrition & Fasting
One is nutrition, with a focus on intermittent fasting. Every three months, I do a week of fasting. I do time-restricted feeding, which means I eat for about eight hours of the day and do not eat for the remaining 16. So that’s pillar number one – nutrition.
Exercise
Pillar number two is exercise. I try to fit in between one and two hours of exercise every day. I think it is the best and safest anti-aging medicine that we have today, and it will remain so for a little while longer, until we discover better medicine.
“I used to be an every-day-one-glass-of-wine drinker. I have seen how much this alters the quality of my sleep and I would now define myself as an occasional rare drinker.”
Sleep
The third pillar is sleep. I think we live in a society that is chronically sleep-deprived. I try to sleep a good amount every day and try to make sure it’s good quality by mitigating all the factors that I’ve discovered actually interfere with my sleep.
I use an Oura ring and the Whoop, which are two wearable devices that allow you to closely monitor your sleep and which have allowed me to determine what the factors are that affect the quality of my sleep.
One of the ones that actually surprised me was the effect of alcohol. I used to be an every-day-one-glass-of-wine drinker. I have seen how much this alters the quality of my sleep and I would now define myself as an occasional rare drinker.
Stress Management
The last thing is stress. I really believe that stress is a significant contributor to aging, so I try to mitigate my stress by doing yoga, by meditating when I have the time and really try to mitigate the factors that stress me. For me, exercise is probably the best anti-stressor.
Supplement Routine
In terms of supplements, I believe in measuring whatever can be measured in your blood.
There are number of companies that provide this type of services and in correcting what seems to be off-balance. So I take a number of supplements, simple ones like vitamin D and vitamin B12.
I also take NMN every day. Having taken it myself at significant doses, I really see the effects, so I’m encouraged to continue.
I take metformin for two reasons. First, it’s been shown to have many properties as an anti-aging medicine. But also my fasting blood sugar was borderline and I thought that, given my age, there were little risks in taking metformin.
So there you are – the pillars of my health or longevity program at this point.
LB: Dr. Verdin, thank you so much for taking the time to answer our questions. It is a true privilege to hear from one of the aging’s sectors most notable and well respected leaders.
Looking for more NMN based content? Look no further
FDA & TGA DISCLAIMER
This information is intended for educational purposes only and is not meant to substitute for medical care or to prescribe treatment for any specific health condition. These blog posts are not intended to diagnose, treat, cure or prevent any disease, and only may become actionable through consultation with a medical professional.
Boosting NAD by down-regulating CD38 and improving the salvage pathway: Nuchido Time+
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Building an NAD boosting supplement stack with Nuchido Time+
In our previous post, Longevity Blog interviewed Nuchido founder and NAD systems expert Dr. Nichola Conlon, on the topic of NAD systems biology and its relative complexity.
Engaging with the challenges one faces when supplementing with an NAD precursor such as NMN or NR alone.
Aging cells see increased activity in an enzyme called CD38 which consumes NAD at an exorbitant rate, as well as a breakdown in the salvage/recycling pathway for the NAD+/NADH redox reaction.
These issues must be solved in order to restore our cells to their youthful state, and while the science in this arena is at quite an early stage - Dr Conlon and her company Nuchido are leading the pack, and have tackled these issues head-on with their Time+ product.
Nuchido Time+ Supplement Review and Discount Code
Longevity Blog used this second part of our Nuchido interview to explore the ingredients included in their Time+ product. For each, we discuss with Dr Conlon a specific ingredient, and its role in solving aging related degradation in the NAD system.
If you’ve not yet read Part 1 - you may find it helpful to review the NAD systems diagram where we focus on each of the major causes for the age-related decline in NAD.
If you’d like to save 10% on Nuchido Time+ use our supplement discount code “LONGEVITYBLOG”!
Lastly, we also briefly discuss the ongoing Time+ supplement clinical trials, which should provide further evidence of the efficacy of this product later this year.
Longevity Blog (LB):
Nichola, let's start thinking about how to repair that NAD factory (reference part 1 of the interview). This is where I see an innovative approach out of Nuchido, which extends from some of your own research background. Tell us what approach you’re taking at Nuchido to restore the NAD ‘factory’ back to youthful levels.
Dr Nichola Conlon (NC):
As you mentioned at the start, we are focused on taking a ‘whole system’ approach. We didn’t want to just put more raw material in, we actually wanted to increase the cell's ability to make NAD by up-regulating those anabolic enzymes. We wanted to increase the cell's ability to recycle NAD, and also inhibit those processes that are breaking down and wasting NAD. From this angle, we knew we were going to have to go beyond simply providing a precursor ingredient.
Nuchido Time+ Ingredients
LB: And you’ve put together quite a unique mix of ingredients in your Nuchido TIME+ product. Could you walk us through these, one by one, and explain what each is doing in terms of the NAD system we’ve discussed?
Saphora Japonica flowers in bloom. Image credit
NC: First of all, I will start with Sophora Japonica. This is an extract from the flowers of a tree, which contain the really powerful flavonoids quercetin, rutin and troxrutin. These were selected based on evidence that they increase expression of the Nampt enzyme - critical for that recycling pathway.
A second ingredient is green tea extract. Everyone's heard of green tea, and how it is a really healthy beverage with lots of powerful actives. But there is one compound in particular that we're really interested in - epigallocatechin-3-gallate, abbreviated to EGCG. The reason that we're really interested in this compound is its ability to inhibit the methylation enzyme NNMT. By inhibiting the methylation process, we promote the recycling pathway for replenishing NAD.
The next ingredient is alpha lipoic acid (ALA), which works in two ways to boost cellular NAD. The first is activation of AMPK, which I'm pretty sure most of you readers will be familiar with. AMPK is a cellular energy sensor, which has another relatively unknown link to NAD. AMPK increases NAD levels in the cell by activating the Nampt enzyme. This is actually the same pathway that promotes the beneficial effects of exercise and fasting.
The second way that ALA works is that it acts on another pathway that promotes the conversion of NADH to NAD+. As a refresher, NAD+ is the oxidized form, and NADH is its reduced form. They are both part of the redox reaction of NAD. In aging cells, the ratio of NAD+ to NADH tends to drift towards NADH, which is unfavorable in terms of the energy status of the cell.
Lastly, an important thing I always like to point out when I talk about ALA, is be really careful what form of ALA you take if you take ALA supplements. There are two forms of ALA - the natural form R-ALA and the unnatural synthetic form S-ALA. Most ALA supplements use the synthetic version (S-ALA) or a 50:50 mix of the two, but there’s evidence to show that S-ALA doesn't work in the body, unlike the natural R-ALA form. So in Nuchido TIME+ we only use pure R-ALA.
