Disosmobacter for the treatment of breast cancer

Abstract

The present invention relates to compositions comprising bacteria of the genus Disosmobacter and / or variants, extracts or fragments thereof for use in preventing and / or treating breast cancer.

Description

[Technical Field]

[0001] The present invention relates to the treatment and / or prevention of breast cancer using bacteria of the genus Dysosmobacter or extracts thereof. [Background technology]

[0002] Cancer has become one of the most important diseases of the last century and is the leading cause of premature death worldwide. It is estimated that in 2020, approximately 19.3 million new cases of cancer were diagnosed worldwide, resulting in 10 million deaths. Of all cancers, breast cancer (BC) is currently the most commonly diagnosed, with an estimated 2.3 million cases, representing 11.7% of all diagnoses and 25% of diagnosed women. However, breast cancer is a highly heterogeneous disease, classified into subtypes based on the (over)expression of progesterone (PR), estrogen (ER), and human epidermal growth factor receptor (HER-2). Early-stage and late-stage patients with hormone receptor-expressing breast tumors, as well as patients with HER-2-enriched tumors, have benefited from targeted therapy with or without conventional chemotherapy. However, patients with triple-negative breast cancer (TNBC), characterized by the absence or overexpression of these receptors, have not benefited from targeted therapy. Due to this unmet need in the field of targeted therapy, TNBC remains the most aggressive and poorest prognostic subtype associated with higher recurrence rates, higher metastatic potential, and poorer survival. Risk factors for BC and TNBC include older age at first birth, physical inactivity, and alcohol consumption, although obesity is increasingly recognized as a major risk factor for BC and TNBC.

[0003] Obesity has been reported to be associated with an increased risk of adverse outcomes and poorer prognosis in breast cancer patients, which is associated with shorter overall and disease-free survival, highlighting the need to explore novel therapeutic strategies for breast cancer in the context of obesity. In obesity, profound changes in the composition of gut microbiota, also known as dysbiosis, occur in clinical and preclinical models (Ley et al., 2006). Furthermore, the gut microbiota is a key regulator of systemic metabolism and low-grade inflammation (Cani et al., 2019), but it has also attracted significant attention in modulating cancer and breast cancer progression and treatment (Sampsell et al., 2020).

[0004] In this study, we investigated the impact of Dysosmobacter welbionis, a novel bacterium discovered, isolated, and named at the Catholic University of Leuven (UCLouvain) (Brussels, Belgium) (Le Roy et al., 2020 and WO 2020011856). Dysosmobacter welbionis is found in 70% of the general population and can therefore be considered a very common bacterium, like Akkermansia muciniphila. As a brief background on this bacterium, Dysosmobacter welbionis is a butyrate-producing bacterium, and Dysosmobacter species and Dysosmobacter welbionis have been found to be underrepresented in the gut of subjects with obesity and type 2 diabetes. In preclinical studies, Dysosmobacter welbionis J115 T has been shown to reduce weight gain and fat mass by acting on mitochondrial activity, and to improve glucose tolerance, lipid and energy metabolism (Le Roy, Moens de Hase et al., 2022).

[0005] In this disclosure, the inventors have identified Disosmobacter welbrionis J115 T demonstrated that it can reduce breast cancer growth. Summary of the Invention

[0006] The present invention relates to a composition comprising at least (a) bacteria of the genus Disosmobacter and / or variants and / or extracts and / or fragments thereof, and / or (b) a culture supernatant of bacteria of the genus Disosmobacter and / or variants thereof, for use in preventing and / or treating breast cancer in a subject in need thereof. In some embodiments, the bacteria belong to the species Disosmobacter welbionis.

[0007] In some embodiments, the bacterium is strain J115, deposited at BCCM / LMG as LMG P-30603 on March 14, 2018. T Belongs to.

[0008] In some embodiments, the breast cancer comprises at least one mutation in a gene encoding a receptor selected from the group comprising or consisting of progesterone receptor (PR), estrogen receptor (ER), and human epidermal growth factor receptor-2 (HER2).

[0009] In some embodiments, the breast cancer is selected from the group comprising or consisting of luminal A breast cancer, luminal B breast cancer, HER2-positive breast cancer, and triple-negative breast cancer (TNBC), preferably said breast cancer is triple-negative breast cancer.

[0010] In some embodiments, the subject further suffers from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity-related disorders, liver diseases, metabolic disorders, adipokine-related disorders, and inflammatory diseases, and combinations thereof.

[0011] In some embodiments, the subject further suffers from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), cirrhosis, diabetes, impaired glucose tolerance, hyperglycemia, dyslipidemia, dyslipidemia, high cholesterol, elevated LDL cholesterol levels, low HDL cholesterol levels, elevated triglyceride levels, and enterocolitis, and combinations thereof.

[0012] In some embodiments, the composition is administered in a therapeutically effective amount, preferably 1×10 2 ~Approx. 1×10 15 CFU of said bacteria.

[0013] In some embodiments, the composition comprises live bacteria.

[0014] In some embodiments, the composition comprises dead or killed bacteria.

[0015] In some embodiments, the composition comprises pasteurized bacteria.

[0016] In some embodiments, the composition further comprises at least another anti-cancer agent.

[0017] In some embodiments, the composition is in the form of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

[0018] The present invention further relates to a prebiotic comprising one or more active ingredients or substances that increase the level of bacteria of the genus Disosmobacter in the microbiome of a subject in need thereof, for use in preventing and / or treating breast cancer in said subject.

[0019] The present invention further relates to a composition comprising at least one bacterium of the genus Disosmobacter and / or a variant, extract or fragment thereof for use as an adjuvant for a therapeutic agent administered to a subject suffering from breast cancer.

[0020] definition In the present invention, the following terms have the following meanings:

[0021] The word "about" before a number means an average of ±10% of the value of said number.

[0022] "Acceptable," as used, for example, in the expressions "pharmaceutically acceptable" or "nutraceutical acceptable," refers to molecular entities and compositions that do not produce side effects, allergic reactions, or other adverse reactions when administered to a subject, particularly a human, as appropriate.

[0023] "Strain" refers to a subtype of a bacterial species.

[0024] "Dysosmobacter" (pronounced Dys.os.mo.bac'ter; Greek masculine adjective "dysosmos" (bad-smelling) + Dutch masculine noun "bacter" (rod) = Dutch masculine noun "Dysosmobacter" (bad-smelling bacillus)) refers to a genus of bacteria described herein characterized by "strictly anaerobic, non-pigmented, non-spore-forming, non-motile, Gram-stain-negative cells." Cells primarily form linear rods measuring 1.8-3.0 μm, although elongated rods up to 20 μm in length are frequently formed regardless of growth phase. Respiratory menaquinones are not produced. The genus belongs to the family Ruminococcaceae. The type species is Dysosmobacter welbiornis. In one embodiment, the diagnostic diamino acid in the cell wall is meso-2,6-diaminopimelic acid.

[0025] "Disosmobacter welbionis" (pronounced wel.bi.o'nis; Dutch common noun "welbionis") refers to a bacterial species described herein that possesses the characteristics of the genus Disosmobacter described above, plus the following characteristics: "After 72 hours of incubation at 37°C under anaerobic conditions, colonies on solid modified YCFA are generally punctate, creamy, translucent, and round, slightly convex, and smooth." Growth is inhibited by the presence of 2% w / v bile or 2% w / v NaCl. Esculin is not hydrolyzed. Indole is not produced. Nitrate is not reduced. Gelatin is not digested. Urease is not produced. Catalase is not produced. Acid is produced from myo-inositol, but not from D-glucose, D-arabinose, D-ribose, and D-xylose. Positive reactions are obtained for arginine dihydrolase and glutamic acid decarboxylase. All other tests (API 20A and Rapid ID 32A) (bioMerieux, Lyon, France) are negative. The main fermentation end product from myo-inositol is butyric acid. The DNA GC content of the type strain is 59.3 mol% by high performance liquid chromatography (HLPC). In one embodiment, the DNA GC content of the type strain is 58.9 mol% based on the genome sequence. The type strain is J115 T (Deposited on March 14, 2018 at the Belgian Coordinated Collections of Microorganisms / Laboratory of Microbiology (BCCM / LMG), Ghent University, KLLedeganckstraat 35, 9000 Gent, Belgium, as LMG P-30603) and isolated from human feces. In one embodiment, the predominant cellular fatty acids are branched-chain saturated fatty acids and DMA. In one embodiment, the predominant DMA fatty acid is C 18:0 DMA, with the main branched chain saturated fatty acid being iso-C 15:0 and Anteiso-C 15:0 is.

[0026] "Fermentation" refers to the metabolic process of consuming sugars in the absence of oxygen. The products are organic acids, gases, or alcohol. It occurs in yeast and bacteria, and also in oxygen-starved muscle cells, as in lactic acid fermentation.

[0027] The terms "gut microbiota" or "gastrointestinal microbiota" are used interchangeably to refer to the complex community of microorganisms that live in the digestive tract of humans and other animals. The composition of the gut microbiota changes over the life of the host organism or when the host's diet changes. It also varies throughout the gastrointestinal tract. The gastrointestinal tract contains up to 10 microorganisms per gram of gut contents. 12 The gut contains a dense microbial ecosystem containing millions of cells. Many bacterial species in the gut are largely unculturable and have never been studied outside their host. The four predominant bacterial phyla in the human gut are Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria. Most bacteria belong to the genera Bacteroides, Clostridium, Faecalibacterium, Eubacterium, Ruminococcus, Peptococcus, Peptostreptococcus, Blautia, Subdoligranulum, Alistipes, Coprococcus, Dialister, Lachnoclostridium, Oscillospira, Parabacteroides, Prevotella, Roseburia, Ruminiclostridium, Sutterella, and Bifidobacterium. Other genera, such as Escherichia, Enterococcus, Barnesiella, Butyricimonas, Butyricicoccus, Lachnospira, Odoribacter, Turicibacter, and Lactobacillus, are present to a lesser extent. The gut microbiota is thought to function in defense against pathogens by competing with potential pathogens, participating in the development and metabolism of the intestinal protective and immune systems, assisting in the digestion of ingested nutrients, aiding in the absorption of nutrients, and synthesizing vitamins. The gut microbiota also interacts with the function of the central nervous system and the neuroendocrine and neuroimmune systems.

[0028] As used herein, "mutant" refers to a biological entity that has undergone a natural or induced (i.e., mutagenesis) change in its genetic structure that does not interfere with the defining characteristics of said biological entity. The change in genetic structure may be an insertion, deletion, or substitution of one or several nucleotides in the genomic sequence. For example, a Disosmobacter wellbionis mutant refers to a Disosmobacter wellbionis strain that has undergone a natural or genetic engineering change in its genetic structure that does not interfere with its belonging to the Disosmobacter wellbionis species.

