Akkermansia species and body weight maintenance

Abstract

The invention is concerned with use of an Akkermansia species for maintaining body weight of a subject after a period of weight loss, wherein the Akkermansia species is administered during and after the period of weight loss. The invention is also concerned with an Akkermansia species for use in maintaining reduced risk of Diabetes mellitus type 2 and / or Diabetes mellitus type 2 complications of a subject after a period of weight loss, wherein the Akkermansia species is administered during and after the period of weight loss, and wherein the reduced risk is relative to not using said Akkermansia species. Preferably, the Akkermansia species has been subjected to pasteurization.

Description

[0001] Akkermansia species and body weight maintenance

[0002] TECHNICAL FIELD

[0003] The present invention relates to the field of interventions for maintaining body weight of a subject after a period of weight loss.

[0004] BACKGROUND OF THE DISCLOSURE

[0005] The worldwide prevalence of obesity, obesity-associated insulin resistance and Type 2 Diabetes Mellitus (T2DM) has grown dramatically in the last decades. In every region of the world, obesity prevalence has more than tripled since 1975. In accordance, about 9% of adults worldwide have the obesity-associated disease T2DM. The economic impact of obesity and T2DM is tremendous, accounting for at least 2 - 6% of total health care costs in developed countries. In addition, obesity is associated with a multitude of other diseases, see e.g. Pi-Sunyer (Postgrad Med. 2009 Nov; 121(6): 21-33).

[0006] Even though obesity treatment strategies such as lifestyle interventions (focused on diet and / or physical activity) and bariatric surgery have improved, there is a large variability in response and the expectation is that the prevalence of obesity and T2DM further rise dramatically in the upcoming decades.

[0007] In the short term, reducing body weight is relatively easy for most people. However, it is normally very challenging to maintain weight loss in the long term. The reality is that weight regain and incremental weight ‘creep’ are very common and even a modest weight loss obtained through lifestyle changes is difficult to maintain. Hence, novel strategies to reduce these pandemics and to support weight maintenance are strongly warranted.

[0008] In the past decades, the gut microbiome has emerged as an important regulator of host energy metabolism, thereby contributing to the etiology of obesity and obesity related insulin resistance. However, scientific evidence is mainly derived from animal experiments and association studies and support for causality in humans using mechanistic studies is still limited. In the last years the research field focuses on the identification of individual microbial species and their role in host metabolism. However, there is a need in the art for new and improved interventions for maintaining body weight of a subject after a period of weight loss. It is an object of the present disclosure, amongst other objects, to address this need.

[0009] SUMMARY OF THE DISCLOSURE

[0010] The present inventors surprisingly found that administration of an Akkermansia species dramatically improves maintenance of body weight of a subject after a period of weight loss (in comparison to not using the same).

[0011] Accordingly, administration of an Akkermansia species may be applied in a strategy for prevention and / or treatment of the so-called yoyo effect after a period of weight loss (refer for information on the yoyo effect e.g. van Baak et al (Nat Rev Endocrinol. 2023 Nov;19(11 ):655- 670); and Contreras et al (Front Genet. 2019 Dec 11 :10:1015). Use of an Akkermansia species dramatically reduces weight regain after weight loss. This is accompanied by an improved whole body insulin sensitivity and tendencies towards improvement in other glycemic parameters, and evidently a positive impact on all obesity-associated comorbidities.

[0012] Surprisingly, administering an Akkermansia species already during the weight loss period, i.e. during the weight loss period as well as during the weight management period, much better maintains weight (as well as lower risk of T2D / T2D complications), as compared to administering an Akkermansia species only during the weight maintenance period.

[0013] In addition, administering the Akkermansia species at a very high dose of more than 1x1010cells (instead of regular dose of ~1x108to ~1x109cells) even better maintained weight (as well as lower risk of T2D / T2D complications). In particular, the combination of receiving the Akkermansia species already during the weight loss period (i.e. during the weight loss period as well as during the weight management period) and dosing the Akkermansia species at above 1x101° cells leads to stable weight during the weight management period. Strikingly, this is particularly evident in case of participants who underwent bariatric surgery before / during the weight loss period.

[0014] The present inventors further found that combined administration of A. muciniphila and a GLP1 receptor agonist has a beneficial and synergistic effect, both during the weight loss period as well as during the weight maintenance period. DETAILED DESCRIPTION OF THE DISCLOSURE

[0015] The present disclosure relates to use of (at least one) Akkermansia species for maintaining body weight of a subject after a period of weight loss (of the same subject), wherein the (at least one) Akkermansia species is administered (to the subject) during and / or after the period of weight loss.

[0016] Akkermansia is a genus in the phylum Verrucomicrobia. It was previously found that Akkermansia species improve intestinal mucosal barrier function, or intestinal barrier function, which refers to the property of the intestinal mucosa that ensures adequate containment of undesirable luminal contents within the intestine while preserving the ability to absorb nutrients. Its role in protecting the mucosal tissues and circulatory system from exposure to pro-inflammatory molecules, such as microorganisms, toxins, and antigens is vital for the maintenance of health and well-being. Accordingly, Akkermansia species have been suggested for preventing or treating intestinal mucosal barrier dysfunction, which has been implicated in numerous health conditions such as: food allergy, microbial infection, irritable bowel syndrome, inflammatory bowel disease, celiac disease, metabolic syndrome, nonalcoholic fatty liver disease, diabetes, and septic shock.

[0017] Considering the different Akkermansia species, in particular Akkermansia muciniphila is promising. A. muciniphila is a gram-negative, strict anaerobe and mucin-degrading bacterium that colonizes the guts of humans and rodents. Although Akkermansia species are conserved in the gut microbiota throughout the animal kingdom, A. muciniphila thrives in the outer mucus layer in proximity to the host’s cells, and it has been shown to rely preferentially on host-derived mucin as an energy source. A. muciniphila is highly abundant in the gut microbiota and represents 1-5% of all intestinal bacteria. Such a successful coevolution between A. muciniphila and its hosts clearly indicates a relevance of this genus to host gut function and physiology.

[0018] The presence of A. muciniphila has been associated with healthy intestine and its abundance has been inversely correlated to several disease states including obesity and insulin resistance. The growth requirements of live A. muciniphila as well as its oxygen sensitivity rendered this bacterium not ideal for human investigations or putative therapeutic opportunities. Therefore, pasteurization, a mild heat inactivation method (30 min at 70 degrees Celsius), and its impact on diet induced metabolic disorders in mice were investigated. Unexpectedly, this method of inactivation did not abolish the effect of A. muciniphila but even exacerbated its beneficial impact. In a study assessing the effects of pasteurized A. muciniphila on diet-induced metabolic disorders in mice, not only was it found that pasteurization did not diminish the beneficial effects, but it unexpectedly enhanced the beneficial impact of A muciniphila (also in human context, see e.g. Depommier et al (Nat Med. 2019 Jul;25(7): 1096-1103). Accordingly, the at least one Akkermansia species according to the present disclosure is pasteurized. For example, the at least one Akkermansia species has been subjected to pasteurization (i.e. heating to 55-99, preferably 65-80 degrees Celsius for 5-60 seconds or 1- 60 minutes, preferably 60-80 degrees Celsius for 20-40 minutes, more preferably 65-75 degrees Celsius for 25-35 minutes).

[0019] The at least one Akkermansia species of the present disclosure preferably is or includes one or more of

[0020] - Akkermansia muciniphila or relative thereof having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity with the 16S rRNA gene sequence of the type strain of Akkermansia muciniphila (NCBI accession code AY271254, SEQ ID NO:1).

