Lactobacillus mucosa and diseases requiring increase in GLP-1

Abstract

The present invention relates to the use of a bacterial strain of the species Lactobacillus mucosa, or a lysate or culture supernatant thereof, for the prevention and / or treatment of a disease in need of an increase in GLP-1 production in a subject in need of GLP-1, selected from (i) a disease associated with insulin sensitivity and / or dysregulation of blood glucose concentration, and (ii) a loss of muscle mass and / or muscle function.

Description

Technical Field

[0001] The present invention relates to new probiotic and prebiotic agents that allow the stimulation of intestinal cells to synthesize GLP-1.

[0002] In particular, the present invention relates to a bacterial strain of the species Limosilactobacillus mucosae, or a lysate or culture supernatant thereof, for the prevention and / or treatment of diseases in a subject in need thereof that require an increase in GLP-1 levels, said diseases being selected from (i) diseases associated with abnormal regulation of insulin sensitivity and / or blood glucose, and / or (ii) loss of muscle mass and / or muscle function. The present invention also relates to the non-therapeutic use of a bacterial strain of the species Limosilactobacillus mucosae for maintaining or increasing muscle mass and / or function in a subject in need thereof, in particular a subject selected from malnourished subjects, elderly subjects (in particular malnourished elderly subjects), and subjects engaged in intense physical sports. Finally, the present invention relates to the bacterial strain Limosilactobacillus mucosae deposited at the CNCM under accession number CNCM I-5661. Background Art

[0003] It is expected that by 2050, the proportion of the elderly (65 years and older) will account for one-third of the French population. This population is very diverse and, at the extreme, is characterized by individuals with good physical health, being active, having a good nutritional status, low sarcopenia (age-related loss of muscle mass and function), and usually living at home. In contrast, there are elderly populations (of the same age) with malnutrition and high sarcopenia, who are usually dependent and living in institutions, and are in a state of chronic malnutrition (Buckinx et al., Burden of frailty in the elderly population: perspectives for a public health challenge. Archives of public health=Archives belges de santé publique 2015; 73(1):19). At the same time, in some Western countries, overweight or obese individuals account for more than half of the population. A part of this population, in addition to having an increased susceptibility to developing diabetes or cardiovascular diseases, also shows increased muscle wasting, which reduces the autonomy of these people, especially restricting the maneuvering space when treating obesity with a low-calorie diet (Barazzoni et al., Sarcopenic Obesity: Time to Meet the Challenge. Obesity Facts. 2018;11(4):294-305). In addition, the loss of muscle mass and function also affects individuals from time to time during their lives, depending on their background and experiences. This is especially the case for subjects with "severe burns", immobile patients, cancer patients, and even those who have undergone bowel resection or show intestinal malabsorption.

[0004] Recent data have shown a causal relationship between the activity of the gut microbiota and various diseases associated with abnormal regulation of nutrient metabolism and insulin resistance (Marchesi et al., The gut microbiota and host health: a new clinical frontier [review]. Gut. February 2016;65(2):330-9). Therefore, these data suggest an interest in developing probiotic strategies. In addition, comparative physiology studies focus on pathophysiological situations in which individuals show metabolic adaptations that favor maintaining insulin sensitivity, muscle mass, and / or better efficiency in the utilization of nutrients.

[0005] It has been observed that certain Lactobacilli are overrepresented in individuals who have undergone intestinal resection (Mayeur et al., Extensive Intestinal Resection Triggers Behavioral Adaptation, Intestinal Remodeling and Microbiota Transition in Short Bowel Syndrome [Review]. Microorganisms. March 8, 2016; 4(1)). The digestive tract and microbiota of these patients are characterized by the fact that, despite a reduced intestinal food absorption capacity, they have adapted metabolically to maintain their energy-nitrogen anabolism (Gillard et al., Enhanced Ghrelin Levels and Hypothalamic Orexigenic AgRP and NPY Neuropeptide Expression in Models of Jejuno-Colonic Short Bowel Syndrome. Scientific reports. June 21, 2016; 6:28345).

[0006] A study that transferred these bacteria to germ-free rats showed that increased concentrations of plasma peptides such as, for example, leptin, ghrelin, and GLP-1 indicate increased insulin sensitivity and increased energy recovery efficiency in these animals (Gillard L, Mayeur C, Robert V, et al. Microbiota Is Involved in Post-resection Adaptation in Humans with Short Bowel Syndrome. Frontiers in Physiology. 2017;8:224). In fact, GLP-1, a digestive peptide produced by enteroendocrine cells in the terminal ileum and colon after ingestion of a meal, has many metabolic effects, including regulation of pancreatic insulin secretion (but also regulation of food intake and transit) (Laurindo et al., GLP-1a: Going beyond Traditional Use. Int J Mol Sci. January 10, 2022;23(2)). GLP-1 analogs are currently also used to improve glucose tolerance in patients, and a role for GLP-1 in the regulation of Parkinson's disease and mood disorders has even been reported. GLP1 receptor agonists are often used to treat type II diabetes and obesity (Laurindo et al., GLP-1a: Going beyond Traditional Use. Int J Mol Sci. January 10, 2022;23(2)).They have also been proposed for many other diseases, such as appetite regulation (Aldawsari et al., “The Efficacy of GLP-1 Analogues on Appetite Parameters, Gastric Emptying, Food Preference and Taste Among Adults with Obesity: Systematic Review of Randomized Controlled Trials.” Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy 16 (2023): 575-595), liver diseases (Newsome et al., “A Placebo-Controlled Trial of Subcutaneous Semaglutide in Nonalcoholic Steatohepatitis.” New England Journal of Medicine 384, no. 12 (March 25, 2021): 1113-1124), or degenerative diseases (such as Alzheimer's disease (Femminella et al., “Evaluating the Effects of the Novel GLP-1 Analogue Liraglutide in Alzheimer's Disease: Study Protocol for a Randomised Controlled Trial (ELAD Study).” Trials 20, no. 1 (April 3, 2019): 191), or Parkinson's disease (Athauda et al., “Exenatide Once Weekly versus Placebo in Parkinson's Disease: A Randomised, Double-Blind, Placebo-Controlled Trial.” Lancet (London, UK) 390, no. 10103 (October 7, 2017): 1664-1675).

[0007] In particular, it has been shown that certain GLP-1 agonists have beneficial effects on body composition, particularly on maintaining muscle mass, including but not limited to obese or diabetic individuals (Osaka et al., "Favorable Appendicular Skeletal Muscle Mass Changes in Older Patients With Type 2 Diabetes Receiving GLP-1 Receptor Agonist and Basal Insulin Co-Therapy," Clinical Medicine Insights: Endocrinology and Diabetes 16 (January 1, 2023): 11795514231161884; or Hong et al., "Amelioration of Muscle Wasting by Glucagon-like Peptide-1 Receptor Agonist in Muscle Atrophy." J Cachexia Sarcopenia Muscle 10, no. 4 (August 2019): 903-918), or older individuals (Abdulla et al., "Glucagon-like Peptide 1 Infusions Overcome Anabolic Resistance to Feeding in Older Human Muscle," Aging Cell 19, no. 9 (2020): e13202).

[0008] If bacteria present in the distal part of the intestine (colon, terminal ileum) can stimulate the synthesis of endogenous GLP-1, all of the above-mentioned metabolic effects associated with GLP-1 can be expected.

[0009] The beneficial effects of certain bacteria of the genus Lactobacillus on humans are known in the art. For example, it is well known that Lactobacillus reuteri can control body weight and obesity, or improve insulin sensitivity and glucose homeostasis, through different mechanisms and metabolites (Abuqwider et al., Limosilactobacillus reuteri in Health and Disease. Microorganisms 2022, 10, 522). In addition, it has been demonstrated that, for example, the bacterial strain Limosilactobacillus fermentum MG4295 can improve hyperglycemia in mice fed a high-fat diet, and this bacterial strain exhibits characteristics favorable for its use as a probiotic (Kim et al., Limosilactobacillus fermentum MG4295 Improves Hyperglycemia in High-Fat Diet-Induced Mice. Foods. 2022, 11, 231).

[0010] In addition, certain strains of the species Limosilactobacillus mucosae, which are known in the art, can exhibit probiotic potential in, for example, the control of lipid metabolism, particularly hyperlipidemia (CN 111979145 A1), also in the treatment of memory disorders, learning disorders, mental disorders and inflammatory diseases (EP 3715449 A2), or even in the prevention of certain cardiovascular diseases (Ryan et al., BMC Microbiology (2019) 19:33).

[0011] However, to the inventors' knowledge, it has never been proposed in the past that bacterial strains of the species Limosilactobacillus mucosae can exhibit probiotic properties that allow the prevention and / or treatment of diseases that require an increase in GLP-1 production in a subject in need thereof.

[0012] Therefore, there is still a need to propose new probiotic compositions that are capable of stimulating the production of GLP-1, depending on the subject concerned, for (i) preventing or treating diseases associated with abnormal regulation of insulin sensitivity and / or blood glucose, or (ii) preventing or treating loss of muscle mass and / or loss of muscle function. Summary of the Invention

[0013] Surprisingly, it is shown in the examples that bacterial strains of the species Limosilactobacillus mucosae, or their lysates or culture supernatants, induce or increase the production of GLP-1 protein, particularly the production of GLP-1 protein by intestinal cells of a human subject.

[0014] According to a first objective, the present invention relates to a bacterial strain of Lactobacillus mucosae, or a lysate or culture supernatant thereof, for preventing and / or treating a disease in a subject in need of an increase in GLP-1 production, said disease being selected from (i) diseases associated with abnormal regulation of insulin sensitivity and / or blood glucose, and (ii) loss of muscle mass and / or muscle function.