[0029] A "nutraceutically effective amount" refers to the amount of a nutraceutical composition, food or dietary supplement or functional food that is necessary and sufficient to provide a physiological benefit or relieve discomfort in a subject.

[0030] "Pasteurized bacteria" refers to bacteria that have been subjected to a heat treatment (or heating process).

[0031] A "pharmaceutically acceptable carrier or excipient" refers to a molecular entity or composition that does not produce side effects, allergic reactions, or other adverse reactions when administered to a subject, particularly a human, as appropriate. It includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. For human administration, formulations must meet pyrogenicity, general safety, and purity standards required by regulatory agencies such as the FDA or EMA. Thus, a pharmaceutically acceptable carrier or excipient may refer to any type of non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, or formulation auxiliary.

[0032] "Prebiotics" refers to substances that cannot be digested by a subject (e.g., a human), but whose metabolism by microorganisms in the intestine modulates the composition and / or activity of the intestinal microbiota, thus conferring a beneficial physiological effect on the host.

[0033] "Probiotics" refers to microbial cell preparations (e.g., live microbial cells) that, when administered in an effective amount, have a beneficial effect on the health or well-being of a subject. By definition, all probiotics are known to be non-pathogenic. In one embodiment, these health benefits are related to improving the balance and / or restoring normal microflora of a human or animal in the gastrointestinal tract.

[0034] "Subject" refers to a warm-blooded animal, preferably a human, a pet, or livestock. As used herein, the terms "pet" and "livestock" include, but are not limited to, dogs, cats, guinea pigs, rabbits, pigs, cows, sheep, goats, horses, and poultry. In some embodiments, the subject is a male or female subject, preferably a female subject. In some embodiments, the subject is an adult or a child. In some embodiments, the subject may be a "patient," i.e., a subject awaiting or undergoing medical care, who has previously been, is currently, or will be a subject for medical treatment according to the methods of the present invention, or who is being monitored for the development of a disease.

[0035] A "therapeutically effective amount" refers to a level or amount of an agent intended to (1) delay or prevent the onset of a disease, disorder, or condition, (2) slow or halt the progression, worsening, or deterioration of one or more symptoms of a disease, disorder, or condition, (3) bring about amelioration of symptoms of a disease, disorder, or condition, (4) reduce the severity or incidence of a disease, disorder, or condition, or (5) cure a disease, disorder, or condition, without causing significant adverse or harmful side effects to the target. A therapeutically effective amount may be administered prior to the onset of a disease, disorder, or condition for prophylactic treatment. Alternatively, or additionally, a therapeutically effective amount may be administered after the onset of a disease, disorder, or condition for therapeutic treatment.

[0036] "Treatment" refers to both therapeutic and prophylactic treatment, the purpose of which is to prevent or slow (alleviate) the targeted pathological condition or disorder, preferably breast cancer. Those in need of treatment include those already suffering from the disorder, as well as those susceptible to the disorder or those in whom the disorder is to be prevented. A subject or mammal is successfully "treated" if, after administration of a therapeutic amount of Disosmobacter wellbionis and / or its variants and / or fragments thereof of the present invention, the patient exhibits one or more of the following observable and / or measurable changes: remission of one or more symptoms associated with the specific disease or condition, preferably breast cancer; reduced morbidity and mortality; and improved quality of life issues. The above parameters for assessing successful treatment and disease improvement are readily measurable using routine procedures familiar to physicians.

[0037] "Type strain" refers to the naming standard for a species and the reference point to which all other strains are compared to determine whether they belong to that species, as defined in the International Code of Prokaryotic Nomenclature. For example, strain J115, isolated from a fecal sample of a healthy 25-year-old woman, T is the type strain of the Disosmobacter welbrionis species.

[0038] "Variant" refers to any genetically or phenotypically distinct strain of a bacterial species that retains species-defining characteristics. The term variant is also used in reference to other phylogenetic groups, such as genera or strains. As used herein, the term "variant" refers to both naturally occurring variants or mutants and specifically developed variants or mutants of the bacteria disclosed and exemplified herein. In one embodiment, variants may or may not possess the same distinguishing biological properties of the bacteria exemplified herein, so long as they share similar advantageous properties with respect to disease treatment or prevention. In one embodiment, variants of the bacteria of the present invention have the same functional and / or therapeutic properties as the bacteria of the present invention. Illustrative examples of suitable methods for preparing variants of the microbial strains exemplified herein include, but are not limited to, gene integration techniques mediated by insertion of elements or transposons or by homologous recombination, other recombinant DNA techniques for modification, insertion, deletion, activation or silencing, intraspecific protoplast fusion, mutagenesis by ultraviolet or X-ray irradiation or treatment with chemical mutagens such as nitrosoguanidine, methyl methanesulfonate and nitrogen mustard, and bacteriophage-mediated transduction. DETAILED DESCRIPTION OF THE INVENTION

[0039] The present invention relates to a composition comprising at least (a) bacteria of the genus Disosmobacter and / or variants, extracts or fragments thereof, and / or (b) a culture supernatant of bacteria of the genus Disosmobacter and / or variants thereof, for use in preventing and / or treating breast cancer in a subject in need thereof.

[0040] In some embodiments, the Disosmobacter bacterium included in a composition for use according to the invention belongs to a species selected from the group including or consisting of: Disosmobacter welbiornis, Dysosmobacter acutus, Dysosmobacter segnis, and Dysosmobacter hominis. In some embodiments, the Disosmobacter bacterium included in a composition for use according to the invention belongs to a species selected from the group including or consisting of: Disosmobacter welbiornis.

[0041] The biological properties of Disosmobacter welbrionis have been previously reported by the present inventors in WO2020011856.

[0042] In some embodiments, the bacterium is J115 T , Disosmobacter actus MSJ-2 T (CGMCC Accession Number: 1.32896T, KCTC Accession Number: 15976T), Disosmobacter segnis BX15 (CGMCC Accession Number: 1.32894), Marseille-Q4140, MM13, and Disosmobacter hominis NSJ-60 (CGMCC Accession Number: 1.32836, KCTC Accession Number: 25148). In some embodiments, the bacterium belongs to strain J115, deposited at BCCM / LMG as LMG P-30603 on March 14, 2018. T and / or its variants. T The strain is the type strain of the Disosmobacter welbrionis species. In a particular embodiment, the present invention provides the strain Disosmobacter welbrionis J115 for use in preventing and / or treating breast cancer in a subject in need thereof. T (Deposit number: LMG P-30603) and / or a variant, extract or fragment thereof.

[0043] In one embodiment, the genome sequence of the bacterium has the sequence of SEQ ID NO:1 or is at least about 65% identical to SEQ ID NO:1, preferably at least about 70%, 75%, 80%, 85%, 90% identical to SEQ ID NO:1, more preferably at least about 91%, 92%, 93%, 94%, 95%, 96%, 96.5%, 97%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 97.10%, 97.11%, 97.12%, 97.13%, 97.14%, 97.15%, 97.16%, 97.17%, 97.18%, 97.19 ... and having a sequence that exhibits 0.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.65%, 98.7%, 98.8%, 98.9%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or more identity.

[0044] In one embodiment, the bacterium has an average nucleotide identity (ANI) score of greater than about 60, preferably greater than about 74, 75, 80, 85, 90, more preferably greater than about 95, even more preferably greater than about 96, 97, 98, 98.5, 98.65, 99 or more when compared to the genome of the sequence of SEQ ID NO:1.

[0045] Techniques for determining ANI values are known to those skilled in the art (such as the method performed in Kim et al., Int J Syst Evol Microbiol. 2014 Feb;64(Pt 2):346-51). Briefly, ANI corresponds to the sum of the identities of each bidirectional best hit (orthologous sequences identified based on their position and sequence identity within the BBH-genome) multiplied by the length of the alignment and divided by the total length of the BBH gene.

[0046] In one embodiment, the bacterium has a hybrid DNA-DNA hybridization value (also referred to as a DDH value) with SEQ ID NO: 1 of greater than about 60%, preferably greater than about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, and more preferably greater than about 70%.

[0047] Techniques for determining DDH values are known to those skilled in the art (such as the methods reviewed in Stackebrandt et al., "Molecular Identification, Systematics, and Population Structure of Prokaryotes", p. 23-50, 2006, Springer, Berlin, Heidelberg) and are based on the following general principles: (i) the genomic DNA (gDNA) of the organism to be assayed and the gDNA of a reference organism (e.g., in the context of the present invention, the type strain J115 T This method relies on (ii) shearing the DNA fragments from both strains (deposited at BCCM / LMG as LMG P-30603 on March 14, 2018) into 600-800 bp fragments, (iii) heating the mixture of DNA fragments from both strains to dissociate the DNA duplex, and (iv) subsequently lowering the temperature until the fragments reanneal. Because the melting temperature of a duplex is determined by the degree of base pairing identity between the two strands, the (dis)similarity of genomes can be inferred from the melting temperature. Hybrid DDH values are usually specified relative to the DDH value obtained by hybridizing a reference genome with itself. A DDH value of 70% or greater may be considered an indication that the test organism belongs to a different species from the reference strain used as a standard. DDH values may also be evaluated based on the genome sequences of the strains being compared using publicly available computer programs.

[0048] In one embodiment, the bacterium has a genomic distance to SEQ ID NO:1 of less than about 0.5, preferably less than about 0.22, 0.21, 0.20, 0.19, 0.18, 0.17, 0.16, 0.15, 0.14, more preferably less than about 0.13, 0.12, 0.11, 0.10 or less.

[0049] Techniques for determining genome-to-genome distance (GGD) are known to those skilled in the art. For example, the method described by Meier-Kolthoff et al. (BMC Bioinformatics 2013;21:14-60; Int J Syst Evol Microbiol 2014;1:352-6) may be used. Such methods may be performed using the Genome Calculator 2.1 (Deutsche Sammlung von Mikroorganismen und Zellkulturen-DSMZ) using BLAST+ as a local alignment tool and the sum of all identities calculated by dividing high-scoring segment pairs (HSPs) by the total HSP length.

[0050] In one embodiment, the nucleotide sequence of the 16S rRNA gene of the bacterium has the sequence of SEQ ID NO:2 (deposited under GenBank / EMBL / DDBJ accession number: MG963288) or has at least about 90% identity to SEQ ID NO:2, preferably at least about 91%, 92%, 93%, 94%, 95%, 95.5%, 96%, 96.5%, 97%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 97.10%, 97.11%, 97.12%, 97.13%, 97.14%, 97.15%, 97.16%, 97.17%, 97.18%, 97.19%, 97.20%, 97.21%, 97.22%, 97.23%, 97.24%, 97.25%, 97.26%, 97.27%, 97.28%, 97.29 ... and having a sequence that exhibits 5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.65%, 98.7%, 98.8%, 98.9%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or more identity.