[0021] - Akkermansia glycanipila or relative thereof having a 16S rRNA gene with at least 90, 95, 97, 98, 99, 100% sequence identity with the 16S rRNA gene sequence of the type strain of Akkermansia glycanipila (NCBI accession code NR152695, SEQ ID NO:2). In the context of the present disclosure, the period of weight loss may be 1-1000, 7-730, or 14-180 days. The period after the weight loss period wherein the Akkermansia species is (continued to be) administered may be 1-000, 7-730, or 14-365 days. It is preferred that the body weight (e.g. obtained directly after the weight loss period) is maintained for at least 10, 50, 100, 200 days, after the period of weight loss.

[0022] The weight loss period is preferably due to adherence to a low-calorie diet. A low-calorie diet, in this regard, refers to a dietary intake that is reduced to less than the typical caloric needs of the respective subject. For a male human subject, this typically implies consuming fewer than about 2,500 kilocalories (kcal) per day, with a typical range being 1000-2400, 1000-2250, 1250-2000, 1500-1800 kcal per day. For a female human subject, this typically entails an intake of fewer than 2,000 kcal per day, typically ranging from 900-1900, 1000-1800, 1250- 1800, 1400-1700 kcal per day. The weight loss preferably may be between 1-250 g per kg of the subject’s weight before the weight loss period, more preferably between 10-250 g per kg or most preferably 100-200 g per kg or at least 1, 5, 10, 100 g / kg.

[0023] Several well-known dietary approaches can facilitate this reduced caloric intake, contributing to weight loss, independent of the exact composition. The (low-calorie) that may be used in the context of the present disclosure may be chosen for example from, as long as they are energy-restricted and result in weight loss:

[0024] 1. Mediterranean Diet: Emphasizes consumption of fruits, vegetables, whole grains, olive oil, and lean proteins such as fish and poultry, while limiting red meat and processed foods.

[0025] 2. WW (Weight Watchers): A point-based system that encourages healthy eating patterns and portion control, often promoting low-calorie food choices.

[0026] 3. MIND Diet: Combines elements of the Mediterranean and DASH diets, focusing on brain-healthy foods that are low in calories and high in nutrients.

[0027] 4. DASH Diet: Stands for Dietary Approaches to Stop Hypertension, it emphasizes fruits, vegetables, whole grains, and lean proteins while minimizing sugar and sodium intake.

[0028] 5. Intermittent Fasting: Alternates periods of eating with periods of fasting, typically resulting in an overall reduction in calorie intake.

[0029] 6. Plant-Based Diets: Focus on vegetables, fruits, legumes, nuts, and seeds, which are generally lower in calories and higher in nutrients.

[0030] 7. Low-Carb Diets: Restrict carbohydrate intake to promote fat loss, often leading to a reduced overall calorie intake. 8. Mayo Clinic Diet: Emphasizes eating low-calorie foods and following a balanced eating plan that includes fruits, vegetables, and whole grains.

[0031] 9. Volumetries Diet: Focuses on eating foods that are low in energy density but high in volume, such as fruits and vegetables, to promote satiety and reduce calorie intake.

[0032] 10. High-Fibre Diets: Increase the intake of dietary fiber from sources like fruits, vegetables, whole grains, and legumes, which can help control hunger and reduce calorie consumption.

[0033] 11. South Beach Diet: A phased low-carb diet that emphasizes lean proteins, healthy fats, and good carbs to achieve a caloric deficit.

[0034] Preferably, a low-calorie diet for use in the present disclosure is not excessively restrictive, such as consuming 800 kcal or less per day, as such extreme caloric restriction can lead to adverse health effects.

[0035] To support weight loss during the weight loss period as disclosed herein, subjects may use an energy-restricted diet (energy-restricted means that the subject’s energy intake is lower than energy expenditure) and / or optionally use various weight loss supporting products. These can include meal replacement shakes (like SlimFast or Cambridge weight plan), which provide balanced nutrition while controlling calorie intake. Other products may include appetite suppressants, dietary supplements, or snacks that are specifically designed to be low in calories but high in nutrients, aiding in adherence to the low-calorie diet and promoting weight loss.

[0036] In line with the above, the term “period of weight loss” in the present disclosure preferably refers to a (sustained and intentional) reduction in body weight, typically resulting in a net loss of at least 3%, 5%, 7%, or 10% of the subject’s body weight over a period of at least 7, 14, 21, or 30 consecutive days (e.g. 1-000, 7-730, 14-365, preferably 7-70 days days) .

[0037] Accordingly, this period can be characterized by a downward trend in body weight attributable to energy intake being lower than energy expenditure, commonly through the use of an energy-restricted diet as outlined above. Preferably, the weight loss period excludes shortterm, non-sustained fluctuations in weight due to diurnal variation, fluid shifts, or temporary changes such as dehydration, or illness. Body weight may be measured using standardized protocols (e.g., in the morning after voiding, under fasting conditions, and using calibrated digital scales) at multiple time points (e.g., weekly) to confirm weight reduction, distinguishing the period from natural day-to-day variability. The weight loss period can be considered completed once the subject has reached a weight plateau, defined for example as <1% change in body weight over at least two consecutive weekly measurements, and has transitioned to a period of weight maintenance. In the present disclosure, maintaining body weight is intended to mean that body weight is maintained within at most 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 20 % of the weight obtained (directly) after the weight loss period. In addition or alternatively, maintaining body weight is intended to mean that body weight does not increase (or decrease) more than 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 20% with respect to the weight obtained (directly) after the weight loss period. In other words, “use for maintaining body weight of a subject after a period of weight loss (of the same period)” is intended to mean that the use is for preventing body weight regain after the period of weight loss, preferably preventing body weight regain with more than 1 , 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, 60, 70, 80% of the weight that was lost during the period of weight loss. The use according to the disclosure may be non-therapeutic (non-medical), e.g. cosmetic use. The present disclosure may also be in the context of preventing or treating overweight (e.g. BMI between 25 and 29.9) or obesity (e.g. BMI 30 or higher). For example, the subject according to the present disclosure may be an overweight subject or obese subject, before and / or after the weight loss period, in particular before (at the start of) the weight loss period. Body Mass Index (BMI) is a subject's weight in kilograms divided by the square of the height in meters (kg / m2).