LB: This is a great chance to interject with a common objection to the Nuchido TIME+ product: if I search around for many of these ingredients, and combine them all together, I can ‘re-create’ the product for myself, possibly at a lower price. However, what I am hearing from you, and S-ALA is a great example, is that it takes a lot of careful decision making and expertise to appropriately source these ingredients. Could you respond to those who think: “I’ll have a go at making my own?”
Dr Conlon is an expert on the bioavailability of molecules and supplements
NC: You know, obviously, we hear that conclusion quite a lot. My background is actually in bioavailability of molecules, drugs and supplements in the body. It’s what I did my PhD thesis on, so I was really passionate about making sure that each of the ingredients in Nuchido TIME+ were included at the optimal level so that it is able to be absorbed with good bioavailability and have maximum efficacy in the body. So the amounts and the ratio of the ingredients in our formulation have been very carefully optimised to give maximum benefit which took a lot of scientific consideration and testing to determine.
The other thing to note is that many people read about active ingredients in scientific papers and presume that they can just go and buy the ingredient in its raw form, and start taking it and it will have the same effect. But there are two things to consider here. The first is that many experiments are performed on isolated human cells and translating the dose of an active ingredient from what works on an isolated cell, to what will work when given orally to a whole human is actually very complex. Also you'll notice, with the green tea extract, parsley and sophora japonica in Nuchido TIME+ - that we don’t actually use the active molecules in their purest form, instead we use them in their raw form because this actually improves bioavailability in the body especially when taken orally.
LB: Thanks for sharing that philosophy and how your research background has informed it. The attention to detail on bioavailability is not to be underestimated. Now, let’s specifically address what I usually consider a salad ingredient, not part of my NAD boosting protocol - parsley.
NC: [Laughs], yes that sounds like a crazy one. But parsley contains very high levels of a molecule called apigenin. Apigenin is really important for inhibiting CD38, and has been shown to increase cellular NAD levels by 50%, just by inhibiting that one enzyme. Apigenin by itself doesn't have the best bioavailability, but you can get more of it into the system through its raw form, parsley.
Parsley is a really critical part of the mix, because CD38 breaks down so much NAD, I can't emphasize that enough. So it's in there (Nuchido TIME+) to provide the apigenin to actually inhibit CD38.
LB: I guess I’ll view parsley in a new light, beyond the salad bowl! Are there some other ingredients to address - zinc, vitamin C, nicotinamide and piperine are all in the mix as well?
Learn more about Nicotinamide and NAD precursor NMN in this Longevity Blog Post
NC: Nicotinamide is used as the raw material/precursor for NAD. We know the cell is very efficient at converting nicotinamide into NAD, as long as it's got all the other pathways working efficiently. Unlike stand-alone precursors like NMN or NR, Nuchido TIME+ not only supplies the cells with the raw material they need to make NAD+ but it also has all the other ingredients to make sure the root causes of NAD+ decline are addressed too.
Piperine is known to be very good at helping increase the uptake of nutrients from the small intestine. This means they can better enter the bloodstream and access the rest of the body. So it's included to help with absorption and bioavailability of the other ingredients.
Then we include vitamin C and zinc as further support ingredients. We did this, as we were conscious of the fact that many people like to have a regime that includes vitamin C and zinc for immunity and other benefits. We believe having it all wrapped up in one supplement is more convenient for the customer.
Nuchido Clinical Trials
LB: Thank you for that thorough walk through. I really appreciate the holistic approach that Nuchido is applying to this problem. I know that you've had some early results from a handful of people whom have tested Nuchido TIME+, and that you are working on a broader clinical trial. Could you comment on the observations you’ve gathered from test subjects to date?
NC: We know from our initial pilot studies, that we can expect around a 242% increase in NAD within two weeks of taking the product. This is quite a significant increase compared to some of the reported values for NAD precursors.
The clinical trial that we are running, should have been finished by now. But it's been delayed due to COVID. This is a 28 person, placebo controlled, double blind crossover study with participants aged from 20 to 80, taking the Nuchido TIME+ supplement. Not only are we looking at NAD levels, but also more deeply at the downstream effects of higher NAD.
This includes mitochondrial biogenesis and sirtuin pathway activation. We’ve collected many samples and are starting the analysis, but due to COVID this is not complete.
LB: So you were able to run the trial and collect multiple data points, but have effectively completed the clinical trial component. Now that you're in the analysis phase, can you comment on when you expect to be able to talk about your results?
Clinical trials are often a key step to proving efficacy
NC: Yeah, we're hoping within the next couple of months. The issue is that in the UK, we're again, back into lockdown. There are different policies between the labs we have to use, and when we thaw the samples, we need to be able to analyse them all at the same time. It's very important that when analysis starts we are confident that we can complete it all in one go, without any spot-start that could negatively impact results.
LB: We'd be very happy to share your results with our audience when it is completed. Pivoting, as we close out the interview - you've been very comprehensive in discussing the NAD system and ways to intervene with respect to aging. But I can’t let you go without asking you about your own personal longevity strategy. This is clearly something you're very knowledgeable on and passionate about. For starters, I noticed you're a competitive runner?
NC: Yes! I absolutely love running. I do a lot of running and a lot of running competitions. I really can't emphasize enough the importance of lifestyle choices that you can do for free. I'm really big on having a good diet and exercise routine. For me, that is a non negotiable. I get up every morning, and at 6am I go for a run or I go to the gym, or I do something physical every day, that's my routine. It boosts your energy levels, but is just good for your wellbeing in general.
I obviously take our supplement, and I take other supplements, but don't normally go into much detail about what I personally take - I'm a firm believer that everyone's an individual. You can't just copy what someone else does. You know, first of all, I'm a woman. So it's very different to what a man should be taken or doing.
LB: In personalising your own longevity strategy, what types of data to you collect?
NC: A key thing that I say to people is - get to know your own blood. Get your blood levels different markers analysed on a regular basis. That way you can keep track of any changes in your body early because what is normal for you might not be what is normal for the rest of the population. It is much better to understand your own body and know what's normal for you.
For example, I have mutations in particular genes that mean I don't absorb certain nutrients as well. So I supplement with things like folate, because my folate levels are always really low. I've seen that's really helped. Understanding if there's any parts of your biology that need a little bit of help by getting your blood drawn, or even analysing your genetics or epigenetics, can really help to track what works for you.
LB: One last thing I have to ask you - how would one describe your lovely accent?
NC: [Laughs] It’s called a Geordie accent. I grew up in the north of England, in a place called Newcastle. It is right up at the top near Scotland. We do have a very distinct accent, which also includes lots of words and phrases, which apparently the rest of the world doesn’t use or understand - but I didn’t actually realise this until I grew up and started travelling outside of Newcastle!
LB: That’s lovely, I’d like to learn some of those phrases over a pint sometime :)
We’ll close it there Nichola, thank you so much for your time on the Longevity Blog today.
