Malodobacter sp. For treatment of breast cancer

Abstract

The present invention relates to a composition comprising bacteria from the genus malodobacter and / or variants, extracts or fragments thereof for use in the prevention and / or treatment of breast cancer.

Description

Field of the Invention

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

[0003] Cancer has become one of the most important diseases of the last century and a major cause of premature death worldwide. It is estimated that there were approximately 19.3 million new cancer cases globally in 2020 and 10 million people died from cancer. Among all cancers, breast cancer (BC) is currently the most diagnosed cancer, with an estimated 2.3 million cases, accounting for 11.7% of all diagnosed cases and 25% of diagnosed women. However, breast cancer is a highly heterogeneous disease and is classified into different subtypes based on the (over)expression of progesterone (PR), estrogen (ER), and human epidermal growth factor receptor (HER-2). Early and late-stage patients with hormone receptor-expressing breast tumors and patients with HER-2-enriched tumors can benefit from targeted therapy, whether used alone or in combination with traditional chemotherapy. However, patients with triple-negative breast cancer (TNBC) are characterized by the absence or overexpression of the aforementioned receptors and do not benefit from targeted therapy. Due to these unmet needs in the field of targeted therapy, TNBC remains the most aggressive and poorest-prognosis subtype, with a high recurrence rate, high metastatic potential, and decreased survival. Risk factors for BC and TNBC include advanced age at first pregnancy, lack of physical exercise, and alcohol consumption, but obesity is increasingly regarded as a major risk factor for BC and TNBC.

[0004] It has been described that obesity is associated with a higher risk of adverse outcomes and poorer prognosis in breast cancer patients with shorter overall disease-free survival, thus highlighting the need to explore new treatment strategies for breast cancer in the context of obesity. In obesity, important alterations in the gut microbiota composition, also known as dysbiosis, occur in clinical and preclinical models (Ley et al., 2006). In addition, the gut microbiota is a key regulator of whole-body metabolism and low-grade inflammation (Cani et al., 2019), but it has also received significant attention in regulating cancer and breast cancer progression and treatment (Sampsell et al., 2020).

[0005] In the present invention, the inventors studied the impact of Dysosmobacter welbionis, a novel bacterium discovered, isolated and named at UCLouvain (Brussels, Belgium) (Le Roy et al., 2020 and WO2020011856). Dysosmobacter welbionis is present in 70% of the general population and can thus be considered a highly prevalent bacterium, similar to Akkermansia muciniphila. As a brief background on this bacterium, Dysosmobacter welbionis is a butyrate producer, and species of Dysosmobacter and Dysosmobacter welbionis have been found to be less abundant in the gut of patients with obesity and type 2 diabetes. In preclinical studies, it has been shown that Dysosmobacter welbionis J115 T reduces weight gain, increases in fat mass and improves glucose tolerance, lipid and energy metabolism by acting on mitochondrial activity (Le Roy, Moens de Hase et al., 2022).

[0006] In the present disclosure, the inventors demonstrate that Dysosmobacter welbionis J115 T can reduce the growth of breast cancer. Summary of the Invention

[0007] The present invention relates to a composition comprising at least (a) bacteria from the genus Dysosmobacter and / or variants and / or extracts and / or fragments thereof, and / or (b) culture supernatants of bacteria from the genus Dysosmobacter and / or variants thereof, for the prevention and / or treatment of breast cancer in a subject in need thereof. In some embodiments, the bacteria belong to the species Dysosmobacter welbionis.

[0008] In some embodiments, the bacteria belong to strain J115 T , which was deposited at BCCM / LMG under the accession number LMG P-30603 on March 14, 2018.

[0009] In some embodiments, the breast cancer comprises at least one mutation in a gene encoding a receptor selected from the group consisting of or comprising a progesterone receptor (PR), an estrogen receptor (ER) and a human epidermal growth factor receptor-2 (HER2).

[0010] In some embodiments, the breast cancer is selected from the group consisting of or comprising luminal A breast cancer, luminal B breast cancer, HER2-positive breast cancer and triple-negative breast cancer (TNBC), preferably the breast cancer is triple-negative breast cancer.

[0011] In some embodiments, the subject also has one or more diseases or conditions that accelerate tumor growth, the diseases or conditions selected from the group consisting of or comprising: obesity-related diseases, liver diseases, metabolic disorders, adipokine-related diseases, and inflammatory diseases, and combinations thereof.

[0012] In some embodiments, the subject also has one or more diseases or conditions that accelerate tumor growth, the diseases or conditions selected from the group consisting of or comprising: obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), cirrhosis, diabetes, glucose intolerance, hyperglycemia, dyslipidemia, lipid metabolism disorder, hypercholesterolemia, elevated LDL-cholesterol, reduced HDL-cholesterol, elevated triglycerides, and intestinal inflammation, and combinations thereof.

[0013] In some embodiments, the composition comprises a therapeutically effective amount of the bacterium, preferably from 1×10 2 to about 1×10 15 CFU.

[0014] In some embodiments, the composition comprises live bacteria.

[0015] In some embodiments, the composition comprises dead or killed bacteria.

[0016] In some embodiments, the composition comprises pasteurized bacteria.

[0017] In some embodiments, the composition further comprises at least one other anti-cancer agent.

[0018] In some embodiments, the composition is in the form of a pharmaceutical composition, which further comprises a pharmaceutically acceptable carrier.

[0019] The present invention also relates to a prebiotic comprising one or more active ingredients or substances that increase the level of bacteria of the genus Odoribacter in the microbiota of a subject in need thereof, for preventing and / or treating breast cancer in the subject.

[0020] The present invention also relates to a composition comprising at least one bacterium from the genus Odoribacter and / or a variant, extract or fragment thereof, for use as an adjuvant to a treatment administered to a subject suffering from breast cancer.

[0021] Definitions

[0022] In the present invention, the following terms have the following meanings:

[0023] "About" following a numerical value means plus or minus 10% of the value of that number.

[0024] "Acceptable", e.g., when used in the context of "pharmaceutically acceptable" or "nutritionally acceptable", means that the molecular entity and composition do not produce adverse, allergic, or other untoward reactions when appropriately administered to a subject, particularly a human.

[0025] "Strain" refers to a subtype of a bacterial species.

[0026] "Dysosmobacter" [Dys.os.mo.bac'ter. Gr. masc. adj. Dysosmos meaning ill-smelling; NL masc. n. bacter meaning rod-shaped bacterium; NL masc. n. Dysosmobacter meaning ill-smelling rod-shaped bacterium] refers to a genus of bacteria described herein having the following characteristics: The cells are obligately anaerobic, non-pigmented, non-spore-forming, non-motile, and Gram-negative. The cells predominantly form straight rods 1.8 - 3.0 μm in length, but at all growth stages, usually form long, slender rods up to 20 μm in length. Respiratory menaquinones are not produced. The genus belongs to the family Ruminococcaceae. The type species is Dysosmobacter welbionis. In one embodiment, the diagnostic diamino acid in the cell wall is meso-2,6-diaminopimelic acid.

[0027] "Dysosmobacter welbionis" [wel.bi.o′nis. NL gen. n. welbionis] refers to a bacterium described herein that, in addition to having the characteristics of the genus Dysosmobacter described above, also has the following characteristics: After incubation at 37 °C for 72 hours under anaerobic conditions, the colonies on solid modified YCFA are punctiform, cream-colored, translucent, round, entire, slightly convex, and smooth. The presence of 2% w / v bile or 2% w / v NaCl inhibits its growth. 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 glutamate decarboxylase. All other tests with API 20A and Rapid ID 32A (bioMérieux, Lyon, France) are negative. The major end product of myo-inositol fermentation is butyrate. By high-performance liquid chromatography (HLPC), the DNA GC content of the type strain is 59.3 mol%. In one embodiment, based on the genomic sequence, the DNA GC content of the type strain is 58.9 mol%. The type strain is J115 T(Deposited with the Belgian Co-ordinated Collections of Micro-organisms / Laboratory of Microbiology (BCCM / LMG), Universiteit Gent, KL Ledeganckstraat 35, 9000 Gent, Belgium under the accession number LMG P-30603 on March 14, 2018), was isolated from human feces. In one embodiment, the major cellular fatty acids are saturated branched-chain fatty acids and DMA. In one embodiment, the major DMA fatty acids are C 18：0 DMA, and the major saturated branched-chain fatty acids are iso-C 15：0 and anteiso-C 15：0 .

[0028] "Fermentation" refers to the metabolic process of consuming sugars in the absence of oxygen. The products are organic acids, gases or alcohols. It occurs in yeast and bacteria, and also in muscle cells deprived of oxygen, as in the case of lactic acid fermentation.