[0015] According to a particular embodiment, the subject in need is a subject indicated for administration of a GLP-1 receptor agonist. In particular, the subjects indicated for administration of a GLP-1 receptor agonist are selected from the list consisting of: insulin-resistant subjects (such as diabetic subjects), overweight subjects (especially obese subjects), subjects who have undergone metabolic surgery, subjects seeking to regulate appetite, subjects suffering from liver disease, subjects suffering from cardiovascular disease (especially subjects suffering from cardiovascular disease associated with diabetes), subjects suffering from inflammation (especially inflammation associated with diabetes and / or metabolic syndrome), subjects suffering from sarcopenia, subjects suffering from cachexia, and subjects suffering from neurodegenerative diseases.

[0016] According to one embodiment, the disease is a disease associated with abnormal regulation of insulin sensitivity and / or blood glucose, selected from prediabetes, type 1 diabetes, and type 2 diabetes.

[0017] According to one embodiment, the disease is loss of muscle mass and / or function, and the subjects in need are selected from elderly subjects suffering from sarcopenia, overweight and obese subjects with sarcopenic obesity on a diet, diabetic or prediabetic subjects, and / or subjects suffering from cachexia (especially cachexia associated with cancer, inflammatory bowel disease or chronic obstructive pulmonary disease, limited mobility, "severe burn" status, sepsis or viral infection, recovery period and / or limited mobility, and / or bowel resection or intestinal malabsorption).

[0018] In particular, the bacterial strain is selected from the strain of Lactobacillus mucosae deposited at the CNCM under the accession number CNCM I-5661, the strain of Lactobacillus mucosae deposited at the DSM under the accession number DSM 13345, the strain of Lactobacillus mucosae deposited at the DSM under the accession number DSM 13346, the strain of Lactobacillus mucosae deposited at the DSM under the accession number DSM 102820, or a mixture thereof, especially the strain of Lactobacillus mucosae deposited at the CNCM under the accession number CNCM I-5661.

[0019] According to one embodiment, the bacteria of Lactobacillus mucosae are in a live or dead form, preferably in a live form.

[0020] In particular, the bacterial strain may be included in a composition containing a physiologically acceptable medium, in particular in an oral composition, and more particularly in an oral composition selected from the group consisting of: foods, beverages, pharmaceuticals, nutraceuticals, food additives, food supplements, dairy products, and live biotherapeutic products (LBPs).

[0021] In particular, the composition further comprises one or more other probiotic bacterial strains, especially selected from the species Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus salivarius, Lactobacillus johnsonii, Lactobacillus salivarius, Lactococcus lactis, Enterococcus faecium, Enterococcus faecalis, Saccharomyces cerevisiae, Saccharomyces boulardii, Faecalibacterium prausnitzii, Akkermansia muciniphila, Blautia faecis, Faecalibacterium prausnitzii, Streptococcus thermophilus, or mixtures thereof, preferably selected from the group consisting of: Bifidobacterium longum NCC3001 (ATCC BAA-999), Bifidobacterium longum NCC2705 (CNCM 1-2618), Bifidobacterium longum NCC490 (CNCM 1-2170), Bifidobacterium lactis NCC2818 (CNCM I-3446), Bifidobacterium breve strain A, Lactobacillus johnsonii NCC533 (CNCM 1-1225), Enterococcus faecium SF 68 (NCC2768; NCIMB10415), Lactobacillus casei (CNCM I-5662 and CNCM I-5663), Streptococcus thermophilus (CNCM I-5334), and combinations thereof.

[0022] According to certain embodiments, the composition does not contain any bacteria of other Lactobacillus species.

[0023] In particular, the composition further comprises one or more prebiotics.

[0024] According to a second object, the present invention relates to a non-therapeutic use of a bacterial strain of Lactobacillus mucosae, or a lysate or culture supernatant thereof (in particular as defined according to the present invention), for maintaining or increasing muscle mass and / or function in a subject in need thereof, in particular a subject selected from undernourished subjects, elderly subjects (in particular undernourished elderly subjects), and subjects engaged in intense physical sports.

[0025] Finally, the present invention relates to the bacterial strain Lactobacillus mucosae deposited at the CNCM under accession number CNCM I-5661. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 shows the concentration of GLP-1 in the supernatant of intestinal neuroendocrine cells of SCT-1 mice after 4 hours of incubation in the presence of various bacteria and test conditions. The x-axis represents the different test conditions, i.e., from left to right: strain 1 (Lactobacillus casei 1), strain 2 (Lactobacillus casei 2), strain 3 (Lactobacillus camelliae), strain 4 (Lactobacillus salivarius), strain 5 (Lactobacillus rhamnosus), strain 6 (Lactobacillus reuteri), strain CNCM I-5661 (Lactobacillus mucosae), and control (negative - in sterile cell culture medium). The y-axis represents the GLP-1 concentration in pg / mL.

[0027] Figure 2 shows the functional relationship between the concentration of GLP-1 in the supernatant of intestinal neuroendocrine cells of SCT-1 mice and the incubation time in the presence of strain CNCM I-5661 (Lactobacillus mucosae). The x-axis represents the incubation time, i.e., from left to right: 3 hours of incubation, 4 hours of incubation, and 5 hours of incubation. The y-axis represents the GLP-1 concentration in pg / mL.

[0028] Figure 3 shows the concentration of GLP-1 in the supernatant of intestinal neuroendocrine cells of SCT-1 mice after 3 hours of incubation in the presence of different bacteria of Lactobacillus mucosae at different bacterial concentrations. The x-axis represents the different test conditions, i.e., from left to right: control (negative control, sterile cell culture medium), strain Lacticaseibacillus casei (strain A - negative control, ~5·10 9 CFU / mL), strain CNCM I-5661 (strain B - positive control, ~5·10 9 ​​​CFU / mL), strain CNCM I-5661 diluted 1 / 10 (strain B dilution - ~5·10 8 CFU / mL), strain DSM 13345 (strain C - ~2·10 9 CFU / mL), strain DSM 13345 diluted 1 / 10 (strain C dilution - ~2·10 8 CFU / mL), strain DSM13346 (strain D - ~4·10 9 CFU / mL), strain DSM 13346 diluted 1 / 10 (strain D dilution - ~4·10 8 CFU / mL), strain DSM 102820 (strain E - ~5·10 9 CFU / mL), and strain DSM 102820 diluted 1 / 10 (strain E dilution - ~5·10 8 CFU / mL). The y-axis represents the GLP-1 concentration in pg / mL. Significant differences (P < 0.05). As in the previous comment, there was a clear difference between the complete absence of secretion (for the negative control condition) and the detectable GLP-1 secretion detected under the conditions of Lactobacillus mucosae bacteria, for all strain combinations, but no statistical data were calculated (insufficient data), and given this result, this did not seem necessary.

[0029] Figure 4 shows the muscle weights (in mg / g of rat weight) of the hind legs of rats in different groups of aged rats (20 months) after one month of study. The groups of rats studied are as follows from left to right on the x-axis: ad libitum-fed rats (AL), rats restricted to 75% to 80% of ad libitum intake (R), rats restricted and supplemented with Lactobacillus mucosae of strain CNCM-I5661 (R + Lactobacillus mucosae I5561), rats restricted and supplemented with Lactobacillus casei (R + Lactobacillus casei). Detailed Description

[0030] The inventors conducted in-depth research to determine the ability of a bacterial species (the species Lactobacillus mucosae) to treat and / or prevent diseases in a subject in need of increased production of GLP-1.

[0031] ​In fact, the inventors unexpectedly showed that Lactobacillus mucosae strains are able to stimulate the synthesis of GLP-1 by intestinal cells in vitro, while strains of other different Lactobacillus species are unable to induce such synthesis of GLP-1 by intestinal cells. In addition, Lactobacillus mucosae exhibits good resistance to the environmental conditions encountered in the intestine (acidic pH of the stomach and bile salts), as well as good adhesion ability to intestinal cells. Lactobacillus mucosae is present in large numbers in patients with short bowel syndrome (Joly et al., Biochemistry 2010; PMID: 20172013), and is able to colonize the original digestive tract (germ-free animals) after fecal transfer (Gillard et al.; Front Physiol. 2017); this indicates that this species is an intestinal symbiont. The ability of Lactobacillus mucosae to increase the synthesis of the intestinal peptide GLP-1 and to be present in the digestive tract is important because this peptide has many metabolic pleiotropic effects in the host; in particular, it improves insulin secretion in the pancreas, promotes glucose tolerance, and thus optimizes / improves the energy and nitrogen metabolism of the host. Finally, the inventors showed that administration of Lactobacillus mucosae strains can increase muscle mass in a sarcopenic rodent model.

[0032] Lactobacillus mucosae

[0033] The present invention relates to the use of bacterial strains of the species Lactobacillus mucosae, or their lysates or culture supernatants.

[0034] For the purposes of the present invention, the term "lysate" can be used interchangeably to refer to the whole lysate obtained by lysing the relevant microorganism or only a part thereof.

[0035] Dead bacterial cells can be obtained by any method known to those skilled in the art.

[0036] Bacterial cell lysates consist of all or part of the intracellular biological components as well as the components of the cell wall and membrane. It particularly contains the cytoplasmic part containing enzymes such as lactate dehydrogenase, phosphatase, phosphoketolase, and transaldolase. For example, the components of the cell wall are in particular peptidoglycan, murein or mucopeptide, and teichoic acid, and the components of the cell membrane consist of glycerophospholipids.

[0037] Bacterial cell lysates can be obtained by different techniques, for example by exposing the bacterial cells to osmotic shock, heat shock or ultrasound. More particularly, the lysate can be obtained according to the technique described in U.S. Patent No. 4,464,362.