[0051] In one embodiment, the nucleotide sequence of the 16S rRNA gene of the bacterium has the sequence of SEQ ID NO:2 or has a sequence that exhibits at least about 99.9% identity to SEQ ID NO:2, preferably at least about 99.91%, 99.92%, 99.93%, 99.94%, 99.95%, 99.96%, 99.97%, 99.98%, 99.99% or more identity to SEQ ID NO:2.

[0052] In one embodiment, the nucleotide sequence of the 16S rRNA gene of the bacterium has the sequence of SEQ ID NO:2 or has at least about 90% identity or greater over the entire length of SEQ ID NO:2, preferably at least about 91%, 92%, 93%, 94%, 95%, 96%, 96.5%, 97%, 97.1%, 97.2%, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1% identity over the entire length of SEQ ID NO:2. , 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.65%, 98.7%, 98.8%, 98.9%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.91%, 99.92%, 99.93%, 99.94%, 99.95%, 99.96%, 99.97%, 99.98%, 99.99% or more identity.

[0053] The term "identity," when used in the context of the sequences of two or more polypeptides or two or more nucleic acid molecules, refers to the degree of sequence relatedness between the polypeptides or nucleic acid molecules as determined by the number of matches between sequences of two or more amino acid or nucleotide residues. "Identity" is a measure of the percent exact match between shorter sequences of two or more sequences, with gap alignments (if any) handled by a particular mathematical model or computer program (i.e., "algorithm"). The identity of related polypeptides can be readily calculated by known methods.

[0054] The compositions for use according to the invention may comprise variants of the bacteria described herein above. Said variants may also be referred to as derivatives of the bacteria. In one embodiment, the variants of the bacteria may be obtained by mutation, diversification or recombination of the bacteria described herein. Within the scope of the present invention, variants may also be referred to as mutants.

[0055] In one embodiment, the bacterial variant of the genus Disosmobacter contained in the composition for use according to the invention is a variant of Disosmobacter welbrionis. In one embodiment, the bacterial variant of the genus Disosmobacter contained in the composition for use according to the invention is the variant strain of Disosmobacter welbrionis, J115. T is.

[0056] In one embodiment, the bacterial variant has a genome that is at least about 70%, preferably at least about 80%, at least about 90%, at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or more identical to the genome of the bacterium from which it is derived.

[0057] In one embodiment, the genomic sequence of the bacterial variant is at least about 65% identical to the genomic sequence of the bacterium from which it was derived, preferably at least about 70%, 75%, 80%, 85%, 90% identical to the genomic sequence of the bacterium from which it was derived, and more preferably at least about 91%, 92%, 93%, 94%, 95%, 96%, 96.5%, 97%, 97.1%, 97.2% identical to the genomic sequence of the bacterium from which it was derived. %, 97.3%, 97.4%, 97.5%, 97.6%, 97.7%, 97.8%, 97.9%, 98.0%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.65%, 98.7%, 98.8%, 98.9%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% or higher identity.

[0058] In one embodiment, the bacterial variant has the same function and / or therapeutic properties as the bacterium from which it is derived.

[0059] In some embodiments, the Disosmobacter bacteria and / or variants thereof included in the compositions for use according to the present invention are live or killed bacteria.

[0060] Within the scope of the present invention, the term "live bacteria" is used interchangeably with the term "viable bacteria" and refers to bacteria that are capable of growth, as opposed to "dead bacteria" (i.e., "nonviable bacteria"), which are unable to grow. Methods for measuring viability and growth rate are known to those skilled in the art. For example, bacterial viability and growth rate may be assessed by spreading a solution containing at least one bacterium of the present invention over an entire petri dish and counting the number of colonies after incubation under optimal growth conditions for a period of time. Alternatively, bacteria may be grown in liquid medium, and growth rate may be measured by measuring the optical density of the bacterial culture after incubation under optimal growth conditions for a period of time. The number of bacteria, including viable and nonviable bacteria, can also be determined by microscopic observation. Phase contrast microscopy is a well-known method for doing so, but the bacteria of a microorganism can be further visualized by specific staining with dyes, fluorescent probes, or antibodies to facilitate microscopic observation or to count the bacteria by flow cytometry.

[0061] In some embodiments, the Disosmobacter bacteria and / or variants thereof included in the compositions for use according to the invention are viable bacteria, hi some embodiments, the bacteria are metabolically active.

[0062] In some embodiments, the Disosmobacter bacteria and / or variants thereof included in the compositions for use according to the invention are killed bacteria. In some embodiments, the bacteria and / or variants thereof are incapable of proliferation. In some embodiments, the bacteria and / or variants thereof are metabolically inactive. In some embodiments, the bacteria and / or variants thereof are heat-inactivated or heat-killed.

[0063] Heat-inactivated or heat-killed bacteria can be obtained by heating to a temperature of at least about 90°C, preferably at least about 100°C, 105°C, 110°C, 115°C, or 120°C, more preferably at least about 121°C, 125°C, 130°C, 135°C, 140°C, or higher. Heating may be carried out for at least about 5 minutes, preferably at least about 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, or more. Heating may be carried out using a saturated steam pressure of at least about 10 psig, preferably at least about 11, 12, 13, 14, 15 psig, or higher.

[0064] In some embodiments, the Disosmobacter bacteria and / or variants thereof included in the composition for use according to the present invention are pasteurized bacteria.

[0065] In one embodiment, the pasteurized bacteria and / or variants thereof have been heated at a temperature ranging from about 50°C to about 100°C, preferably from about 60°C to about 95°C, more preferably from about 70°C to about 90°C. In one embodiment, the pasteurized bacteria and / or variants thereof have been heated at a temperature of about 50, 51, 52, 53, 54, 55, 56, 57, 58 or 59°C. In another embodiment, the pasteurized bacteria and / or variants thereof have been heated at a temperature of about 60, 61, 62, 63, 64, 65, 66, 67, 68 or 69°C. In yet another embodiment, the pasteurized bacteria and / or variants thereof have been heated at a temperature of about 70, 71, 72, 73, 74, 75, 76, 77, 78 or 79°C. In yet another embodiment, the pasteurized bacteria and / or variants thereof have been heated at a temperature of about 80, 81, 82, 83, 84, 85, 86, 87, 88 or 89°C. In yet another embodiment, the pasteurized bacteria and / or variants thereof have been heated to a temperature of about 90, 91, 92, 93, 94, 95, 96, 97, 98, 99°C or 100°C.

[0066] In one embodiment, the pasteurized bacteria and / or variants thereof have not been heated to a temperature of about 100°C or higher. In certain embodiments, the pasteurized bacteria and / or variants thereof have not been heated to an ultra-high temperature, e.g., a temperature in the range of about 110°C to about 140°C. In one embodiment, the pasteurized bacteria and / or variants thereof have not been heated to a temperature of about 90°C or higher. Thus, in one embodiment of the present invention, the bacteria and / or variants thereof have not been sterilized. Sterilization is a process intended to destroy, kill, or inactivate all living organisms and other biological agents. This includes microorganisms and their spores as well as viruses and prions. Unlike sterilization, pasteurization does not aim to kill all microorganisms, but is typically used in foods to reduce the number of viable pathogens.

[0067] In one embodiment of the invention, the pasteurized bacteria and / or variants thereof have been heated for at least about 10 minutes. In another embodiment of the invention, the pasteurized bacteria and / or variants thereof have been heated for at least about 15, 20, 25, 30, 35, or 45 minutes. In one embodiment, the pasteurized bacteria and / or variants thereof have been heated for about 10 to about 45 minutes.

[0068] In one embodiment, the pasteurized bacteria and / or variants thereof have not been heated for a short period of time. In particular embodiments, the pasteurized bacteria and / or variants thereof have not been heated for less than about 30 seconds, less than about 60 seconds, less than about 90 seconds, or less than about 120 seconds. In preferred embodiments, the pasteurized bacteria and / or variants thereof have not been heated for less than about 1 minute, preferably less than about 5, 6, 7, 8, or 9 minutes.

[0069] In one embodiment, the pasteurized bacteria and / or variants thereof have been heated to a temperature ranging from about 50°C to about 100°C for at least about 10 minutes. In a particular embodiment, the pasteurized bacteria and / or variants thereof have been heated to about 60°C for about 20 minutes or about 30 minutes. In another particular embodiment, the pasteurized bacteria and / or variants thereof have been heated to about 70°C for about 20 minutes or about 30 minutes. In another particular embodiment, the pasteurized bacteria and / or variants thereof have been heated to about 80°C for about 20 minutes or about 30 minutes. In another particular embodiment, the pasteurized bacteria and / or variants thereof have been heated to about 90°C for about 20 minutes or about 30 minutes.

[0070] In certain embodiments, the pasteurized bacteria and / or variants thereof have not been heated to a temperature of about 110° C. or greater for about 1 to about 120 seconds. In another particular embodiment, the pasteurized bacteria and / or variants thereof have not been heated to a temperature of about 100° C. or greater for about 1 to about 120 seconds. In another particular embodiment, the pasteurized bacteria and / or variants thereof have not been heated to a temperature of about 90° C. or greater for about 1 to about 120 seconds.

[0071] In one embodiment, the bacterium and / or variants thereof have been treated with ultra-high temperature (UHT) treatment.

[0072] As used herein, "UHT" treatment refers to ultra-high temperature treatment or ultra-heat treatment (both abbreviated as UHT) that at least partially sterilizes a composition by heating it to a temperature of at least about 135°C for a short period of time, such as from about 1 to about 60 seconds, preferably from about 1 to about 30 seconds, and more preferably from about 1 to about 10 seconds.

[0073] There are two main types of UHT systems: direct and indirect. In direct systems, the product is treated by steam injection, while indirect systems heat treat the product using plate, tubular, or scraped surface heat exchangers. A combination of UHT systems may be applied at any step or steps in the product preparation process.

[0074] In one embodiment, the bacteria and / or variants thereof are flash killed, i.e., in one embodiment, the bacteria and / or variants thereof are treated at a temperature in the range of about 71.5°C to about 74°C for a period in the range of about 15 to about 30 seconds.

[0075] In some embodiments, the Disosmobacter bacteria and / or variants thereof included in the compositions for use according to the invention are fresh, the term "fresh" as used herein meaning that the bacteria have not been frozen between the last amplification step and use.