[0038] The subject preferably has a (baseline) level of Akkermansia species below 1.5X107cells / g of fecal matter (of the subject) or a relative abundance of Akkermansia species below approximately 0.3 % of the total bacterial cell count in the fecal sample. Relative abundance (Akkermansia species cell count) can be determined by quantitative PCR, metagenomic sequencing, or flow- cytometric cell counting, each relative to total bacterial abundance (i.e. relative abundance refers to Akkermansia species cell count relative to total bacterial cell count) in the same sample. For example, the subject has a (baseline) Akkermansia species level in fecal matter of below a reference level of Akkermansia species, e.g. preferably of between 1 x 106and 1 x 108, preferably between 5 x 106and 5 x 107, more preferably between 8 x 106and 5 x 107cells per gram feces. Particularly, the subject may have a (baseline) Akkermansia species level in feces (fecal matter) of below 1 x 1011, 7 x 101°, 5 x 101°, 4 x 1010, 4.2 x 1010, 4.5 x 101°, 3 x 101°, 1 x 101°, 7 x 109, 5 x 109, 3 x 109, 1 x 109, 7 x 108, 5 x 108, 3 x 108, 1 x 10s, 7 x 107, 5 x 107, 3 x 107, 1 x 107, 7 x 106, 5 x 106, 3 x 1Q6, 1 x 106, 7 x 105, 5 x 105, 3 x 105, or 1 x 105cells per gram of feces. Alternatively, the following relative abundance thresholds can be used, e.g. below a reference level of Akkermansia species of between 0.05% and 0.6% relative abundance, preferably between 0.1% and 0.4%, more preferably between 0.2% and 0.4%, more preferably between 0.25 and 0.35%, or approximately 0.3% 2.0 %, 1.5 %, 1.0 %, 0.9 %, 0.7 %, 0.5 %, 0.3 %, 0.2 %, 0.1 %, relative to total bacterial abundance (i.e. relative abundance refers to Akkermansia species cell count relative to total bacterial cell count) in the same (feces) sample. This can be determined prior to the use. Accordingly, the use according to the present disclosure preferably comprises: a) selecting a subject having a level of Akkermansia species below a reference level of Akkermansia species, e.g. preferably of between 1 x 106and 1 x 108, preferably between 5 x 106and 5 x 107, more preferably between 8 x 106and 5 x 107cells per gram feces. Particularly, the subject may have a (baseline) Akkermansia species level in feces (fecal matter) of below 1 x 1O11, 7 x 1O10, 5 x 1O10, 4 x 1O10, 4.2 x 1O10, 4.5 x 1O10, 3 x 1O10, 1 x 1O10, 7 x 109, 5 x 109, 3 x 109, 1 x 109, 7 x 108, 5 x 108, 3 x 108, 1 x 10s, 7 x 107, 5 x 107, 3 x 107, 1 x 107, 7 x 1 o6, 5 x 106, 3 x 1 o6, 1 x 106, 7 x 1 o5, 5 x 105, 3 x 1 o5, or 1 x 105of fecal matter, or alternatively, the following relative abundance thresholds can be used, e.g. selecting a subject having a level of Akkermansia species below a reference level of Akkermansia species of between 0.05% and 0.6% relative abundance, preferably between 0.1% and 0.4%, more preferably between 0.2% and 0.4%, more preferably between 0.25 and 0.35%, or approximately 0.3% 2.0 %, 1.5 %, 1.0 %, 0.9 %, 0.7 %, 0.5 %, 0.3 %, 0.2 %, 0.1 %, relative to total bacterial abundance (i.e. relative abundance refers to Akkermansia species cell count relative to total bacterial cell count in the same (feces) sample) For example, the selection may be among a group of subjects including subjects having a level of Akkermansia species above said threshold and including subjects having a level of Akkermansia species below said threshold); and b) administering the Akkermansia species to the selected subject for maintaining body weight after a period (e.g. 1-1000 day) of weight loss caused by following an energy-restricted diet (and excluding not selected subjects).

[0039] The Akkermansia species refers to any species of Akkermansia, and includes in particular Akkermansia muciniphila.

[0040] The present disclosure preferably excludes (cesarian section born) infants (i.e. a subject, preferably a human, under the age of 1).

[0041] The present disclosure envisions use for reducing blood pressure, particularly for reducing systolic blood pressure and / or diastolic blood pressure. In addition or alternatively, in the present disclosure the subject may have has a systolic blood pressure above 120 mmHg or diastolic blood pressure above 80 mmHg (prior to the use / intervention according to the present disclosure). This can be additionally selected for in step a) above. Blood pressures can be measured by Automatic oscillometric monitors (e.g., Omron). The present disclosure may also comprise monitoring fecal energy content as a marker of treatment response. Preferably the subject in the present disclosure is selected (e.g. in stap a) above) based on a fecal energy content below 4000, 3750, 3500, 3250, 3000 kJ / day (prior to the use / intervention according to the disclosure). The present disclosure also provides for (at least one) Akkermansia species for use in treating or preventing (or maintaining reduced risk of) Diabetes mellitus type 2 and / or Diabetes mellitus type 2 complications and / or insulin resistance of a subject after a period of weight loss, wherein preferably the Akkermansia species is administered (to the subject) during and / or after the period of weight loss (and wherein the reduced risk is relative to not using said Akkermansia species). It is preferred that the reduced risk of Diabetes mellitus type 2 and / or Diabetes mellitus type 2 complications (e.g. obtained directly after the weight loss period) is maintained for at least 10, 50, 100, 200 days, after the period of weight loss. As used herein, the term “Type 2 Diabetes complications” refers to medical conditions that arise as a consequence of prolonged hyperglycemia and insulin resistance associated with T2D, including but not limited to microvascular complications (e.g., diabetic nephropathy, neuropathy, retinopathy), macrovascular complications (e.g., cardiovascular disease, stroke, hypertension), and metabolic complications (e.g., dyslipidemia, non-alcoholic fatty liver disease).

[0042] The risk of Diabetes mellitus type 2 and / or Diabetes mellitus type 2 complications of a subject may be determined by measuring fasting glucose, HBA1c, disposition index, Matsuda index, and / or HOMA IR. Accordingly, also provided is for the (at least one) Akkermansia species for use in maintaining (e.g. within at most 1, 2, 3, 4, 5% margin in the respective units) of fasting glucose level, HBA1c level, disposition index, insulin secretion, Matsuda index, and / or HOMA IR of a subject after a period of weight loss, wherein the Akkermansia species is administered during and / or after the period of weight loss (and wherein the reduced risk is relative to not using said Akkermansia species).

[0043] Fasting glucose refers to the level of glucose (sugar) in the blood after a person has not eaten for at least 8 hours (8-12 hours). It is a common measure used to assess blood sugar control and diagnose diabetes or prediabetes. A blood sample is taken from the subject after an overnight fast. Fasting glucose is typically measured in milligrams per deciliter (mg / dL) or millimoles per liter (mmol / L), and a normal range, for most individuals, should be between 70 to 100 mg / dL (3.9 to 5.6 mmol / L). Levels between 100 and 125 mg / dL (5.6 to 6.9 mmol / L) indicate prediabetes, while levels of 126 mg / dL (7.0 mmol / L) or higher suggest diabetes.

[0044] HbA1c (Hemoglobin A1c) is a measure of the average blood glucose levels over the past two to three months. It reflects the percentage of hemoglobin (the protein in red blood cells that carries oxygen) that is coated with sugar (glycated). For measurement, a blood sample is analyzed in a laboratory to determine the percentage of glycated hemoglobin, i.e. it is reported as a percentage (%). The normal HbA1c level is below 5.7%. Levels between 5.7% and 6.4% indicate prediabetes, and levels of 6.5% or higher are diagnostic for diabetes. The disposition index is a measure of beta-cell function in the pancreas relative to insulin sensitivity. It reflects how well the beta cells can compensate for insulin resistance to maintain normal glucose levels. Method: Calculated using data from glucose tolerance tests or other dynamic tests of insulin secretion and insulin sensitivity. Formula: Disposition Index = Insulin Secretion x Insulin Sensitivity. It is a dimensionless number derived from the product of insulin secretion (measured as the acute insulin response to glucose) and insulin sensitivity (measured by various indices like HOMA-IR or the Matsuda index).

[0045] The Matsuda index is an estimate of whole-body insulin sensitivity derived from the oral glucose tolerance test (OGTT). It is calculated from glucose and insulin levels measured during an OGTT, where a subject consumes a glucose solution and blood samples are taken at multiple time points. Formula: Matsuda Index = 10,0001 > / (Fasting Glucose x Fasting Insulin) x (Mean Glucose x Mean Insulin during OGTT). It is a dimensionless number. Higher values indicate better insulin sensitivity.