[0038] "Culture supernatant" in the context of the present invention refers to the culture medium in which a bacterial strain is present during the culturing of the bacterial strain, and may also be referred to in this specification by the term "extracellular medium". In particular, the culture supernatant may include metabolites produced and secreted by the bacteria, also known as extracellular metabolites. Examples of metabolites are peptides, glycopeptides or lipopeptides produced by the bacteria. The culture supernatant may be raw or may be subjected to one or more steps of filtration, concentration, lyophilization, heating, etc. These techniques for transforming the culture supernatant of bacteria are well known to those skilled in the art.

[0039] The composition of the culture supernatant varies depending on the conditions of the bacterial strain and the cell culture method. It mainly includes extracellular metabolites of the biomass during growth or only when active, but may also include intracellular metabolites of the culture at the end of the stationary phase and when microbial lysis is significantly greater. A portion of the cells die and other cells lose their membrane integrity during lysis, so the supernatant will include cell walls from dead cells as well as intracellular metabolites.

[0040] In addition, the culture supernatant may also include bacterial cells of the species Lactobacillus mucosae in live or dead form.

[0041] Lactobacillus mucosae is a rod-shaped bacterial species of lactic acid bacteria that was first isolated from the porcine intestine. It exhibits mucin adhesion activity.

[0042] Lactobacillus mucosae is a Gram-positive, obligate anaerobic bacterium, however, it can reproduce to some extent in the presence of oxygen. This bacterial species was specifically described by Roos et al. (2000, International Journal of Systematic and Evolutionary Microbiology, Vol. 50, No. 1: 251-258). This bacterial species has been described in the human microbiota of patients with short bowel syndrome (Drastic changes in fecal and mucosa-associated microbiota in adult patients with short bowel syndrome. Joly F, Mayeur C, Bruneau A, Noordine ML, Meylheuc T, Langella P, Messing B, Duée PH, Cherbuy C, Thomas M. Biochemistry. July 2010; 92(7):753-61).

[0043] Regarding the names of certain probiotics, especially bacteria of the genus Lactobacillus, it is important to note the most recent changes in their taxonomic classification, as reported in the article by Zheng et al. (2020, Int. J. Syst. Evol. Microbiol, Vol. 70: 2782 - 2858). Thus, certain probiotics of the genus Lactobacillus with these taxonomic changes recorded in previously published literature are now named according to the new nomenclature in force.

[0044] Thus, the bacterial species now known as Lactobacillus mucosae may also be designated in the literature as Lactobacillus mucosae.

[0045] Bacterial strains of Lactobacillus mucosae suitable according to the present invention may be derived from the feces of a subject suffering from short bowel syndrome.

[0046] According to a particular embodiment, the bacterial strains according to the present invention are selected from the strains of the species Lactobacillus mucosae deposited at the CNCM under the accession number CNCM I - 5661, the strains of the species Lactobacillus mucosae deposited at the DSM under the accession number DSM 13345, the strains of the species Lactobacillus mucosae deposited at the DSM under the accession number DSM 13346, the strains of the species Lactobacillus mucosae deposited at the DSM under the accession number DSM 102820, or a mixture thereof, especially the strains of the species Lactobacillus mucosae deposited at the CNCM under the accession number CNCM I - 5661.

[0047] According to a particular embodiment, the bacterial strains of the species Lactobacillus mucosae are in a live or dead form, preferably in a live form.

[0048] In the context of the present invention, a "dead bacterial strain" means bacterial cells that are absolutely no longer able to reproduce and thus form colonies in culture. Different from lysates, dead bacterial strains may retain their entire membrane integrity.

[0049] Dead bacterial strains can be obtained by any known method of cell death. According to a particular embodiment of the present invention, the dead bacterial cells according to the present invention can in particular be obtained by exposure to high temperature (e.g., by exposure to a UHT protocol).

[0050] Thus, for the purposes of the present invention, especially in the examples herein, unless otherwise stated, the term "live" refers to live bacterial cells, including stable bacterial cells, i.e., viable and resuscitable bacterial cells (by all known stabilization methods, such as by freezing, freeze - drying or spray - drying).

[0051] Composition

[0052] This specification also relates to a composition comprising a bacterial strain of Lactobacillus mucosae species, or a lysate or culture supernatant thereof.

[0053] Thus, according to a particular embodiment, the bacterial strain according to the invention is comprised in a composition containing a physiologically acceptable medium.

[0054] The term "physiologically acceptable medium" is intended to designate a medium that is compatible with the organism of the individual to whom the composition must be administered. For example, this can be a non-toxic solvent such as water. The medium can also be food, particularly when the bacterial strain according to the invention is comprised in a nutritional composition, as defined below. The medium can also be mucus, such as snail mucus. In particular, mucus suitable as a medium according to the invention can be obtained as described by Gillard et al. (Enhanced Ghrelin Levels and Hypothalamic Orexigenic AgRP and NPY Neuropeptide Expression in Models of Jejuno-Colonic Short Bowel Syndrome, Sci Rep. June 21, 2016; 6:28345).

[0055] In particular, the medium is compatible with oral administration.

[0056] The composition described can be a nutritional composition.

[0057] For example, an athletic subject who needs to maintain or increase muscle mass without presenting any medical condition only requires nutritional supplementation. According to another example, the composition according to this specification can be a nutritional composition intended for elderly or overweight individuals suffering from malnutrition, such as because they are on a diet.

[0058] This specification also relates to a pharmaceutical composition comprising a bacterial strain of Lactobacillus mucosae species, or a lysate or culture supernatant thereof, which is intended for a subject having or likely to have a medical condition for which the composition has a preventive or therapeutic effect. For example, the composition according to this specification consists of a pharmaceutical composition intended to prevent or treat abnormalities in insulin sensitivity and / or blood glucose regulation, as in the case of diabetes or prediabetes.

[0059] However, unless otherwise specified, compositions containing bacterial strains of the species Lactobacillus mucosae do not differ in the general characteristics set forth in the specification depending on whether they are nutritional or therapeutic compositions. Thus, in essence, the difference between a pharmaceutical composition and a nutritional composition lies in that the pharmaceutical composition has a preventive and / or therapeutic effect on diseases in the subject to which it is administered. Each of these compositions also complies with its own regulations and is characterized by its mode of action. In fact, the pharmaceutical composition exerts a "pharmacological, immunological or metabolic action"; while the nutritional composition has a "nutritional or physiological" action.

[0060] In certain embodiments of the composition according to the specification, the bacterial strain of the species Lactobacillus mucosae is used as the only bacterial strain present in the composition.

[0061] In certain other embodiments of the composition according to the specification, the bacterial strain of the species Lactobacillus mucosae is combined with bacteria of one or more other probiotic strains, which include symbiotic probiotic strains.

[0062] Non-limiting examples of probiotics include bacterial strains belonging to the following genera: Bifidobacterium, Lactobacillus, Lactococcus, Enterococcus, Streptococcus, Kluyveromyces, Saccharomyces, Candida, Blautia, Faecalibacterium, Akkermansia, and combinations thereof.

[0063] The probiotics can be selected from the group consisting of the following bacterial species: Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei (now Lacticaseibacillus casei), Lactobacillus paracasei (now Lacticaseibacillus paracasei), Lactobacillus salivarius (now Ligilactobacillus salivarius), Lactobacillus lactis (now called Lactobacillus delbrueckii subsp. Lactis), Lactobacillus rhamnosus (now Lacticaseibacillus rhamnosus), Lactobacillus johnsonii, Lactobacillus plantarum (now called Lactiplantibacillus plantarum subsp. Plantarum), Lactococcus lactis, Enterococcus faecium, Enterococcus faecalis, Saccharomyces cerevisiae, Saccharomyces boulardii, Akkermansia muciniphila, Blautia obeum, Faecalibacterium prausnitzii, Streptococcus thermophilus, or a mixture thereof.

[0064] According to certain embodiments, the composition as described further comprises one or more other probiotic strains, particularly selected from the species Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus salivarius, Lactobacillus johnsonii, Lactobacillus salivarius, Lactococcus lactis, Enterococcus faecium, Enterococcus faecalis, Saccharomyces cerevisiae, Saccharomyces boulardii, Faecalibacterium prausnitzii, Akkermansia muciniphila, Blautia coccoides, Streptococcus thermophilus, or mixtures thereof, preferably selected from the group consisting of: Bifidobacterium longum NCC3001 (ATCC BAA-999), Bifidobacterium longum NCC2705 (CNCM 1-2618 cited in CA 2761573A1), Bifidobacterium longum NCC490 (CNCM 1-2170 cited in WO2006037922 A1), Bifidobacterium lactis NCC2818 (CNCM I-3446 cited in WO 2008116916 A1), Bifidobacterium breve strain A, Lactobacillus johnsonii NCC533 (CNCM 1-1225 cited in WO 2017060468), Enterococcus faecium SF 68 (NCC2768; NCIMB10415), Lactobacillus casei (CNCM I-5662 and CNCM I-5663 cited in Front Nutr. 2022; 9:928798, published online on August 10, 2022 by Giron et al.), Streptococcus thermophilus (CNCM I-5334 cited in EP 3826655 B1), and combinations thereof.

[0065] In certain embodiments, the composition does not contain any other bacteria of the species Lactobacillus.

[0066] In some embodiments, the composition further comprises one or more prebiotics. The term "prebiotic" is used in its conventional meaning in the prior art. Prebiotics consist of food substances that promote the growth of probiotics, including the bacteria contained in the microbiota.