[0076] In some embodiments, the Disosmobacter bacteria and / or variants thereof included in the compositions for use according to the invention are not fresh. In some embodiments, the bacteria and / or variants thereof have been frozen at least once. In some embodiments, the bacteria and / or variants thereof have been frozen.

[0077] As used herein, the term "frozen (has been frozen)" refers to bacteria that have been cooled to a temperature at or below that allows for a liquid to solid phase transition in the bacteria. In one embodiment, the temperature is about -5°, -20°C, -70°C, -80°C, or -190°C.

[0078] In one embodiment, the recovered frozen bacteria and / or variants thereof are viable, in other words, in one embodiment, the Disosmobacter bacteria and / or variants thereof included in the composition for use according to the invention are frozen and viable.

[0079] In some embodiments, the composition for use according to the present invention comprises a fragment of bacteria of the genus Disosmobacter, preferably a fragment of Disosmobacter welterbionis, more preferably Disosmobacter welterbionis strain J115. T Contains fragments of.

[0080] The term "fragment" as used herein refers to bacterial components, metabolic products, secreted molecules and / or vesicles, and compounds obtained by metabolism of bacteria and / or variants thereof that are included in the compositions for use according to the present invention. Examples of bacterial components include, but are not limited to, bacterial cell wall components such as peptidoglycan, bacterial nucleic acids such as DNA and RNA, bacterial membrane components, bacterial structural components such as proteins, carbohydrates, and lipids, and combinations thereof such as lipoproteins, glycolipids, and glycoproteins, bacterial metabolites, metabolic products including organic acids, inorganic acids, bases, peptides, enzymes, coenzymes, amino acids, carbohydrates, lipids, glycoproteins, lipoproteins, glycolipids, vitamins, biologically active compounds, and inorganic components. The fragment may be obtained by collecting the supernatant of a culture of the bacterium and / or its variants, or by extracting cellular components or cell fractions, metabolites or secreted compounds from a culture of the bacterium and / or its variants, degradation products, components in isolated form, any mixture of one or more components obtained from the bacterium and / or its variants, or one or more components present in the bacterium and / or its variants produced otherwise, for example using recombinant DNA technology, in a microbial host or any other (bio)synthetic process. In one embodiment, the fragment is contained in the culture medium of the bacterium, i.e., the fragment is not purified or isolated. In another embodiment, the fragment has been purified by methods known in the art, for example by affinity purification.

[0081] In some embodiments, compositions for use according to the invention comprise an extract of the supernatant of a culture medium of bacteria of the genus Disosmobacter and / or variants thereof.

[0082] Suitable media for bacterial culture are known in the art. In some embodiments, particularly for the culture of bacteria of the genus Disosmobacter, the medium is yeast extract-casein hydrolysate-fatty acid (YCFA) medium. In some embodiments, the YCFA medium is modified.

[0083] In some embodiments, the YCFA medium contains yeast extract in an amount of about 8 g / L, KH2PO4 in an amount of about 2 g / L, sodium propionate in an amount of about 1 g / L, cysteine in an amount of about 0.5 g / L, hemin in an amount of about 1 g / L, L-cysteine in an amount of about 1 g / L, and a vitamin solution as described below. In some embodiments, the modified YCFA medium additionally or alternatively contains soy peptone in an amount ranging from greater than 0 g / L to about 20 g / L, preferably from about 2 g / L to about 6 g / L, and more preferably about 4 g / L. In some embodiments, the modified YCFA medium contains wheat peptone in an amount ranging from greater than 0 g / L to about 20 g / L, preferably from about 2 g / L to about 6 g / L, and more preferably about 4 g / L. In some embodiments, the modified YCFA medium contains NaCO in an amount ranging from greater than 0 g / L to about 20 g / L, preferably from about 2 g / L to about 6 g / L, and more preferably about 3 g / L. In some embodiments, the modified YCFA medium contains MgCl in an amount ranging from greater than 0 mg / L to about 500 mg / L, preferably from about 25 mg / L to about 75 mg / L, and more preferably about 50 mg / L. In some embodiments, the modified YCFA medium contains glutathione in an amount ranging from greater than 0 g / L to about 5 g / L, preferably from about 0.5 g / L to about 1.5 g / L, and more preferably about 1 g / L. In some embodiments, the modified YCFA medium contains ascorbic acid in an amount ranging from greater than 0 g / L to about 1 g / L, preferably from about 0.25 g / L to about 0.75 g / L, and more preferably about 0.5 g / L. In some embodiments, the modified YCFA medium comprises soy peptone in an amount of about 4 g / L, wheat peptone in an amount of about 4 g / L, Na2CO3 in an amount of about 3 g / L, glutathione in an amount of about 1 g / L, ascorbic acid in an amount of about 1 g / L, and ascorbic acid in an amount of about 0.5 g / L.

[0084] In one embodiment, modified YCFA medium contains 8 g yeast extract, 4 g soy peptone, 4 g wheat peptone, 5 g KH2PO4, 3 g Na2CO3, 50 mg MgCl2, 50 mg CaCl2, 1 mg hemin, 1 mL resazurin solution (1 g / L), 10 g myo-inositol, 1 g cysteine, 1 g glutathione (reduced), 0.5 g ascorbate, 0.3 g uric acid, 1 mL vitamin solution, and qs 1000 mL HO. In one embodiment, the vitamin solution used to prepare modified YCFA medium contains 2 mg biotin, 2 mg folic acid, 10 mg pyridoxine-HCl, 2 mg thiamine-HCl x 2H, 5 mg riboflavin, 5 mg nicotinic acid, 5 mg D-Ca-pantothenate, 12 mg vitamin B, 5 mg p-aminobenzoic acid, 5 mg lipoic acid, and qs to 1000 mL HO.

[0085] In some embodiments, the modified YCFA medium comprises soy peptone in an amount of about 4 g / L, wheat peptone in an amount of about 4 g / L, Na2CO3 in an amount of about 3 g / L, glutathione in an amount of about 1 g / L, ascorbic acid in an amount of about 1 g / L, ascorbic acid in an amount of about 0.5 g / L, MgCl2 in an amount of 0.5 g / L, yeast extract in an amount of 8 g / L, KH2PO4 in an amount of 2 g / L, sodium propionate in an amount of 1 g / L, cysteine in an amount of 0.5 g / L, hemin in an amount of 1 g / L, L-cysteine in an amount of 1 g / L, and a vitamin solution as described hereinabove.

[0086] In one embodiment, the medium does not contain viable bacteria, i.e., the medium is sterile. Means for sterilizing a medium are known in the art and include, for example, heating and irradiation. In another embodiment, the medium contains a substantially pure population of viable bacteria, preferably bacteria of the genus Disosmobacter and / or variants thereof.

[0087] In some embodiments, the medium is frozen and stored at a temperature of about -5°, -20°C, -70°C, -80°C, or -190°C for future use.

[0088] In some embodiments, the medium is fermented. In some embodiments, the fermentation of the medium comprises: 1) About 10 8 ~about 10 9 of the genus Disosmobacter, preferably Disosmobacter welbrionis, more preferably Disosmobacter welbrionis strain J115 T incubating the live bacteria in 50 mL of modified YCFA medium under anaerobic conditions at a temperature of 37°C for 48 hours; 2) incubating the culture of step 1) in 3.5 L of modified YCFA medium under anaerobic conditions at a temperature of 37°C for 48 hours; 3) recovering the fermentation medium from step 2); and 4) Optionally, removing the viable bacteria from the medium collected in step 3) (i.e., sterilizing the medium). Includes:

[0089] In some embodiments, the extract from a bacterium of the genus Disosmobacter and / or a variant thereof or a culture supernatant included in a composition for use according to the present invention comprises at least one metabolite secreted by the bacterium.

[0090] In some embodiments, the composition for use according to the present invention comprises a strain of the genus Disosmobacter, preferably Disosmobacter welbrionis, more preferably Disosmobacter welbrionis strain J115. T These include metabolic products produced and / or secreted by bacteria.

[0091] In some embodiments, the at least one metabolite is selected from the group comprising or consisting of short chain fatty acids (SCFAs), such as butyrate and acetate.

[0092] In some embodiments, the at least one metabolite is selected from the group consisting of butyrate, acetate, C8-3OH, C10-3OH, C12-3OH, C16-2OH, C16-3OH, C18-2OH, C18-3OH, C18-130H, C18, C18:2n-6, C22:2n-6, C22:6n-3, C18:1n-9c, C22:1n-9, C18:2n-6, 10-TriHOME, 10-TriHOME, C12asn, C12asnGABAOH, 9,10-DiHOME, 12,13-DiHOME, 13-oxoODE, 8-HETE, and 9-HODE. In some embodiments, the at least one metabolite is butyrate. In some embodiments, the at least one metabolite is acetate.

[0093] The present inventors have identified the genus Disosmobacter, preferably Disosmobacter welbrionis, more preferably Disosmobacter welbrionis strain J115. T We have demonstrated that the effect of the bacteria is not limited to the metabolic products produced and / or secreted by the bacteria. In particular, we have demonstrated that this effect is not limited to the presence or secretion of short-chain fatty acids (SCFAs), such as butyrate and acetate. Indeed, our results show that the medium obtained after fermentation of the bacteria appears to completely inhibit cell growth at a concentration of 20% (corresponding to a 10- to 20-fold higher potency than SCFAs) (see Figures 5B and 5D, "SCFA-matched medium" vs. "fermented medium").

[0094] In some embodiments, the composition for use according to the invention comprises an active substance from the culture supernatant of a bacterium of the genus Disosmobacter and / or a variant thereof, hi one embodiment, the active substance is a metabolite produced and / or secreted by the bacterium of the invention as described herein above.

[0095] Another object of the present invention is a culture supernatant of bacteria of the genus Disosmobacter and / or variants thereof for use in preventing and / or treating breast cancer in a subject in need thereof.

[0096] In one embodiment, the culture supernatant is obtained by collecting the culture medium of the bacteria of the genus Disosmobacter and / or variants thereof. In one embodiment, the culture supernatant is the fermentation medium obtained after culturing the bacteria of the invention. In one embodiment, the culturing of the bacteria of the genus Disosmobacter and / or variants thereof is carried out as disclosed herein above.

[0097] In certain embodiments, the culture supernatant is subjected to the following steps: 1) About 10 8 ~about 10 9 Disosmobacter spp., preferably Disosmobacter welbrionis, more preferably Disosmobacter welbrionis strain J115 T incubating the live bacteria in 50 mL of modified YCFA medium under anaerobic conditions at a temperature of 37°C for 48 hours; 2) incubating the culture of step 1) in 3.5 L of modified YCFA medium under anaerobic conditions at a temperature of 37°C for 48 hours; 3) recovering the fermentation medium from step 2); and 4) Optionally, removing the viable bacteria from the harvested medium of step 3). Obtained by.