[0029] "Gut microbiota" or "gastrointestinal microbiota" are used interchangeably and refer to the complex microbial community living in the digestive tract of humans and other animals. The composition of the gastrointestinal microbiota changes over the lifetime of the host organism or with changes in the host diet. It also varies throughout the digestive tract. The digestive tract contains a dense microbial ecosystem with up to 10 12cells. Many species in the gut have not been studied outside their hosts because most are unculturable. The four major bacterial phyla in the human gut are Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria. Most bacteria belong to the genera Bacteroides, Clostridium, Faecalibacterium, Eubacteria, Ruminococcus, Peptococcus, Peptostreptococcus, Blautia, Subdoligranulum, Alistipes, Coprococcus, Dialister, Lachnoclostridium, Oscillospira, Parabacteroides, Prevotella, Roseburia, Ruminiclostridium, Sutterella, and Bifidobacteria. Other genera, such as Escherichia, Enterococcus, Barnesiella, Butyricimonas, Butyricicoccus, Lachnospira, Odoribacter, Turicibacter, and Lactobacillus, are present in smaller amounts. The gut microbiota is thought to play a role in defending against pathogens by competing with potential pathogens and participating in the development of gut protection and the immune system, metabolism, assisting in the digestion of ingested food, helping with nutrient absorption and vitamin synthesis. The gut microbiota also interacts with the functions of the central nervous system, neuroendocrine, and neuroimmune systems.

[0030] As described herein, a "mutant" refers to a biological entity whose genetic structure has undergone natural or induced (i.e., by mutagenesis) changes that do not affect its defined properties. The changes in the genetic structure of the biological entity may be insertions, deletions, or substitutions of one or more nucleotides in the genomic sequence. For example, a mutant of Bacillus foetidus is a strain of Bacillus foetidus whose genetic structure has undergone changes that do not affect its belonging to the species Bacillus foetidus, either naturally or through genetic engineering techniques.

[0031] "Nutritionally effective amount" refers to the amount of a nutritional composition, food, or dietary supplement or functional food that is necessary and sufficient to provide a physiological benefit or relieve discomfort to a subject.

[0032] "Pasteurized bacteria" refers to bacteria that have undergone a heat treatment (or heating process).

[0033] "Pharmaceutically acceptable carrier or excipient" refers to molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to a subject (especially a human) under appropriate circumstances. It includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, and absorption delaying agents, etc. For human administration, the formulations should meet the pyrogenicity, general safety, and purity standards required by regulatory agencies (such as the FDA Office or EMA). Thus, a pharmaceutically acceptable carrier or excipient can refer to any type of non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, or formulation aid.

[0034] "Prebiotic" refers to a substance that may not be digested by a subject (such as a human), but modulates the composition and / or activity of the gut microbiota through the metabolism of the gut microbiota, thereby conferring beneficial physiological effects on the host.

[0035] "Probiotic" refers to a preparation of microbial cells (such as live microbial cells) that, when administered in an effective amount, has a beneficial effect on the health or well-being of a subject. By definition, all probiotics have proven non-pathogenic properties. In one embodiment, these health benefits are related to improving the balance of the gastrointestinal microbiota of a human or animal and / or restoring the normal microbiota.

[0036] "Subject" refers to a warm-blooded animal, preferably a human, 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 can be a "patient", i.e., a subject who is waiting to receive or is receiving medical care or who has been / will be the subject of a medical procedure according to the methods of the present invention or who is being monitored for the development of a disease.

[0037] "Therapeutically effective amount" means the level or amount of an agent that, without causing significant negative or adverse side effects to the target, is intended to achieve one or more of the following purposes: (1) delaying or preventing the onset of a disease, disorder or condition; (2) slowing or halting the progression, aggravation or worsening of one or more symptoms of a disease, disorder or condition; (3) bringing about an improvement in the symptoms of a disease, disorder or condition; (4) reducing the severity or incidence of a disease, disorder or condition; or (5) curing a disease, disorder or condition. A therapeutically effective amount may be administered before the onset of a disease, disorder or condition to provide prophylaxis or prevention. Alternatively or additionally, a therapeutically effective amount may be administered after the onset of a disease, disorder or condition to provide treatment.

[0038] "Treatment" refers to therapeutic treatment and prophylactic or preventive measures that are intended to prevent or slow down (reduce) a target pathological condition or disorder, preferably, the disorder is breast cancer. Those in need of treatment include those who already have the disorder and those who are predisposed to having the disorder or in need of preventing the disorder. A patient is successfully "treated" if the subject or mammal exhibits one or more of the following observable and / or measurable changes after receiving a therapeutic amount of Bacillus foetidus of the present invention and / or its variants and / or its fragments: improvement in one or more symptoms associated with a specific disease or disorder (preferably breast cancer), reduction in morbidity and mortality, and improvement in the quality of life issues. The above parameters for assessing successful treatment and improvement of a disease can be easily measured by conventional procedures familiar to a physician.

[0039] "Type strain" refers to the named type and reference point of a species as defined in the International Code of Nomenclature of Bacteria, against which all other strains are compared to determine whether they belong to that species. For example, strain J115T isolated from a fecal sample of a 25-year-old healthy female is the type strain of the species Bacillus foetidus.

[0040] "Variant" refers to all genetically or phenotypically distinct strains of a species that retain the defining characteristics of the species. The term variant is also used to refer to other phylogenetic taxa, such as genera or strains. The term "variant" as used herein refers to naturally occurring and specially developed variants or mutants of the bacteria disclosed and exemplified herein. In one embodiment, a variant may or may not have the same identifying biological characteristics as the bacteria exemplified herein, provided that they have similar advantageous properties in the treatment or prevention of disease. In one embodiment, variants of the bacteria of the present invention have the same functions and / or therapeutic properties as the bacteria of the present invention. Exemplary examples of methods for generating variants of the microbial strains exemplified herein include, but are not limited to, gene integration techniques such as those mediated by insertion elements or transposons or by homologous recombination, other recombinant DNA techniques for modifying, inserting, deleting, activating or silencing genes, intraspecific protoplast fusion, mutagenesis by irradiation with ultraviolet or X-rays, or treatment with chemical mutagens such as nitrosoguanidine, methyl methanesulfonate, nitrogen mustard, etc., and phage-mediated transduction. DETAILED DESCRIPTION OF THE INVENTION

[0042] The present invention relates to a composition comprising at least (a) bacteria and / or variants, extracts or fragments thereof from the genus Dysosmobacter, and / or (b) culture supernatants of bacteria and / or variants thereof from the genus Dysosmobacter, for the prevention and / or treatment of breast cancer in a subject in need thereof.

[0043] In some embodiments, the bacteria from the genus Dysosmobacter comprised in the composition used in the present invention belong to a species selected from the group consisting of or comprising Dysosmobacter welbicki, Dysosmobacter acutus, Dysosmobacter segnis, and Dysosmobacter hominis. In some embodiments, the bacteria from the genus Dysosmobacter comprised in the composition used in the present invention belong to a species selected from the group consisting of or comprising Dysosmobacter welbicki.

[0044] The present inventors previously reported the biological characteristics of Dysosmobacter welbicki in WO2020011856.

[0045] In some embodiments, the bacteria belong to a strain selected from the group consisting of or comprising J115 T , Dysosmobacter acutus MSJ-2 T(CGMCC Accession No.: 1.32896T; KCTC Accession No.: 15976T), Bacteroides putredinis BX15 (CGMCC Accession No.: 1.32894), Marseille-Q4140, MM13, and Bacteroides putredinis NSJ-60 (CGMCC Accession No.: 1.32836; KCTC Accession No.: 25148). In some embodiments, the bacterium belongs to strain J115 T , which was deposited with BCCM / LMG under the accession number LMG P-30603 on March 14, 2018, and / or a variant thereof. J115 T The strain is a type strain of the species Bacteroides wexlerae. In certain aspects, the present invention relates to a composition comprising a bacterium from Bacteroides wexlerae strain J115 T (deposit number LMG P-30603) and / or a variant, extract, or fragment thereof, for preventing and / or treating breast cancer in a subject in need thereof.

[0046] In one embodiment, the genomic sequence of the bacterium has sequence SEQ ID NO: 1, or has a sequence that exhibits at least about 65% identity to SEQ ID NO: 1, preferably a sequence that exhibits at least about 70%, 75%, 80%, 85%, 90% identity to SEQ ID NO: 1, more preferably a sequence that exhibits at least about 91%, 92%, 93%, 94%, 95%, 96%, 96.5%, 97%, 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.

[0047] In one embodiment, when compared to the genome of sequence SEQ ID NO: 1, the average nucleotide identity (ANI) score of the bacterium is higher than about 60, preferably higher than about 74, 75, 80, 85, 90, more preferably higher than about 95, even more preferably higher than about 96, 97, 98, 98.5, 98.65, 99 or higher.

[0048] Techniques for determining ANI values are known to those skilled in the art (e.g., the method implemented 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 match (BBH - orthologous sequences determined based on their positions in the genome and sequence identity) multiplied by the alignment length divided by the total length of the BBH genes.

[0049] In one embodiment, the heterologous DNA-DNA hybridization value (also known as the DDH value) of the bacterium with SEQ ID NO: 1 is higher than about 60%, preferably higher than about 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, more preferably higher than about 70%.