[0067] Non-limiting examples of prebiotics include: oligosaccharides optionally containing fructose, galactose, mannose; dietary fiber, especially fermentable fiber, soy fiber; inulin; human milk oligosaccharides (HMO); polyphenols; chicory, mucilage, and combinations thereof. Preferred prebiotics are fructooligosaccharides (FOS), galactooligosaccharides (GOS), isomaltooligosaccharides (IMO), xylooligosaccharides (XOS), arabinoxylooligosaccharides (AXOS), mannanoligosaccharides (MOS), soy oligosaccharides, glycosyl sucrose (GS), lactosucrose (LS), lactulose (LA), palatinose oligosaccharides (PAO), maltodextrin, resistant starch, gums and / or their hydrolysates, pectins and / or their hydrolysates, or combinations thereof.

[0068] Prebiotics can also include peptides, proteins, and complex secretions of peptides, sugars, or sulfur, such as mucilage (e.g., intestinal mucilage). In pigs, the positive effects of mannanoligosaccharides and Lactobacillus mucosae have been tested (PMID: 34879142; J Anim Sci. December 1, 2021; 99(12)). It has also been shown that a high-fiber diet can induce the cultivation of Lactobacillus mucosae (Lactobacillus Mucosae Strain Promoted by a High-Fiber Diet. Microorganisms. August 12, 2020; 8(8):1225. PMID: 32806628).

[0069] In certain embodiments, the composition comprises a combination of a probiotic and a prebiotic, which is subsequently referred to as a "synbiotic". The term "synbiotic" is used in its conventional meaning in the prior art. The purpose of a synbiotic is to increase the survival of the probiotic and enhance its biological properties.

[0070] Non-limiting examples of synbiotics include the Bifidobacterium / fructooligosaccharide combination, the Lactobacillus / lactilol combination, or even the Bifidobacterium / galactooligosaccharide combination.

[0071] In some embodiments, the composition further comprises one or more vitamins. The vitamins can be folic acid, vitamin B12, and vitamin B6, especially folic acid and vitamin B12, especially folic acid. In some embodiments, the composition comprises one or more fat-soluble vitamins, such as one or more vitamins selected from vitamin A, vitamin D, vitamin E, and vitamin K, or comprises one or more water-soluble vitamins, such as vitamin C.

[0072] In some embodiments, the composition comprises one or more polyphenols, such as flavanols, flavanones, flavonols, hydroxycinnamic acids, and anthocyanins.

[0073] In some embodiments, the composition further comprises one or more minerals. The minerals can be selected from sodium, potassium, chloride, calcium, phosphate, magnesium, iron, zinc, copper, selenium, manganese, fluorine, iodine, chromium, or molybdenum. The minerals are typically added in the form of salts. The minerals can be added individually or in combination.

[0074] In some embodiments, the composition according to the present specification generally comprises a support or a vehicle. "Support" or "vehicle" refers to a material suitable for administration, including any material known in the prior art, such as any non-toxic liquid, gel, solvent, liquid diluent, solubilizer, or others that do not interact with the components of the composition in a harmful manner. Examples of nutritionally acceptable carriers include, for example, water, saline, alcohol, silicone, wax, petroleum jelly, vegetable oil, polyethylene glycol, propylene glycol, liposomes, sugars, gelatin, lactose, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, perfume oil, glycerol monoesters and diesters of fatty acids, fatty acid esters of petroleum, hydroxyethyl cellulose, polyvinylpyrrolidone, and the like.

[0075] In certain embodiments, the composition further comprises any other ingredients or excipients known for the type of composition under consideration. Non-limiting examples of such ingredients include: proteins, amino acids, carbohydrates, oligosaccharides, lipids, nucleotides, nucleosides, other vitamins, minerals, bacterial metabolites, bioactive molecules, and other micronutrients.

[0076] For example, the bacterial strain according to the present invention can be combined with a protein to limit muscle wasting in the subject to which it is administered.

[0077] According to another example, the bacterial strain according to the present invention can be combined with n-3 polyunsaturated fatty acids to increase insulin sensitivity in a subject.

[0078] In certain embodiments, the composition contains a source of carbohydrates, for example in the form of a prebiotic, or prebiotics (when they are present in the composition). Any source of carbohydrates commonly used in infant formula can be used, such as lactose, sucrose, maltodextrin, starch, and mixtures thereof, although the preferred source of carbohydrates is lactose.

[0079] In some embodiments, the composition according to the present specification consists of a nutritional composition.

[0080] In some embodiments, the nutritional composition is selected from complete food compositions, food supplements, nutraceutical compositions, etc. The compositions of the present specification can be used as food ingredients and / or animal feed ingredients. The food ingredient can be in the form of a solution or a solid, depending on the use and / or the method of application and / or the method of administration. The term "food" as used herein refers to liquid (i.e., beverage), solid or semi-solid dietary compositions, particularly complete food (food substitute) compositions that do not require additional nutrients or food supplement compositions. Food supplement compositions do not completely replace the nutrients provided by other means. As used in this specification, the term "food ingredient" includes formulations for functional foods or feeds or that can be added to functional foods or feeds as food supplements. "Nutritional food" or "nutraceutical" or "functional food" refers to a feed containing ingredients that have a beneficial effect on health or can improve physiological functions. "Food supplement" refers to a feed intended to supplement a normal diet. A food supplement is a concentrated source of nutrients or other substances that have a nutritional or physiological effect when administered alone or in small combinations. According to this specification, "functional food" is used to refer to such feeds and corresponding products: these feeds and corresponding products are valued not only for their nutritional and taste values, but also because they contain ingredients with beneficial physiological effects.

[0081] In some embodiments, the composition is a fermented dairy product or a milk-based product, which is preferably orally administered or ingested once or more times a day. Fermented dairy products include milk-based products such as (but not limited to) desserts, yogurt, yogurt drinks, cottage cheese, kefir, fermented milk drinks, buttermilk, cheese, salad dressings, low-fat spreads, cream cheese, soy drinks, ice cream, etc.

[0082] In certain embodiments, the composition is a fermented product based on a plant matrix.

[0083] As a variant, in some embodiments, the nutritional composition and / or the nutritional supplement composition can be non-dairy or non-fermented dairy products. Unfermented dairy products can include ice cream, nutritional bars, and seasonings, etc. Non-dairy products can include powdered beverages and nutritional bars, etc. These products can be manufactured using known methods, such as adding an effective amount of a bacterial strain of Lactobacillus mucosae or a bacterial combination comprising a bacterial strain of Lactobacillus mucosae to a food matrix (such as skim milk, or milk, or milk-based composition) and fermenting using any known technique. In some embodiments, the composition is a beverage, which can be a functional beverage or a therapeutic beverage, a thirst-quenching beverage, or a conventional beverage. For example, the composition according to the present specification can be used as an ingredient in carbonated beverages, fruit juices or beverages containing whey protein, tea, cocoa beverages, milk beverages, yogurt (including drinking yogurt), cheese, ice cream, ice pops and desserts, candies, cookies, cakes and cake mixes, snacks, health foods and beverages, frostings, acidified soy beverages / juices, aseptic / reduced chocolate beverages, stick seasonings, powdered beverage mixes, calcium-rich soy milk and chocolate, calcium-rich coffee beverages.

[0084] In some embodiments, the composition includes any other ingredients or excipients known for the type of composition under consideration. Non-limiting examples of such ingredients include: proteins, amino acids, carbohydrates, oligosaccharides, lipids, prebiotics or probiotics, nucleotides, nucleosides, other vitamins, minerals, and other micronutrients.

[0085] In certain other embodiments, the bacterial strain of Lactobacillus mucosae species is optionally combined with one or more other strains of probiotics and administered to a subject in the form of a pharmaceutical composition, which can correspond to a product of the live biotherapeutic product (LBP) type, reference: Front Med (Lausanne) Rouanet et al., Jun 19, 2020; 7:237. doi:10.3389 / fmed.2020.00237. For example, the bacteria of interest can be combined with a pharmaceutically acceptable excipient and an optional sustained-release matrix, such as a biodegradable polymer, to form a therapeutic composition. The term "pharmaceutically" or "pharmaceutically acceptable" refers to molecular entities and compositions that do not cause adverse reactions, allergic reactions, or other reactions when administered to a mammal, especially a human, if applicable. A pharmaceutically acceptable carrier or excipient refers to any type of non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, or formulation aid. In the pharmaceutical compositions of the present specification for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, topical, or rectal administration, the active ingredient or combination of active ingredients can be administered to animals and humans in unit dosage forms mixed with conventional pharmaceutical carriers. Suitable unit dosage forms include oral dosage forms for administration, such as tablets, gel capsules, powders, granules, and oral suspensions or solutions, sublingual and buccal dosage forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subcutaneous, transdermal, intrathecal, and intranasal dosage forms, and rectal dosage forms. Generally, the pharmaceutical composition contains a pharmaceutically acceptable vehicle for injection dosage forms. These can especially be isotonic sterile saline (monosodium phosphate or disodium phosphate, sodium chloride, potassium chloride, calcium chloride, or magnesium chloride, etc., or a mixture of these salts) or dry compositions, especially lyophilized compositions, which, depending on the circumstances, can form an injection solution by adding sterile water or physiological serum. Pharmaceutical forms suitable for injection include sterile aqueous solutions or dispersions; preparations containing sesame oil, peanut oil, or aqueous propylene glycol solutions; and sterile powders for the temporary preparation of sterile injection solutions or dispersions. In all cases, the form must be sterile and must be fluid so that it can be easily administered under control.