[0046] HOMA-IR (Homeostasis Model Assessment of Insulin Resistance) is an estimate of insulin resistance. It quantifies how much insulin is needed to control blood glucose levels. It is calculated from fasting glucose and fasting insulin levels. Formula: HOMA-IR = (Fasting Insulin x Fasting Glucose) 122.5. It is a dimensionless number. Higher values indicate greater insulin resistance. Typically, a HOMA-IR value above 2.5 suggests insulin resistance.

[0047] Understanding the above measures is important in evaluating the metabolic health and risk of diabetes in a subject. Each measure provides specific insights into different aspects of glucose metabolism and insulin function.

[0048] In the present disclosure, maintaining reduced risk of Diabetes mellitus type 2 and / or Diabetes mellitus type 2 complications is intended to mean that said risk is maintained within at most 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10% of the reduced risk obtained (directly) after the weight loss period.

[0049] In line with the above, the present disclosure also provides for use of Akkermansia species according to the disclosure for maintaining (e.g. within 5% of the value (directly) after the weight loss period) or improving (enhancing) insulin sensitivity (e.g. within 5% of the value (directly) after the weight loss period), e.g. as measured by Matsuda Index, for example in context of preventing / treating insulin resistance. Similarly, the present disclosure also provides for use of Akkermansia species according to the disclosure for maintaining (a healthy) blood pressure (e.g., within 5% of the value obtained after the weight loss period, and / or within a clinically normal range) or improving blood pressure when deviating from healthy values, e.g., as measured by standard clinical metrics such as systolic and diastolic readings. For example the present disclosure may be in the context of preventing / treating hypertension, e.g. systolic blood pressure above 130, or 140 mmHg, and / or diastolic blood pressure above 80 or 90 mmHg.

[0050] In the present disclosure, the Akkermansia species may be administered, e.g. during and / or after the period of weight loss, in (daily) dosage(s) comprising at least 1 x 109, 2 x 109, 3 x 109, 4 x 109, 5 x 109, 6 x 109, 7 x 109, 8 x 109, 9 x 109, 1 x 101°, 2 x 101°, 3 x 101°, 4 x 101°, 5 x 1010, 6 x 1010, 7 x 1010, 8 x 1010, 9 x 1010, 10 x 1O10Akkermansia species cells per dosage. In addition or alternatively in dosage(s) comprising at most 4 x 1010, 5 x 1010, 6 x 1010, 7 x 1010, 8 x 1010, 9 x 1010, 10 x 1O10Akkermansia species cells. In addition or alternatively, the dosage(s) may be administered at least monthly or at least weekly, preferably daily or twice daily. The administration may involve at least 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 50, 100 separate administrations of the (at least one) Akkermansia species.

[0051] In addition or alternatively, the at least one Akkermansia species may be comprised in the composition in an amount ranging from 104to 1015cells. For instance, the at least one Akkermansia species may be comprised in an amount of 106cells to 1013cells, preferably 107cells to 1012cells, preferably 108cells to 1011cells, more preferably 109cells to 1011cells, e.g. per dose or per ml or per g of formulation or composition comprising said. Alternatively, the amount of the at least one Akkermansia species and / or administration frequency is chosen such that it is between, 106to 1013, preferably 107to 1012, preferably 108to 1011, more preferably 109to 1011, all in cells per day.

[0052] Flow Cytometry can be used to determine the amount of (pasteurized) Akkermansia species cells. The number of cells may be represented by Active Fluorescent Units (AFU) and / or Total Fluorescent (TFU), if cells are alive or pasteurised, respectively. Flow cytometry analysis is used in many industrial processes and can be done according to the ISO 19344 of the International Dairy Federation.

[0053] In a particularly preferred embodiment, the subject to whom the Akkermansia species is to be administered, is a subject who underwent a bariatric procedure / surgery. In this embodiment, the weight loss (period) may (in part) be due to the bariatric surgery. For example, the bariatric surgery is performed prior to (or during) the weight loss period.

[0054] Bariatric surgery encompasses surgical procedures aimed at helping individuals lose weight by making changes to their digestive system. In the present disclosure, the subject preferably underwent (bariatric) procedure / surgery to e.g. receive a gastric bypass, gastric sleeve and / or gastric balloon.

[0055] Gastric Bypass (Roux-en-Y Gastric Bypass)

[0056] This is one of the most common types of bariatric surgery. It involves creating a small pouch from the stomach and connecting it directly to the small intestine. The procedure typically involves the following steps:

[0057] Stomach Pouch Creation: The surgeon divides the stomach into a small upper section (pouch) and a larger lower section. The small pouch is about the size of a walnut and can hold only about an ounce of food.

[0058] Bypassing: The small intestine is then divided, and the lower part is connected to the small stomach pouch. This bypasses most of the stomach and the first part of the small intestine.

[0059] Reconnecting: The upper part of the small intestine is reconnected further down to allow digestive juices to mix with food.

[0060] Benefits: Significant weight loss, improvement or resolution of obesity-related conditions (e.g., type 2 diabetes, hypertension).

[0061] Risks: Nutrient deficiencies, dumping syndrome (rapid gastric emptying), and complications from surgery.

[0062] Gastric Sleeve (Sleeve Gastrectomy)

[0063] This procedure involves removing a large portion of the stomach, resulting in a tube-like structure or "sleeve." The procedure typically involves:

[0064] Stomach Reduction: About 75-80% of the stomach is removed, leaving a sleeveshaped stomach that can hold a smaller amount of food.

[0065] Functionality: The stomach still functions normally but is significantly smaller, reducing the amount of food intake and promoting satiety with less food.

[0066] Benefits: Significant weight loss, lower risk of nutrient deficiencies compared to gastric bypass, and improvement in obesity-related conditions.

[0067] Risks: Possibility of leakage from the staple line, nutrient deficiencies, and potential need for further surgery.

[0068] Gastric Balloon (Intragastric Balloon)

[0069] This is a non-surgical, temporary weight loss procedure. A deflated balloon is placed into the stomach and then filled with saline. The procedure typically involves the following steps:

[0070] Placement: The balloon is inserted through the mouth and esophagus into the stomach using an endoscope. Inflation: Once in place, the balloon is filled with saline, occupying space in the stomach.

[0071] Duration: The balloon is typically left in place for six months to help patients feel fuller with less food.

[0072] Benefits: Non-surgical, reversible, and can be an option for those who are not candidates for other types of bariatric surgery.

[0073] Risks: Balloon deflation, gastric discomfort, nausea, vomiting, and risk of balloon migration or obstruction.

[0074] In addition or alternatively, the (at least one) Akkermansia species according to the present disclosure may be combined, e.g. during and / or after the period of weight loss, with administration of a therapeutic (e.g. agonist or antagonist) that targets one or more chosen from glucagon-like peptide 1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP), glucagon (GOG), amylin, oxyntomodulin, dipeptidyl peptidase IV, and / or peptide YY receptor (see e.g. Ref. Annu Rev Med 2023;74:125- 39). The combination may be sequential or simultaneous, or first the (at least one) Akkermansia species may be administered and then the (at least one) therapeutic, or vice versa.

[0075] The mentioned peptides and hormones play crucial roles in regulating metabolism, appetite, and glucose homeostasis.

[0076] Glucose-dependent insulinotropic polypeptide (GIP) is an incretin hormone, which means it enhances insulin secretion in response to food intake. A therapeutic targeting GIP can improve insulin secretion and glucose control.

