Longevity in the laboratory: Key biomarkers for healthy aging

Oliver Schmidt: Welcome to today’s session on longevity in the laboratory.

My name is Oliver Schmidt. I’m Product Manager at Tecan for this product line, and I would really like to take the chance to take up a very hot topic in current social media, news, whatever there is. I want to make you aware that you can also help as a laboratory.

Therefore, I want to take you on an imaginary route. Imagine yourself sitting at your parents’ home, looking at a photo book. You’re looking at the photo book and you see pictures of your grandparents, and they look quite old. You ask your parents, “How old were they when this picture was taken?”

Your parents tell you, “They were around 50 at that time.” You think, “Whoa, they look old when I compare them to you. You’re now 70, and you look the same.”

This really gives the introduction to the topic of longevity, because with today’s healthcare, lifestyle possibilities, interventions and what we know today, it is possible to influence our looks, our health and everything that is associated with aging.

These are the topics. I will tell you a little about what is meant by the term longevity. I will talk a little about aging-associated diseases. Then I want to show you, which is really interesting, that there is a discrepancy between chronological and biological age.

Then, how do you get the samples? I also want to show you the possibilities for obtaining samples at home so that you send them directly to the patient. I will talk about saliva, urine and, nowadays, also blood.

Then I will talk about the biomarkers that you will need to measure and that you can use to help the patient. I will talk about the hormones cortisol, DHEA, melatonin and testosterone in saliva. I will also talk about leptin and adiponectin, which are adipokines.

I will talk about titin N-fragment. I will talk about alpha-Klotho. I will talk about sIL-2R and free 25-hydroxyvitamin D. Then I will try to sum it up for you.

We start with this background: definitions, diseases and aging, which I already mentioned in the topics.

Longevity, what does it really mean in the current context? It means expanding the healthspan, living healthily for longer alongside our chronological age. Longevity, in total, means embracing a holistic approach to healthy aging, improving biological age through thoughtful lifestyle choices and understanding the optimal choices via advanced diagnostic tools.

Longevity is also influenced by diseases that are connected to aging, lifestyle, diet and nutritional supplementation.

This leads us to aging-associated diseases, diseases and conditions that are linked to aging. Conditions that usually have an increasing incidence with age are typically osteoporosis, sarcopenia, chronic inflammation, atherosclerosis, cardiovascular diseases, arthritis, type 2 diabetes, hypertension and several neurodegenerative diseases, for example, Alzheimer’s disease.

Then we go to chronological versus biological age. There is usually a discrepancy between chronological and biological age, and that can be overcome. It can also be monitored by the laboratory.

I found this very nice graph from an anti-aging clinic in India, which describes very well what influences our biological or chronological age and either leads to accelerated aging or decelerated aging.

We can see negative influences and positive influences. Negative influences include sedentary behavior, lack of sleep, smoking, fast or junk food, and stress. All these factors make sure that we age faster than we actually are.

But there are possibilities to influence this and age more slowly, so decelerated aging. This can be done with exercise, everyday activity, sleep time and quality, appropriate nutrition and relaxation techniques. Everything that we know and like, actually.

These are the biomarkers linked to it. Soluble alpha-Klotho, for example, is linked to sedentary behavior. We have melatonin and cortisol, which we can measure for lack of sleep, but also, on the other side, for sleep time and quality.

The sIL-2 receptor, or sIL-2R, can be considered for smoking. Leptin, adiponectin and sIL-2R can be considered for fast and junk food. Cortisol and DHEA are typical hormones that are part of the stress profile.

Then we can control exercise, because not every exercise is good exercise. We can measure titin N-fragment, for example, in urine to see whether we are doing good exercise. We can also measure something, as you will learn later, with testosterone in saliva.

Again, we can check our everyday activity with soluble alpha-Klotho. Cortisol and melatonin, as I already said, can be considered for sleep. Leptin, adiponectin and sIL-2R can also be considered for appropriate nutrition.