[0086] The pharmaceutical compositions must be stable under the manufacturing and storage conditions and must be protected against the contaminating action of microorganisms such as, for example, bacteria and fungi. Solutions containing the compounds of the present disclosure in the form of the free base or a pharmaceutically acceptable salt can be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and their mixtures, and oils. Under normal storage and use conditions, these preparations contain preservatives to prevent the growth of microorganisms. At least one bacterial strain of Lactobacillus mucosae according to the species of the present specification can be formulated in a composition in neutral form or in salt form. Pharmaceutically acceptable salts include acid addition salts (formed from the free amino groups of proteins), which are formed from inorganic acids such as hydrochloric acid or phosphoric acid, or from organic acids such as acetic acid, oxalic acid, tartaric acid, mandelic acid and similar acids. Salts formed from free carboxyl groups can also be derived from inorganic bases such as sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide or ferric hydroxide, and from organic bases such as isopropylamine, trimethylamine, histidine, procaine and the like.

[0087] According to a particular embodiment, the bacterial strain according to the invention is included in an oral composition, more particularly in an oral composition selected from the group consisting of: foods, beverages, pharmaceuticals, nutraceuticals, food additives, food supplements, dairy products, and live biotherapeutic products (LBPs).

[0088] In the context of the present specification, the term "therapeutically effective amount" is an equivalent term and refers to a therapeutic amount (e.g., a prophylactic or therapeutic agent) that is sufficient to reduce the severity and / or duration of a disease, alleviate one or more symptoms of the disease, prevent the progression of the disease, or cause regression of the disease, or is sufficient to cause prevention of the development, recurrence, onset, or progression of a disease or one or more of its symptoms, or is sufficient to enhance or improve the prophylactic and / or therapeutic effects of another therapy (e.g., another therapeutic agent) that can be used to treat the disease (e.g., a prophylactic or therapeutic agent). Generally, in the pharmaceutical compositions according to the present specification, the amount of the bacterial strain of Lactobacillus mucosae species present is sufficient to induce a reduction in insulin resistance in a subject being treated. Any technique known to those skilled in the art can be used to measure the insulin resistance of a subject. Preferably, the insulin resistance of a subject is measured by calculating the HOMA-IR index, as shown in the examples. The HOMA-IR method (for "Homeostatic Model Assessment of Insulin Resistance") was developed from the mathematical modeling of the quantitative responses of the major organs of glucose metabolism. The HOMA-IR index value is obtained using plasma insulin or C-peptide values and fasting blood glucose (Sheen, 2007, Therapy, Vol. 62: 311-318). The occurrence and / or level of insulin resistance can also be determined by measuring fasting blood glucose, by measuring fasting insulin, or by an OGTT test (for "Oral Glucose Tolerance Test" or "Induced Hyperglycemia Test" - see, for example, "Measurement of insulin resistance and glucose tolerance", 2006, French Ministry of Health, Haute Autorité de Santé (HAS)).

[0089] In the compositions according to the present specification, whether nutritional or food compositions (food supplements where appropriate) or pharmaceutical compositions, the bacteria can be present in various forms, such as in liquid form or in powder form. The bacteria can be in lyophilized form.

[0090] The number of bacteria of the Lactobacillus mucosae species administered to a subject can be variable and depends on the physiological state of the subject, in particular on the level of nutritional intake imbalance related to the nutritional requirements of the subject. The number of bacteria of the Lactobacillus mucosae species to be administered to the subject can be easily adjusted by those skilled in the art.

[0091] In a preferred embodiment, regardless of whether the composition is a nutritional composition or a pharmaceutical composition, the composition contains an amount of bacteria of the Lactobacillus mucosae species suitable for a daily intake of at least 10 3 colony forming units (or "CFU"), preferably a daily oral intake.

[0092] In these preferred embodiments, the daily intake, preferably the daily oral intake, of the bacteria of Lactobacillus mucosae species is at most 10 13 colony forming units (or "CFU").

[0093] In embodiments where the composition further comprises other probiotics, the amounts of these probiotics are determined by those skilled in the art based on their common general knowledge. The amounts of these other probiotics can vary from 10 3 other probiotics to 10 13 other probiotics.

[0094] In some embodiments, the composition according to the present specification generally comprises a carrier or vehicle. A "carrier" or "vehicle" refers to a material suitable for administration, including any material known in the prior art, such as any liquid, gel, solvent, liquid diluent, solubilizer or the like that is non-toxic and does not interact with the components of the composition in a harmful manner. Examples of nutritionally acceptable carriers include, for example, water, saline, alcohols, silicones, waxes, petrolatum, vegetable oils, polyethylene glycols, propylene glycols, liposomes, sugars, gelatin, lactose, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, perfume oils, glycerol monoesters and diesters of fatty acids, fatty acid esters of petroleum, hydroxyethylcellulose, polyvinylpyrrolidone and the like.

[0095] In other embodiments of the composition according to the present specification, the composition is in the form of a pharmaceutical composition comprising one or more pharmaceutically acceptable excipients.

[0096] Such a pharmaceutical composition may be presented in a packaging form comprising a plurality of dosage units.

[0097] The term "dosage unit" is used in its conventional meaning in pharmacy (e.g. the contents of a pill, capsule, tablet, ampoule, etc.).

[0098] Use

[0099] GLP-1 is a peptide that has many metabolic effects in a host; in particular, it improves insulin secretion by the pancreas and promotes glucose tolerance, thereby optimizing the energy and nitrogen metabolism of the host. The inventors have surprisingly shown that bacteria of the Lactobacillus mucosae type can increase the production of GLP-1 in vitro, thereby preventing or limiting the metabolic effects of diseases or conditions associated with defects in the action of GLP-1: insulin resistance (e.g. type II diabetes), metabolic diseases, difficulties in regulating food intake, regulation of transport disorders, etc.

[0100] As will be presented in the present specification, the bacterial strains of Lactobacillus mucosae species according to the invention or compositions comprising it are mainly used for the prevention and / or treatment of:

[0101] (i) Diseases associated with abnormal regulation of insulin sensitivity and / or blood glucose, or

[0102] (ii) Loss of muscle mass and / or muscle function.

[0103] According to certain embodiments, the loss of muscle mass and / or muscle function is associated with insulin resistance.

[0104] These diseases are associated with defects in the action of GLP-1.

[0105] The bacterial strain of Lactobacillus mucosae of the present invention or a composition comprising it can be used in particular for the prevention and / or treatment of diseases related to transport or diseases of appetite regulation.

[0106] According to certain embodiments, the bacterial strain is comprised in a composition containing a physiologically acceptable medium as defined above.

[0107] In the context of the present specification, the term "prevention" means reducing the risk or likelihood of a given phenomenon (i.e., in the present invention, a disease that requires an increase in GLP-1 levels) to a lesser extent.

[0108] In the present context, the term "treatment" means the alleviation or attenuation of a pathological process, or the alleviation or attenuation of one or more symptoms associated with the pathological process, in particular the alleviation or attenuation of one of the diseases described in the present application that requires an increase in GLP-1 levels.

[0109] The present specification relates to the nutritional and therapeutic uses of the bacterial strain or composition as defined in the present application.

[0110] The compositional features are described in detail in the present specification, including the compositional features of the nutritional and therapeutic compositions included in various embodiments, including the amounts of the active ingredients, in particular the amount of the bacterial strain of Lactobacillus mucosae comprised in these compositions.

[0111] (i) Diseases associated with abnormal regulation of insulin sensitivity and / or blood glucose

[0112] Dysregulation of insulin sensitivity, such as the occurrence of insulin resistance or insulin insensitivity, in particular, describes a situation where the liver, muscles, and fat cells, for example, become resistant to insulin. Then, the amount of glucose entering these cells decreases, and the glucose remains in the blood. In response to insulin resistance, the pancreatic insulin-secreting cells tend to produce more insulin (hyperinsulinemia) and will eventually become exhausted. Then, insufficient insulin is produced, and the blood glucose level becomes too high (hyperglycemia). Insulin resistance can also involve other types of metabolism regulated by insulin, such as protein metabolism. In fact, insulin combined with a meal is the main stimulator of muscle protein anabolism. Therefore, insulin resistance contributes to muscle wasting in this regard because of the lack of anabolism in the muscles, especially during meals.

[0113] Blood glucose regulation is the process of keeping the level of glucose in the blood (referred to as blood sugar) close to a value beneficial to the body. This regulation is part of the process of maintaining homeostasis. Statistically, the normal fasting blood glucose in humans is between 0.70 g / L and 1.10 g / L. When the blood glucose level is abnormally higher or lower than this threshold, this is called dysregulation.

[0114] According to this article, the main diseases associated with dysregulation of insulin sensitivity and / or blood glucose are prediabetes, type 1 diabetes, and type 2 diabetes. They may or may not be caused by obesity.

[0115] However, GLP-1 activates receptors expressed at the level of insulin cells (β) and certain peripheral tissues, stimulates insulin secretion (insulinotropic effect), also activates the transcription of the insulin gene, increases insulin biosynthesis, and inhibits the release of glucagon (tonic glucagon effect).

[0116] "Prediabetes" refers to a physiopathological state characterized, in particular, by a blood glucose level higher than the normal level but lower than the threshold defining type 2 diabetes. The fasting blood glucose level is considered to be (i) a normal level between 0.70 g / l and 1.10 g / l, (ii) indicating prediabetes between 1.10 g / l and 1.25 g / l, and (iii) indicating diabetes when >1.25 g / l. Prediabetes usually does not cause symptoms but is usually associated with obesity, dyslipidemia, and hypertension. Prediabetes is a risk factor for cardiovascular diseases. Prediabetes is characterized, in particular, by glucose intolerance. GLP1 stimulates the pancreas to secrete insulin and glucagon and controls blood glucose fluctuations by reducing fasting blood glucose. GLP1 induces glucose uptake and liver storage and inhibits gluconeogenesis. In addition, in addition to increasing insulin secretion, GLP1 also plays a role in improving insulin sensitivity in many tissues, including muscles (Muller et al., 2019, Mol Metab).