[0077] Glucagon (GOG) is a hormone that raises blood glucose levels by promoting glycogen breakdown in the liver. Targeting glucagon can be used to treat hypoglycemia (low blood sugar) or to control hyperglycemia by modulating its effects. In the context of weight loss, glucagon has been combined with other hormones to reduce appetite and increase energy expenditure.

[0078] Amylin is co-secreted with insulin by pancreatic beta cells and helps regulate blood glucose by slowing gastric emptying, promoting satiety, and inhibiting glucagon secretion.

[0079] Amylin analogs (like pramlintide) are used to improve glycemic control in diabetes by mimicking these effects.

[0080] Oxyntomodulin is a hormone that suppresses appetite and increases energy expenditure. It can be used for weight loss therapies due to its dual action on appetite suppression and metabolic rate enhancement.

[0081] Peptide YY (PYY) Receptor. PYY is a gut hormone that reduces appetite and inhibits gastric motility. PYY receptor agonists can help reduce food intake and manage obesity by promoting feelings of fullness.

[0082] In a particularly preferred embodiment, the (at least one) Akkermansia species according to the present disclosure may be combined, e.g. during and / or after the period of weight loss, with administration of a GLP1 receptor agonist and / or a Dipeptidyl Peptidase IV (DPP IV) inhibitor. The combination may be sequential or simultaneous, or first the (at least one) Akkermansia species may be administered and then the (at least one) GLP1 receptor agonist, or vice versa.

[0083] GLP-1 receptor agonists are a class of medications used primarily to treat type 2 diabetes and, more recently, for weight management (weight loss). GLP-1 stands for glucagon-like peptide-1 , a hormone that plays a crucial role in blood sugar regulation and appetite control.

[0084] GLP-1 receptor agonists mimic the action of the natural hormone GLP-1:

[0085] 1. Increase Insulin Secretion: GLP-1 receptor agonists typically enhance the secretion of insulin from the pancreas in response to meals, which helps lower blood sugar levels.

[0086] 2. Suppress Glucagon Secretion: They typically reduce the release of glucagon, a hormone that increases blood sugar levels, especially after meals.

[0087] 3. Slow Gastric Emptying: These medications typically slow down the emptying of the stomach, leading to a feeling of fullness and reduced appetite.

[0088] 4. Promote Weight Loss: by typically decreasing appetite and increasing satiety, they help in weight loss, which is beneficial for people with type 2 diabetes and obesity.

[0089] Examples of GLP-1 Receptor Agonists and DPP IV inhibitors include:

[0090] 1. Semaglutide

[0091] Uses: semaglutide is primarily used for the treatment of type 2 diabetes. It helps control blood sugar levels and promotes weight loss.

[0092] - Administration: It can be administered as a subcutaneous injection, typically once a week. Typical dosage: 0.1 - 5 mg.

[0093] Mechanism: semaglutide mimics GLP-1, enhancing insulin release, suppressing glucagon, and reducing appetite.

[0094] 2. Liraglutide Uses: While liraglutide is also used for type 2 diabetes, it is additionally approved for weight management in individuals with obesity or overweight with weight-related conditions.

[0095] - Administration: It can be administered as a (daily) subcutaneous injection. Typical dosage: 0.1 - 5 mg.

[0096] Mechanism: Similar to other GLP-1 receptor agonists, liraglutide increases insulin secretion, decreases glucagon release, and slows gastric emptying.

[0097] 3. Tirzepatide

[0098] Uses: Tirzepatide is used for the treatment of type 2 diabetes and is being investigated for weight management due to its significant impact on weight loss.

[0099] - Administration: It is administered as a subcutaneous injection, typically once a week.

[0100] Mechanism: Tirzepatide is a dual agonist that targets both GLP-1 and GIP receptors. This dual action enhances insulin release, suppresses glucagon, reduces appetite, and improves overall glucose control more effectively than targeting GLP-1 alone.

[0101] 4. Dipeptidyl Peptidase IV (DPP IV) inhibitor

[0102] Uses: DPP4 inhibitors are primarily used for the treatment of type 2 diabetes. They help improve blood sugar control by prolonging the action of incretin hormones.

[0103] - Administration: DPP4 inhibitors are typically administered orally, usually once daily.

[0104] Mechanism: DPP4 inhibitors work by inhibiting the enzyme dipeptidyl peptidase IV, which is responsible for breaking down incretin hormones such as GLP-1. By preventing the degradation of incretins, DPP4 inhibitors enhance the levels of these hormones, leading to increased insulin secretion, decreased glucagon release, and improved blood glucose regulation.

[0105] The present disclosure also provides for the use of Akkermansia species according to the present disclosure, wherein the use is (further) for maintaining visceral fat mass after the period of weight loss, preferably within at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10% of the visceral fat mass obtained after the weight loss period. Visceral fat typically refers to the amount of fat stored within the abdominal cavity, surrounding internal organs such as the liver, pancreas, and intestines. In addition, the present disclosure provides for use of Akkermansia species according to the present disclosure, wherein the use is (further) for maintaining waist circumference within at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10% of the waist circumference obtained after the weight loss period. Waist circumference can be measured at the midpoint between the lowest rib and the top of the iliac crest (hip bone).

[0106] The present disclosure also provides for the use of Akkermansia species (further) for maintaining fat percentage or fat mass within at most 5% of the values obtained after the weight loss period. Fat percentage or fat mass can be determined by e.g. Bioelectrical Impedance Analysis (BIA) or Skinfold Thickness (Calipers), e.g. triceps.

[0107] The (at least one) Akkermansia species according to the present disclosure is preferably comprised in

[0108] - a pharmaceutical composition, preferably in solid dosage form, such as a capsule, a tablet, or a powder; or

[0109] - a food composition, preferably a dairy or plant-based milk product, more preferably a fermented dairy product, most preferably a yogurt or a yogurt drink. The food composition may be a plant-based milk, such as almond based milk (almond comprising milk) or oat based milk (oat comprising milk).

[0110] The composition according to the present disclosure may be used as medicament and / or accompanied by a physiologically acceptable carrier which may be any inert carrier. For instance, non-limiting examples of suitable physiologically or pharmaceutically acceptable carriers include any well-known physiological or pharmaceutical carriers, buffers, diluents, and excipients. It will be appreciated that the choice for a suitable physiological carrier will depend upon the intended mode of administration of the composition as taught herein (e.g., oral). The skilled person knows how to select a physiologically acceptable carrier, which is suitable for or compatible with the compositions for use as taught herein.

[0111] The (at least one) Akkermansia species according to the present disclosure may be in lyophilized and / or microencapsulated form (to protect from gastric environment). It is also envisaged that the (at least one) Akkermansia species or the composition according to the present disclosure is comprised in and / or encapsulated by an (enteric) coating, preferable wherein said coating does not dissolute and / or disintegrate in the gastric environment of the recipient. Such coating may help the composition to reach the intended site for delivery without suffering breakdown due to the acidic environment of the stomach. Preferred (enteric) coatings work by presenting a surface that is stable at the highly acidic pH found in the stomach, but breaking down more rapidly at a lower pH. For example, it will not dissolve in the gastric acids of the stomach (pH ~3), but it will dissolve in the alkaline (pH 7-9) environment present in the small intestine, or duodenum.

[0112] The present inventors furthermore surprisingly found that micro-encapsulation of the at least one Akkermansia species according to the present disclosure, may provide a further synergistic therapeutic effect in the maintenance of body weight and / or reduced risk of Diabetes mellitus type 2 and / or Diabetes mellitus type 2 complications.