NC: It’s been a pleasure, I’ve really enjoyed it!
FDA & TGA DISCLAIMER
This information is intended for educational purposes only and is not meant to substitute for medical care or to prescribe treatment for any specific health condition. These blog posts are not intended to diagnose, treat, cure or prevent any disease, and only may become actionable through consultation with a medical professional.
Do NAD precursors work? It may be the wrong question. Interview with Nuchido founder Dr Nichola Conlon
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Does Nicotinamide Mononucleotide Work in Humans?
Readers of the blog are well aware, and in rapt attention, as our Founder Nick personally delves into the self-experimentation required to answer this question - Does NMN work? In just under two weeks, he will re-test his biological age after a 6-month NMN led protocol, using the most advanced and detailed epigenetic aging clock available.
The aim - get to the bottom of what many thousands of you are asking - should I be taking a NAD precursor?
As Longevity Blog has continued to review the latest research on NAD boosting strategies for enhanced longevity, a number of important discoveries have emerged, which can broadly be grouped into two categories.
First, the absorption and effectiveness of Nicotinamide Mononucleotide in humans is still hotly contested.
If you cite the research from advocates of Nicotinamide Riboside (NR), you’ll note objections about the lack of clinical research for NMN and how NMN ultimately must first be converted into NR prior to entering into the cell.
Now most of these folks are financially involved in NR’s success (which is a patented molecule), so we must consider the potential role this will play in their personal biases.
Ultimately, we owe it to NMN to allow it to pass through the rigour of clinical trials, of which a growing number are now underway.
The list of NMN clinical trials continues to expand. Review them yourself on clinicaltrials.gov
Secondly, it is clear that the NAD precursor approach to NAD boosting in the cell is actually a bit myopic.
The NAD system, like all things in biology, is much more complex than our favourite supplement company’s marketing bullet points.
The layperson could be forgiven for overlooking this complexity, but we think it is incredibly important to explore it in an approachable form.
Enter NAD systems expert Dr Nichola Conlon
As Longevity Blog explored the complexities of NAD systems biology in greater detail, the expertise, charming accent and straight-forward explanations of Nuchido founder Dr Nichola Conlon emerged as a bright spot in the perhaps oversimplified NAD precursor debate (NMN vs. NR vs. Nicotinamide, etc).
One particularly valuable point of reference was Dr Conlon’s explanation of the ‘multi-targeted approach’ her company has developed (led by her own expert background) to the NAD boosting challenge, presented at Undoing Aging 2019:
To dive deeper into this complexity, including how a more nuanced understanding of NAD biology can help us enhance our approach to boosting it to more youthful levels, Longevity Blog conducted a zoom interview with Nichola in February 2021. Let’s dive into Part 1…
Interview with Dr Nichola Conlon of Nuchido: Part 1 - NAD Systems Biology
Longevity Blog (LB):
Readers of the Longevity Blog are familiar with NAD precursors, however, what may be new information is how taking precursors can be equated with a single targeted approach. You’ve previously spoken about this on many occasions, could you introduce the reader to this concept? What do you mean by a single targeted approach?
Dr Nichola Conlon (NC):
So first of all, biology is extremely complex and complicated. And nothing in biology ever exists alone, or in isolation. Everything that is going on within our bodies and in our cells, is actually very interconnected and intertwined. For example, there are multiple pathways that are running alongside each other performing the same job, and there are multiple pathways that are talking to each other. There are pathways, proteins and genes that are feeding information to each other all the time, and may be also activating or inhibiting each other and other processes.
Another very important concept for your readers to appreciate, is that, in order for scientists to understand and analyse biology, it is normal for them to pick these complex processes, for example, they might do experiments on a particular protein or a particular pathway, and then write a scientific paper on it. Overall, this is fine, because it helps us to understand that small piece of biology. But problems arise when you try to interpret those results in isolation without the wider context - you've got to appreciate that a single protein or pathway in the body is a very small part of a much bigger picture.
LB: What you are describing is precisely what science has done with NAD to date. What is in our mind is the classic chart of our NAD levels declining with with age - the natural conclusion becomes “We need to boost NAD levels”, and that's where much of the early research has focused. But what you’re very clearly pointing out is - it's not that simple.
NC: Yeah, that's it. All of the science says, ‘OK, NAD declines over age’, so the most obvious response is ‘how do we boost NAD?’. So at first sight, it would seem that taking a NAD precursor is a good idea.
These NAD precursors are basically the raw material that the cell needs to make NAD. But the problem is, it's now known that NAD doesn't decline because your cells have a shortage of the raw material to make NAD. Rather, it's actually declining because there are multiple other things going on in the cell causing its decline. For example, the cell is losing its ability to manufacture and recycle NAD.
Image credit: Nuchido
LB: You have kindly provided a really nice graphic (pictured) showing why NAD declines with age, within which this ability to make and recycle NAD is declining over time. Let’s focus on the major components in this graphic. Can we walk through each of those points and have you provide the reader with the basic concepts?
NC: The first point is that old cells use up more NAD. The reason for this is that they have more damage and inflammation, and so they need more repair. NAD is also a substrate for important repair pathways in the cell that are trying to fix this damage. What this means is that the enzymes and repair processes are turned right up, and whilst performing their function they’re consuming huge amounts of NAD.
LB: This consumption of NAD by repair processes is discussed broadly in the anti aging community, including compounds such as resveratrol, and its role in activating sirtuins. Could you elaborate on how aging impacts how these repair processes consume NAD?
NC: DNA repair enzymes use NAD as a substrate to drive their activity, and these activities start to increase as our cells get older - which means aging cells are actually consuming more NAD. In a young cell, this NAD consumption is not actually a problem, as young cells have a very good ability to make their own NAD via recycling. So when NAD is used up in young cells it is broken down into nicotinamide and they have the ability to recycle this straight back into fresh NAD again. This means the cells are simply recycling the same NAD time and time again.
LB: This recycling process, also commonly known as the ‘salvage pathway’, is #2 in the provided image. There are a few key inputs to this pathway, one of those is Nampt - walk us through it.
NC: Correct, this is the salvage pathway. And that's why it's called the salvage pathway - it is salvaging NAD. This pathway relies on an enzyme called Nampt, which is known as a ‘rate limiting enzyme’. This means that it is the bottleneck in the process. When the levels of this enzyme go down, it slows down the recycling of nicotinamide back into NAD.
What's been discovered recently, is that one of the main reasons that NAD is declining in cells as we get older, is because the Nampt enzyme declines with age. This means that older cells are less able to recycle NAD. And this is problematic because, as I have already mentioned, in older cells NAD is getting used up more quickly (#1, as discussed previously), because of increased activity of the damage repair systems. So large amounts of NAD is being used up and broken down in nicotinamide, which has the capacity to be recycled - but its not.