[0117] "Type 1 diabetes" or "T1D" refers to a chronic disease characterized by a complete absence of insulin production in an individual. Therefore, individuals with type 1 diabetes rely on insulin injections or insulin pumps daily to ensure survival. As elaborated above for prediabetes, GLP1 plays many roles in blood glucose control, including controlling the stimulation of insulin and glucagon secretion, glucose uptake and liver storage, inhibiting gluconeogenesis, and improving insulin sensitivity in many tissues.

[0118] "Type 2 diabetes" or "T2D" refers to a chronic disease that occurs when the pancreas does not produce enough insulin (a blood glucose regulating hormone) or when the body cannot effectively utilize the insulin it produces.

[0119] According to a particular embodiment, the disease associated with abnormal regulation of blood glucose is type 2 diabetes.

[0120] (ii) Loss of muscle mass and / or muscle function

[0121] According to a particular embodiment, the disease to be prevented and / or treated is loss of muscle mass and / or function, characterized in that the subject in need is selected from among elderly subjects with sarcopenia, overweight and obese subjects with sarcopenic obesity due to dieting, subjects with cachexia (especially cachexia associated with cancer, inflammatory bowel disease or chronic obstructive pulmonary disease), subjects with limited mobility, diabetic or prediabetic subjects, "severely burned" subjects, septic or virus-infected subjects, convalescent and / or immobile subjects, and subjects who have undergone bowel resection or have intestinal malabsorption.

[0122] Subjects with muscle mass loss in the above situations are usually resistant to the anabolic effects of diet and / or the effects of insulin. Although subjects experiencing severe muscle wasting usually have insulin resistance, not all insulin-resistant or prediabetic subjects are necessarily in a state of muscle wasting, which is a phenomenon that gradually develops over time, and the development of muscle wasting and insulin resistance is strongly correlated metabolically (Daily and Park, 2022 Cells - DOI: 10.3390 / cells11030338).

[0123] In fact, GLP-1 is known to have beneficial properties that can optimize the energy use of nutrients and limit muscle wasting (Hong et al., Journal of Cachexia, Sarcopenia and Muscle 2019; 10:903-918), particularly in individuals with insulin resistance (Massimo et al., The Impact of Glucose-Lowering Drugs on Sarcopenia in Type 2 Diabetes: Current Evidence and Underlying Mechanisms. Cells 2021, 10, 1958).

[0124] Loss of muscle mass can be measured by different techniques known to those skilled in the art, such as upper arm circumference, bioelectrical impedance, bioenergetic X-ray absorptiometry (DXA), and CT scan (Mareschal et al., 2019, Journal of Clinical Medicine, doi:10.3390 / jcm8071040).

[0125] Loss of muscle function (strength, physical fitness, physical performance) can be measured by different techniques known to those skilled in the art, such as grip strength test (grip strength test using a dynamometer), 6-minute walking speed, 30-second chair stand test, SPPB (Short Physical Performance Battery) test (which combines walking speed, balance, and sit-to-stand performance), and "Timed Up and Go" test (consisting of getting up from a chair, walking 3 meters, turning around, and sitting down again) (Beaudart et al., 2019, Calcified Tissue International, doi:0.1007 / s00223-019-00545-w).

[0126] In the meaning of this specification, an "elderly subject" refers to a human subject or a non-human mammalian subject, including pet animals such as dogs or cats, which show signs of aging, such as impaired metabolic function (such as absorption, digestion, excretion, tissue and organ dysfunction), difficulty in movement, and reduced resistance to external aggression. In the case of humans, an "elderly subject" or "elderly person" refers to a subject aged 65 years or older. In the case of canines, particularly dogs, an "elderly subject" refers to (i) a subject over 12 years of age for small dogs (especially small dogs), or (ii) a subject over 9 years of age for medium-sized dogs (especially medium-sized dogs), or (iii) a subject over 7 years of age for large dogs (especially large dogs). In the case of felines, particularly cats, an "elderly subject" refers to a subject over 13 years of age.

[0127] Sarcopenia (a word from Greek that can be translated as "lack of flesh"), initially defined as the loss of skeletal muscle mass, is currently characterized by the loss of muscle mass associated with functional decline (European Working Group on Sarcopenia in Older People (EWGSOP) - Cruz-Jentoft et al., 2019, Age and Aging). After growing until 20 to 30 years old, muscle mass decreases by about 1% per year, even in healthy individuals. This process accelerates between 50 and 60 years old. Sarcopenia affects physical function, promotes walking disorders, and constitutes a factor for frailty and increased risk of dependency, especially in the elderly. Note that sarcopenia affects not only the elderly but also insulin-resistant populations such as those with obesity, a condition known as sarcopenic obesity (European Society for Clinical Nutrition and Metabolism (ESPEN) and European Association for the Study of Obesity (EASO) - Donini et al., 2022, Obesity Facts). This muscle wasting can also occur over a shorter period, especially in strongly catabolic situations, such as cancer and related intensive treatments, such as chemotherapy. In this case, it refers to cachexia (see below). Sarcopenia is associated with an increased risk of activity limitation due to falls and fractures, which in turn exacerbates sarcopenia. Thus, a snowball effect is observed, amplifying the age-related loss of muscle mass and function.

[0128] "Obesity" refers to a pathophysiological state in which an individual particularly exhibits weight gain and excessive adipose tissue, usually caused by obesogenic diets, especially including overconsumption of high-calorie foods, genetic susceptibility, or insufficient or absent physical activity. The body mass index (BMI) of an individual called obese is greater than 30. According to the official definition of the World Health Organization (WHO), the body mass index is an indicator of the health risk associated with overweight or underweight. BMI is calculated by dividing an individual's weight (in kilograms) by the square of their height (in meters). According to the classification given by the WHO, BMI values are associated with specific body types.

[0129] Unlike an "obese" individual, an "overweight individual" refers to an individual whose condition is not pathophysiological. Overweight individuals usually also have excessive adipose tissue. An individual is generally considered overweight when their body mass index (BMI) is between 25 and 30.

[0130] An individual is called sarcopenic obese when they have excessive fat mass and reduced lean muscle mass. This clinical condition most commonly affects the elderly.

[0131] "Cachexia" is a severe debilitation of the body, characterized in particular by the depletion of adipose tissue and muscle. It is common in many diseases, including cancers that are difficult (if not impossible) to control or cure, and as mentioned above, occurs over a relatively short period of time (days to weeks). Cancers, especially pancreatic and gastric cancers, cause severe cachexia. Patients can lose 10% to 20% of their body weight. Cachexia is also associated with other catabolic diseases (such as inflammatory bowel disease or chronic obstructive pulmonary disease, viral infections or sepsis).

[0132] It has been shown that decreased mobility is associated with a decrease in muscle mass index and muscle strength. Vice versa, since in general, decreased mobility and physical activity worsen or cause sarcopenia. Therefore, an object with limited mobility may experience loss of muscle mass and / or function. In particular, in the context of this article, it will relate to an object with limited mobility and / or recovery after a fall.

[0133] An object in a case of "severe burn", such as in the case of a disease that induces hypermetabolism (sepsis, viral attack, even cancer), is characterized by a significant need for macronutrients and micronutrients to meet the increased needs associated with the pathology, especially due to the healing process (in "severe burn victims") and the inflammatory response (Knuth et al., 2021, American Journal of Physiology. Cell Physiology). In addition, due to high inflammation, resistance to the anabolic action of the diet and the above-mentioned insulin resistance are generally considered determinants of obesity and sarcopenia in elderly objects.

[0134] For an object with intestinal malabsorption or who has undergone intestinal resection, GLP-1 can help optimize the utilization of nutrients. In fact, in patients who have undergone intestinal resection, an increase in the production of GLP1 can be seen (Jeppesen et al., 2000, Gut; Gillard et al., 2017, Front Physiol), enabling adaptation to intestinal motility and optimization of the host's metabolism with the lowest supply of nitrogen and energy nutrients. The effect on the energy (adipose mass) and nitrogen (lean mass and muscle mass) stores of the object will be particularly evident. Therefore, increasing the intestinal secretion of GLP-1 in these objects, especially by administering GLP-1-secreting probiotics, is beneficial to them.

[0135] According to a specific embodiment, the object to be treated, or the object in need, is an object showing an indication for administering a GLP-1 receptor agonist. An individual with an indication for administering a GLP-1 receptor agonist is an individual suffering from a treatable disorder or disease, or whose symptoms can be alleviated by administering one or more GLP-1 receptor agonists to the said individual.

[0136] GLP-1 receptor agonists are most commonly synthetic peptides, whose polypeptide sequences are close to those of GLP-1 and act by binding to GLP-1 receptors, and generally have increased stability compared to endogenous GLP-1.

[0137] Examples of GLP-1 receptor agonists include exenatide, liraglutide, dulaglutide, semaglutide, tirzepatide, lixisenatide, albiglutide, exenatide QW, BI 456906 mazdutide (IBI362; LY3305677), retatrutide (LY3437943), danuglipron, exendin-4, or cotadutide.

[0138] According to certain embodiments, the subject showing an indication for administration of a GLP-1 receptor agonist is selected from the list consisting of: insulin-resistant subjects (such as diabetic subjects), overweight subjects (especially obese subjects), subjects who have undergone metabolic surgery, subjects seeking to regulate appetite, subjects suffering from liver diseases, subjects suffering from cardiovascular diseases (especially subjects suffering from cardiovascular diseases related to diabetes), subjects suffering from inflammation related to diabetes and / or metabolic syndrome, subjects suffering from sarcopenia, subjects suffering from cachexia, and subjects suffering from neurodegenerative diseases.

[0139] "Metabolic surgery" refers to a surgery aimed at treating metabolic diseases by surgical methods. Metabolic surgery is also called diabetes or sugar surgery and includes surgical procedures for controlling diabetes and obesity. These can include surgical interventions that regulate appetite, gastric emptying, food preference, and / or taste.