Relaxation techniques are very interesting. You can also measure their effects using cortisol.

Then we come to how the laboratory can actually sell this to the patient. You have to give patients a holistic overview. You have to be patient-centric. You have to have a purpose behind it. It needs to be convenient and preventive. All these topics are what patients want to have. It needs to be very easy for them to provide their samples.

Of course, first we need to obtain the samples, and that is very simple. At-home sampling is becoming common.

Nowadays, it is very easy to obtain samples, even blood samples, at home. You don’t have to go and see your doctor anymore to get a body fluid out of your body and send it to the laboratory.

For example, obtaining a saliva sample is really easy. The biggest advantage of saliva is that serial sampling is possible. The only thing you have to make sure of is that you have sufficient volume to perform enough measurements, with many more parameters than one.

It is also important that you take preanalytical measures to make sure that everything is correctly measured in saliva. This includes freezing, thawing and centrifugation.

You should be aware of patient medication because it can falsify the sample, and you should also be aware of the influence of the collection device.

On the right-hand side, you can see a graph from a colleague of mine. We researched several collection devices and checked how they influenced the results for the five main steroid hormones.

Urine self-sampling has become routine practice since the late 1970s. First of all, it is easy to collect. The main sample you usually provide is spontaneous urine.

It is important to avoid the first stream to avoid contamination with cellular debris, microbes and urogenital biomarkers.

During the last decade, stabilizers were also invented to ensure urine stability during transport. Nowadays, it can be sent by post to the laboratory and then be analyzed.

The most recent development is that, nowadays, you can even obtain blood samples at home. For some years now, several blood self-sampling devices have come onto the market, such as the five you see on the right-hand side.

These include the PreciHealth SA EasyDraw blood sampler, the Loop Medical SA Onflow, YourBio Health, Tasso Inc.’s Tasso+ or Tasso Mini, and RedDrop Dx with its RedDrop device.

All these devices are designed to be used with standard blood collection tubes from companies such as Becton Dickinson, Sarstedt and others.

What is also quite useful is that you can collect up to 3,000 microliters of blood per draw, which leaves you with sufficient sample to process.

That brings us to the biomarkers. I want to explain a little about their current use because I think this is the most important part for you. You want to understand what biomarkers you can offer and what consequences can be drawn from them.

I think the oldest and most interesting part is the hormones. This is really interesting because the hormonal picture, in the end, tells the truth.

The idea of measuring different hormones such as cortisol, DHEA, estradiol and testosterone is to monitor stress. But you can also monitor other things such as hair loss, menopause, erectile dysfunction, fertility and sleep.

I think the most important ones for longevity are stress and sleep in this case.

That leads us to why cortisol in saliva is useful. What does it really tell us? It is very simple. Cortisol is the stress hormone, and it is linked to stress.

Here, you can see three graphs showing typical daily profiles of cortisol. The green area shows where cortisol should usually be throughout the day.

In the first example, the cortisol levels throughout the entire day are always above the upper limit. Saliva is particularly useful here because, as I said before, you can perform serial measurements. In this case, we have chronic adrenal stress.

This person is really not feeling well. The person is overstressed.

In the middle picture, the values first thing in the morning are in the upper range, and then they drop to the lower range, but they are all in the normal area.

However, you see this steep drop, and this is a sign of a fatigue or stress pattern. This person is not completely stressed out yet, but could be very close and should be monitored closely to make sure that the patient does not, for example, develop fatigue.

Then there is the other pattern, where you have adrenal exhaustion. It is really fatigue or even burnout. You can see this when all the values throughout the day simply do not go up. The cortisol levels do not go up anymore.

The person feels all the time that they want to stay in bed. That is usually what you have. You have a suppressed pattern.

What does DHEA in saliva tell us? First of all, DHEA is considered the anti-aging hormone because it is age dependent. You can see in the picture below that DHEA declines with age, and that led to it being named the anti-aging hormone.

It is quite common, especially in some Western countries, to supplement DHEA to feel younger. It is very simple, and that is why they call it the anti-aging hormone.