This is because with Nampt enzyme levels declining, the recycling process can’t keep up. So the nicotinamide starts to build up and rather than being recycled back into fresh NAD. So right at a time when our cells need this recycling process to work really well, it's actually declined. This creates a deficit in NAD, and from there, quite an exponential drop in NAD levels.
LB: That’s a great segue into part #3, which relates to methylation. Methylation plays a collaborative role with the recycling pathway of NAD.
NC: The body wants to keep a very tight handle on the levels of anything the cell considers a waste product. In the cell, when NAD is used up and broken down, for example by PARPs, sirtuins, etc the output is nicotinamide. If the recycling pathway is working well, it can be recycled straight back into NAD. But when it doesn’t work well (i.e. in aging), nicotinamide can build up and the cell essentially says “Oh my goodness, and there’s a buildup of nicotinamide in the cell, we need to get rid of it!”
The cell does this by increasing expression of another enzyme called NNMT, which adds a methyl group to nicotinamide, creating methyl nicotinamide. This acts as a signal to tell the cell to excrete it, which helps the cell to get rid of this excess nicotinamide from the NAD breakdown.
So what you find is that in older cells, there is increased expression of the NNMT enzyme because the cell is actively trying to remove excess nicotinamide, because it is no longer being recycled. The consequence of this is that your cells end up using a lot of methyl groups to process it. Methyl groups are also very important in other areas of biology such as epigenetics, so this has a knock on effect and the result is methyl donor depletion, meaning that the cell doesn’t have available methyl groups for those other important processes.
This is a great example of how everything's really interlinked in biology, and you can't look at any of these processes in isolation.
LB: This complexity is one of the reasons why Longevity Blog was so interested to speak with you about the NAD system in more depth. One of the things that you had spoken about previously, was this idea of there being anabolic and catabolic elements of the NAD cycle. As a reminder for the reader, anabolic means building up, and catabolic means breaking down. How do these terms relate to this image that we're looking at now?
NC: The main anabolic enzyme is Nampt, because that is the main way that cells replenish their NAD - by recycling the breakdown product, which is nicotinamide. The main driver on the catabolic side, or breakdown of NAD, which we haven't spoken about yet is CD38. CD38 is another really important part of the puzzle.
LB: Discussion in the longevity community on CD38 is certainly picking up, with more compounds for inhibiting CD38 becoming available. Talk to us about CD38, our readers are quite curious to dive into this topic as well.
CD38 via Wikipedia
NC: CD38 is a membrane protein, which basically acts as an enzyme that uses NAD as a substrate to produce cADPR, which is a secondary messenger for the cell. CD38 expression is found to massively increase with age and the main problem with this is that CD38 uses a lot of NAD. To create just one molecule of its downstream messenger, it has to metabolize around 100 molecules of NAD. This means that even relatively small increases in the levels of CD38, result in a massive decrease in the amount of NAD available to the cell. Inhibiting CD38, can increase cellular NAD levels by 50%, which shows what a huge impact CD38 actually has.
LB: Now we're starting to really understand the difference between the ‘single targeted approach’ of adding more NAD precursor (e.g. NR, NMN) into the cellular system, and a ‘multi targeted approach’ which considers CD38 levels, as well as NNMT, Nampt, etc.
Clearly, we need to think about this in any NAD boosting strategy, as well as managing methylation. How do we engage with each of these components to raise cellular NAD to more youthful levels, as we grow older and the systems we discussed are starting to break down?
NC: Exactly, a multi targeted approach is what Nuchido is all about, because nothing in biology exists in isolation. I think just one thing just to point out, which will probably be interesting for your readers is just looking at it from the point of view of what happens if you are only taking a precursor?
LB: Let’s absolutely discuss this, Nichola, at the moment, Longevity Blog is running a self-experiment with the NAD precursor NMN. This includes a before and after biological age test with UK based Chronomics, and will conclude at the end of February 2021.
Longevity Blog’s self-experiment with NMN is powered by DoNotAge. Use the code ‘longevityblog’ to save 10% on any order.
The supplement stack I am testing is the popular NMN + Resveratrol + Tri-Methyl Glycine combo from David Sinclair's Lifespan book, which many folks around the world are now taking. This is why Longevity Blog is running this experiment - to see if it can impact biological age, as Dr. Sinclair seems to suggest.
NC: We'll use that exact protocol as an example. You are adding NMN into the system, which will be converted in the cell to NAD. This NAD will then be used up in the cell by processes that rely on NAD such as your DNA repair enzymes and also the sirtuins, which you're also activating with the resveratrol that you mentioned. It will also be consumed by CD38 (#1), and all of these processes cause the NAD to be broken down into nicotinamide.
This is where the problem begins, especially in older people, because the levels of the Nampt enzyme which would usually recycle this nicotinamide back into NAD (#2) have declined. So by taking an NAD precursor, you've put NAD into the system, but it only has the opportunity to be used once, when really the cell could keep recycling that precursor back into new NAD again, if it had an efficient salvage pathway, but in older cells, it doesn't.
NAD precursor supplementation can deplete the Methyl donor pool - learn more about this risk in our previous post on NMN safety and risk management.
The outcome of this failure to recycle results in a buildup of nicotinamide in the cell. Now we’re back to the methylation problem (#3) because the cell has all this new nicotinamide hanging around. It can't convert it back into NAD because Nampt isn't working as well as it should be so it needs to get rid of it, so it increases expression of the NNMT enzyme which methylates nicotinamide to help excrete it from the cell. And the resulting problem is a reduction in methyl groups in the cell. Which is exactly why you are taking tri-methyl glycine (TMG) alongside NMN. Because it's known that people taking precursors have a big problem with methyl donor depletion, and TMG contains three methyl groups and a glycine to try to replenish this.
The key point is that you're never addressing the root of the problem and instead by effectively trying to paper over the cracks you are creating other issues. And actually, if you just fixed the recycling problem, then you wouldn't have that issue with methylation because the levels of nicotinamide would never build up so big that they need to be methylated and excreted, it would just get recycled.
LB: Brilliant explanation Nichola. So would you say that with the precursor supplement approach that we're on the right track, but we're missing some components? Or is the precursor led approach fundamentally flawed?
NC: Precursors were the first solution that was available based on the best available information at the time. But the scientific understanding has developed since then. We now know the underlying reasons why NAD declines with age, and we know how we can fix them.
People who are experimenting should always keep re-evaluating what they're doing - asking themselves ‘is this still a sensible approach based on what the science is saying?’