[0050] Techniques for determining DDH values are known to those skilled in the art (e.g., the methods reviewed in Stackebrandt et al., Molecular Identification, Systematics, and Population Structure of Prokaryotes, p23-50, 2006, Springer, Berlin, Heidelberg) and rely on the following general principles: (i) cutting the genomic DNA (gDNA) of the test organism and the gDNA of the reference organism (e.g., the type strain J115 T (deposited as LMG P-30603 at BCCM / LMG on March 14, 2018) into small fragments of 600-800 bp; (ii) heating the DNA fragment mixture of the two strains to dissociate the DNA double strands; and (iii) subsequently lowering the temperature until the fragments re-anneal. Since the melting temperature of the double strand depends on the degree of base pairing match between the two strands, genomic (dis)similarity can be inferred from the melting temperature. The heterologous DDH value is usually determined relative to the DDH value obtained by hybridizing the reference genome with itself. A DDH value ≤ 70% can be considered as indicating that the test organism belongs to a different species from the type strain used as a reference. The DDH value can also be evaluated using publicly available computer programs based on the genomic sequences of the strains to be compared.

[0051] In one embodiment, the intergenomic distance of the bacterium from SEQ ID NO: 1 is lower than about 0.5, preferably lower than about 0.22, 0.21, 0.20, 0.19, 0.18, 0.17, 0.16, 0.15, 0.14, more preferably lower than about 0.13, 0.12, 0.11, 0.10 or less.

[0052] Techniques for determining the inter-genomic distance or genome-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) can be used. This method can be implemented using the Genome Calculator 2.1 (Deutsche Sammlung von Mikroorganismen und Zellkulturen - DSMZ), using BLAST+ as the local alignment tool, and dividing the sum of all identities found in the high-scoring segment pairs (HSPs) by the total length of the HSPs.

[0053] In one embodiment, the nucleotide sequence of the 16S rRNA gene of the bacterium has the sequence SEQ ID NO: 2, or has a sequence that exhibits at least about 90% identity with 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%, 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 with SEQ ID NO: 2 (deposited in GenBank / EMBL / DDBJ, accession number MG963288).

[0054] In one embodiment, the nucleotide sequence of the 16S rRNA gene of the bacterium has the sequence SEQ ID NO: 2, or has a sequence that exhibits at least about 99.9% identity with 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 higher identity with SEQ ID NO: 2.

[0055] In one embodiment, the nucleotide sequence of the 16S rRNA gene of the bacterium has SEQ ID NO: 2, or has a sequence that exhibits at least about 90% identity or more identity 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%, 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 higher identity over the entire length of SEQ ID NO: 2.

[0056] When used to describe the relationship between two or more polypeptide sequences or two or more nucleic acid molecule sequences, the term "identity" refers to the degree of sequence relatedness between the polypeptides or nucleic acid molecules, determined by the number of matches between fragments of two or more amino acid or nucleotide residues. "Identity" measures the percentage of identical matches between the smaller of two or more sequences, where gap alignments (if any) are treated by a particular mathematical model or computer program (i.e., "algorithm"). The identity of related polypeptides can be readily calculated by known methods.

[0057] The compositions used in the present invention may contain variants of the bacterium as described above. The variants may also be referred to as derivative strains of the bacterium. In one embodiment, the variant of the bacterium can be obtained by mutation, variation or recombination of the bacterium described herein. Within the scope of the present invention, the variant may also be referred to as a mutant.

[0058] In one embodiment, the variant of the bacterium from the genus Odoribacter contained in the composition used in the present invention is a variant of Odoribacter winogradskyi. In one embodiment, the variant of the bacterium from the genus Odoribacter contained in the composition used in the present invention is a variant of Odoribacter winogradskyi strain J115 T of.

[0059] In one embodiment, the bacterial variant has a genome that has 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 higher identity to the genome of the bacterium from which it is derived.

[0060] In one embodiment, the genomic sequence of the variant of the bacterium has at least about 65% identity to the genomic sequence of the bacterium from which it is derived, preferably at least about 70%, 75%, 80%, 85%, 90% identity to the genomic sequence of the bacterium from which it is derived, more 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%, 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 to the genomic sequence of the bacterium from which it is derived.

[0061] In one embodiment, the variant of the bacterium has the same function and / or therapeutic properties as the bacterium from which it is derived.

[0062] In one embodiment, the bacteria of the genus Odoribacter and / or variants thereof contained in the composition used in the present invention are live bacteria or killed bacteria.

[0063] 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 proliferation, as opposed to "killed bacteria" (or "non-viable bacteria") that are not capable of proliferation. Methods for measuring viability and proliferation are known to those skilled in the art. For example, the viability and proliferation of bacteria can be evaluated by spreading a solution containing at least one bacterium of the present invention on a culture dish and counting the number of colonies after incubation for a determined time under optimal growth conditions. Alternatively, the bacteria can be grown in a liquid medium, and proliferation can be measured by measuring the optical density of the bacterial culture after incubation for a determined time under optimal growth conditions. The number of bacteria, including live bacteria and non-live bacteria, can also be determined by microscopic observation. While phase-contrast microscopy is a well-known method, the microbial bacteria can be further visualized by specific staining with dyes, fluorescent probes or antibodies for easy microscopic observation or counting of bacteria by flow cytometry.

[0064] In one embodiment, the bacteria and / or variants thereof from the genus Odoribacter contained in the composition used in the present invention are live bacteria. In some embodiments, the bacteria have metabolic activity.

[0065] In one embodiment, the bacteria and / or variants thereof from the genus Odoribacter contained in the composition used in the present invention are killed bacteria. In some embodiments, the bacteria and / or variants thereof cannot proliferate. In some embodiments, the bacteria and / or variants thereof have no metabolic activity. In some embodiments, the bacteria and / or variants thereof are heat-inactivated or heat-killed.

[0066] Heat-inactivated or heat-killed bacteria can be obtained by heating at 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. The heating can be carried out for at least about 5 minutes, preferably at least about 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes or longer. The heating can be carried out using a saturated steam pressure of at least about 10 psig, preferably at least about 11, 12, 13, 14, 15 or higher psig.

[0067] In some embodiments, the bacteria and / or variants thereof from the genus Odoribacter contained in the composition used in the present invention are pasteurized bacteria.

[0068] In one embodiment, the pasteurized bacteria and / or variants thereof are heated at a temperature of about 50 °C to about 100 °C, preferably about 60 °C to about 95 °C, more preferably about 70 °C to about 90 °C. In one embodiment, the pasteurized bacteria and / or variants thereof are 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 are 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 are 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 are 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 are heated at a temperature of about 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 °C or 100 °C.

[0069] In one embodiment, the pasteurized bacteria and / or its variants are not heated at a temperature above about 100 °C. In a particular embodiment, the pasteurized bacteria and / or its variants are not heated at ultra-high temperatures, such as at a temperature of about 110 °C to about 140 °C. In one embodiment, the pasteurized bacteria and / or its variants are not heated at a temperature above about 90 °C. Thus, in one embodiment of the present invention, the bacteria and / or its variants are not sterilized. Sterilization is a process intended to destroy, kill, or inactivate all forms of life and other biological agents. This includes microorganisms and their spores, as well as viruses and prions. Unlike sterilization, pasteurization is not intended to kill all microorganisms, but is typically applied to food to reduce the number of live pathogens.

[0070] In one embodiment of the present invention, the pasteurized bacteria and / or its variants are heated for at least about 10 minutes. In another embodiment of the present invention, the pasteurized bacteria and / or its variants are heated for at least about 15, 20, 25, 30, 35, or 45 minutes. In one embodiment, the pasteurized bacteria and / or its variants are heated for a time of about 10 to about 45 minutes.

[0071] In one embodiment, the pasteurized bacteria and / or its variants are not heated for a short time. In a particular embodiment, the pasteurized bacteria and / or its variants are not heated for less than about 30 seconds, less than about 60 seconds, less than about 90 seconds, or less than about 120 seconds. In a preferred embodiment, the pasteurized bacteria and / or its variants are not heated for less than about 1 minute, preferably not heated for less than about 5, 6, 7, 8, or 9 minutes.

[0072] In one embodiment, the pasteurized bacteria and / or its variants are heated to at least about 10 minutes at a temperature of about 50 °C to about 100 °C. In a particular embodiment, the pasteurized bacteria and / or its variants are heated to about 60 °C for about 20 minutes or about 30 minutes. In another particular embodiment, the pasteurized bacteria and / or its variants are heated to about 70 °C for about 20 minutes or about 30 minutes. In another particular embodiment, the pasteurized bacteria and / or its variants are heated to about 80 °C for about 20 minutes or about 30 minutes. In another particular embodiment, the pasteurized bacteria and / or its variants are heated to about 90 °C for about 20 minutes or about 30 minutes.