It is known that DHEA has immunomodulatory, antidiabetic and neuroprotective properties. Studies have also shown that low DHEA and an increased cortisol-to-DHEA ratio are linked to cognitive and immunological impairments.

This can be read very simply in the two publications at the bottom. As a consequence, it is useful to tell the patient that a low cortisol-to-DHEA ratio is an indicator of good brain and immunological health. That is the typical consequence drawn from it.

Then we have testosterone in saliva. Among other things, it can be used to monitor sports activity.

Here, you can see a typical hormonal profile of a person who goes jogging. You can see that when the person starts to jog, cortisol levels increase immediately. At the beginning of the jog, cortisol goes up. The person is full of energy and is actually experiencing positive stress.

Stress does not have to be negative. It can also be positive.

Then you see that, during the jog, testosterone levels also increase with exercise. If this is working correctly, then you have a balanced testosterone-to-cortisol ratio.

But if you have a depressed testosterone-to-cortisol ratio, which is very low, then you have an indication of overtraining syndrome. You have trained too much, which is also not so good.

This is described in both publications at the bottom.

Then we have melatonin in saliva. Melatonin is the sleep hormone, and it directly influences our sleep. During the day, melatonin levels are very low and start to rise with the dim-light melatonin onset.

That means that when the sun goes down, our melatonin goes up and indicates to our body that it is time to sleep.

Melatonin secretion is regulated by a pathway that starts in the retina of the eye. The eye is the starting point of our melatonin production. Changes in melatonin levels and alterations to its secretion patterns have been reported to coincide with sleep quality, insomnia, daytime sleepiness, jet lag and many more negative influences.

You can read about this in the publications below.

I have shown a typical pattern of what it looks like when you have a delayed sleep onset. In this graph, you see the daily profile of a person. Usually, melatonin levels go up when the light fades, and then the person goes to sleep.

But when you have a person with a delayed melatonin onset, you can see that it occurs two to three hours later, and perhaps the person’s quality of sleep is not as good either.

That leads us to leptin. Leptin is another biomarker in the field that is linked to aging because it is important for food intake.

Leptin is usually an appetite regulator via a very complex pathway. Leptin is directly linked to body fat mass, and leptin levels increase exponentially with body fat mass, measured using BMI.

I think it is very important to have control over leptin because it is also very interesting. With leptin, you can determine the optimal training stimulus in, for example, patients with obesity.

At a specific point, leptin goes to the optimal level, and you can say that the training is now perfect. This is described in the literature below.

Adiponectin is another of these adipokines. Leptin and adiponectin are both called adipokines.

This biomarker is predominantly released by white adipose tissue, which is, by the way, the good fat. Adiponectin is stored and released there. It usually positively modulates the endocrine system and increases insulin sensitivity in healthy individuals, as well as in glucose and lipid disorders.

Adiponectin is reduced in individuals with obesity because of low physical activity and a sedentary lifestyle.

With adiponectin, you can also optimize training intensity, especially in patients with obesity, to make sure that they train correctly and get the body into the right condition. This is described in the literature below.

Another really interesting biomarker is the titin N-fragment. Titin is, by the way, one of the biggest proteins in our body and is part of our muscular structure.

As it is part of our muscular structure, when titin N-fragment levels increase, we can link that directly to muscle damage.

Titin N-fragment levels also increase in patients with sarcopenia when compared with healthy controls because, as you might know, sarcopenia is muscle loss associated with aging.

Titin N-fragment levels also increase with eccentric muscle training compared with concentric muscle training. This means that we can even control how we train our body because the body likes one form of muscle training more than the other.

With correct exercise, we can also prolong muscle health because we make sure that our muscles stay healthy. This is also described in the literature below.

One really interesting biomarker in the context of aging is alpha-Klotho. The name already says it all because Klotho is one of the Greek Fates who spins the thread of life.

Since its discovery, this has led to the suggestion that the more alpha-Klotho we have, the longer a person potentially lives. Only potentially.