As we age, the NAD factory is losing function
I often ask people who are taking NAD precursors to think of it in this way - imagine that your cells are little NAD factories, and and all of a sudden, the production of NAD in this factory declines. You find that the reason that NAD production has gone down is because the machines have become old and broken and the factory pipes are leaking, wasting material. In this scenario, do you think it would be a good idea to try and boost production in this factory, simply by ordering more raw material to be delivered to the factory gates?
Of course not! If you want to boost production again you are going to have to fix the factory first, otherwise the raw material will just pile up. But this is exactly what the precursor approach to boosting NAD does - it ignores that the cell’s NAD enzymes aren’t working like they used to and that proteins such as CD38 are wasting NAD and just piles more raw material into the cell. But if you want to have any real impact you need to address the root causes.
LB: It's a wonderful illustration. And I'm glad you shared that with us. In doing my background research for this interview, I’d heard you use it before, and I thought it was a very appropriate way to visualise the problem.
Nichola, let's start thinking about how to repair that factory. This is where I see an innovative approach out of Nuchido, which extends from some of your own research background. Tell us what approach you’re taking at Nuchido to restore the NAD ‘factory’ back to youthful levels…
FDA & TGA DISCLAIMER
This information is intended for educational purposes only and is not meant to substitute for medical care or to prescribe treatment for any specific health condition. These blog posts are not intended to diagnose, treat, cure or prevent any disease, and only may become actionable through consultation with a medical professional.
Choosing an NMN Supplier for Self-Experimentation
What is the Best NMN Supplement?
Who should you choose for your NMN supplement?
Fast-take: I recommend DoNotAge, who offer a third party tested pure product, have a bulk supply option (100g) with further savings when you subscribe to regular shipments. Use the code ‘longevityblog’ to save a further 10%!
Want the details and the ‘why’ I make this recommendation? Do read on!
Today on Longevity Blog, I wrap up my three part series on self-experimenting with Nicotinamide Mononucleotide. Complete with NMN before and after testing.
As discussed in the first post of the series on NMN risk management, there are three key questions that Longevity Blog will engage with on the topic of NMN. These are:
What NMN supplement should I use?
In this third post of the three-part series, we’re going to zoom right in on the question - how do I choose a high quality NMN supplier?
Choosing the best NMN supplement: Potential Pitfalls
NMN is a supplement, meaning its production and distribution are largely unregulated. As such, this potentially exposes the self-experimenter to several potential pitfalls when purchasing.
The most important of which are:
Imposters (i.e. not actually NMN)
Contaminants (e.g. heavy metals)
Impurities (e.g. fillers, extras)
Unnecessarily high cost per dose (e.g. low potency)
Naturally this raises the question - How can we avoid these potential disruptors to our self-experimentation?
For those currently searching for their own NMN supplier, I personally understand where you are coming from.
First up - you are aiming to purchase a product with confidence in knowing it is NMN, but fake supplements are common and are lurking amongst in the available NMN suppliers on the web right now.
Even if you can become confident the product in question is NMN (not an imposter), you still must consider the risk of contaminants such as heavy metals showing up in your supply.
Furthermore, we don’t want to waste our money on a product that has low potency and/or uses fillers to push more product (impurities).
And then there is cost. Once satisfied on the above three - imposters, contaminants, impurities - we still need an NMN supply that fits our budget. This is complicated by varying levels of potency across the options:
what is a ‘dose’?
how much NMN per dose?
how many doses per container?
shipping costs?
The above long list of factors can make selecting an NMN supplier quite daunting, and perhaps overwhelming!
No one individual can take on the challenge of sorting through these points on their own. Thankfully, the NMN self-experimentation community is working together on this one…
NMN Supplement Review Useful Sources
One of my favourite aspects of the self-experimentation community, is the collective spirit of sharing knowledge and helping each other navigate the pitfalls of sourcing supplements.
Despite its relative novelty, NMN suppliers have been usefully reviewed in several formats - the most valuable of which, at this stage, are two YouTube videos.
In the first, we find that once again, my mate Dr. Brad Stanfield continues to be one of the top sources of information on NMN.
In his videos, he reviews several brands - and parallels my above selection criteria - analysing imposters, impurities, contaminants and cost per dose.
Dr. Stanfield’s endeavour to find 3rd party testing data reveals a key resource in our journey to select our NMN supplier.
The second YouTube video is from Vince and his channel “My NMN Experiment”.
Vince has several relevant videos for the NMN self-experimenter on the topic of NMN sourcing, self-experimentation and review of available third party testing.
Of particular use is his video on ‘Testing Heavy Metals” which you can view below:
Choosing the Best NMN Supplier
Use the code ‘longevityblog’ to save 10% on any order with DoNotAge
Amongst the growing number of choices for supplying NMN, is UK based company DoNotAge.
I recently became a strategic advisor to this company, owing to our shared interest in making longevity supplements like NMN widely available.
Simply put, if NMN can deliver even a fraction of the longevity benefits to humans that are observed in mice, then I firmly believe in the democratisation of access to the molecule.
By democratisation, I mean - making it widely available in both supply and cost. Quality is of course pre-requisite.
Before we move on, it is important to disclose that I stand to benefit from the sales of NMN by DoNotAge due to my advisory role.
However, this monetary benefit is modest and frankly, is not part of what motivates me to self-experiment with NMN, nor to recommend the company to you personally.
Instead, what my position as an advisor has allowed me to do is more clearly understand the NMN global supply chain and access certain confidential information about its production, proprietary competitor testing, distribution and profit margin.
Many NMN options are low quality
Through the clarity this position has provided me, I have a heightened level of concern on the practices of many of the other NMN suppliers.
The issues I am now privy to are numerous and commonplace. I am aware of products sold on Amazon, claiming to be NMN, which contain nicotinamide instead (imposters).
Suppliers who claim to be ‘manufactured in the USA’, source their products from China (lack of integrity) and package it in the United States in order to ‘cheat’ their way to this label.
Others have degradation of the product due to poor packaging and storage practices (potency), while many others provide no third party certification regarding heavy metals (contaminants).
Moreover, many sell NMN at low potency, including doses at 50 - 150mg, which are well below even the conservative doses being used in early clinical trials.
DoNotAge is a High Quality Supplier of NMN
In addition to the “insider scoop” on the NMN supply landscape, my advisory position has also provided me with a high degree of confidence on DoNotAge’s Pure NMN product.
I know with certainty it is NMN (not an imposter), is appropriately free of contaminants or impurities, and it is amongst the highest potency options available on the market (500g NMN per capsule).
This combination of factors given me the confidence to personally consume the DoNotAge product, using it in my current NMN self-experiment, selecting it from the ‘Wild West’ of NMN suppliers.
Despite the potential conflict of interest I hold in monetarily benefiting from the sales of this product, it is my hope that this vote of confidence may contribute to your own decision making process.
Simply put - in my longevity journey, I am very disciplined in choosing the supplements I consume! DoNotAge passes all my checkpoints.