[0073] In certain embodiments, the pasteurized bacteria and / or variants thereof are not heated at a temperature above about 110 °C for about 1 to about 120 seconds. In another particular embodiment, the pasteurized bacteria and / or variants thereof are not heated at a temperature above about 100 °C for about 1 to about 120 seconds. In another particular embodiment, the pasteurized bacteria and / or variants thereof are not heated at a temperature above about 90 °C for about 1 to about 120 seconds.

[0074] In one embodiment, the bacteria and / or variants thereof are treated with ultra-high temperature (UHT) treatment.

[0075] As used herein, "UHT" treatment refers to ultra-high temperature processing or ultra-heat treatment (both abbreviated as UHT), which involves at least partial sterilization of a composition by short-time heating at a temperature of at least about 135 °C (e.g., about 1 to about 60 seconds, preferably about 1 to about 30 seconds, more preferably about 1 to about 10 seconds).

[0076] There are mainly two types of UHT systems: direct systems and indirect systems. In a direct system, the product is treated by steam injection or steam infusion, while in an indirect system, the product is heat-treated using a plate heat exchanger, a tubular heat exchanger, or a scraped surface heat exchanger. Combinations of UHT systems can be applied to any step or multiple steps in the product preparation process.

[0077] In one embodiment, the bacteria and / or variants thereof are subjected to flash pasteurization. Thus, in one embodiment, the bacteria and / or variants thereof are treated at a temperature of about 71.5 °C to about 74 °C for a time of about 15 to about 30 seconds.

[0078] In some embodiments, the bacteria and / or variants thereof from the genus Odoribacter contained in the composition used in the present invention are fresh. As used herein, the term "fresh" means that the bacteria have not been frozen between the last amplification stage and use.

[0079] In some embodiments, the bacteria and / or variants thereof from the genus Odoribacter contained in the composition used in the present invention are not fresh. In some embodiments, the bacteria and / or variants thereof are frozen at least once. In some embodiments, the bacteria and / or variants thereof are frozen.

[0080] As used herein, the term "frozen" means cooling the bacteria to or below the temperature that allows it to change from the liquid phase to the solid phase. In one embodiment, the temperature is about -5 °C, -20 °C, -70 °C, -80 °C, or -190 °C.

[0081] In one embodiment, the recovered frozen bacteria and / or variants thereof are viable. In other words, in one embodiment, the bacteria and / or variants thereof from the genus Odoribacter contained in the composition used in the present invention are frozen and viable.

[0082] In some embodiments, the composition used in the present invention comprises a fragment of a bacterium from the genus Odoribacter, preferably a fragment of Odoribacter wuertzii, more preferably a fragment of the Odoribacter wuertzii strain J115 T of the fragment.

[0083] As used herein, the term "fragment" refers to bacterial components, metabolites, secreted molecules and / or vesicles, and compounds produced by the metabolism of bacteria and / or their variants contained in the composition used in the present invention, etc. 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, and bacterial structural components (such as proteins, carbohydrates, lipids and their combinations, such as lipoproteins, glycolipids and glycoproteins), bacterial metabolites, organic acids, inorganic acids, bases, peptides, enzymes and coenzymes, amino acids, carbohydrates, lipids, glycoproteins, lipoproteins, glycolipids, vitamins, bioactive compounds and metabolites containing inorganic components. The fragment can be obtained by recovering the supernatant of the culture of bacteria and / or their variants or by extracting cell components or cell fractions, metabolites or secreted compounds, degradation products, separated forms of components, any mixture of one or more components of bacteria and / or their variants, or one or more components produced in another way (such as using recombinant DNA technology) in a microbial host or produced in any other (bio)synthesis process in bacteria and / or their variants. In one embodiment, the fragment is contained in the culture medium of the bacteria, i.e., the fragment is not purified or isolated. In another embodiment, the fragment is purified by methods known in the art, such as affinity purification.

[0084] In some embodiments, the composition used in the present invention comprises an extract of the supernatant of the culture medium of bacteria from the genus Odoribacter and / or their variants.

[0085] Media suitable for bacterial culture are known in the art. In some embodiments, particularly for the culture of bacteria from the genus Odoribacter, the medium is yeast extract-casein hydrolysate-fatty acid (YCFA) medium. In some embodiments, the YCFA medium is modified.

[0086] 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 further or alternatively contains soy peptone in an amount greater than 0 g / L to about 20 g / L, preferably about 2 g / L to about 6 g / L, more preferably about 4 g / L. In some embodiments, the modified YCFA medium contains wheat peptone in an amount greater than 0 g / L to about 20 g / L, preferably about 2 g / L to about 6 g / L, more preferably about 4 g / L. In some embodiments, the modified YCFA medium contains Na2CO3 in an amount greater than 0 g / L to about 20 g / L, preferably about 2 g / L to about 6 g / L, more preferably about 3 g / L. In some embodiments, the modified YCFA medium contains MgCl2 in an amount greater than 0 mg / L to about 500 mg / L, preferably about 25 mg / L to about 75 mg / L, more preferably about 50 mg / L. In some embodiments, the modified YCFA medium contains glutathione in an amount greater than 0 g / L to about 5 g / L, preferably about 0.5 g / L to about 1.5 g / L, more preferably about 1 g / L. In some embodiments, the modified YCFA medium contains ascorbic acid in an amount greater than 0 g / L to about 1 g / L, preferably about 0.25 g / L to about 0.75 g / L, more preferably about 0.5 g / L. In some embodiments, the modified YCFA medium contains 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, and.

[0087] In one embodiment, the modified YCFA medium contains 8 g of yeast extract, 4 g of soy peptone, 4 g of wheat peptone, 5 g of KH2PO4, 3 g of Na2CO3, 50 mg of MgCl2, 50 mg of CaCl2, 1 mg of hemin, 1 mL of resazurin solution (1 g / L), 10 g of myo-inositol, 1 g of cysteine, 1 g of reduced glutathione, 0.5 g of ascorbic acid, 0.3 g of uric acid, 1 mL of vitamin solution, and H2O to make up to 1000 mL. In one embodiment, the vitamin solution for preparing the modified YCFA medium contains 2 mg of biotin, 2 mg of folic acid, 10 mg of pyridoxine hydrochloride, 2 mg of thiamine dihydrochloride, 5 mg of riboflavin, 5 mg of nicotinic acid, 5 mg of calcium D-pantothenate, 12 mg of vitamin B, 5 mg of p-aminobenzoic acid, 5 mg of lipoic acid, and H2O to make up to 1000 mL.

[0088] In some embodiments, the modified YCFA medium comprises about 4 g / L of soy peptone, about 4 g / L of wheat peptone, about 3 g / L of Na2CO3, about 1 g / L of glutathione, about 1 g / L of ascorbic acid, about 0.5 g / L of ascorbic acid, 0.5 g / L of MgCl2, 8 g / L of yeast extract, 2 g / L of KH2PO4, 1 g / L of sodium propionate, 0.5 g / L of cysteine, 1 g / L of hemin, 1 g / L of L-cysteine, and the vitamin solution as described above.

[0089] In one embodiment, the medium is free of live bacteria, i.e., the medium is sterilized. Methods for sterilizing the medium are known in the art and include, for example, heating and radiation. In another embodiment, the medium contains live bacteria, preferably a substantially pure population of bacteria from the genus Odoribacter and / or its variants.

[0090] In some embodiments, the medium is frozen and stored at a temperature of about -5°C, -20°C, -70°C, -80°C, or -190°C for later use.

[0091] In some embodiments, the medium is fermented. In some embodiments, the fermentation of the medium comprises the following steps:

[0092] 1) Incubate about 10 T to about 10 8 live bacteria from the genus Odoribacter, preferably from Odoribacter wieringae, more preferably from Odoribacter wieringae strain J115 9 in 50 mL of the modified YCFA medium at 37°C under anaerobic conditions for 48 hours;

[0093] 2) Incubate the culture from step 1) in 3.5 L of the modified YCFA medium at 37°C under anaerobic conditions for 48 hours;

[0094] 3) Collect the medium obtained from step 2), wherein the medium is fermented;

[0095] 4) And optionally, remove live bacteria from the medium collected in step 3) (i.e., sterilize the medium).

[0096] In some embodiments, the extract from bacteria of the genus Odoribacter and / or its variants, or the extract from the culture supernatant, contained in the composition used in the present invention, contains at least one metabolite secreted by said bacteria.

[0097] In some embodiments, the compositions used in the present invention comprise metabolites produced and / or secreted by bacteria from the genus Odoribacter, preferably from Odoribacteraceae, more preferably from Odoribacteraceae strain J115 T and / or secreted metabolites.

[0098] In some embodiments, the at least one metabolite is selected from the group consisting of or consisting of short-chain fatty acids (SCFAs), such as butyrate and acetate.

[0099] In some embodiments, the at least one metabolite is selected from the group consisting of or consisting of butyrate, acetate, C8-3OH, C10-3OH, C12-3OH, C16-2OH C16-3OH, C18-2OH C18-3OH, C18-130H, C18, C18:2 n-6, C22:2 n-6, C22:6 n-3, C18:1 n-9c, C22:1 n-9, C18:2 n-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.