It is also important to know that the more alpha-Klotho we have, the more resilient the brain is against damage. But alpha-Klotho can also be an indicator of kidney problems.

We know that alpha-Klotho decreases with age. What is really good is that alpha-Klotho can be increased through moderate exercise.

Alpha-Klotho also plays an important role in vitamin D metabolism. It is a very broad protein that plays a role in many different conditions that are linked to aging.

Then we have a very interesting marker, which I would call the inflammaging marker. It is called suPAR, which stands for soluble urokinase plasminogen activator receptor.

It is shed from cells and can then enter the circulation. suPAR indicates chronic low-grade inflammation, and low-grade inflammation is really interesting.

It is symptomless. You cannot say anything about it, but it drives accelerated aging.

There has been a really interesting publication about life stressors. You can find it below. These stressors might include divorce, bankruptcy, the loss of relatives or even job loss. They are all life stressors.

You might not consider that bad, but it is really interesting that, under these conditions, you have increased suPAR levels.

Elevated suPAR levels are therefore directly linked to accelerated aging.

People with higher suPAR levels showed signs of looking older and exhibited older brain structure according to MRI. That was also another very interesting study.

Lifestyle interventions can positively or negatively influence suPAR levels because you can decide to change your lifestyle for the worse.

You can find an excerpt from these publications below, but there are many more.

Then we have sIL-2R. A lot of these sIL-2R publications that link the marker to aging are almost 30 years old, but it is very important.

First of all, we should know what sIL-2R is. It is an indicator of T-cell activation in the immune system. When the immune system becomes activated, something is ongoing. Of course, that is another factor that influences our aging.

We know that sIL-2R is slightly elevated because of smoking or poor dietary habits. This is described in the literature below, but I think it is very well known. All these publications are already from the 1990s.

sIL-2R should be part of the entire panel of biomarkers that you can offer to your patients because you are monitoring one small part of their health, immune system activation.

That leads us to one more and final biomarker I want to talk about: free 25-hydroxyvitamin D.

First of all, what we consider is that measuring the free fraction of 25-hydroxyvitamin D gives the true status of vitamin D supply.

Sometimes, we have people who have very low total vitamin D levels, but they feel very well and everything seems to be okay. Everything is okay because they still have sufficient vitamin D for the work that needs to be done.

It only means that they have little stored vitamin D.

Another factor that influences the availability of vitamin D is vitamin D-binding protein. The concentration of this protein may vary under different conditions and therefore influences total 25-hydroxyvitamin D levels.

This has been demonstrated by more recent studies indicating that lower total 25-hydroxyvitamin D levels do not necessarily show insufficient vitamin D supply.

You should measure free vitamin D, at least as an add-on, to make sure that everything is okay. There is a nice overview by the person below, who has published a really good summary of this factor.

That leads me to a summary. I have tried to summarize many of the biomarkers that you could measure and offer to your patients.

First of all, longevity isn’t a goal; it’s a lifestyle.

This has been driven by many things. We now have AI-generated predictions that address health deviations before they happen and negatively affect health. This drives change towards proactive preventive care, rather than modern medicine, which is usually reactive care.

It is a global phenomenon. There has been a worldwide reset around preventive care, functional medicine and wellness.

This is because of social media, which has taught the next generation that self-care is the new care.

Conventional healthcare professionals and private laboratories that do not service this growing demand for functional medicine will watch their patients purchase collection kits online and send samples out of the country.

Saliva samples are the best example of how this is not limited by geography because saliva, for example, is stable during temperature changes and transit.

Therefore, start helping people by confirming lifestyle choices with thoughtful laboratory measurements so that, in the end, when the next generation opens the photo books again, they ask their parents, “How old were you when you were in those pictures?”

They say, “I was 80.”

And they say, “Oh, Granddad was 70, and the generation before that looked so old.”

Then the parents can say, “Yes, but you need to take care of your life.”

Thank you very much.