[0140] Liver diseases for which administration of a GLP-1 agonist may be required are, for example, non-alcoholic steatohepatitis (NASH).

[0141] Cardiovascular diseases for which administration of a GLP-1 agonist may be required particularly include atherosclerosis, myocardial infarction, stroke, or changes in lipid profiles (total cholesterol, LDL cholesterol, HDL / LDL ratio).

[0142] Neurodegenerative diseases for which administration of a GLP-1 agonist may be required particularly include Alzheimer's disease or Parkinson's disease.

[0143] According to certain embodiments, the subject to be given a prophylactic and / or therapeutic treatment is selected from elderly subjects suffering from sarcopenia, overweight subjects, and obese subjects with sarcopenic obesity on a diet.

[0144] The present specification also describes a method for preventing and / or treating a disease in a subject in need thereof that requires an increase in GLP-1 levels, said use being selected from (i) preventing or treating a disease associated with insulin resistance and / or abnormal regulation of blood glucose, and (ii) preventing or treating loss of muscle mass and / or loss of muscle function, the method being characterized by comprising at least one step of administering a bacterial strain of Lactobacillus gasseri.

[0145] Furthermore, the present invention relates to the non-therapeutic use of a bacterial strain of Lactobacillus gasseri according to the present invention or a composition comprising the same for maintaining or increasing muscle mass and / or function in a subject in need thereof, said subject being in particular a subject selected from the following: a malnourished subject, an elderly subject (in particular a malnourished elderly subject), and a subject engaged in intense physical exercise.

[0146] In the meaning of the present specification, "malnutrition" means "the state of nutritional imbalance of an organism" (see "Diagnosis of undernutrition in children and adults - Method recommendations for clinical practice", November 2019, National Health Service and French Nutrition Federation), said imbalance being characterized by a negative energy and / or protein balance. This definition includes various situations that can lead to a state of malnutrition, such as insufficient individual nutritional intake, increased consumption, or losses that cause imbalance. The term also covers subjects with increased nutritional requirements. The imbalance inherent in malnutrition causes harmful effects on the body, accompanied by measurable changes in body functions and / or body composition, which may be related to the worsening of the disease progression that may affect the subject.

[0147] "Elderly subject" is defined as above.

[0148] "Intense physical exercise" means a physical activity that requires great effort, which results in rapid breathing and an increased heart rate. Intense physical exercise is characterized by a MET (task metabolic equivalent - 1 MET is equivalent to an oxygen uptake of 3.5 mL per kilogram of body weight per minute) greater than 6. MET represents the ratio between the energy consumption during physical exertion and the energy consumption at rest. During physical exercise, especially in insulin-resistant subjects, GLP-1 can constitute an interesting complementary strategy to help optimize blood glucose management and enhance insulin sensitivity. Intense physical exercise also leads to the stimulation of GLP-1 production.

[0149] The present invention also describes a method for maintaining or increasing muscle mass and / or function in a subject in need thereof, said subject being in particular a subject selected from the group consisting of: malnourished subjects, elderly subjects (in particular malnourished elderly subjects), and subjects practicing intense physical sports, said method comprising at least the step of administering to the subject in need thereof at least one bacterial strain of Lactobacillus mucosae.

[0150] Finally, the invention relates to a strain of Lactobacillus mucosae, which is deposited at the CNCM under the accession number CNCM I-5661.

[0151] The present invention is described in more detail below using the following examples, which are presented by way of illustration.

[0152] Examples

[0153] 1. Selection of bacterial strains capable of secreting GLP-1

[0154] 61 Lactobacillus isolates were tested, which were derived from the feces of germ-free animals previously transferred together with the feces of short bowel syndrome patients. These Lactobacilli have a strong tropism for the intestine because these strains are abundant in the feces of short bowel syndrome patients and are able to colonize the digestive tract of germ-free rats.

[0155] The inventors sequenced the 16S gene and divided the isolates into 6 species, and selected 1 to 2 isolates from each species, a total of 7 strains, for preliminary morphological, phenotypic and genomic characterization.

[0156] The 6 species are:

[0157] Lactobacillus casei (strains 1 and 2)

[0158] Lacticaseibacillus camelliae (strain 3)

[0159] Lactobacillus salivarius (strain 4)

[0160] Lactobacillus rhamnosus (strain 5)

[0161] Lactobacillus reuteri (strain 6)

[0162] Lactobacillus mucosae (strain CNCM I-5661, also called strain I-5661)

[0163] The strains were characterized by functional tests at the INRAE in Aurillac (Unité Mixte de Recherche sur le Fromage Aurillac, UMR 0545).

[0164] All the isolated strains described above were incubated on the intestinal neuroendocrine cells of SCT-1 mice. 10 9 bacteria were incubated with 2 x 10 6 SCT-1 cells at 80% confluence for 4 hours at 37 °C and 5% CO2. The cell supernatant was then recovered and the peptide GLP-1 was assayed in this supernatant (by ELISA).

[0165] The results are shown in Figure 1 .

[0166] Among all the strains tested, none of the strains showed significant synthesis of GLP-1 in the culture medium, except for strain CNCM I-5661, which led to a very significant production of GLP-1 in the culture medium (P < 0.05).

[0167] In fact, among all the strains from the same donor, only the strain of the species Lactobacillus mucosae CNCM I-5661 specifically increased the production of GLP-1 by 25-fold. When lactobacilli of the species Lactobacillus casei isolated from the same patient were incubated under the same conditions, no release of GLP-1 was observed (results not provided here - see Example 4 below and related results).

[0168] This strain corresponds to the species Lactobacillus mucosae.

[0169] The control group corresponded to the culture of intestinal cells in a sterile medium (no GLP-1 production).

[0170] Results of inducing GLP-1 secretion through STC-1 cells

[0171] STC-1 cells were from the ATCC collection center (number STC-1, CRL-3254 TM ) and were cultured and prepared for co-incubation one day before the experiment. On the day of the experiment, the bacteria frozen in dry pellets were thawed and cultured in a medium without FCS or antibiotics. The bacteria were prepared by serial dilution to achieve the following MOI (multiplicity of infection): 1:10, 1:100, and 1:1000.

[0172] The STC-1 cells were washed and then the medium containing the bacteria was added. After 4 hours of co-culture, the supernatant was collected and counted. Then the supernatant and the cells were stored at -80 °C for subsequent assay of GLP-1. The amount of GLP-1 was assayed by ELISA according to the instructions of the supplier Assay Genie (ELISA GLP-1 mouse (reference: MOFI00854)).

[0173] The production of GLP-1 was expressed as pg / ml.

[0174] The definitions of the terms used in Table 1 below are as follows:

[0175] Control: STC-1 cells in the presence of the culture medium.

[0176] MOI: 10 (number of bacteria added per STC-1 cell).

[0177] C: Indicates that the CNCM-I5661 bacteria were cultured in MRS medium (supplier BD, reference: 288130, reconstituted according to the supplier's instructions), then centrifuged and frozen in the form of dry pellets. The dry pellets were then resuspended in STC cell medium.

[0178] V: Indicates that the CNCM-I5661 bacteria were cultured in "vegan MRS" medium (supplier BIOKAR, reference: BK176HA, reconstituted according to the supplier's instructions), then centrifuged and frozen in the form of dry pellets. The dry pellets were then resuspended in STC-1 cell medium.

[0179] 61C-10: STC-1 cells were incubated in the presence of the CNCM-I5661 strain. MOI: 10 (the bacteria are 10 times the number of STC-1 cells).

[0180] 61C-100: STC-1 cells were incubated in the presence of the CNCM-I5661 strain. MOI: 100 (the bacteria are 100 times the number of STC-1 cells).

[0181] 61C-1000: STC-1 cells were incubated in the presence of the CNCM-I5661 strain. MOI: 1000 (the bacteria are 1000 times the number of STC-1 cells).

[0182] [Table 1]

[0183] Experimental conditions GLP1 (pg / ml) Control 0.0 61C-10 256.0 61C-100 454.2 61C-1000 1193.7 61V-10 253.6 61V-100 405.5 61V-1000 1240.5

[0184] The results shown in the above table indicate the dose effect of the strain and also indicate that the strain retains its function even when cultured in another medium.

[0185] Conclusion: The presence of the CNCM-5661 strain induces the production of the intestinal hormone GLP-1 in STC-1 cells. As the number of bacteria increases, this production becomes more significant. This production was not observed in the presence of a strain of Lactobacillus casei from the same patient. The GLP-1 promoting effect of the CNCM I-5661 strain is not dependent on the medium used to culture the strain.

[0186] In addition, the inventors showed that the production of GLP-1 increases with the increase in the contact time between the bacteria of the CNCM I-5661 strain and intestinal cells (seeFigure 2 )。

[0187] Survival ability of strain 2-CNCMI-5661 in the intestinal environment

[0188] To determine whether strain CNCM I-5661 is able to survive in the intestinal environment, various tests were conducted.

[0189] - Tolerance of bacteria to gastric (stomach) acidity

[0190] The bacteria were placed in acidic media (pH 2.5 and 3) for 45 minutes and 90 minutes, corresponding to the average residence time of the food bolus in the stomach before entering the duodenum.

[0191] The bacteria showed very good resistance to the gastric medium (composition: HCl, pepsin, NaCl), and there was no significant difference between the incubation times and the two pH values tested (p > 0.05, Fisher's exact test).

[0192] The results are shown in Table 2 below.