Reviewing NMN for Self-experimentation Protocol:
Use the code ‘longevityblog’ to save 10% on any order with DoNotAge
Zooming out from the details of choosing the best NMN supplier, let’s come back to my NMN self-experiment.
I am presently wrapping up week 4 of the protocol, which will run for a total of 6 months.
The experiment will rely on several different data points, one of which is before & after biological age tests from Chronomics, and will wrap up at the end of February.
Unlike many of the other self-experimenters out there, I am holding constant my daily habits, diet and supplement routine, in the interest of generating the most robust scientific outcome.
When complete, I will finalise the results and protocol, in order to support the self-experimentation community.
The self-experimentation protocol is as follows:
NMN 500mg per day (updated)
TMG 500mg, every day (1:1 ratio to support methylation)
All of which is being supplied by DoNotAge.
You can find the relevant DoNotAge products in each of the above hyperlinks.
Use the discount code ‘longevityblog’ to save 5%!
Follow Longevity Blog on Twitter for the latest #Longevity news!
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FDA & TGA DISCLAIMER
This information is intended for educational purposes only and is not meant to substitute for medical care or to prescribe treatment for any specific health condition. These blog posts are not intended to diagnose, treat, cure or prevent any disease, and only may become actionable through consultation with a medical professional.
Self Experimentation with NMN Nicotinamide Mononucleotide
NMN Before and After with Biological Age
Given the introduction of nicotinamide riboside (NR) and more recently nicotinamide mononucleotide (NMN) as supplements over the past four years, many in the longevity community are self-experimenting with these molecules which boost NAD+ levels.
Looking for an NMN supplier? I recommend DoNotAge, who offer a third party tested pure product, have a bulk supply option (100g) with further savings when you subscribe to regular shipments. Use the code ‘longevityblog’ to save a further 10%.
Get the details in Part 3 of this NMN self experiment series
As discussed in the previous post on NMN risk management, there are three key questions that Longevity Blog will engage with on the topic of NMN. These are:
NMN
How can we self-experiment to determine efficacy?
In this post, I’ll discuss how we can self-experiment to determine the efficacy of NMN supplementation. Let’s do it!
NMN Self-experimentation: A first draft “how to” guide
Let’s science this and see what happens
As with any good self-experiment, we should design a solid experimental framework based on the principles of the scientific method.
This has caveats however, owing to our inherent complexity as a biological system.
As you up your self-experimentation game (what a biohacker you are!), you’ll find out that self experiments are:
Difficult to control
Often hard to measure directly
Prone to bias and placebo effect
Important to make repeatable
Through this post and others that follow, we will do our best to continually consider these issues with self-experimentation (an example of this can type of thinking can be seen in previous posts).
In your endeavour to self-experiment with NMN, getting the experiment right is particularly important.
It should be our goal to provide insight to the broader self-experimentation community, and facilitate others to effectively self-experiment so we can benefit from collective knowledge sharing.
So let’s aim to get this right, in the name of science!
NMN Before and After, Our Hypothesis: What are we testing?
After thorough consideration, I believe our hypothesis for NMN supplementation can be summarised as follows:
NMN supplementation will boost intracellular NAD+ levels
Boosting NAD+ levels with NMN will measurably improve select characteristics of youthfulness
This hypothesis is formed upon on a large volume of work around NAD+ boosting supplements such as Nicotinamide, Nicotinamide Riboside (NR) and NMN (nicotinamide mononucleotide), which indicate 1) the importance of NAD+ levels on a plethora of cellular processes, 2) the known decline of NAD+ levels with age and 3) the apparently causal relationship in this drop in NAD+ levels with a wide array of age-related diseases.
It also formulated in such a way that we can test it, by gathering related data and analysing them to draw conclusions.
Does NMN Work in Humans? How We Can Test the Hypothesis:
In order to test our hypothesis, we need to determine:
An appropriate NMN dose for raising NAD+ levels
How we will measure the NAD+ levels
What controls we can apply, and adopt defensive positions against bias/placebo
How much NMN should you take?
In order to determine the appropriate dose for the experiment, I suggest we look to the two FDA approved clinical trials which are currently evaluating NMN safety and efficacy:
You can search for clinical trials on any topic you like at clinicaltrials.gov
Clinical Trial #1: Dosing for participants 40-65 years olds set at 300mg/day
Clinical Trial #2: Dosing for post-menopausal women set at 250mg/day
Anecdotes from elsewhere suggest this a good level for supplementation. Consider this 27 year old who was totally ‘buzzed’ on 500mg/day and this discussion thread for a 34 year old (same age as me at the time of writing) settling on ~250mg day.
Setting the NMN supplementation level should be done based on age and moderated based on one’s experience with the dose (e.g. insomnia, flushing, etc can be inferred to mean one should titrate downwards).
Measure: Data Collection for the NMN Self-Experimenter
From first principles, our goal here will be to measure related biometrics before NMN supplementation, and then again after sustained NMN supplementation. However, the details here are challenging. As, unfortunately sampling intracellular NAD+/NADH levels presents a few issues.
So for widespread, democratised access to NAD+/NADH testing, we face these two barriers:
First, direct testing of NAD+/NADH is limited to professional assay test kits (like this one)
This violates our goal to make the experiment repeatable for other self-experimenters.
Secondly, it turns out they are fairly ‘noisy’ data anyway, making interpretation difficult and making representativeness of the measurement problematic
NAD+/NADH levels change dramatically across the day with activity, eating, circadian rhythm (Interestingly, blood glucose exhibits these same characteristics; which is perhaps unsurprising as they are both part of our body’s ‘energy system’ );
even if we could test it directly, we would also need to sample it semi-continuously (many measurements)
However, given the widespread interest, research funding and commercial development of NAD+ boosters, measuring NAD+/NADH is likely to become more widely available in the near future. My research found at least two promising options,:
Liquid chromatography - “high-performance liquid chromatography (HPLC) to accurately measure the levels of NAD+ in cells and tissues”
A bioluminescent biosensor - “for the rapid quantification of NAD+ levels in biological samples, which can be used either in laboratories or at the point of care”
Establishing an NAD+ Proxy
In science, where we can’t measure something directly, we use a suitable proxy. In atmospheric science, to study the past climate, we can use bubbles captured in glacial ice cores to estimate past CO2 concentrations and global temperatures.
Climate researchers do this because they inconveniently didn’t have thermometers sampling the atmosphere 100,000 years ago. Here, our issue is we don't have the ‘thermometer’ for NAD+/NADH levels yet. So what’s our proxy?
I have spent many hours researching and considering this. It is my suggestion is that we leverage several different biomarkers that are representative of youthfulness as our proxies. The reasoning for this reaches back to the hypothesis and the purpose of boosting NAD+ : to restore youthfulness.