[0100] The inventors have demonstrated that the action of bacteria from the genus Odoribacter, preferably from Odoribacteraceae, more preferably from Odoribacteraceae strain J115 T is not limited to the metabolites produced and / or secreted by the bacteria. Specifically, they have demonstrated that this action is not limited to the presence or secretion of short-chain fatty acids (SCFAs) such as butyrate and acetate. In fact, from the results obtained by the inventors, it can be seen that the culture medium obtained after bacterial fermentation completely inhibits cell proliferation at a concentration of 20%, thus equivalent to a potency 10 to 20 times higher than that of SCFAs (see Figure 5B and 5D , SCFA-matched compared to fermented).

[0101] In some embodiments, the compositions used in the present invention comprise an active agent from the culture supernatant of bacteria from the genus Odoribacter and / or its variants. In one embodiment, the active agent is a metabolite produced and / or secreted by the bacteria of the present invention as described above.

[0102] Another object of the present invention is the culture supernatant of bacteria from the genus Odoribacter and / or its variants for the prevention and / or treatment of breast cancer in a subject in need thereof.

[0103] In one embodiment, the culture supernatant is obtained by harvesting the culture medium of bacteria from the genus Odoribacter and / or its variants. In one embodiment, the culture supernatant is the fermentation medium obtained after culturing the bacteria of the present invention. In one embodiment, the culture of bacteria from the genus Odoribacter and / or its variants is carried out as disclosed above.

[0104] In a specific embodiment, the culture supernatant is obtained by the following steps:

[0105] 1) Approximately 10 T to approximately 10 8 live bacteria from the genus Odoribacter, preferably from Odoribacter wuellerstorfii, more preferably from Odoribacter wuellerstorfii strain J115 9 are incubated in 50 mL of modified YCFA medium at a temperature of 37 °C under anaerobic conditions for 48 hours;

[0106] 2) The culture of step 1) is incubated in 3.5 L of modified YCFA medium at a temperature of 37 °C under anaerobic conditions for 48 hours;

[0107] 3) The culture medium obtained in step 2) is collected, wherein the culture medium is fermented;

[0108] 4) And optionally, live bacteria are removed from the culture medium collected in step 3).

[0109] As used herein, the term "breast cancer" refers to cancer of the breast that is histologically or cytologically confirmed. In some embodiments, breast cancer is carcinoma. In some embodiments, breast cancer is adenocarcinoma. In some embodiments, breast cancer is sarcoma.

[0110] In certain embodiments, breast cancer is hormone receptor-positive (HR+) breast cancer or hormone receptor-negative (HR-) breast cancer. In one embodiment, hormone receptor-positive breast cancer is estrogen receptor-positive (ER+) breast cancer and / or progesterone receptor-positive (PR+) breast cancer. In some embodiments, ER+ breast cancer is luminal A breast cancer. In some embodiments, ER+ breast cancer is luminal B breast cancer.

[0111] In some embodiments, breast cancer is human epidermal growth factor receptor 2-positive (HER2+) breast cancer. In some embodiments, breast cancer is human epidermal growth factor receptor 2-negative (HER2-) breast cancer.

[0112] In some embodiments, breast cancer is a cancer with combined expression of receptors, such as hormone receptor-positive and HER2-negative (HR+HER2-).

[0113] In some embodiments, breast cancer is a cancer of Group 1 (luminal A), Group 2 (luminal B), Group 3 (HER2+), or Group 4 (basal-like). Group 1 includes tumors that are ER+ and 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- and PR-, but HER2+. Group 4, also known as TNBC, includes tumors that are ER-, PR-, and HER2-.

[0114] In some embodiments, breast cancer is HER2+ breast cancer or triple-negative breast cancer (TNBC). In some embodiments, breast cancer is HER2+ breast cancer, preferably HER2+ breast adenocarcinoma. In some embodiments, breast cancer is triple-negative breast cancer (TNBC).

[0115] In some embodiments, breast cancer is non-invasive breast cancer, particularly ductal carcinoma in situ or lobular carcinoma in situ. In certain embodiments, breast cancer is invasive breast cancer, particularly selected from the group consisting of or comprising: 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.

[0116] In some embodiments, the tissue type in which breast cancer occurs is the milk duct, milk-producing lobule, or connective tissue.

[0117] In some embodiments, breast cancer is metastatic or locally advanced breast cancer. The term "locally advanced breast cancer" refers to a cancer that has spread from its origin in the breast to nearby tissues or lymph nodes, but has not spread to other parts of the body.

[0118] In certain embodiments, the cancer is a metastatic cancer or a cancer prone to metastasis, particularly metastatic breast cancer or a breast cancer prone to metastasis.

[0119] The term "metastatic breast cancer" refers to a cancer that has spread from the breast to other parts of the body (such as the bone, liver, lung, or brain). Metastatic breast cancer may also be referred to as stage IV breast cancer. As used herein, "a cancer prone to metastasis" refers to an invasive cancer in which cancer cells may break away from the primary tumor and spread in other organs, where the cancer cells can grow to form secondary tumors, also known as metastases.

[0120] Many pathophysiological factors that can affect tumor growth are known in the art, for example, conditions related to diet, obesity, glucose and lipid metabolism, and inflammation, in particular. These conditions are relevant to the scope of the present invention because commensal bacteria (including bacteria from the genus Odoribacter) can affect these conditions.

[0121] In some embodiments, the subject also has one or more diseases or disorders that accelerate tumor growth, and the diseases or disorders are selected from the group consisting of or comprising: obesity-related disorders, liver diseases, metabolic disorders, adipokine-related disorders, and inflammatory diseases, and combinations thereof. In some embodiments, the one or more diseases or disorders that accelerate tumor growth are selected from the group consisting of or comprising: obesity-related disorders, metabolic disorders, and inflammatory diseases, and combinations thereof. In some embodiments, the one or more diseases or disorders that accelerate tumor growth are selected from the group consisting of or comprising: obesity-related disorders and metabolic disorders.

[0122] In some embodiments, the subject also has an obesity-related disorder. In some embodiments, the subject also has a liver disease. In some embodiments, the subject also has a metabolic disorder. In some embodiments, the subject also has an adipokine-related disorder. In some embodiments, the subject also has an inflammatory disease.

[0123] In some embodiments, the subject does not have an obesity-related disorder. In some embodiments, the subject has a normal BMI index.

[0124] In some embodiments, the subject also has one or more diseases or disorders that accelerate tumor growth, which are selected from the group consisting of or comprising: obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), cirrhosis, diabetes, glucose intolerance, hyperglycemia, dyslipidemia, hyperlipidemia, hypercholesterolemia, elevated LDL-cholesterol, reduced HDL-cholesterol, elevated triglycerides, and intestinal inflammation, and combinations thereof.

[0125] In some embodiments, the subject also has obesity. In some embodiments, the subject also has NASH. In some embodiments, the subject also has NAFL. In some embodiments, the subject also has cirrhosis. In some embodiments, the subject also has diabetes. In some embodiments, the subject also has glucose intolerance. In some embodiments, the subject also has hyperglycemia. In some embodiments, the subject also has dyslipidemia. In some embodiments, the subject also has abnormal lipid metabolism. In some embodiments, the subject also has hypercholesterolemia. In some embodiments, the subject also has elevated LDL-cholesterol. In some embodiments, the subject also has reduced HDL-cholesterol. In some embodiments, the subject also has elevated triglycerides. In some embodiments, the subject also has intestinal inflammation.

[0126] In some embodiments, the subject has an imbalance of the gut microbiota. As used herein, an imbalance of the gut microbiota refers to an overrepresentation or underrepresentation of at least one microbial species, preferably at least one bacterial species. In some embodiments, the population of bacteria from the genus Odoribacter is underrepresented. In some embodiments, the population of the species Odoribacter wuellerstorfii is underrepresented. In some embodiments, in subjects with obesity and / or obesity-related disorders, the population of Odoribacter wuellerstorfii is reduced.

[0127] In some embodiments, the population of Odoribacter wuellerstorfii is negatively correlated with the body mass index of the subject.

[0128] In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacterium, preferably from about 1×10 2 to about 1×10 15 CFU.

[0129] As used herein, "CFU" means "colony forming unit".

[0130] In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacterium, preferably from about 1×10 4 to about 1×10 12 CFU, more preferably from about 1×10 5 to about 1×10 10 CFU, even more preferably from about 1×10 6 to about 5×10 9 CFU.

[0131] In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacterium, preferably from about 1×10 2 to about 1×10 15 CFU, from about 1×10 4 to about 1×1012 CFU, about 1×10 5 to about 1×10 10 CFU, about 1×10 6 to about 5×10 9 CFU. In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacteria, preferably about 1×10 4 to about 1×10 14 CFU, about 1×10 5 to about 1×10 13 CFU, about 1×10 6 to about 1×10 12 CFU, about 1×10 7 to about 1×10 11 CFU, about 1×10 8 to about 1×10 10 CFU, about 2×10 8 to about 6×10 9 CFU.

