Human Milk Oligosaccharides (HMOs) and Gut Microbiota, Immune System in Antarctica

Healthy Adults Clinical Trial

Official title:

Effects of Human Milk Oligosaccharides (HMOs) on Gut Microbiota, Immune System and Metabolism in Adults Wintering Over in Antarctica

Clinical Trial Details — Status: Enrolling by invitation

Administrative data

• NCT number: NCT06133530
• Other study ID #: PreGlu
• Status: Enrolling by invitation
• Phase: N/A
• Start date: September 24, 2023
• Est. completion date: August 2027

Study information

• Verified date: November 2023
• Source: IU University of Applied Sciences
• Contact: n/a
• Is FDA regulated: No
• Study type: Interventional

Clinical Trial Summary

Human milk oligosaccharides (HMOs) are the third-most abundant component in mothers' milk and are an important prebiotic factor for the development of the gut microbiota of infants, promoting the growth of certain beneficial bacterial strains and providing protection against many bacterial and viral infections. HMOs induce immunomodulatory activity by affecting immune cell populations and functions. In a simulator of the adult human intestinal microbial ecosystem, fermentation of HMOs led to an increase of bifidobacteria in parallel with an increase in short-chain fatty acids as well as a reduction in inflammation markers, supporting the potential of HMOs to provide health benefits also in adults. Long-term stay in microgravity induces many physiological responses, including diminished immune function and impaired glucose tolerance which may lead to rather severe consequences. Similarly, hypoxia conditions as in the Concordia station, affects the immune system and may lead to impaired glucose tolerance and insulin resistance. The hypothesis is that HMOs as a prebiotic supplement will mitigate changes in immune function, glucose tolerance, lipid homeostasis, and neurotransmitter production. It is expected that HMO supplementation will

• Modulate gut microbiota composition and function

• Improve inflammation status

• Improve immune function

• Improve glucose tolerance

• Improve nutritional status

• Prevent changes in neurotransmitters associated with anxiety and depression. During the stay in Antarctica an HMO blend will be supplemented to the verum group of volunteers. The control group will receive a placebo. Experiment days with blood drawing, an oral glucose tolerance test, saliva sampling, and feces samples are planned once before, about every second month in Concordia, and once after return.

Description:

The aim of this study is to investigate the effect of supplementation with prebiotics, a specific mixture of carbohydrates found in breast milk (HMOs), as a measure to improve intestinal flora, inflammatory processes in the gut, general immune status, carbohydrate regulation, fat and bone metabolism and neurological changes in Antarctica. The approximately one-year stay at Concordia Station (Antarctica) at an altitude of about 3200 m includes a long journey and subsequent extreme environmental conditions as well as corresponding adaptation mechanisms and is therefore a very good terrestrial model (analogue model) for investigating changes during space stays. Long-term (i.e. > 6 months) exposure to microgravity leads to numerous physiological responses, including impaired glucose tolerance, reduced immune function and bone loss, which can lead to quite severe health consequences. In this analogue model, similar changes, albeit less severe, are observed with regard to the immune system or glucose tolerance as in space flight.

The composition of the diet significantly influences the composition of the intestinal flora. In particular, dietary carbohydrates provide the substrate for certain bacteria that could have a positive effect on health. Prebiotics are defined as substrates that are selectively used by host microorganisms and that provide health benefits. Prebiotics are digested only by gut microorganisms and stimulate the growth or activity of bacteria in the gut that have shown health benefits or improved well-being. The main nutrients for bacterial growth are non-digestible carbohydrates, which by definition are not digested by the host, in this case the human organism (host). Non-digestible carbohydrates include, for example, resistant starch and dextrins, non-starch polysaccharides (NSPs) (e.g. pectins) and non-digestible oligosaccharides (e.g. Raffinose, inulin, etc.). These carbohydrates are fermented by the intestinal bacteria to short-chain fatty acids (SCFAs), mainly acetic acid, propionic acid and butyric acid. This process takes place mainly in the large intestine, but also in the small intestine. SCFAs are rapidly absorbed by the mucosa of the large intestine and exert whole-body effects, contributing, for example, to the host's energy needs or mediating gut-brain communication, which has a major impact on the physiology and health of the host.