There are a few proxies we can draw from the clinical trials mentioned above, these are:
Walking speed tests
Blood pressure
Lipid profile
I suggest we eliminate the walking speed test, as it is really only valuable in advancing aging or for sedentary individuals. The latter three are widely accessible, so we’ll adopt them.
But I suggest we go a bit further in our assessment of expressing a youthful phenotype, and adopt measurements of biological aging. My reasoning for using biological age (BA) is the following:
BA is our target for longevity optimisation
BA measurement and calculation is accessible to others, therefore can be repeated
BA is a wide reaching measurement of overall bodily youthfulness (that is its purpose)
BA has multiple modes of testing available to us; so we can work with more than one
Epigenetic test kits for before and after the Longevity Blog NMN based planned biological age reversal experiment. Read about what I made this choice!
For the NMN self-experimenter, I suggest the following two biological age computation methods.
The first is the Phenotypic Age test, which I cover in this post.
The second is a biological age test based on measurement of DNA methylation (aka epigenetic age)
While the prior test is a blood test, the latter is direct measurement of methylation at CpG sites in the epigenome.
There is a fundamental connection here between NAD+ levels, which are the ‘fuel’ for the SIRT enzymes which repair DNA and regulate expression of the genome.
In theory, increasing NAD+ levels could improve our bodily ability to regulate the epigenome, so potential improvements could be realised in the epigenetic age.
Epigenetic age kits are also becoming widely accessible very quickly, and therefore are repeatable.
Analysis: How should the self-experimenter analyse their data?
At the most fundamental level, at the very least, one should statistically compare the before & after measurements of our proxy measurements.
An added benefit would be the inclusion of additional baseline measurements from before the experiment begins.
This inherently needs to include some basic corrections for the non-stationary nature of the data. In layperson terms - chronological time is passing over our measurement period.
So the ‘after’ measurements should be corrected in our analysis, considering the amount of time that has passed over the course of the experiment.
Lastly, and perhaps more importantly, we must employ purposeful management of potential confirmation bias.
Anyone who is running an NMN self-experiment and is looking after their longevity wants and likely expects the supplement to work.
Since we know the result we want, therefore we need to be defensive against our bias when we draw conclusions based on this result.
This is particularly relevant for bloggers and social media focussed folks - we inherently want the Ben Greenfield headline to draw clicks and eyeballs.
I refuse to do that to you, and will revisit these issues when we analyse the data and draw our conclusions.
NMN Self-experimentation: What’s Next for Longevity Blog?
We’ve nearly designed our experimental framework, but we’ve not yet discussed the dosing protocol. So here’s what I personally plan to do:
Baseline
Measurements of:
Blood lipids
Blood pressure
Estimation of:
Phenotypic Age (established here)
Epigenetic Age (via Chronomics)
Complete the SF-36 questionnaire
Post Protocol
Measurements of:
Blood lipids
Blood pressure
Estimation of:
Phenotypic Age (established here)
Epigenetic Age (via Chronomics)
Complete the SF-36 questionnaire
So, what will the exact dosing protocol be? It is still in draft form, as I am awaiting potential additional insights from first Chronomics test.
These may provide additional insights on what to add to the protocol to further improve my biological age.
This does step outside of the experimental design for NMN self-experimentation, but let’s delay that discussion until further posts.
NMN Self-experimentation Protocol (DRAFT):
NMN dose will be set at 250mg per day (based on clinical trial dosing levels)
Update: Two months into the protocol, I raised to 500mg per professional advice from a knowledgeable third-party
For risk management, we’ll include 1:1 TMG (as discussed in my last post)
Resveratrol at 500mg/day, taken with a high fat meal (I’ve been doing this for 2+ years)
Inclusion of Resveratrol is based on its role as a SIRTuin activator, which pairs well with NAD+ boosting and potential benefits to the ‘health’ of the epigenome
Will you experiment with NMN? You’ll need a trusted supplier - I recommend DoNotAge, who offer a third party tested pure product, have a bulk supply option (100g) with further savings when you subscribe to regular shipments.
Use the code ‘longevityblog’ to save a further 10%.
Get the details in Part 3 of this NMN self experiment series
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FDA & TGA DISCLAIMER
This information is intended for educational purposes only and is not meant to substitute for medical care or to prescribe treatment for any specific health condition. These blog posts are not intended to diagnose, treat, cure or prevent any disease, and only may become actionable through consultation with a medical professional.
NMN - Nicotinamide Mononucleotide Safety and Risk Management
NAD boosters are a HOT topic. NMN is the hottest 🔥
One of my favourite ways to analyse the latest health action in longevity technologies is to use Google’s keyword planner and trends tools. They form a great way to track the ‘pulse’ of human interest on healthspan. And as I planned my next upcoming series of blog posts on NAD boosting with NMN and diving deep into epigenetics and biological age, I certainly found plenty of evidence of our collective curiosity on the topic.
Given the introduction of nicotinamide riboside (NR) and more recently nicotinamide mononucleotide (NMN) as supplements over the past four years, many in the longevity community are self-experimenting with these molecules. And while the requisite clinical trials (those in humans which reveal their effectiveness) are still ongoing, it is clear from the data and trends I analysed that these molecules, particularly NMN, are moving into the mainstream. Heck, I’ve even run into many folks in the energy sector (where I run my day job - Solcast), taking NR and/or NMN!
Trends in NMN and NR over time
Here is a great illustration on how the search analysis for NR and NMN have changed over time. While NR got an early boost with the launch of Elysium Health and their product “Basis”, searches quickly returned to baseline within a matter of weeks.
NMN however, after an outstanding boost from David Sinclair’s appearance on the Joe Rogan Experience podcast, and likely supported by Dr. Sinclair’s impressively wide-reaching Lifespan book launch tour, has clearly achieved a wider and more sustained appeal. NMN is clearly the hottest topic in the world of NAD+ boosters, and this is supported further by the search results present in many English speaking countries, with thousands of searches for NMN and related topics each month (and growing steadily).
What is NMN? What are its benefits?
It is hardly my role to explain the basics of NMN and NAD+ boosting supplementation, as that is not the focus of the Longevity Blog (here we focus on decision making, actionable insights). Thankfully however, my mate from across the pond Dr Brad Stanfield (New Zealand) runs a fantastic and educational YouTube channel covering these topics. You should be able to find the resources you need to become more familiar with NAD+ boosting supplementation by watching his videos (be sure to SMASH that like button 😂).
NMN Supplementation: Safety, Strategy, Sourcing
There are however three key questions that Longevity Blog will engage with on the topic of NMN. These are:
Is NMN safe for self-experimentation?
How can we self-experiment to determine efficacy?
What NMN supplement should I use?