[0193] [Table 2]

[0194]

[0195] - Tolerance of bacteria to gastric acidity (bile salts)

[0196] The inventors also tested the survival rate of the bacteria under the conditions encountered downstream of the stomach (including the presence of bile salts). Therefore, the survival rate of strain CNCM I-5661 was tested in the presence of bile salts after incubation at 37 °C for 1 hour, 2 hours, 3 hours, and 4 hours.

[0197] The bacteria showed very good resistance to bile salts, and there was no significant difference between the incubation times (p > 0.05, Fisher's exact test).

[0198] The results are shown in Table 3 below.

[0199] [Table 3]

[0200]

[0201] 3-Adhesion ability of strain CNCMI-5661 to intestinal cells

[0202] Finally, to evaluate the probiotic potential of strain CNCM I-5661, the inventors determined its ability to adhere to human intestinal Caco-2 cells. The viable cell count of lactic acid bacteria adhering to intestinal cells was determined after 3 hours of contact with Caco-2 cells at MOI (multiplicity of infection) values of 0.1, 1, 10, and 100.

[0203] The strain CNCM I-5661 showed satisfactory adhesion percentages similar to other microorganisms, with no significant differences between the tested MOIs (p > 0.05, Fisher's exact test).

[0204] The results are shown in Table 4 below.

[0205] [Table 4]

[0206]

[0207] 4 - Extend the properties described for the strain CNCM I-5661 to the entire species Lactobacillus mucosae

[0208] Other strains of the species Lactobacillus mucosae were tested for their ability to allow SCT-1 cells to secrete GLP-1 in culture to determine whether the beneficial effects observed on the strain Lactobacillus mucosae CNCM I-5661 also apply to other strains of the same species.

[0209] Thus, three other strains were tested: DSM 13345, DSM 13346, and DSM 102820. The strain CNCM I-5661 (as a positive control) and a strain of Lactobacillus casei (negative control, does not induce GLP-1 synthesis) were also used:

[0210] - Control: Sterile cell culture medium;

[0211] - Lactobacillus casei strain (negative control) ~ 5·10 9 CFU / mL (strain A);

[0212] - Lactobacillus mucosae CNCM I-5661 (positive control) ~ 5·10 9 CFU / mL (strain B);

[0213] - Lactobacillus mucosae DSM 13345 (S32T) ~ 2·10 9 CFU / mL (strain C);

[0214] - Lactobacillus mucosae DSM 13346 ~ 4·10 9 CFU / mL (strain D);

[0215] - Lactobacillus mucosae DSM 102820 ~ 5·10 9 CFU / mL (strain E);

[0216] Under "diluted" conditions, strains B to E were diluted 1 / 10.

[0217] Figure 3The results shown indicate that in the presence of all strains of the tested species Lactobacillus mucosae, the synthesis of GLP-1 by SCT-1 cells was observed. Moreover, this production of GLP-1 was dose-dependent.

[0218] Effect of strain 5-CNCM I-5661 on muscle mass in a frail elderly rat model

[0219] To evaluate the effect of anabolic probiotics on GLP-1 targets, the inventors determined the ability of the Lactobacillus mucosae strain CNCM I-5661 according to the invention (shown to be able to induce the synthesis of GLP-1 by SCT-1 cells in culture) and a strain of the species Lactobacillus casei outside the present invention (not inducing GLP-1 production) on muscle mass in a frail elderly rat model.

[0220] The inventors conducted a 1-month in vivo study on elderly rats (20 months old) according to the following groups:

[0221] - A group fed ad libitum (n = 15);

[0222] - A group fed 75% to 80% of ad libitum intake (simulating one of the factors of geriatric malnutrition and frailty) (n = 16);

[0223] - A group fed 75% to 80% of ad libitum intake supplemented daily with the bacterium CNCM I-5661 (10 9 CFU) (n = 13); and

[0224] - A group fed 75% to 80% of ad libitum intake supplemented daily with a bacterium of the species Lactobacillus casei (10 9 CFU) (n = 16).

[0225] At the end of the experiment, the animals were sacrificed, weighed, the hind leg muscles (gastrocnemius, extensor digitorum longus, soleus, tibialis anterior) were excised and weighed. The sum of the weights of these 4 muscles was calculated and reduced to the weight of the animal.

[0226] The results, as Figure 4 shown, indicate that supplementation with only Lactobacillus mucosae CNCM-I5661 allowed restricted rats to increase their muscle mass relative to ad libitum-fed animals (P = 0.01) and restricted animals (trend, P = 0.092).

[0227] Moreover, the muscle mass in the AL group, R group, and R + Lactobacillus casei group was similar, indicating that, unlike Lactobacillus mucosae CNCM-I5661, Lactobacillus casei did not increase muscle mass. This indicates that, unlike other lactobacilli, Lactobacillus mucosae CNCM I-I5661 has a specific effect of increasing muscle mass in a frail elderly rodent model.

[0228]

Claims

1. Use of a bacterial strain of Lactobacillus mucosae, or a lysate or culture supernatant thereof, for the prevention and / or treatment of a disease in a subject in need thereof, which disease requires an increase in GLP-1 production, said disease being selected from (i) diseases associated with abnormal regulation of insulin sensitivity and / or blood glucose, and (ii) loss of muscle mass and / or muscle function.

2. Use of the bacterial strain according to claim 1, characterized in that, The subject in need thereof is a subject showing an indication for administration of a GLP-1 receptor agonist.

3. Use of the bacterial strain according to claim 2, characterized in that, The subjects showing an indication for administration of a GLP-1 receptor agonist are selected from the list consisting of: insulin-resistant subjects, such as diabetic subjects, overweight subjects, especially obese subjects, subjects who have undergone metabolic surgery, subjects seeking to regulate appetite, subjects suffering from liver diseases, subjects suffering from cardiovascular diseases, especially subjects suffering from cardiovascular diseases associated with diabetes, subjects suffering from inflammation, especially subjects suffering from inflammation associated with diabetes and / or metabolic syndrome, subjects suffering from sarcopenia, subjects suffering from cachexia, and subjects suffering from neurodegenerative diseases.

4. Use of the bacterial strain according to any one of claims 1 to 3, characterized in that, The disease is a disease associated with abnormal regulation of insulin sensitivity and / or blood glucose, and is selected from prediabetes, type 1 diabetes and type 2 diabetes.

5. Use of the bacterial strain according to any one of claims 1 to 3, characterized in that The disease is loss of muscle mass and / or muscle function, and the subject in need thereof is selected from elderly subjects suffering from sarcopenia, overweight and obese subjects with sarcopenic obesity on a diet, diabetic or prediabetic subjects, and / or subjects suffering from cachexia, said cachexia being especially associated with cancer, inflammatory bowel disease or chronic obstructive pulmonary disease, limited mobility, "severe burn" status, sepsis or viral infection, convalescence and / or limited mobility, and / or bowel resection or intestinal malabsorption.

6. Use of the bacterial strain according to any one of claims 1 to 5, characterized in that The bacterial strain is selected from the strains of Lactobacillus mucosae deposited at the CNCM under the accession number CNCM I-5661, the strains of Lactobacillus mucosae deposited at the DSM under the accession number DSM 13345, the strains of Lactobacillus mucosae deposited at the DSM under the accession number DSM13346, the strains of Lactobacillus mucosae deposited at the DSM under the accession number DSM 102820, or a mixture thereof, especially the strain of Lactobacillus mucosae deposited at the CNCM under the accession number CNCMI-5661.

7. Use of the bacterial strain according to any one of claims 1 to 6, characterized in that, The bacteria of Lactobacillus mucosae are in a live or dead form, preferably in a live form.

8. Use of the bacterial strain according to any one of claims 1 to 7, characterized in that The bacterial strain is contained in a composition containing a physiologically acceptable medium, especially in an oral composition, more particularly in an oral composition selected from the group consisting of: food, beverage, medicine, nutraceutical, food additive, food supplement, dairy product and live biotherapeutic product (LBP).

9. Use of the bacterial strain according to claim 8, characterized in that, The composition further comprises one or more other probiotic strains, in particular selected from the species Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus salivarius, Lactobacillus johnsonii, Lactobacillus salivarius, Lactococcus lactis, Enterococcus faecium, Enterococcus faecalis, Saccharomyces cerevisiae, Saccharomyces boulardii, Faecalibacterium prausnitzii, Akkermansia muciniphila, Blautia obeum, Faecalibacterium prausnitzii, Streptococcus thermophilus, or mixtures thereof, preferably selected from the group consisting of: Bifidobacterium longum NCC3001 (ATCC BAA-999), Bifidobacterium longum NCC2705 (CNCM 1-2618), Bifidobacterium longum NCC490 (CNCM 1-2170), Bifidobacterium lactis NCC2818 (CNCM I-3446), Bifidobacterium breve strain A, Lactobacillus johnsonii NCC533 (CNCM 1-1225), Enterococcus faecium SF 68 (NCC2768; NCIMB10415), Lactobacillus casei (CNCM I-5662 and CNCM I-5663), Streptococcus thermophilus (CNCM I-5334), and combinations thereof.

10. Use of the bacterial strain according to claim 8, characterized in that, The composition does not contain any bacteria of other Lactobacillus species.

11. Use of a bacterial strain according to any one of claims 8 to 10, characterized in that, The composition further comprises one or more prebiotics.

12. Non-therapeutic use of a bacterial strain of the species Lactobacillus mucosae, or a lysate or culture supernatant thereof, in particular a bacterial strain of the species Lactobacillus mucosae as defined in any one of claims 6 to 11, or a lysate or culture supernatant thereof, for maintaining or increasing muscle mass and / or function in a subject in need thereof, in particular said subject in need thereof being selected from malnourished subjects, elderly subjects, in particular malnourished elderly subjects, and subjects engaged in intense physical sports.

13. A bacterial strain of Lactobacillus mucosae deposited at the CNCM under accession number CNCM I-5661.

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