HMOs are also prebiotics and are the third most abundant solid component in breast milk. Most of the research published to date on the effects of HMOs has therefore been conducted on infants. HMOs are not digested in the intestine and are mainly utilised in the large intestine by the bacteria that reside there. In infants, it has been shown that HMOs not only promote a healthy composition of the intestinal flora, but also impair the growth of pathogens and modulate the expression of genes involved in inflammation. Several in vitro and in vivo studies have shown antiviral properties of HMOs against various viruses such as rotavirus, norovirus, HIV and influenza. HMOs play a significant role in the immune system by enhancing the immune response. Initial applications of HMOs in adults have supported these findings.

Therefore, this study aims to investigate how daily administration of 5.5 g of an HMO mixture during hibernation at Concordia Station in Antarctica affects:

• glucose tolerance

• inflammatory processes

• immune function

• fat and bone metabolism and

• Well-being

Study procedure This is a so-called placebo-controlled, randomised intervention study in parallel design. This means that subjects will be randomly selected to receive either the mixture of HMOs daily or a placebo. The study duration for a test person is approx. 20 months. It is divided into an examination phase before departure to Antarctica, approximately in September of a year, six phases during the wintering in Antarctica and one phase approximately 6 months after return.

Each individual phase is identical, one example shown in Figure 1.

Day 1 (Weekend day) Day 2 Day 3 Day 4 Day 5 Fasting blood collection X Oral glucose tolerance-test X Saliva sample X Feces sample X X X (X) Diet log X X X HMO supplementation Daily, onset after first session at Concordia

Figure 1: Representation of each phase (U as in figure 1) of testing

Intervention The intervention, i.e. the treatment measure used in this study, in one group is the daily administration of 5.5 g of a carbohydrate mixture analogous to human milk but produced by bacteria. This amount is certified by the European Control Authority as safe for human consumption. The other group receives a corresponding placebo, Maltose.

Measurement method Blood test and venous access Blood sampling from the peripheral vein is associated with the usual risks of venipuncture. These are haematomas or infections at the puncture site. When a peripheral venous access is placed, in rare cases there may be injuries to the vein (sclerotherapy) and the peripheric nerves. Blood collection and venous access are performed by an experienced doctor, which minimises the risk.

Fasting blood collections Fasting blood samples for analysis of various blood parameters are taken on the same day as the oral glucose tolerance test (OGTT) (see figure 1). Since several blood samples are taken for the OGTT, also after the drink has been taken, a venous access in the form of an indwelling venous cannula (small plastic tube) will be placed in the area of the forearm in order to avoid repeated puncturing of the arm veins.

Glucose tolerance by means of an oral glucose tolerance test The so-called oral glucose tolerance test (OGTT) can be used to determine differences in glucose metabolism (sugar metabolism) between the intervention and control phases. On approximately day 3 of each study phase, a drink will be consumed in the morning, fasting state, which is prepared from 75 g glucose (Medicalfox Gluko 75 powder) and 300 ml drinking water. Blood is taken every 15 minutes in the first hour after the drink is given, and every 30 minutes in the second hour.

Saliva sample Saliva samples will be taken to test certain hormones that reflect stress levels, as well as the reactivity of certain viruses (herpes virus, rotavirus). For this purpose, a small cotton wool roll from a prepared container (salivette) is chewed in the mouth for several minutes and returned to the collection tube once during the examination phase. The saliva obtained is frozen and later analysed in the laboratory.

Faecal collection In order to examine the changes in the intestinal flora, faeces samples are taken on 3 consecutive days in each phase. For this purpose, a so-called 'Feces Catcher' is placed over the toilet to better collect the faeces. Three samples are then taken from these faeces in 3 different tubes. The small spoon in each tube is filled with faeces, put back into the tube, closed and frozen.