[0113] The term ‘micro-encapsulation’ is used to describe the encapsulation of bacteria in a matrix, coating, or membrane, generally a protective matrix or protective membrane. The (average) diameter of the microcapsules may be between 50 nm and 2 mm, preferably between 100 nm and 1 mm. The matrix, coating or membrane is typically comprised of milk, milk protein, and / or a polymer. The purpose of micro-encapsulation, among other possible purposes, may be to protect bacteria and their components against destruction by the surrounding environment, such as the gastrointestinal environment. The micro-encapsulation of bacteria may also support improved incorporation of bacteria into dairy products, food products, pharmaceutical formulations, and / or pharmaceutical compositions.

[0114] Various materials may be used for the micro-encapsulation of bacteria, such as pea protein, milk, milk protein, whey protein, casein, xanthan gum, alginate, gelatin, chitosan, carboxymethyl cellulose, starch, and / or carrageenan, and combinations thereof. In a preferred embodiment, the Akkermansia species according to the present disclosure is micro- encapsulated in one or more polymers.

[0115] The subject receiving the (at least one) Akkermansia species according to the present disclosure or composition as taught herein may be selected from the group consisting of animal (e.g. dog or cat), human being, non-human primate, mouse, rat, dog, cow, and pig. In a preferred embodiment, the subject is a human.

[0116] In an embodiment, the (at least one) Akkermansia species or composition as taught herein may comprise one or more ingredients which are suitable for promoting survival and / or viability of the bacterium or strain derived therefrom as taught herein during storage and / or during exposure to bile and / or during passage through the Gl tract of a mammal (e.g. a human being). Non-limiting examples of such ingredients include an enteric coating, and controlled release agents allowing passage through the stomach. The skilled person knows how to select suitable ingredients for maintaining a bacterium as taught herein viable and functional i.e. able to carry out intended function(s).

[0117] It may be advantageous to add one or more prebiotic ingredients to the combination as taught herein, for example, to supplement the effects (e.g. production of propionic acid / propionate and / or butyric acid / butyrate or a derivative thereof) of the bacterium as taught herein. The prebiotic ingredients may also enhance the activity and / or stimulate the growth of the bacterium, or a strain derived therefrom, as taught herein. A ‘prebiotic’ as used herein generally refers to a non-digestible food ingredient that promotes the growth of beneficial microorganisms in the intestines. Prebiotics or prebiotic products consist mainly of fermentable fibres or non-digestible carbohydrates. The fermentation of these fibres by probiotics promotes the production of beneficial end products, such as SCFAs, particularly butyrate. Non-limiting examples of suitable prebiotics include fibres such as inulin, pectin, and resistant starch, as well as cellobiose, maltose, mannose, salicine, trehalose, amygdalin, arabinose, melibiose, sorbitol, rhamnose and / or xylose. The skilled person is well-acquainted with the field of prebiotics and knows how to select ingredients endowed with prebiotic activity.

[0118] Without wishing to be bound by any theories, it is believed that the bacterial strain(s) according to the present disclosure, when administered to a human being or when ingested by a human being in an adequate amount, is / are able to survive and at least transiently colonize the gastrointestinal tract of said human being.

[0119] In an embodiment, the at least one Akkermansia species may be comprised in a food formulation, feed formulation, feed supplement formulation, food supplement formulation or pharmaceutical formulation. At the same time or alternatively, the at least one Akkermansia species and / or the at least one Lactobacillus species may be comprised in a liquid, liquid beverage (including dairy beverage and fermented beverage), yogurt, cheese, gel, gelatine, gelatine capsule, powder, paste, tablet, or a capsule.

[0120] The food or food supplement formulation is preferably a dairy product, more preferably a fermented dairy product, most preferably a yogurt or a yogurt drink.

[0121] The pharmaceutical formulation may be for example a liquid or solid form, more preferably a solid form solid dosage form, e.g., may be a capsule, a tablet, or a powder. Preferably, a pharmaceutical formulation does not relate to pure water or aqueous medium comprising more than 99 wt.% water. The formulations as taught herein comprising the combination for use according to the present disclosure may further comprise any acceptable carrier that is suitable for keeping the Akkermansia species as according to the present herein viable or stable until consumption by a subject (e.g. human or animal). For instance, non-limiting examples of acceptable carriers that are suitable for this purpose include any of well-known physiological or pharmaceutical carriers, buffers, and excipients. It will be appreciated that the choice for a suitable physiological or pharmaceutical carrier will depend upon the intended mode of administration of the formulations as taught herein (e.g. oral) and the intended form of the formulations (e.g. beverage, yogurt, powder, capsules, and the like). The skilled person knows how to select a physiological or pharmaceutical carrier, which is suitable for the formulations as taught herein.

[0122] The composition according to the present disclosure may be administered by enteral, preferably by oral, nasal and / or rectal administration. Preferably, the (at least one) Akkermansia species according to the present disclosure is not comprised in fecal matter.

[0123] In an embodiment, the present disclosure is concerned with the composition for use as a probiotic. Accordingly, ‘probiotics’ as used herein refers to microorganisms such as intestinal bacteria, which - when administered or ingested in effective amounts - confer health benefits to the host (e.g. humans or mammals). Preferably, probiotics should be alive or viable when administered to a subject so as to allow the probiotics to colonize the large intestine of the host. However, under certain conditions, probiotics may also be dead (e.g. pasteurized) when administered provided that substances produced by the probiotics still exert probiotic, beneficial effects on the host.

[0124] In an embodiment, the present disclosure is concerned with the composition for use as a postbiotic (refer Salminen et al (Nat Rev Gastroenterol Hepatol. 2021 Sep;18(9):649-667). A postbiotic refers to metabolic (by)products and / or compounds produced by probiotics. Unlike probiotics, which are live microorganisms, postbiotics typically are non-living (or pasteurized) and may include a wide range of (by)products and / or compounds.

[0125] The terms ‘comprising’ or ‘to comprise’ and their conjugations, as used herein, refer to a situation wherein said terms are used in their non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. It also encompasses the more limiting verb ‘to consist essentially of’ and ‘to consist of’. Reference to an element by the indefinite article ’a’ or ‘an’ does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article ‘a’ or ‘an’ thus usually means ‘at least one’.

[0126] As used herein, the term “identity" refers to a measure of the identity of nucleotide sequences or amino acid sequences. In general, the sequences are aligned so that the highest order match is obtained. "Identity" perse has an art-recognized meaning and can be calculated using published techniques. See, e.g.: (COMPUTATIONAL MOLECULAR BIOLOGY, Lesk, A. M., ed., Oxford University Press, New York, 1988; BIOCOMPUTING: INFORMATICS AND GENOME PROJECTS, Smith, D. W., ed., Academic Press, New York, 1993; COMPUTER ANALYSIS OF SEQUENCE DATA, PART I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; SEQUENCE ANALYSIS IN MOLECULAR BIOLOGY, von Heinje, G., Academic Press, 1987; and SEQUENCE ANALYSIS PRIMER; Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991). While there exist a number of methods to measure identity between two polynucleotide or polypeptide sequences, the term "identity" is well known to skilled artisans (Carillo, H., and Lipton, D., SIAM J. Applied Math (1988) 48:1073). Methods commonly employed to determine identity or similarity between two sequences include, but are not limited to, those disclosed in GUIDE TO HUGE COMPUTERS, Martin J. Bishop, ed., Academic Press, San Diego, 1994, and Carillo, H., and Lipton, D., SIAM J. Applied Math (1988) 48:1073. Methods to determine identity and similarity are codified in computer programs. For example NCBI Nucletide Blast with standard settings (blastn, https: / / blast.ncbi.nlm.nih.gov / ). Preferred computer program methods to determine identity and similarity between two sequences include, but are not limited to, GCS program package (Devereux, J., et al., Nucleic Acids Research (1984) 12(1):387), BLASTP, BLASTN, FASTA (Atschul, S. F. et al., J. Molec. Biol. (1990) 215:403).