[0098] As used herein, the term "breast cancer" refers to a histologically or cytologically confirmed cancer of the breast. In some embodiments, the breast cancer is a carcinoma. In some embodiments, the breast cancer is an adenocarcinoma. In some embodiments, the breast cancer is a sarcoma.

[0099] In certain embodiments, the breast cancer is hormone receptor positive (HR+) breast cancer or hormone receptor negative (HR-) breast cancer. In one embodiment, the hormone receptor positive breast cancer is estrogen receptor positive (ER+) breast cancer and / or progesterone receptor positive (PR+) breast cancer. In some embodiments, the ER+ breast cancer is luminal A breast cancer. In some embodiments, the ER+ breast cancer is luminal B breast cancer.

[0100] In some embodiments, the breast cancer is human epidermal growth factor receptor 2 positive (HER2+) breast cancer. In some embodiments, the breast cancer is human epidermal growth factor receptor 2 negative (HER2-) breast cancer.

[0101] In some embodiments, the breast cancer is a cancer with mixed receptor expression, eg, a hormone receptor positive, HER2 negative (HR+HER2-) cancer.

[0102] In some embodiments, the breast cancer is a Group 1 (Luminous A), Group 2 (Luminous B), Group 3 (HER2+), or Group 4 (Basal-like) cancer. Group 1 includes tumors that are ER+, progesterone receptor positive (PR+), but HER2 negative (HR+HER2-). Group 2 includes tumors that are ER+, PR-, and HER2+. Group 3 includes tumors that are ER-, PR-, but HER2+. Group 4, also known as TNBC, includes tumors that are ER-, PR-, and HER2-.

[0103] In some embodiments, the breast cancer is HER2+ breast cancer or triple-negative breast cancer (TNBC). In some embodiments, the breast cancer is HER2+ breast cancer, preferably HER2+ breast adenocarcinoma. In some embodiments, the breast cancer is triple-negative breast cancer (TNBC).

[0104] In some embodiments, the breast cancer is non-invasive breast cancer, particularly ductal carcinoma in situ or lobular carcinoma in situ. In particular embodiments, the breast cancer is an invasive breast cancer, particularly selected from the group including or consisting of invasive ductal carcinoma, invasive lobular carcinoma, Paget's disease of the nipple, inflammatory breast cancer, phyllodes tumor of the breast, locally advanced breast cancer, and metastatic breast cancer.

[0105] In some embodiments, the tissue types in which breast cancer arises are the milk ducts, milk-producing lobules, or connective tissue.

[0106] In some embodiments, the breast cancer is metastatic or locally advanced breast cancer. The term "locally advanced breast cancer" refers to cancer that has spread from where it began in the breast to nearby tissues or lymph nodes, but has not spread to other parts of the body.

[0107] In a particular embodiment, the cancer is a metastatic cancer, ie, a cancer prone to metastasis, particularly metastatic breast cancer or breast cancer prone to metastasis.

[0108] The term "metastatic breast cancer" refers to cancer that has spread from the breast to other parts of the body, such as the bone, liver, lung, or brain. Metastatic breast cancer is sometimes referred to as stage IV breast cancer. As used herein, the term "cancer prone to metastasis" refers to an invasive cancer whose cancer cells can break away from the primary tumor and spread to other organs, where they can grow and form secondary tumors, also called metastases.

[0109] Many pathophysiological factors are known in the art that can influence tumor growth, particularly conditions related to diet, obesity, glucose and lipid metabolism, and inflammation. The ability of commensal bacteria, including bacteria of the genus Disosmobacter, to influence these conditions makes them suitable within the scope of the present invention.

[0110] In some embodiments, the subject further suffers from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity-related disorders, liver diseases, metabolic disorders, adipokine-related disorders, and inflammatory disorders, and combinations thereof. In some embodiments, the one or more diseases or conditions that accelerate tumor growth are selected from the group including or consisting of obesity-related disorders, metabolic disorders, and inflammatory disorders, and combinations thereof. In some embodiments, the one or more diseases or conditions that accelerate tumor growth are selected from the group including or consisting of obesity-related disorders and metabolic disorders.

[0111] In some embodiments, the subject further suffers from an obesity-related disorder. In some embodiments, the subject further suffers from liver disease. In some embodiments, the subject further suffers from a metabolic disorder. In some embodiments, the subject further suffers from an adipokine-related disorder. In some embodiments, the subject further suffers from an inflammatory disease.

[0112] In some embodiments, the subject does not suffer from an obesity-related disorder, hi some embodiments, the subject has a normal BMI index.

[0113] In some embodiments, the subject further suffers from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFL), cirrhosis, diabetes, impaired glucose tolerance, hyperglycemia, dyslipidemia, dyslipidemia, high cholesterol, elevated LDL cholesterol levels, low HDL cholesterol levels, elevated triglyceride levels, and enterocolitis, and combinations thereof.

[0114] In some embodiments, the subject is further affected by obesity. In some embodiments, the subject is further affected by NASH. In some embodiments, the subject is further affected by NAFL. In some embodiments, the subject is further affected by cirrhosis. In some embodiments, the subject is further affected by diabetes. In some embodiments, the subject is further affected by impaired glucose tolerance. In some embodiments, the subject is further affected by hyperglycemia. In some embodiments, the subject is further affected by dyslipidemia. In some embodiments, the subject is further affected by dyslipidemia. In some embodiments, the subject is further affected by high cholesterol. In some embodiments, the subject is further affected by elevated LDL cholesterol levels. In some embodiments, the subject is further affected by reduced HDL cholesterol levels. In some embodiments, the subject is further affected by elevated triglyceride levels. In some embodiments, the subject is further affected by enteritis.

[0115] In some embodiments, the subject suffers from an imbalance in the gut microbiota. As used herein, an imbalance in the gut microbiota refers to the over- or under-representation of at least one microbial species, preferably at least one bacterial species. In some embodiments, the population of bacteria of the genus Disosmobacter is under-represented. In some embodiments, the population of Disosmobacter welterbionis is under-represented. In some embodiments, the population of Disosmobacter welterbionis is reduced in subjects suffering from obesity and / or obesity-related disorders.

[0116] In some embodiments, the population of Disosmobacter welterbionis is inversely correlated with the body mass index of the subject.

[0117] In some embodiments, the compositions for use according to the present invention are administered in a therapeutically effective amount, preferably about 1×10 2 ~Approx. 1×10 15 CFU of said bacteria.

[0118] As used herein, "CFU" means "colony forming unit."

[0119] In some embodiments, the compositions for use according to the present invention are administered in a therapeutically effective amount, preferably about 1×10 4 ~Approx. 1×10 12 CFU, more preferably about 1 x 10 5 ~Approx. 1×10 10 CFU, and even more preferably about 1 x 10 6 ~Approx. 5×10 9 CFU of said bacteria.

[0120] In some embodiments, the compositions for use according to the present invention are administered in a therapeutically effective amount, preferably about 1×10 2 ~Approx. 1×10 15 CFU, approximately 1 x 10 4 ~Approx. 1×10 12 CFU, approximately 1 x 10 5 ~Approx. 1×10 10 CFU, approximately 1 x 10 6 ~Approx. 5×10 9 In some embodiments, the compositions for use according to the invention comprise a therapeutically effective amount, preferably about 1 x 10 CFU of said bacteria. 4 ~Approx. 1×10 14 CFU, approximately 1 x 10 5 ~Approx. 1×10 13 CFU, approximately 1 x 10 6 ~Approx. 1×10 12 CFU, approximately 1 x 10 7 ~Approx. 1×10 11 CFU, approximately 1 x 10 8 ~Approx. 1×10 10 CFU, approximately 2 x 10 8 ~Approx. 6×10 9 CFU of said bacteria.

[0121] In some embodiments, the compositions for use according to the present invention are administered in a therapeutically effective amount, preferably about 1×10 4 ~Approx. 1×10 12 CFU / mL, more preferably about 1 x 10 5 ~Approx. 1×1010 CFU / mL, and even more preferably about 1 x 10 6 ~Approx. 5×10 9 CFU / mL of the bacteria.

[0122] In some embodiments, the compositions for use according to the present invention are administered in a therapeutically effective amount, preferably about 1×10 4 ~Approx. 1×10 14 CFU / mL, preferably about 1 x 10 5 ~Approx. 1×10 13 CFU / mL, more preferably about 1 x 10 6 ~Approx. 1×10 12 CFU / mL, and even more preferably about 1 x 10 7 ~Approx. 1×10 11 CFU / mL, 1 × 10 8 ~Approx. 1×10 10 CFU / mL, and even more preferably about 2 x 10 8 ~Approx. 6×10 9 CFU / mL of the bacteria.

[0123] In some embodiments, the compositions for use according to the present invention are administered in a therapeutically effective amount, preferably about 1×10 2 ~Approx. 1×10 15 CFU / g, preferably about 1 x 10 4 ~Approx. 1×10 12 CFU / g, more preferably about 1 x 10 5 ~Approx. 1×10 10 CFU / g, and even more preferably about 1 x 10 6 ~about 5.10 9 In some embodiments, the compositions for use according to the invention contain a therapeutically effective amount of the bacteria, preferably about 1 x 10 CFU / g. 4 ~Approx. 1×10 14 CFU / g, preferably about 1 x 10 5 ~Approx. 1×10 13 CFU / g, more preferably about 1 x 10 6 ~Approx. 1×10 12 CFU / g, and even more preferably about 1 x 10 7 ~Approx. 1×10 11 CFU / g, approximately 1 x 10 8~Approx. 1×10 10 CFU / g, and even more preferably about 2 x 10 8 ~Approx. 6×10 9 CFU / g of said bacteria.

[0124] In some embodiments, the compositions for use according to the present invention are administered in a therapeutically effective amount, preferably about 1×10 6 ~Approx. 1×10 10 CFU / g or CFU / mL, preferably about 1 x 10 8 ~Approx. 1×10 10 CFU / g or CFU / mL, more preferably about 1 x 10 9 ~Approx. 1×10 10 In some embodiments, the compositions for use according to the invention contain a therapeutically effective amount of the bacteria, preferably about 1 x 10 CFU / g or CFU / mL. 6 ~Approx. 1×10 11 CFU / g or CFU / mL, preferably about 1 x 10 8 ~Approx. 1×10 11 CFU / g or CFU / mL, more preferably about 1 x 10 10 ~Approx. 1×10 11 Contains CFU / g or CFU / mL of the bacteria.