[0132] In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacteria, preferably about 1×10 4 to about 1×10 12 CFU / mL, more preferably about 1×10 5 to about 1×10 10 CFU / mL, even more preferably about 1×10 6 to about 5×10 9 CFU / mL.

[0133] In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacteria, preferably about 1×10 4 to about 1×10 14 CFU / mL, preferably about 1×10 5 to about 1×10 13 CFU / mL, more preferably about 1×10 6 to about 1×10 12 CFU / mL, even more preferably about 1×10 7 to about 1×10 11 CFU / mL, 1×10 8 to about 1×10 10 CFU / mL, and even more preferably about 2×10 8 to about 6×10 9 CFU / mL.

[0134] In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacteria, preferably about 1×10 2 to about 1×1015 CFU / g, preferably about 1×10 4 to about 1×10 12 CFU / g, more preferably about 1×10 5 to about 1×10 10 CFU / g, and even more preferably about 1×10 6 to about 5.10 9 CFU / g. In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacteria, preferably about 1×10 4 to about 1×10 14 CFU / g, preferably about 1×10 5 to about 1×10 13 CFU / g, more preferably about 1×10 6 to about 1×10 12 CFU / g, even more preferably about 1×10 7 to about 1×10 11 CFU / g, about 1×10 8 to about 1×10 10 CFU / g, and even more preferably about 2×10 8 to about 6×10 9 CFU / g.

[0135] In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacteria, preferably about 1×10 6 to about 1×10 10 CFU / g or CFU / mL, preferably about 1×10 8 to about 1×10 10 CFU / g or CFU / mL, more preferably about 1×10 9 to about 1×10 10 CFU / g or CFU / mL. In some embodiments, the composition used in the present invention comprises a therapeutically effective amount of said bacteria, preferably about 1×10 6 to about 1×10 11 CFU / g or CFU / mL, preferably about 1×10 8 to about 1×10 11 CFU / g or CFU / mL, more preferably about 1×10 10 to about 1×10 11 CFU / g or CFU / mL.

[0136] In one embodiment of the present invention, the amount of the fragment of the bacteria and / or its variant comprised in the composition used in the present invention corresponds to about 1×10 2 to about 1×10 15 cells / g, preferably about 1×10 4to about 1×10 12 cells / g, more preferably about 1×10 5 to about 1×10 10 cells / g, even more preferably about 1×10 6 to about 1×10 9 cells / g of the amount of bacteria and / or its variants. In one embodiment of the present invention, the amount of the fragment of the bacteria and / or its variants contained in the composition used in the present invention corresponds to about 1×10 4 to about 1×10 14 cells / g, preferably about 1×10 5 to about 1×10 13 cells / g, more preferably about 1×10 6 to about 1×10 12 cells / g, even more preferably about 1×10 7 to about 1×10 11 cells / g of the amount of bacteria and / or its variants. In one embodiment of the present invention, the amount of the fragment of the bacteria and / or its variants contained in the composition used in the present invention corresponds to about 1×10 8 to about 1×10 10 cells / g, preferably about 1×10 9 to about 1×10 10 cells / g of the amount of bacteria and / or its variants.

[0137] In one embodiment of the present invention, the amount of the fragment of the bacteria and / or its variants contained in the composition used in the present invention corresponds to about 1×10 2 to about 1×10 15 cells / mL, preferably about 1×10 4 to about 1×10 12 cells / mL, more preferably about 1×10 5 to about 1×10 10 cells / mL, even more preferably about 1×10 6 to about 1×10 9 cells / mL of the amount of bacteria and / or its variants. In one embodiment of the present invention, the amount of the fragment of the bacteria and / or its variants contained in the composition used in the present invention corresponds to about 1×10 4 to about 1×10 14 cells / mL, preferably about 1×10 5 to about 1×10 13 cells / mL, more preferably about 1×10 6 to about 1×10 12 cells / mL, even more preferably about 1×10 7 to about 1×10 11The amount of bacteria and / or its variants at [X] cells / mL. In one embodiment of the present invention, the composition used in the present invention contains an amount of fragments of bacteria and / or its variants corresponding to about 1×10 8 to about 1×10 10 cells / mL, preferably about 1×10 9 to about 1×10 10 cells / mL of bacteria and / or its variants.

[0138] In one embodiment of the present invention, the composition used in the present invention contains an amount of fragments of bacteria and / or its variants corresponding to about 1×10 6 to about 1×10 10 cells / g or cells / mL, preferably about 1×10 8 to about 1×10 10 cells / g or cells / mL, more preferably about 1×10 9 to about 1×10 10 cells / g or cells / mL of bacteria and / or its variants.

[0139] In one embodiment of the present invention, the composition used in the present invention contains an amount of fragments of bacteria and / or its variants corresponding to about 1×10 6 to about 1×10 11 cells / g or cells / mL, preferably about 1×10 8 to about 1×10 11 cells / g or cells / mL, more preferably about 1×10 10 to about 1×10 11 cells / g or cells / mL of bacteria and / or its variants.

[0140] In some embodiments, the composition used in the present invention is in the form of a pharmaceutical composition and further contains a pharmaceutically acceptable carrier.

[0141] It should be understood that the carrier must be "acceptable", that is, compatible with the composition used in the present invention and will not produce harmful effects when administered to an individual. Generally, the carrier will not produce adverse, allergic or other adverse reactions when administered to an individual (preferably a human individual).

[0142] In some embodiments, the composition used in the present invention or the pharmaceutical composition used in the present invention further contains at least one other anti-cancer agent.

[0143] Anticancer agents are known in the prior art. Non-limiting examples of anticancer agents include acalabrutinib, alectinib, alemtuzumab, anastrozole, avapritinib, avelumab, belinostat, bevacizumab, bleomycin, blinatumomab, bosutinib, brigatinib, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, copanlisib, cytarabine, daunorubicin, decitabine, dexamethasone, docetaxel, doxorubicin, encorafenib, erdafitinib,) Etoposide, everolimus, exemestane, 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, zanubrutinib, etc.

[0144] In a preferred embodiment, the anticancer agent is an anticancer agent suitable for the treatment of breast cancer.

[0145] In certain embodiments, the anticancer agent will be administered simultaneously or sequentially in combination with the composition used in the present invention or the pharmaceutical composition used in the present invention.

[0146] The present invention also relates to a drug comprising the composition or pharmaceutical composition used in the present invention. In some embodiments, the drug is used for the treatment and / or prevention of breast cancer.

[0147] The present invention also relates to a composition comprising at least: (a) bacteria from the genus Odoribacter, preferably from the species Odoribacter westerdijkiae, more preferably from the strain Odoribacter westerdijkiae J115 T and / or its variants and / or fragments, and / or (b) the culture supernatant of bacteria from the genus Odoribacter, preferably from the species Odoribacter westerdijkiae, more preferably from the strain Odoribacter westerdijkiae J115 T for the manufacture of a drug for the treatment and / or prevention of breast cancer in a subject in need thereof.

[0148] Another object of the present invention is bacteria from the genus Halomonas, preferably from the species Halomonas ventosae, more preferably from the Halomonas ventosae strain J115 T and / or its variants and / or fragments, for use in the manufacture of a medicament for treating and / or preventing breast cancer in a subject in need thereof.

[0149] Another object of the present invention is the culture supernatant of bacteria from the genus Halomonas, preferably from the species Halomonas ventosae, more preferably from the Halomonas ventosae strain J115 T and / or its variants, for use in the manufacture of a medicament for treating and / or preventing breast cancer in a subject in need thereof.

[0150] In some embodiments, the composition used in the present invention is in the form of a nutritional product or nutritional composition, which further comprises a pharmaceutically acceptable agent. In one embodiment, the nutritional composition comprises a nutritionally effective amount of bacteria from the genus Halomonas and / or its variants, extracts or fragments.

[0151] In some embodiments, the nutritional composition is intended for ingestion by human users and may be useful due to its prophylactic and medicinal properties. In one embodiment, the nutritional composition of the present invention is used for preventing and / or treating breast cancer in a subject in need thereof.

[0152] In one embodiment, the nutritional composition is in the form of a food or dietary supplement. In another embodiment, the nutritional composition is in the form of a functional food. In some embodiments, the nutritional composition is formulated as a food additive, beverage, tablet, capsule, caplet, lozenge, syrup, suspension or emulsion.

[0153] In some embodiments, the nutritional composition further comprises one or more secondary nutritional components, wherein the secondary nutritional components have nutritional or therapeutic activity in preventing and / or treating breast cancer. In some embodiments, the secondary nutritional components are vitamins, minerals.

[0154] In some embodiments, the pharmaceutically acceptable agent is selected from one or more of the group consisting of talc, titanium dioxide, starch, corn starch, modified corn starch, kaolin, microcrystalline cellulose and powdered cellulose.