[0127] As an illustration, by a nucleotide sequence having at least, for example, 95% "identity" to a reference nucleotide sequence, it is intended that the nucleotide sequence is identical to the reference sequence except that there may be up to five point mutations per each 100 nucleotides of the reference polypeptide sequence. In other words, to obtain a nucleotide sequence being at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted and / or substituted with another nucleotide, and / or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. In a sequence listing, a “n” may denote a, t, g, or c. Should there be an inconsistency between the sequences disclosed in the description and the sequences disclosed in the sequence listing, the sequences disclosed in the description are preferred. Alternatively, the sequences of the sequence listing may be used.

[0128] Figure descriptions

[0129] Figure 1 - use of A. muciniphila allowed for much better maintaining of body weight, as compared to use of placebo.

[0130] Figure 2 - A. muciniphila allowed for much better maintaining improved Matsuda index value (indicative of insulin sensitivity), as compared to use of placebo.

[0131] Figure 3 - use of A. muciniphila allowed for much better maintaining low fasting glucose level, as compared to use of placebo.

[0132] Figure 4 -use of A. muciniphila allowed for much better maintaining low visceral fat, as compared to use of placebo.

[0133] Figure 5 - use of A. muciniphila allowed for much better improvement of disposition index value, as compared to use of placebo.

[0134] Figure 6 - use of A. muciniphila allowed for much better maintaining low HOMA-IR value, as compared to use of placebo.

[0135] Figure 7 - use of A. muciniphila allowed for much better maintaining low HOMA-beta value, as compared to use of placebo.

[0136] Figure 8 - Effect of baseline Akkermansia abundance on body weight regain during intervention. In the MucT group, participants with low baseline A. muciniphila abundance showed much better maintenance of body weight compared to those with high abundance.

[0137] Figure 9 - Effect of baseline Akkermansia abundance on waist circumference change. Participants in the MucT group with low Akkermansia levels showed a much better maintenance in waist circumference reduction compared to those with high abundance.

[0138] Figure 10 - Effect of baseline Akkermansia abundance on fat mass regain.

[0139] In the MucT group, participants with low Akkermansia abundance showed a much showed much better maintenance of fat mass loss compared to those with high abundance.

[0140] Figure 11 - Effect of baseline Akkermansia abundance on body fat percentage regain. Participants in the MucT group with low Akkermansia abundance showed a much better maintenance of body fat percentage compared to those with high abundance.

[0141] Figure 12 - Effect of baseline Akkermansia abundance on visceral adipose tissue.

[0142] In the MucT group, participants with low Akkermansia abundance showed a much greater reduction in visceral adipose tissue compared to those with high abundance. Figure 13 - Effect of baseline Akkermansia abundance on Matsuda Index (insulin sensitivity). Within the MucT group, participants with low Akkermansia abundance showed a much better improvement in Matsuda Index compared to those with high abundance.

[0143] Figure 14 - Effect of baseline Akkermansia abundance on systolic blood pressure.

[0144] Within the MucT group, participants with low Akkermansia abundance showed a much better maintaining low systolic blood pressure compared to those with high abundance.

[0145] Figure 15 - Effect of baseline Akkermansia abundance on diastolic blood pressure. Within the MucT group, participants with low Akkermansia abundance in the MucT group experienced a much better maintaining low diastolic blood pressure compared to those with high abundance.

[0146] Figure 16 - Effect of baseline Akkermansia abundance on fecal energy content.

[0147] In the MucT group, participants with low Akkermansia abundance exhibited a much higher fecal energy content.

[0148] EXPERIMENTAL EXAMPLE 1

[0149] The present inventors studied the efficacy of pasteurized A. muciniphila for protection against weight regain in individuals with overweight / obesity, and aimed to address the following key objectives:

[0150] 1. To investigate the effects of pasteurized A. muciniphila on the maintenance of body weight after a phase of weight loss.

[0151] 2. To investigate the effects of pasteurized A. muciniphila on body composition and body fat distribution, glucose homeostasis and insulin sensitivity and metabolic health.

[0152] 3. To investigate the effects of pasteurized A. muciniphila on the faecal bacterial composition and functionality, systemic inflammation, gut barrier function and identify relevant biomarkers

[0153] Study design: Double blind, controlled, randomized, parallel design.

[0154] Study population: 108 males and females with overweight and / or obesity (age 20-70 y, BMI > 28 kg / m2 < 40 kg / m2)

[0155] Intervention: The participants will first undergo a period of weight loss with a low caloric diet (LCD, ~800kcal / day) for 8 weeks. Participants were randomized and either received pasteurized A. muciniphila or a placebo (only during LCD or during both LCD as WM period). The weight maintenance intervention diet is to be consumed ad libitum with respect to energy, i.e. the participants are not asked to count the energy content, and they are not provided with an individual target for energy intake. Main study parameters / endpoints:

[0156] Primary parameters: Before and post LCD and after the 6-months weight maintenance (WM) period changes in body weight will be assessed as primary outcome

[0157] Secondary parameters: Changes in body composition, glucose metabolism, insulin sensitivity and metabolic health as well as in the gut microbiome by evaluation the following parameters:

[0158] - BMI, waist and hip circumference and body composition (DEXA scan)

[0159] - fasting blood glucose and HbA1c

[0160] - postprandial glucose response by 7 pts OGTT

[0161] - postprandial insulin response by OGTT

[0162] - Lipid profile: total cholesterol, TG, LDL, HDL, NEFA.

[0163] - adipose tissue gene / protein expression

[0164] - faecal and circulating SCFA

[0165] - faecal microbiota composition

[0166] - circulating markers of renal and liver function and inflammatory markers

[0167] - Markers of gut barrier function: serum / faecal zonulin, faecal calprotectin, serum LBP, serum LPS

[0168] - Three-day food record, physical activity, well-being and sleep questionnaires

[0169] - Gastrointestinal Symptom Rating Scale questionnaire and Bristol Stool Charts

[0170] - vital signs.

[0171] Results

[0172] As shown in Figure 1 (including all participants and all dosages), use of A. muciniphila allowed for much better maintaining of body weight, as compared to use of placebo.

[0173] Figure 2 (including all participants and all dosages), shows that use of A. muciniphila allowed for much better maintaining improved Matsuda index value (indicative of insulin sensitivity), as compared to use of placebo.

[0174] Figure 3 (including all participants and all dosages), shows that use of A. muciniphila allowed for much better maintaining low fasting glucose level, as compared to use of placebo.

[0175] Figure 4 (including all participants and all dosages), shows that use of A. muciniphila allowed for much better maintaining low visceral fat, as compared to use of placebo.

[0176] Figure 5 (including all participants and all dosages), shows that use of A. muciniphila allowed for much better improvement of disposition index value, as compared to use of placebo. Figure 6 (including all participants and all dosages), shows that use of A muciniphila allowed for much better maintaining low Homa-IR value, as compared to use of placebo.

[0177] Figure 7 (including all participants and all dosages), shows that use of A. muciniphila allowed for much better maintaining low Homa-beta value, as compared to use of placebo.