[0125] In one embodiment of the invention, the composition for use according to the invention comprises about 1 x 10 2 ~Approx. 1×10 15 cells / g, preferably about 1 x 10 4 ~Approx. 1×10 12 cells / g, more preferably about 1 x 10 5 ~Approx. 1×10 10 cells / g, even more preferably about 1 x 10 6 ~Approx. 1×10 9 In one embodiment of the invention, the composition for use according to the invention comprises fragments of the bacterium and / or its variants in an amount corresponding to an amount of the bacterium and / or its variants in the range of cells / g. 4 ~Approx. 1×10 14 cells / g, preferably about 1 x 10 5 ~Approx. 1×10 13cells / g, more preferably about 1 x 10 6 ~Approx. 1×10 12 cells / g, even more preferably about 1 x 10 7 ~Approx. 1×10 11 In one embodiment of the invention, the composition for use according to the invention comprises fragments of the bacterium and / or its variants in an amount corresponding to an amount of the bacterium and / or its variants in the range of cells / g. 8 ~Approx. 1×10 10 cells / g, preferably about 1 x 10 9 ~Approx. 1×10 10 It contains fragments of said bacteria and / or variants thereof in an amount corresponding to an amount of said bacteria and / or variants thereof in the range of cells / g.

[0126] In one embodiment of the invention, the composition for use according to the invention comprises about 1 x 10 2 ~Approx. 1×10 15 cells / mL, preferably about 1 x 10 4 ~Approx. 1×10 12 cells / mL, more preferably about 1 x 10 5 ~Approx. 1×10 10 cells / mL, even more preferably about 1 x 10 6 ~Approx. 1×10 9 In one embodiment of the invention, the composition for use according to the invention comprises fragments of the bacterium and / or its variants in an amount corresponding to an amount of the bacterium and / or its variants in the range of cells / mL. 4 ~Approx. 1×10 14 cells / mL, preferably about 1 x 10 5 ~Approx. 1×10 13 cells / mL, more preferably about 1 x 10 6 ~Approx. 1×10 12 cells / mL, even more preferably about 1 x 10 7 ~Approx. 1×10 11 In one embodiment of the invention, the composition for use according to the invention comprises fragments of the bacterium and / or its variants in an amount corresponding to an amount of the bacterium and / or its variants in the range of cells / mL. 8 ~Approx. 1×10 10cells / mL, preferably about 1 x 10 9 ~Approx. 1×10 10 It contains fragments of the bacterium and / or its variants in an amount corresponding to an amount of the bacterium and / or its variants in the range of cells / mL.

[0127] In one embodiment of the invention, the composition for use according to the invention comprises about 1 x 10 6 ~Approx. 1×10 10 cells / g or cells / mL, preferably about 1 x 10 8 ~Approx. 1×10 10 cells / g or cells / mL, more preferably about 1 x 10 9 ~Approx. 1×10 10 It contains fragments of the bacteria in an amount corresponding to the amount of bacteria in the range of cells / g or cells / mL.

[0128] In one embodiment of the invention, the composition for use according to the invention comprises about 1 x 10 6 ~Approx. 1×10 11 cells / g or cells / mL, preferably about 1 x 10 8 ~Approx. 1×10 11 cells / g or cells / mL, more preferably about 1 x 10 10 ~Approx. 1×10 11 It contains fragments of the bacteria in an amount corresponding to the amount of bacteria in the range of cells / g or cells / mL.

[0129] In some embodiments, the composition for use according to the invention is in the form of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

[0130] Of course, the carrier must be "acceptable" in the sense of being compatible with the composition for use according to the invention and not harmful when administered to an individual: typically, the carrier does not produce an adverse, allergic or other untoward reaction when administered to an individual, preferably a human individual.

[0131] In some embodiments, the composition for use according to the present invention or the pharmaceutical composition for use according to the present invention further comprises at least another anti-cancer agent.

[0132] Anticancer drugs are known from the state of the art. Non-limiting examples of anticancer drugs include acalabrutinib, alectinib, alemtuzumab, anastrozole, avapritinib, avelumab, belinostat, bevacizumab, bleomycin, blinatumomab, bosutinib, brigutinib, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, copanlisib, cytarabine, daunorubicin, decitabine, dexamethasone, docetaxel, doxorubicin, encorafenib, erdafitinib, etoposide, everolimus, exe These include mestan, fludarabine, 5-fluorouracil, gemcitabine, ifosfamide, imatinib mesylate, leuprolide, lomustine, mechlorethamine, melphalan, methotrexate, mitomycin, nelarabine, paclitaxel, pamidronate, panobinostat, pralatrexate, prednisolone, ofatumumab, rituximab, temozolomide, topotecan, tositumomab, trastuzumab, vandetanib, vincristine, vorinostat, and zanubrutinib.

[0133] In a preferred embodiment, the anti-cancer agent is an anti-cancer agent suitable for treating breast cancer.

[0134] In certain embodiments, the anti-cancer agent may be combined with or administered simultaneously or sequentially with a composition for use according to the present invention or a pharmaceutical composition for use according to the present invention.

[0135] The present invention further relates to a medicament comprising the composition or pharmaceutical composition for use according to the present invention, in some embodiments, the medicament is for treating and / or preventing breast cancer.

[0136] The present invention provides at least (a) a strain of the genus Disosmobacter, preferably the species Disosmobacter wellbionis, more preferably the strain Disosmobacter wellbionis J115, for use in the manufacture of a medicament for treating and / or preventing breast cancer in a subject in need thereof. Tand / or variants and / or fragments thereof, and / or (b) bacteria of the genus Disosmobacter, preferably the species Disosmobacter wellbionis, more preferably the strain Disosmobacter wellbionis J115. T The present invention also relates to a composition comprising the supernatant of a culture of the bacterium and / or a variant thereof.

[0137] Another object of the present invention is to provide a strain of the genus Disosmobacter, preferably the species Disosmobacter wellbionis, more preferably the strain Disosmobacter wellbionis J115, for use in the manufacture of a medicament for treating and / or preventing breast cancer in a subject in need thereof. T and / or variants and / or fragments thereof.

[0138] Yet another object of the present invention is to provide a strain of the genus Disosmobacter, preferably the species Disosmobacter wellbionis, more preferably the strain Disosmobacter wellbionis J115, for use in the manufacture of a medicament for treating and / or preventing breast cancer in a subject in need thereof. T and / or variants thereof.

[0139] In some embodiments, the compositions for use according to the present invention are in the form of a nutraceutical composition further comprising a nutraceutically acceptable agent. In one embodiment, the nutraceutical composition comprises a nutraceutically effective amount of bacteria of the genus Disosmobacter and / or a variant, extract, or fragment thereof.

[0140] In some embodiments, the dietary supplement compositions are for consumption by human users and may be useful for their prophylactic and pharmaceutical qualities. In one embodiment, the dietary supplement compositions of the present invention are for use in preventing and / or treating breast cancer in a subject in need thereof.

[0141] In one embodiment, the dietary supplement composition is in the form of a food or dietary supplement. In another embodiment, the dietary supplement composition is in the form of a functional food. In some embodiments, the dietary supplement composition is formulated as a food additive, a beverage, a tablet, a capsule, a caplet, a lozenge, a syrup, a suspension, or an emulsion.

[0142] In some embodiments, the nutraceutical composition further comprises one or more secondary nutraceutical ingredients that are nutritionally or therapeutically active in the prevention and / or treatment of breast cancer, hi some embodiments, the secondary nutraceutical ingredients are vitamins or minerals.

[0143] In some embodiments, the nutraceutically acceptable agent is selected from one or more of the group including talc, titanium dioxide, starch, corn starch, modified corn starch, kaolin, microcrystalline cellulose, and powdered cellulose.

[0144] The present invention further provides a method for the detection of the genus Disosmobacter, preferably the species Disosmobacter welbrionis, more preferably the strain Disosmobacter welbrionis J115, in the microbiome of a subject in need thereof. T The present invention relates to prebiotics comprising one or more active ingredients or substances that increase the levels of bacteria and / or variants and / or fragments thereof.

[0145] In one embodiment, the prebiotic is for use in preventing and / or treating breast cancer in said subject.

[0146] In some embodiments, the prebiotic is selected from the group comprising or consisting of myo-inositol, inulin and inulin-type fructans, oligofructose, beta-glucan, xylose, arabinose, arabinoxylan, ribose, phytic acid, galactose, rhamnose, cellobiose, fructose, lactose, salicin, sucrose, glucose, esculin, tween 80, trehalose, maltose, mannose, melibiose, mucus or mucin, raffinose, fructooligosaccharides, galactooligosaccharides, amino acids, alcohols, fermentable carbohydrates, water-soluble cellulose derivatives (e.g., methylcellulose, methylethylcellulose, hydroxyethylcellulose, ethylhydroxyethylcellulose, cationic hydroxyethylcellulose, hydroxypropylcellulose, hydroxyethylmethylcellulose, hydroxypropylmethylcellulose, and carboxymethylcellulose), water-insoluble cellulose derivatives, raw oatmeal, metamucil, all-bran, polyphenols, and any combination thereof.

[0147] The present invention also relates to therapeutic combination products for their separate, simultaneous or sequential administration.

[0148] As used herein, the term "therapeutic combination product" (sometimes referred to as a therapeutic kit-of-parts) refers to a product comprising or consisting of at least two of the following parts: a first part comprising a composition for use according to the invention, a pharmaceutical composition for use according to the invention, or a drug according to the invention, and a second part comprising a prebiotic. In one embodiment, the therapeutic combination product is for treating and / or preventing breast cancer.

[0149] In some embodiments, the prebiotic is included in a composition for use according to the invention, a pharmaceutical composition for use according to the invention, or a medicament according to the invention.

[0150] The present invention further provides a method for the treatment of breast cancer comprising administering to a subject a therapeutic agent comprising at least (a) a strain of the genus Disosmobacter, preferably the species Disosmobacter wellbionis, more preferably the strain Disosmobacter wellbionis J115, for use as an adjuvant in a therapeutic agent administered to a subject suffering from breast cancer. T and / or (b) a bacterium of the genus Disosmobacter, preferably the species Disosmobacter welbionis, more preferably the strain Disosmobacter welbionis J115. T The present invention relates to a composition comprising a culture supernatant of the bacterium and / or a variant thereof.

[0151] As used herein, "adjuvant" refers to an agent that enhances the effect of a therapeutic agent, typically a breast cancer therapeutic agent. "Enhancing" means that the positive effect of a therapeutic agent used with an adjuvant on a disease or condition, preferably breast cancer, exceeds the combined effect of the therapeutic agent and adjuvant used individually.