[0155] The present invention also relates to a prebiotic, which comprises one or more active ingredients or substances that can increase the level of bacteria from the genus Halomonas, preferably from the species Halomonas ventosae, more preferably from the Halomonas ventosae J115 T strain and / or its variants and / or fragments in the microbiota of a subject in need thereof.

[0156] In one embodiment, the prebiotic is used for preventing and / or treating breast cancer in said subject.

[0157] In some embodiments, the prebiotic is selected from the group consisting of or comprising: myo-inositol, inulin and inulin-type fructans, fructooligosaccharides, β-glucans, xylose, arabinose, arabinoxylans, ribose, phytates, galactose, rhamnose, cellobiose, fructose, lactose, salicin, sucrose, glucose, aesculin, Tween-80, trehalose, maltose, mannose, melibiose, mucus or mucin, raffinose, fructooligosaccharides, galactooligosaccharides, amino acids, alcohols, fermentable carbohydrates, water-soluble cellulose derivatives (such as methylcellulose, methyl ethylcellulose, hydroxyethylcellulose, ethyl hydroxyethylcellulose, cationic hydroxyethylcellulose, hydroxypropylcellulose, hydroxyethyl methylcellulose, hydroxypropyl methylcellulose, and carboxymethylcellulose), water-insoluble cellulose derivatives, unprocessed oats, Metamucil, whole bran, polyphenols, and any combination thereof.

[0158] The present invention also relates to a therapeutic combination product for separate, simultaneous or sequential administration.

[0159] As used herein, the term "therapeutic combination product" (which may also be referred to as a therapeutic kit) refers to a product comprising or consisting of at least the following two parts: a first part comprising the composition used in the present invention, the pharmaceutical composition used in the present invention, or the medicament of the present invention, and a second part comprising a prebiotic. In one embodiment, the therapeutic combination product is for the treatment and / or prevention of breast cancer.

[0160] In some embodiments, the prebiotic is included in the composition used in the present invention, the pharmaceutical composition used in the present invention, or the medicament of the present invention.

[0161] The present invention also relates to a composition comprising at least (a) a bacterium from the genus Odoribacter, preferably from the species Odoribacteraceae, more preferably from the strain Odoribacteraceae J115 T and / or a variant, extract or fragment thereof, and / or (b) a culture supernatant of a bacterium from the genus Odoribacter, preferably from the species Odoribacteraceae, more preferably from the strain Odoribacteraceae J115 T as an adjuvant for use in the treatment of a subject suffering from breast cancer.

[0162] As used herein, "adjuvant" refers to an agent that enhances the therapeutic effect, typically in the treatment of breast cancer. "Enhanced" means that the treatment used in combination with the adjuvant has a positive effect on the disease or condition (preferably breast cancer) that is greater than the sum of the effects of the treatment and the adjuvant used alone.

[0163] In some embodiments, the breast cancer of the subject is treated by methods known in the art, such as hormone therapy, chemotherapy, and immunotherapy.

[0164] In some embodiments, the subject also has one or more diseases or conditions that accelerate tumor growth, and the diseases or conditions are selected from the group consisting of or comprising: obesity-related diseases, liver diseases, metabolic diseases, adipokine-related diseases, and inflammatory diseases, and combinations thereof. In some embodiments, the subject also has one or more diseases or conditions that accelerate tumor growth, and the diseases or conditions are selected from the group consisting of or comprising: obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), cirrhosis, diabetes, glucose intolerance, hyperglycemia, dyslipidemia, abnormal lipid metabolism, hypercholesterolemia, elevated LDL-cholesterol, reduced HDL-cholesterol, elevated triglycerides, and intestinal inflammation, and combinations thereof.

[0165] In some embodiments, the subject has an imbalance of the gut microbiota. In some embodiments, compared to a healthy subject, the gut microbiota of the subject comprises a reduced bacterial flora from the genus Odoribacter, preferably from Odoribacteraceae.

[0166] 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 composition, the composition comprising at least (a) bacteria from the genus Odoribacter, preferably from the species Odoribacteraceae, more preferably from Odoribacteraceae J115 T strain, and / or variants, extracts or fragments thereof, and / or (b) a culture supernatant of bacteria from the genus Odoribacter, preferably from the species Odoribacteraceae, more preferably from Odoribacteraceae J115 T strain and / or variants thereof.

[0167] The therapeutically effective amount of the composition has been described above.

[0168] 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 have been described 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, the pharmaceutical composition comprising: (i) a composition comprising bacteria from the genus Odoribacter and / or variants, extracts or fragments thereof, and (ii) another therapeutic agent, wherein the another therapeutic agent is an anti-cancer agent.

[0169] In one embodiment, the method is used when the subject has breast cancer, with the aim of slowing down or reversing the progression of tumor growth. In another embodiment, the method is used when the subject does not have breast cancer, with the aim of preventing the occurrence of breast cancer. In one embodiment, the method is used acutely. In another embodiment, the method is used chronically.

[0170] In some embodiments, the subject also has one or more diseases or disorders that accelerate tumor growth, and the diseases or disorders are selected from the group consisting of or comprising: obesity-related diseases, liver diseases, metabolic diseases, adipokine-related diseases, and inflammatory diseases, and combinations thereof. In some embodiments, the subject also has one or more diseases or disorders that accelerate tumor growth, and the diseases or disorders are selected from the group consisting of or comprising: obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), cirrhosis, diabetes, glucose intolerance, hyperglycemia, dyslipidemia, lipid metabolism disorder, hypercholesterolemia, elevated LDL-cholesterol, reduced HDL-cholesterol, elevated triglycerides, and intestinal inflammation, and combinations thereof.

[0171] In some embodiments, the subject has an imbalance of the gut microbiota. In some embodiments, compared to a healthy subject, the gut microbiota of the subject contains a reduced population of bacteria from the genus Odoribacter, preferably Odoribacter wuellerstorfii.

[0172] The present invention also relates to a composition comprising at least (a) bacteria of the genus Odoribacter, preferably of the species Odoribacter wuellerstorfii, more preferably of the strain Odoribacter wuellerstorfii J115 T and / or its variants, and / or (b) the culture supernatant of bacteria of the genus Odoribacter, preferably of the species Odoribacter wuellerstorfii, more preferably of the strain Odoribacter wuellerstorfii J115 T and / or its variants, for use as a probiotic. In certain embodiments, the bacteria comprised in the composition are the strain J115 T and / or its variants, which are beneficial for improving the gastrointestinal environment of the subject.

[0173] The present invention also relates to a method of increasing the population of bacteria of the genus Odoribacter, preferably of the species Odoribacter wuellerstorfii, more preferably of the strain Odoribacter wuellerstorfii J115 T in the gut microbiota of a subject, the method comprising administering the bacteria to the subject, wherein the bacteria are comprised in a composition, a pharmaceutical composition, or a prebiotic. In a preferred embodiment, the subject is at risk of developing breast cancer or has breast cancer.

[0174] The present invention also relates to a method of treating an imbalance of the gut microbiota, the method comprising administering to the subject a composition or a prebiotic, the composition or prebiotic comprising at least (a) bacteria of the genus Odoribacter, preferably of the species Odoribacter wuellerstorfii, more preferably of the strain Odoribacter wuellerstorfii J115 T and / or (b) bacteria of the genus Odoribacter, preferably of the species Odoribacter wuellerstorfii, more preferably of the strain Odoribacter wuellerstorfii J115 TCulture supernatant of the bacterium of the strain and / or its variants.

[0175] Brief Description of the Drawings

[0176] Figures 1A - 1G A set of figures shows Bacillus foetidus J115 T Partially alleviated tumor growth induced by a high-fat diet. Figure 1A Shows the body weight changes under normal diet (ND) and high-fat diet (HFD). Arrows indicate tumor induction. Figure 1B Shows the changes in E0771 tumor volume. Figure 1C Shows the changes in tumor volume, concentrated in the ND and HFD groups. Figure 1D Shows the changes in tumor volume, concentrated in the group treated with J115 T or not treated with J115 T in the HFD group. Figure 1E Shows the survival curve derived from linear regression by calculating the time required for each tumor to reach 300 mm 3 Figure 1F Shows the survival curve concentrating on the difference between the ND and HFD groups. Figure 1G Shows the survival curve concentrating on the difference between the HFD and HFDJ115 T groups. Sample size: n = 9 for ND, n = 8 for HFD, and n = 11 for HFDJ115 T Excluded mice with tumors smaller than 100 mm 3 Statistical analysis: ( Figure 1A , 1B ) Two-way ANOVA analysis, followed by Tukey's multiple comparison test. ***p < 0.001 (HFD vs ND) and $p < 0.05 (HFDJ115 T vs HFD). ( Figure 1C , 1D ) Two-way ANOVA analysis, followed by Sidak's multiple comparison test. *p < 0.05: ***p < 0.001. ( Figure 1E , 1F , 1G) Log - rank (Mantel-Cox) test. *p < 0.05.