[0178] In addition, it was clear that use of A. muciniphila allowed for much better maintaining low HbA1c value, as compared to use of placebo. Furthermore, glycated hemoglobin (HbA1c) levels appear to decrease over time when fasting glucose levels decrease (half life of erythrocytes around 3-4 months). With respect to effect on insulin secretion, an effect was observed relating to decrease in iAUC in particular for insulinaemia.

[0179] Surprisingly, participants who receive pasteurized A. muciniphila already during the weight loss period, i.e. during the weight loss period as well as during the weight management period, much better maintained weight (as well as lower risk of T2D / T2D complications), as compared with participants who receive pasteurized A. muciniphila only during the weight maintenance period. The difference is >1 average percent body weight change from baseline.

[0180] In addition, participants who receive pasteurized A. muciniphila at an extremely high dose of 3x1010cells (instead of regular dose of ~1x109cells) dramatically better maintained weight (as well as lower risk of T2D / T2D complications). The difference is >0.5 average percent body weight change from baseline. In particular, the combination of receiving pasteurized A. muciniphila already during the weight loss period (i.e. during the weight loss period as well as during the weight management period) and dosing A. muciniphila at above 1x1010cells leads to stable weight during the weight management period. Strikingly, this is particularly evident in case of participants who underwent bariatric surgery before the weight loss period.

[0181] Implications

[0182] Oral administration of A. muciniphila dramatically reduces weight regain after weight loss. This is accompanied by an improved whole body insulin sensitivity and tendencies towards improvement in other glycemic parameters.

[0183] EXPERIMENTAL EXAMPLE 2

[0184] The present inventors found that combined administration of A. muciniphila and a GLP1 receptor agonist has a beneficial and synergistic effect, both during the weight loss period as during the weight maintenance period. Intervention

[0185] After a weight loss period, 12 subjects are treated for 6 months according to the single or combinatorial treatment arms shown in Table 1.

[0186] Table 1 : treatment scheme and effect on weight maintenance and T2D biomarkers (as reported by subjects without knowing administered therapeutic / placebo)

[0187] It is expected that results similar to the putative effects as shown in Table 1 can be obtained with larger patient cohorts.

[0188] EXPERIMENTAL EXAMPLE 3

[0189] Low vs high Akkermansia spp.

[0190] The subjects in this trial were segmented after their weight loss period into two groups having either low or high level of fecal Akkermansia spp. as determined by qPCR analysis, as detailed below. The high Akkermansia group related to subjects having a high level of Akkermansia (more than 1.5x107Akkermansia cells per gram feces) and the low Akkermansia group related to subjects with a low level of Akkermansia (less than 1.5x107Akkermansia cells per gram feces).

[0191] Body weight regain

[0192] In the MucT group, participants with low baseline A. muciniphila abundance showed much better maintenance of body weight compared to those with high abundance.

[0193] See Figure 8. Waist circumference

[0194] Participants in the MucT group with low Akkermansia levels showed a much better maintenance in waist circumference reduction compared to those with high abundance. See Figure 9.

[0195] Body fat mass and percentage

[0196] In the MucT group, participants with low Akkermansia abundance showed a much showed much better maintenance of fat mass loss and fat percentage ompared to those with high abundance.

[0197] Fat mass, see Figure 10.

[0198] Fat percentage, see Figure 11.

[0199] Visceral adipose tissue

[0200] In the MucT group, participants with low Akkermansia abundance showed a much greater reduction in visceral adipose tissue compared to those with high abundance See Figure 12.

[0201] Matsuda Index

[0202] Within the MucT group, participants with low Akkermansia abundance showed a much better improvement in Matsuda Index compared to those with high abundance.

[0203] See Figure 13.

[0204] Blood pressure

[0205] Within the MucT group, participants with low Akkermansia abundance showed a much better maintaining low systolic and diastolic blood pressure compared to those with high abundance. Systolic, see Figure 14.

[0206] Diastolic, see Figure 15.

[0207] Fecal energy content

[0208] In the MucT group, participants with low Akkermansia abundance exhibited a much higher fecal energy content.

[0209] See Figure 16.

Claims

CLAIMS1. Cosmetic use of Akkermansia species for maintaining body weight of a subject after a 1- 1000 day period of weight loss, wherein the weight loss is due to following an energy- restricted diet, wherein the Akkermansia species is administered during and / or after the period of weight loss.

2. Akkermansia species for use in preventing Diabetes mellitus type 2, Diabetes mellitus type 2 complications and / or insulin resistance of a subject after a 1-1000 day period of weight loss, wherein the weight loss is due to following an energy-restricted diet, wherein the Akkermansia species is administered during and / or after the period of weight loss.

3. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the preceding claims, wherein the use comprises: a) selecting a subject having a level of Akkermansia species below a reference level of Akkermansia species; and b) administering the Akkermansia species to the selected subject for maintaining body weight after a 1-1000 day period of weight loss, wherein the weight loss is due to following an energy-restricted diet, wherein the reference level of Akkermansia species has a value between 1 x 106and 1 x 108cells per gram feces.

4. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the Akkermansia species is administered during and after the period of weight loss.

5. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the Akkermansia species is administered during and / or after the period of weight loss in dosages comprising at least 1 x 101° cells per dosage.

6. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the subject underwent bariatric procedure, preferably the subject underwent a bariatric procedure to receive a gastric bypass, gastric sleeve and / or gastric balloon.

7. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein during and / or after the period of weight loss, theAkkermansia species is administered in combination with administration of a GLP-1 receptor agonist preferably chosen from Semaglutide, Liraglutide, Tirzepatide and / or a Dipeptidyl Peptidase IV (DPP IV) Inhibitor.

8. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the Akkermansia species is administered during and / or after the period of weight loss at least weekly, preferably twice weekly, more preferably daily.

9. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the period of weight loss is 7-730 days.

10. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the Akkermansia species is administered, after the period of weight loss, for 7-730 days.

11. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein maintaining body weight maintains body weight within at most 5% of the weight obtained after the weight loss period.

12. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the claims 3-11, wherein in step a) the subject is additionally selected for having a systolic blood pressure above 120 mmHg or diastolic blood pressure above 80 mmHg.

13. Akkermansia species for use according to any one of the claims 2-12, wherein the use is additionally for- maintaining or enhancing insulin sensitivity; and / or- maintaining or reducing blood pressure, particularly for reducing systolic blood pressure and / or diastolic blood pressure.

14. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the use is further for maintaining waist circumference within at most 5% of the waist circumference obtained after the weight loss period.

15. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the use is further for maintaining fat percentage or fat mass within at most 5% of the values obtained after the weight loss period.

16. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the use is further for maintaining visceral fat mass after the period of weight loss, preferably within at most 5% of the visceral fat mass obtained after the weight loss period.

17. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein said at least one Akkermansia species has been subjected to pasteurization.

18. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein said Akkermansia species is Akkermansia muciniphila or relative thereof having a 16S rRNA sequence with at least 97% sequence identity with SEQ ID NO:1.

19. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, which is in micro-encapsulated or lyophilized form.

20. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, which is comprised in a composition preferably comprising a physiologically acceptable carrier.

21. Cosmetic use of Akkermansia species or Akkermansia species for use according to any one of the previous claims, wherein the Akkermansia species is comprised in- a pharmaceutical composition, preferably in solid dosage form, such as a capsule, a tablet, or a powder; or- a food composition, preferably a dairy or plant-based milk product, more preferably a fermented dairy product, most preferably a yogurt or a yogurt drink.

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