[0152] In some embodiments, the breast cancer in the subject is treated by methods known in the art, such as hormone therapy, chemotherapy, and immunotherapy.

[0153] In some embodiments, the subject further suffers from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity-related disorders, liver disease, metabolic disorders, adipokine-related disorders, and inflammatory diseases, and combinations thereof. In some embodiments, the subject further suffers from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), cirrhosis, diabetes, impaired glucose tolerance, hyperglycemia, dyslipidemia, dyslipidemia, high cholesterol, elevated LDL cholesterol levels, low HDL cholesterol levels, elevated triglyceride levels, and enterocolitis, and combinations thereof.

[0154] In some embodiments, the subject suffers from an imbalance in the gut microbiota, hi some embodiments, the subject's gut microbiota comprises a reduced population of bacteria of the genus Disosmobacter, preferably Disosmobacter welbrionis, compared to a healthy subject.

[0155] The present invention further provides a method for treating a disease comprising administering to a subject a therapeutically effective amount of at least (a) the genus Disosmobacter, preferably the species Disosmobacter welbrionis, more preferably the strain Disosmobacter welbrionis J115 T and / or a bacterium of the genus Disosmobacter, preferably the species Disosmobacter wellbionis, more preferably the strain Disosmobacter wellbionis J115. T The present invention relates to a method for treating and / or preventing breast cancer in a subject in need thereof, comprising administering to the subject a composition comprising the supernatant of a culture of the bacterium and / or a variant thereof.

[0156] Therapeutically effective amounts of the compositions are described herein above.

[0157] In some embodiments, the method further comprises administering to the subject a therapeutically effective amount of another anti-cancer agent. Examples of other anti-cancer agents are described herein above. Accordingly, the present invention also relates to a method for treating and / or preventing breast cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising (i) a composition comprising a bacterium of the genus Disosmobacter and / or a variant, extract, or fragment thereof, and (ii) another therapeutic agent that is an anti-cancer agent.

[0158] In one embodiment, the method is used when the subject has breast cancer and is intended to slow or reverse the progression of tumor growth. In another embodiment, the method is used when the subject does not have breast cancer and is intended to prevent the onset of breast cancer. In one embodiment, the method is used acutely. In another embodiment, the method is used chronically.

[0159] In some embodiments, the subject further suffers from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity-related disorders, liver disease, metabolic disorders, adipokine-related disorders, and inflammatory diseases, and combinations thereof. In some embodiments, the subject further suffers from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), cirrhosis, diabetes, impaired glucose tolerance, hyperglycemia, dyslipidemia, dyslipidemia, high cholesterol, elevated LDL cholesterol levels, low HDL cholesterol levels, elevated triglyceride levels, and enterocolitis, and combinations thereof.

[0160] In some embodiments, the subject suffers from an imbalance in the gut microbiota, hi some embodiments, the subject's gut microbiota comprises a reduced population of bacteria of the genus Disosmobacter, preferably Disosmobacter welbrionis, compared to a healthy subject.

[0161] The present invention further provides a strain of at least (a) the genus Disosmobacter, preferably the species Disosmobacter welbrionis, more preferably the strain Disosmobacter welbrionis J115, for use as a probiotic. T and / or variants thereof, and / or (b) a bacterium of the genus Disosmobacter, preferably the species Disosmobacter wellbionis, more preferably the strain Disosmobacter wellbionis J115. T In a specific embodiment, the bacteria contained in the composition is strain J115. T and / or variants thereof, which are advantageous for improving the intestinal environment of a subject.

[0162] The present invention relates to the intestinal microbiota of a subject, and to the intestinal microbiota of the genus Disosmobacter, preferably Disosmobacter welbrionis, more preferably Disosmobacter welbrionis strain J115.T The present invention also relates to a method for increasing the population of a bacterium of the present invention, comprising administering the bacterium to a subject, wherein the bacterium is contained in a composition, pharmaceutical composition, or prebiotic. In a preferred embodiment, the subject is at risk of developing or has breast cancer.

[0163] The present invention relates to at least (a) a strain of the genus Disosmobacter, preferably Disosmobacter welbrionis, more preferably Disosmobacter welbrionis strain J115 T and / or (b) a bacterium of the genus Disosmobacter, preferably the species Disosmobacter welbrionis, more preferably the strain Disosmobacter welbrionis J115 T The present invention also relates to a method for treating an imbalance in the gut microbiota, comprising administering to a subject a composition or prebiotic comprising the supernatant of a culture of bacteria derived therefrom and / or variants thereof. [Brief explanation of the drawings]

[0164] [Figure 1]Figures 1A-1G are a set of graphs showing that Disosmobacter welbrionis J115T partially attenuates tumor growth induced by a high-fat diet. Figure 1A shows the progression of body weight in mice fed a normal diet (ND) or a high-fat diet (HFD). Arrows indicate tumor induction. Figure 1B shows the progression of E0771 tumor volume. Figure 1C shows the progression of tumor volume focusing on the ND and HFD groups. Figure 1D shows the progression of tumor volume focusing on the HFD group with and without J115T treatment. Figure 1E shows the survival curves estimated by linear regression, calculating the time required for each tumor to reach 300 mm3. Figure 1F shows the survival curve focusing on the difference between the ND and HFD groups. Figure 1G shows the survival curve focusing on the difference between the HFD and HFD J115T groups. Sample sizes: n = 9 for ND, n = 8 for HFD, and n = 11 for HFD J115T. Mice with tumors smaller than 100 mm3 were excluded. Statistical analysis: (Figure 1A, Figure 1B) Two-way ANOVA followed by Tukey's multiple comparison test. ***p<0.001 (HFD vs. ND) and $p<0.05 (HFD J115T vs. HFD). (Figure 1C, Figure 1D) Two-way ANOVA followed by Sidak's multiple comparison test. *p<0.05; ***p<0.001. (Figure 1E, Figure 1F, Figure 1G) Log-rank (Mantel-Cox) test. *p<0.05. [Figure 2]Figures 2A-2E are a set of graphs showing that Disosmobacter welbrionis J115T reduces tumor growth induced by a high-fat diet and confirm the results in Figure 1. Figure 2A shows the progression of body weight over time in mice fed a normal diet (ND) or a high-fat diet (HFD). Arrows indicate tumor induction. Figure 2B shows the progression of E0771 tumor volume. Figure 2C shows survival curves estimated by linear regression, calculating the time required for each tumor to reach 400 mm3. Figure 2D shows the survival curves focusing on the differences between the ND and HFD groups. Figure 2E shows the survival curves focusing on the differences between the HFD and HFD J115T groups. Sample sizes: n = 9 for ND, n = 10 for HFD, and n = 8 for HFD J115T. Mice with tumors less than 100 mm3 and showing less than 50% growth (compared to day 6) were excluded. Statistical analysis: (Figures 2A and 2B) Two-way ANOVA followed by Tukey's multiple comparison test. *p<0.05; **p<0.01; ***p<0.001 (HFD vs. ND) and $$p<0.01 (HFD J115T vs. HFD) (Figures 2C, 2D, and 2E). Log-rank (Mantel-Cox) test. *p<0.05. [Figure 3] Figures 3A-3B are a set of graphs showing that Disosmobacter welterbionis J115T attenuates high-fat diet-induced tumor growth and confirms the results of Figures 1 and 2. Figure 3A shows the progression of body weight on a high-fat diet (HFD). Arrows indicate tumor induction. Figure 3B shows the progression of E0771 tumor volume. Statistical analysis: Two-way ANOVA followed by Sidak's multiple comparison test. **p<0.01. [Figure 4] Figures 4A-4B are a set of graphs showing that Disosmobacter welterbionis J115 reduces tumor growth in non-obesity-related settings. Figure 4A shows the progression of body weight in Balb / c mice on a normal diet (ND). Figure 4B shows the progression of 4T1 tumor volume. Statistical analysis: Two-way ANOVA followed by Sidak's multiple comparison test. **p<0.01. [Figure 5]Figures 5A-5D are a set of histograms showing that Disosmobacter welbrionis J115T supernatant reduces cell density and proliferation in an in vitro triple-negative breast cancer (TNBC) model. Figures 5A-5B show cell density (A) and cell proliferation (B) of PY8119 cells after 24 h of treatment with either non-fermented bacterial medium, fermented bacterial medium, or phosphate buffered saline (PBS) matched for pH and short-chain fatty acid (SCFA) content (to the fermentation medium) at the indicated concentrations. Figures 5C-5D show cell density (C) and cell proliferation (D) of E0771 cells after 24 h of treatment with either non-fermented bacterial medium, fermented bacterial medium, or phosphate buffered saline (PBS) matched for pH and short-chain fatty acid (SCFA) content (to the fermentation medium) at the indicated concentrations. The pH of the fermentation medium was 6.1, and the SCFA content was equal to 25 mM butyrate and 60 mM acetate. All data are presented as the mean ± SEM of three independent experiments. Statistical analysis: Nested one-way ANOVA followed by Dunnett's multiple comparison test. *p<0.05; **p<0.01; ***p<0.001. [Example]

[0165] The present invention is further illustrated by the following examples.

[0166] Example 1: material and method bacterial culture Disosmobacter welbrionis J115 T were cultured anaerobically in modified YCFA medium supplemented with 10 g / L inositol. The cultures were centrifuged at 5000 g and 4°C for 15 minutes, and the supernatant was removed. The cells were resuspended in anaerobic PBS-carbonate buffer supplemented with 15% (vol / vol) trehalose, then immediately frozen in anaerobic vials and stored at -80°C. For mouse administration, the total and culturable bacterial numbers to be administered to mice were calculated by plating the bacterial cultures after freezing but before centrifugation and bacterial suspension.

[0167] cell culture E0771 and PY8119 mouse mammary adenocarcinoma cell lines were obtained from the American Type Culture Collection (ATCC) and maintained according to the supplier's instructions. 4T1 cells were obtained from the mammary gland of a BALB / c mouse strain. E0771 and 4T1 cells were maintained in Dulbecco's modified Eagle's medium (GIBCO, Thermo Fisher Scientific) containing 25 mM glucose, 4 mM glutamine, and 25 mM HEPES, supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Thermo Fisher Scientific). PY8119 cells were maintained in culture in F-12K medium (GIBCO, Thermo Fisher Scientific) containing 7 mM glucose and 2 mM glutamine, supplemented with 5% heat-inactivated FBS (Thermo Fisher Scientific). They were cultured at 37°C and 5% CO2 in a humidified atmosphere.