In this post, we’re looking into NMN safety and risk management. We’ll come back to #2 and #3 in future posts.
NMN Supplementation: Is it safe? (Key Question #1)
When exploring NMN supplementation safety, there are two important sub-questions to explore:
1) Has the molecule been found to be safe by reputable third-parties?
2) Are there known risks with supplementing the molecule?
NMN Safety
Perhaps the most compelling safety certifications a molecule can receive is the GRAS accreditation from the FDA. GRAS - Generally Regarded As Safe status has been awarded to NR, as well as other forms of Vitamin B3 such as Niacin and Nicotinamide. NMN however has not yet reached this hurdle as of mid 2020.
However, in my opinion, this is inevitable, will arrive soon, and there are currently no reasons other than timing and resourcing as to why this has not yet occurred. NMN supplements are just entering the market en masse (see the above analysis!), and the FDA will be certain to react to this. To be clear, I am not asserting its safety independently, this is the role of the FDA. What I am saying is that closely related molecules have received GRAS status, and the lack of GRAS status for NMN is related to its relative newness in the world of supplementation not due to any known risks.
In summary, NMN is likely to become Generally Recognized as Safe (GRAS), and the other members of the Nicotinamide family (NR, Niacin) have GRAS status. We presently have no reasons to doubt it is a safe molecule to ingest.
NMN Supplementation Risks
However, GRAS status doesn’t ensure it will yield us a positive longevity based outcome. It more or less means it won’t poison us. Safety in this context does not mean there are not risks.
Nicotinamide. Source: Wikipedia
I have spent many tens of hours studying scientific publications available on the topic of Vitamin B3 supplementation and other closely related molecules to NMN (NR, Nicotinamide). What I have found clearly suggests supplementation of Vitamin B3 and its related molecules at high doses. At the most fundamental level, NAD booster supplementation of any form, results in adding significantly higher amounts of B vitamins into the body than it is prepared to metabolise. This is what I would refer to as ‘high doses’ in this context - specifically, consuming 1, 2 or even 3 orders of magnitude greater than the RDA for the vitamin group.
NMN! Source: Wikipedia
Conversely, my research also cemented the claims behind high dose supplementation, as there are a wide array of purported benefits for NMN (limited to mouse studies at the time of writing) and even greater supporting evidence for the benefits of NR (which NMN is converted into prior to entering cells) including early studies in humans. I won’t delve into all of these benefits - if you’re reading this post, you’re likely already sold on the benefits!
In making a judgment call, one is left with a risk management scenario. The possible benefits are significant. However, there are risks associated with high doses of B vitamin intake. To gauge how we make a decision, we turn to the fundamentals of risk management. Understand the motivations for the activity, diagnose the risks of the activity, apply a risk management strategy, and make a decision based on the residual risk.
This means, we next need to understand the risk better, after which we look at the options to managing the risk.
From the abstract:
“However, possible adverse effects and their mechanisms are poorly understood. High-level NAM (Nicotinamide) administration can exert negative effects through multiple routes. For example, NAM by itself inhibits poly(ADP-ribose) polymerases (PARPs), which protect genome integrity. Elevation of the NAD+ pool alters cellular energy metabolism. Meanwhile, high-level NAM alters cellular methyl metabolism and affects methylation of DNA and proteins, leading to changes in cellular transcriptome and proteome. Also, methyl metabolites of NAM, namely methylnicotinamide, are predicted to play roles in certain diseases and conditions.”
NMN Supplementation Primary Risk - Depletion of Methyl Pool
In my review, the primary risk of NMN supplementation (or NR), is the depletion of the methyl group ‘pool’ on which many aspects of your metabolism interact and modulating your DNA expression depend.
A recent study in the journal Biomolecules shares the work of South Korean duo Eun Seong Hwang & Seon Beom Song, who approach this topic from the same place that we opened this blog post - Vitamin B supplementation is growing dramatically in all forms, what are the possible consequences?
Of the many possible negative effects of NMN supplementation, only one of these has been determined to be highly probable. In fact, based on the fundamentals of organic chemistry, it is nearly certain to occur.
Taking high doses of Vitamin B3 (NAM, NMN or NR) will increase the demand on your available methyl pool. This means there are reduced methyl groups available for the methylation of DNA (which, for example, operates in the epigenetic expression of DNA via histones). This could have the deleterious effect of altering your patterns of gene expression and thereby impacting the supply of proteins your cells need to encode for important processes such as cellular repair or replication.
I have adapted the below image from the Biomolecules paper in order to add a bit of clarifying sign-posting on the diagram. From left to right, we have the flow of potential consequences of elevated NAM intake. At far left, the methylation cycle is represented, and at the nexus of it and the NAM cycle, we can see the methylation of NAM (to metNAM) occur. This is what will need to occur in high doses of Vitamin B (NR, NMN or any other form of NAM).
NMN Supplementation Risk Management
Having understood the risk, analysis reveals a critical system (DNA expression, protein encoding) depends on the methyl groups which high doses of Vitamin B are likely to deplete.
So now the question is, can we employ a risk management strategy to minimise this risk?
I believe the answer is yes. Thankfully, it is possible to replenish the cellular methyl pool exogenously via supplementation. Allow me to explain a bit further.
At left in the above image, is an input to the methylation cycle via betaine. Betaine is a metabolite of choline (note how the methylation cycle flows from betaine to choline) and is one of the most important methyl ‘donors’ (another being folate). Betaine can also be called ‘Trimethylglycine’ (TMG). More broadly betaine actually refers to a class of molecules, but in the context of metabolism TMG and betaine are used interchangeably.
TMG is our risk management strategy when it comes to high dose NAM supplementation (including NR and NMN). This molecule is formed of an amino acid (glycine) that has three methyl groups attached to it. It is these three methyl groups that provide the extra inputs we need to support our ramped up methylation cycle when supplementing with NAM. It is possible that different forms of methyl groups, not provided by TMG, could also be required. This will be the subject of future research and updates to featured on the Longevity Blog.
nmn and nr Supplementation gets the green light
Based on the deep dive of research I have been completing on the topics of NAD+ boosters and epigenetics + biological age, I believe NMN supplementation to be risk-appropriate and a potentially effective tool in my longevity supplement stack.
I will soon be embarking upon my own NMN self-experimentation, and as I do so, I will be supplementing with Tri-methyl glycine (TMG) on a 1:1 basis with my NMN intake. In my view, the supplementation of TMG provides self-experimenters with an appropriate risk management strategy for long-term (i.e. more than 1-2 months) NMN (or NR) supplementation.
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FDA & TGA DISCLAIMER
This information is intended for educational purposes only and is not meant to substitute for medical care or to prescribe treatment for any specific health condition. These blog posts are not intended to diagnose, treat, cure or prevent any disease, and only may become actionable through consultation with a medical professional.