Documentation of the diet In order to be able to estimate the influence of the food composition on the intestinal flora, the food consumed is documented on 2 working days and one weekend day. This is done in a prepared dietlog.

Recruitment information / eligibility

• Status: Enrolling by invitation
• Enrollment: 26
• Est. completion date: August 2027
• Est. primary completion date: May 2027
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: All
• Age group: 18 Years to 65 Years

Eligibility

Inclusion Criteria:

• Physically and mentally healthy subjects

• Volunteers that are able and declare their willingness to participate in the entire study

• Fasting blood glucose concentration: <120 mg/dL

• Willing to be assigned randomly either to the treatment or the control group

• Successfully pass the medical screening

• Signed informed consent

• Social insurance

Exclusion Criteria:

• Medication that may interfere with the interpretation of the results

• Recent sub-standard nutritional status

• Abuse of drugs, medicine or alcohol

• Participation in another study up to two months before study onset

• Cannot clear a criminal background check

• No signed consent form before the onset of the experiment

• Blood donors in the past three months before the onset of the experiment

• Vegetarian and Vegans

Related Conditions & MeSH terms

• Healthy Adults
• High Altitude
• Isolation, Social

Intervention

Dietary Supplement:
• Human milk oligosaccharides
Prebiotic
• Maltose
Control

Locations

Country	Name	City	State
Germany	IU International University of Applied Sciences	Erfurt

Sponsors (10)

Lead Sponsor	Collaborator
IU University of Applied Sciences	Chr Hansen, DLR German Aerospace Center, European Space Agency, German Institute of Food Technologies, MVZ Institute for Microecology, National Aeronautics and Space Administration (NASA), University of Bologna, University of Bonn, University of Lorraine