[0168] cell density Cell density was assessed using PrestoBlue reagent (ThermoFisher Scientific) according to the manufacturer's instructions. Briefly, cells were treated with either non-fermented or fermented YCFA medium as described above for 24 hours. To distinguish between the contribution of short-chain fatty acids and the effect of pH, phosphate buffered saline (PBS) containing 25 mM butyrate and 60 mM acetate at pH 6.1 was used. PrestoBlue reagent was added to each well at a concentration of 10%. After 2 hours of incubation, fluorescence intensity was measured (λex / λem = 560 / 590 nm) using a plate reader (SpectraMax M2e, Molecular Devices). All data were normalized to the fluorescence intensity of untreated wells and expressed as a percentage of the control.

[0169] Cell proliferation Cell proliferation was measured using a 5-bromo-2-deoxyuridine (BrDu) incorporation ELISA-based kit (Roche) according to the supplier's protocol. After 24 hours of incubation with the indicated treatments described above, BrDu was added to the medium for 2 hours. Cells were then fixed with a solution provided by the manufacturer and bound with a peroxidase-conjugated anti-BrDu antibody. Cell proliferation was then assessed by measuring absorbance at 370 nm using a plate reader (SpectraMax M2e, Molecular Devices). All data were normalized to the absorbance of untreated wells and expressed as a percentage of the control.

[0170] mouse All mouse experiments were approved by the Animal Experimentation Ethics Committee of the Faculty of Medicine at KU Leuven under specific number 2021 / UCL / MD / 04 and its amendment 2022 / UCL / MD / A10 and were conducted in accordance with the local ethical committee guidelines and the Belgian Act of May 29, 2013 on the Protection of Laboratory Animals (agreement number LA1230467). Specific pathogen-free (SPF) certified 8-week-old female C57BL / 6JRj mice (Janvier Labs) were used for the experiments. Cages were randomly assigned to experimental groups to ensure weight matching at the start of feeding on a normal diet (ND) (D10012M; Research Diets) and a 60% kcal high-fat diet (HFD) (D12492; Research Diets) and before tumor induction. Body weight was assessed weekly.

[0171] Oral administration of bacteria Freshly thawed Disosmobacter welbrionis J115 in the late postprandial stage T Each mouse was administered at least 1.0 × 10 9 ND and HFD control groups were given an oral gavage of an equivalent volume of PBS-carbonate buffer supplemented with 15% (wt / vol) trehalose.

[0172] Tumor growth experiments Six weeks after the start of HFD treatment, 1 × 10 6 E0771 tumors were induced in the fifth mammary fat pad of C57Bl / 6JRj female mice by subcutaneous injection of 2 × 10 cells. 5 4T1 tumors were induced in the fifth mammary fat pad of 8-week-old BALB / c mice by subcutaneous injection of E0771 cells. Freshly passaged E0771 cells were prepared in a 1:1 mixture of PBS and Matrigel (Corning) immediately prior to injection and injected within 30 minutes. Freshly passaged 4T1 cells were prepared in PBS immediately prior to injection and injected within 30 minutes. Tumor size was monitored at least twice a week (preliminary experiments) or daily (confirmation) and measured using electronic calipers in a simple, blinded manner.

[0173] statistical analysis All statistical analyses were performed using GraphPad Prism software version 9.1.2.

[0174] result In the present invention, we investigated the effects of Disosmobacter welbrionis J115 in vivo and in vitro. T was tested for its ability to reduce or delay cancer development under obese conditions.

[0175] More specifically, we used triple-negative breast cancer cells as the first model. Obesity was induced in mice using a high-fat diet, and then TNBC cells were injected into the mammary glands of the mice (Figure 1A, arrow = tumor cell injection). Mice were then treated with either placebo or Disosmobacter welbrionis J115. T On day 15, placebo-treated obese mice showed significantly higher tumor growth than lean mice, with tumor volumes 80% larger (Figure 1B and Figure 1C), and Disosmobacter welbrionis J115. T Obese mice treated with α-glucan showed a significantly lower tumor incidence, similar to that of lean mice (Figures 1B and 1D).

[0176] Obese mice were also characterized by faster tumor growth, as shown in Figure 1E and Figure 1F, and the time to reach the same tumor volume was shorter in obese mice compared to lean mice; in other words, obese mice reached 300 mm in only 19 days. 3 When tumors reach a volume of 1000 mg / kg, tumors in lean mice reach this size in 27 days (Figure 1E and Figure 1F). T This effect on tumor size delay induced by Disosmobacter welterbionis J115 significantly prolonged the time required for tumors to reach the same volume as observed in obese mice. T Mice treated with 300mm 3 The spleen volume reached 100 mcg, which was similar to that of lean mice and significantly slower than that of obese (weight-matched) mice receiving placebo (Figure 1E and Figure 1G). Surprisingly, this beneficial effect was not observed in Disosmobacter welbrionis J115. T This was not associated with weight loss in mice treated with α-glucan (Fig. 1A).

[0177] This first dataset was generated from Disosmobacter welbrionis J115 T Surprisingly, non-invasive techniques such as the use of Disosmobacter welbrionis J115 can be associated with such a significant delay in cancer development. T These results convincingly demonstrate that the anti-obesity effects of cereals containing glutathione are not due to the

[0178] In a second set of independent experiments (Figure 2), the effect of Disosmobacter welbrionis J115 on tumor growth reduction was T The same beneficial effect of was fully reproduced (Figure 2B and Figure 2C), with a threshold of 300 mm 3 instead of 400mm 3 This effect remained significant regardless of the volume chosen (Figures 2D and 2E).

[0179] Finally, a third set of independent experiments (Figure 3) demonstrated the efficacy of Disosmobacter welbrionis J115 for reducing tumor growth in obesity-related settings. T The beneficial effect of α-glucan was confirmed (Figure 3B).

[0180] In the present invention, the in vivo expression of Disosmobacter welbrionis J115 T was tested for its ability to reduce or delay cancer development in non-obesity related situations.

[0181] The cells were subcutaneously injected into the fifth mammary fat pad of mice (Figure 4A). Mice were then treated with placebo or Disosmobacter welbrionis J115. T On day 16, the mice were treated daily with either Disosmobacter welbrionis J115 or T Mice treated with Disosmobacter welbrionis J115 showed significantly lower tumor growth compared to untreated mice (Figure 4B). T has been shown to delay the onset of cancer independently of obesity.

[0182] Next, Disosmobacter welbrionis J115 T In vitro testing using a medium fermented by Disosmobacter welbrionis J115 T The effects of potential metabolites produced by Disosmobacter welbrionis J115 were investigated (Figure 5). T Fermented medium significantly reduced cell density in two different models of triple-negative breast cancer (E0771 and PY8119 cells) in a dose-dependent manner after 24 hours of treatment (Figure 5A and Figure 5C). Conversely, non-fermented bacterial medium had no effect on this parameter.

[0183] Disosmobacter welbrionis J115 Ttested non-fermenting bacterial media with adjusted pH and supplemented with SCFAs (thus matching the content) to produce short-chain fatty acids (SCFAs) (butyrate, acetate). Neither the non-fermenting medium nor the SCFA-enriched non-fermenting medium was able to affect cell density (Figure 5, SCFA-matched medium).

[0184] Disosmobacter welbrionis J115 T The fermented medium reduced cell growth of both E0771 and PY8119 cells after 24 hours of treatment, while the non-fermented medium had no effect on this parameter. T Although not reaching the same level as that obtained by , cell growth was reduced in the SCFA-matched medium. T The medium obtained by fermentation completely inhibited cell growth already at a concentration of 20% (corresponding to a 10- to 20-fold higher potency than SCFAs) (Figure 5B and Figure 5D, "SCFA-matched medium" vs. "fermented medium").

[0185] These data indicate that the antitumor effects of specific metabolites produced by Disosmobacter welterbionis J115T, distinct from SCFAs, mediated the bacterium's antitumor effects.

Claims

1. A composition for use in preventing and / or treating breast cancer in a subject in need thereof, comprising at least (a) bacteria of the genus Disosmobacter and / or variants and / or extracts and / or fragments thereof, and / or (b) a culture supernatant of bacteria of the genus Disosmobacter and / or variants thereof.

2. The composition for use according to claim 1, wherein the bacterium belongs to the species Disosmobacter welbrionis.

3. The bacterium was strain J115, deposited at BCCM / LMG as LMG P-30603 on March 14, 2018. Ｔ 3. A composition for use according to claim 1 or 2, which belongs to the

4. 4. The composition for use according to any one of claims 1 to 3, wherein the breast cancer comprises at least one mutation in a gene encoding a receptor selected from the group comprising or consisting of progesterone receptor (PR), estrogen receptor (ER) and human epidermal growth factor receptor-2 (HER2).

5. 5. The composition for use according to any one of claims 1 to 4, wherein said breast cancer is selected from the group comprising or consisting of luminal A breast cancer, luminal B breast cancer, HER2-positive breast cancer and triple-negative breast cancer (TNBC), preferably said breast cancer is triple-negative breast cancer.

6. 6. The composition for use according to any one of claims 1 to 5, wherein the subject is further suffering from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity-related disorders, liver diseases, metabolic disorders, adipokine-related disorders and inflammatory diseases, and combinations thereof.

7. 7. The composition for use of any one of claims 1 to 6, wherein the subject further suffers from one or more diseases or conditions that accelerate tumor growth selected from the group including or consisting of obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), cirrhosis, diabetes, impaired glucose tolerance, hyperglycemia, dyslipidemia, dyslipidemia, high cholesterol, elevated LDL cholesterol levels, low HDL cholesterol levels, elevated triglyceride levels, and enterocolitis, and combinations thereof.

8. The composition is administered in a therapeutically effective amount, preferably 1×10 ２ ~Approx. 1×10 １５ A composition for use according to any one of claims 1 to 7, comprising CFUs of said bacteria.

9. The composition for use according to any one of claims 1 to 8, wherein the composition comprises live bacteria.

10. The composition for use according to any one of claims 1 to 8, wherein the composition comprises dead or killed bacteria.

11. The composition for use according to any one of claims 1 to 8, wherein the composition comprises pasteurized bacteria.

12. The composition for use according to any one of claims 1 to 11, wherein said composition further comprises at least another anti-cancer agent.

13. The composition for use according to any one of claims 1 to 12, wherein the composition is in the form of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

14. A prebiotic for use in preventing and / or treating breast cancer in a subject in need thereof, comprising one or more active ingredients or substances that increase the level of bacteria of the genus Disosmobacter in the microbiota of said subject.

15. A composition for use as an adjuvant to a therapeutic agent administered to a subject suffering from breast cancer, comprising at least (a) a bacterium of the genus Disosmobacter and / or a variant, extract or fragment thereof, and / or (b) a culture supernatant thereof.

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