Country where clinical trial is conducted

Germany, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Other	Viral activation	Latent herpes virus shedding changes over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Other	Vitamin D status	Serum concentration changes of 25-OH and 1,25 OH vitamin D over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Other	GABA and BDNF	Changes in serum concentrations of GABA and BDNF (pg/ml) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Other	Serotonin in blood	Changes in serum concentrations of serotonin (ng/ml) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Other	Kynurenine in blood	Changes in serum concentrations of Kynurenine (µmol/l) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Other	Changes in further tryptophan metabolites	Changes in serum concentrations of Kynurenic acid, Quinolinic acid, Picolinic acid, Methylnicotinamide, Quinaldic acid, 3-hydroxykynurenine, Anthranilic acid, Nicotinamide (nmol/l) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Other	Bone formation marker	Changes in serum concentrations of PINP in µg/L	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Other	Bone resorption marker	Changes in serum concentrations of ß-CTX in ng/L	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Other	Body weight	Changes in body weight (kg) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Other	Body height	Measurement of body height to derive changes in body mass index	pre-Antarctica
Other	Profile of mood states	The profile of mood states is a psychological rating scale to measure changes in six different dimensions of mood (tension, depression, anger, vigour, fatigue, confusion) measured by a questionnaire. It is a five point scale ranging from "not at all" to "extremely".	5 consecutive days at baseline, pre-Antarctica; 5 consecutive days every month during the 12 month stay at Antarctica; 5 consecutive days 6-7 month after return from Antarctica
Other	Positive and negative affect schedule	The Positive and Negative Affect Schedule (PANAS) is a self-report questionnaire that consists of two 10-item scales to measure both positive and negative affect. Each item is rated on a 5-point scale of 1 (not at all) to 5 (very much).	5 consecutive days at baseline, pre-Antarctica; 5 consecutive days every month during the 12 month stay at Antarctica; 5 consecutive days 6-7 month after return from Antarctica
Other	Appraisal of life events scale	The Appraisal of life events scale (ALE) measures three factors of appraisal: threat, challenge and loss. It is an adjective check list of 16 adjectives rated on a 6-point scale (1 = disagree very strongly, 6= agree very strongly) that can be used to assess appraisals of retrospectively recalled stressful events as well as ongoing stressful encounters.	baseline, pre-Antarctica; about 6-7 month after return from Antarctica
Other	Subjective assessment of performance by a Visual Analogue Scale	The subjective Assessment of Performance Visual Analogue Scale measures factors such as stress, pain, and anxiety using a visual analog scale. The scale is a numerical rating scale ranking the respective factors from 1 to 10. A higher score indicates greater intensity of the respective factor.	baseline, pre-Antarctica; every month during the 12 month stay at Antarctica; about 6-7 month after return from Antarctica
Other	General health Questionnaire (GHQ)	The General health Questionnaire (GHQ) focuses on two major areas - the inability to carry out normal functions and the appearance of new and distressing phenomena. It is a 28 item questionnaire and assesses the respondent's current state and asks if that differs from his or her usual state. Each item is accompanied by four possible responses, typically being 'not at all', 'no more than usual', 'rather more than usual' and 'much more than usual', scoring from 0 to 3, respectively.	baseline, pre-Antarctica; every month during the 12 month stay at Antarctica; about 6-7 month after return from Antarctica
Other	Global Physical Activity Questionnaire	The Global Physical Activity Questionnaire (GPAQ) comprises 19 questions grouped to capture physical activity undertaken in different behavioral domains: work, transport, discretionary and time spent in sedentary activities. It asks for whether at all and if, how many days and how long the respective activity will usually be carried out.	baseline, pre-Antarctica
Other	Depression symptoms	Center for Epidemiologic Studies Depression Scale is a 20-item measure that asks the test subjects to rate how often over the past week they experienced symptoms associated with depression, such as restless sleep, poor appetite, and feeling lonely. Response options range from 0 to 3 for each item (0 = Rarely or None of the Time, 1 = Some or Little of the Time, 2 = Moderately or Much of the time, 3 = Most or Almost All the Time). Scores range from 0 to 60, with high scores indicating greater depressive symptoms.	Mid-term of the 12 month stay in Antarctica
Other	Nutrient intake	Food frequency questionnaire measuring food habits and dietary intake during 3 consecutive days including one weekend day.	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Primary	Glucose tolerance	Area under the serum glucose concentration curve (2 hours) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Primary	Insulin resistance	Area under the serum insulin concentration curve (2 hours) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Fecal calprotectin	Concentration changes in feces (µg/g) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Fecal zonulin	Concentration changes in feces (ng/ml) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Fecal short chain fatty acids	Concentration changes in feces (mmol/l) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Gut microbiota profiling	Feces feces will be analyzed for microbiota composition by 16S rRNA gene-based next-generation sequencing; Alpha and beta diversity, taxonomic assignment and relative abundance of bacterial groups will be characterized by bioinformatics.	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Saliva cortisol	Cortisol concentration changes in saliva over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Saliva DHEA	DHEA concentration changes in saliva over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Inflammation markers in blood	Changes in blood cytokines concentration (EGF, FGF-2, Eotaxin, TGF-a, G-CSF, Flt-3L, GM-CSF, Fractalkine, IFNa2, IFNy, GRO, IL-10, MCP-3), IL-12P40, MDC, IL-12P70, IL-13, IL-15, sCD40L, IL-17, IL-1RA, IL-1a, IL-9, IL-1ß, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IP-10, MCP-1, MIP-1a, MIP-1ß, TNFa, TNFß, VEGF, PDGF-AA, PDGFAB-BB, RANTES) pg/ml changes over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	CRP changes in blood	Changes (mg/L) in blood C-reactive protein concentrations over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Lipid metabolism	Serum concentration changes of triglycerides, total-, HDL-, LDL-cholesterol over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Glycated albumin	Serum concentration changes of blood glycated albumin over time in g/l and %	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	GLP-1	Serum concentration changes of GLP-1 (pmol/l) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica
Secondary	Fetuin-A	Serum concentration changes of Fetuin-A (µg/ml) over time	baseline, pre-Antarctica; every 1-2 month from month 4 to 10 during the stay in Antarctica (total stay about 12 month); about 6-7 month after return from Antarctica