[0177] Figures 2A - 2E A set of figures shows Bacillus foetidus J115 T Alleviated tumor growth induced by a high-fat diet and confirmed the results of Figure 1. Figure 2A Shows the body weight changes under normal diet (ND) and high-fat diet (HFD). Arrows indicate tumor induction. Figure 2B Shows the changes in E0771 tumor volume. Figure 2C ​Shows the survival curves derived by linear regression of the time required for each tumor to reach 400 mm 3 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 HFDJ115 T groups. Sample sizes: n = 9 for ND, n = 10 for HFD, and n = 8 for HFDJ115 T Mice with tumors smaller than 100 mm 3 and with growth less than 50% (compared to day 6) were excluded. Statistical analysis: ( Figure 2A , 2B ) Two-way ANOVA analysis followed by Tukey's multiple comparison test. *p < 0.05; **p < 0.01; ***p < 0.001 (HFD vs ND) and $$p < 0.01 (HFD J115 T vs HFD). ( Figure 2C , 2D , 2E) Log-rank (Mantel-Cox) test. *p < 0.05.

[0178] Figures 3A - 3B The group graph of T shows that B. wexlerae J115 Figure 3A alleviated tumor growth induced by a high-fat diet and confirmed the results of Figures 1 and 2. Figure 3B Shows the body weight changes under a high-fat diet (HFD). The arrow indicates tumor induction.

[0179] Figures 4A - 4B The group graph of Figure 4A shows that B. wexlerae J115 alleviated tumor growth in a non-obesity-related context. Figure 4B Shows the body weight changes of Balb / c mice under a normal diet (ND).

[0180] Figures 5A - 5D Is a set of histograms showing that the supernatant of B. wexlerae J115 T reduced cell density and proliferation in a triple-negative breast cancer (TNBC) model in vitro. Figures 5A - 5B ​Shows the cell density (A) and cell proliferation (B) after treating PY8119 cells with non-fermenting bacterial medium, fermenting bacterial medium, and phosphate-buffered saline (PBS) matched for pH and short-chain fatty acid (SCFA) content (for fermentation) at the indicated concentrations for 24 hours. Figures 5C - 5D Shows the cell density (C) and cell proliferation (D) after treating E0771 cells with non-fermenting bacterial medium, fermenting bacterial medium, and phosphate-buffered saline (PBS) matched for pH and short-chain fatty acid (SCFA) content (for the fermenting medium) at the indicated concentrations for 24 hours. The pH of the fermenting 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 analysis followed by Dunnett's multiple comparison test. *p < 0.05; **p < 0.01; ***p < 0.001. Examples

[0181] The present invention is further illustrated by the following examples.

[0182] Example 1 :

[0183] Materials and Methods

[0184] Bacterial Culture

[0185] Bacteroides putridus J115 T was anaerobically cultured in modified YCFA medium supplemented with 10 g / L inositol. The culture was centrifuged at 5000 g for 15 minutes at 4 °C, and then the supernatant was removed. The cells were resuspended in anaerobic PBS-carbonate buffer supplemented with 15% (vol / vol) trehalose and then immediately frozen in an anaerobic vial and stored at -80 °C. The number of total, culturable bacteria administered to mice was calculated by plating the bacterial culture before centrifugation and the bacterial suspension used for mouse administration after freezing.

[0186] Cell Culture

[0187] The E0771 and PY8119 murine mammary adenocarcinoma cell lines were obtained from the American Type Culture Collection (ATCC) and stored according to the supplier's instructions. The 4T1 cells were derived from the mammary gland of BALB / c mouse strain. The E0771 and 4T1 cells were maintained in Dulbecco's Modified Eagle Medium (GIBCO, Thermo Fisher Scientific) containing 25 mM glucose, 4 mM glutamine, and 25 mM HEPES and supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Thermo Fisher Scientific). The PY8119 cells were maintained in F-12K Medium (GIBCO, Thermo Fisher Scientific) containing 7 mM glucose and 2 mM glutamine and supplemented with 5% heat-inactivated FBS (Thermo Fisher Scientific). They were cultured in a humidified environment at 37 °C and 5% CO2.

[0188] Cell density

[0189] Cell density was evaluated using the PrestoBlue reagent (Thermo Fisher Scientific) according to the manufacturer's instructions. Briefly, cells were treated with unfermented or fermented YCFA medium for 24 h as described above. To distinguish the contribution of short-chain fatty acids and the effect of pH, phosphate-buffered saline (PBS) at pH 6.1 containing 25 mM butyrate and 60 mM acetate was used. A 10% concentration of PrestoBlue reagent was added to each well. After a 2-h incubation, the fluorescence intensity (λex / λem = 560 / 590 nm) was measured using a plate reader (SpectraMax M2e, Molecular Devices). All data were normalized to the fluorescence intensity of the untreated wells and expressed as a percentage of the control.

[0190] Cell proliferation

[0191] Cell proliferation was measured using a kit (Roche) based on 5-bromo-2'-deoxyuridine (BrDu) incorporation ELISA according to the supplier's protocol. After a 24-h incubation with the treatments shown above, BrDu was added to the medium for 2 h. Then, after fixation of the cells with the solution provided by the manufacturer and binding with peroxidase-conjugated anti-BrDu antibody, the absorbance at 370 nm was measured using a plate reader (SpectraMax M2e, Molecular Devices) to evaluate cell proliferation. All data were normalized to the absorbance of the untreated wells and expressed as a percentage of the control.

[0192] Mouse

[0193] All mouse experiments were approved by the Animal Care Ethics Committee of the Health Department of the Catholic University of Leuven, with the specific number 2021 / UCL / MD / 04, and its amendment 2022 / UCL / MD / A10, and were conducted in accordance with the guidelines of the local ethics committee and the Belgian law on the protection of experimental animals of May 29, 2013 (protocol number LA1230467). Female C57BL / 6JRj mice (Janvier Labs) at 8 weeks of age certified pathogen-free (SPF) were used in the experiments. Cages were randomly assigned to experimental groups to ensure that the body weights of each group were matched at the start of normal diet (ND) (D10012M; Research Diets) and high-fat diet (HFD) 60% kcal (D12492; Research Diets) feeding, and before tumor induction. Body weight was evaluated once a week.

[0194] Oral administration of bacteria

[0195] Every day, later in the postprandial period, freshly thawed Bacteroides foetidus J115 T was administered to the mice by oral gavage. Each mouse received at least 1.0 × 10 9 live (culturable) bacteria in 0.2 mL. The ND and HFD control groups were orally gavaged with an equal volume of PBS-carbonate buffer supplemented with 15% (weight / volume) trehalose.

[0196] Tumor growth experiment

[0197] Six weeks after the start of HFD treatment, E0771 tumors were induced by subcutaneous injection of 1 × 10 6 cells into the fifth mammary fat pad of C57B1 / 6JRj female mice. 4T1 tumors were induced by subcutaneous injection of 2 × 10 5 cells into the fifth mammary fat pad of 8-week-old BALB / c mice. E0771 cells were freshly passaged before injection, prepared as a 1:1 mixture of PBS and Matrigel (Corning) and injected within 30 minutes. 4T1 cells were freshly passaged before injection, prepared in PBS solution and injected within 30 minutes. Tumor size was monitored at least twice a week (pilot experiment) or daily (validation), and measured using an electronic caliper in a simple blind method.

[0198] Statistical analysis

[0199] All statistical analyses were performed using GraphPad Prism software version 9.1.2.

[0200] Results

[0201] In the present invention, the ability of Bacillus foetidus J115 to reduce or delay cancer development under obese conditions was tested in vivo and in vitro. T

[0202] More precisely, as a first model, triple-negative breast cancer cells were used. Obesity was induced in mice by treatment with a high-fat diet, and then TNBC cells were injected into the mammary glands of the mice ( Figure 1A , see arrow = injection of tumor cells). Mice were treated daily with placebo or Bacillus foetidus J115 T . On day 15, the tumor growth rate of obese mice treated with placebo was significantly higher, and the tumor volume was 80% larger than that of lean mice ( Figure 1B and 1C ), while the tumor development of obese mice treated with Bacillus foetidus J115 T was significantly reduced and was similar to that of lean mice ( Figure 1B and 1D ).

[0203] As Figure 1E and 1F shown, obese mice were also characterized by faster tumor growth, and the time required for obese mice to reach the same tumor volume was shorter than that of lean mice. In other words, the tumors of lean mice reached a volume of 300 mm 3 in 27 days, while obese mice reached this size in only 19 days ( Figure 1E and 1F ). In summary, the effect of Bacillus foetidus J115 T in inducing a delay in tumor size significantly prolonged the time required to reach the same volume observed in obese mice. Therefore, the mice treated with Bacillus foetidus J115 T will reach a volume of 300 mm 3 in 27 days, which is similar to that of lean mice and much later than that of obese (weight-matched) mice receiving placebo ( Figure 1E and 1G ). Surprisingly, this beneficial effect was not associated with weight loss in the mice treated with Bacillus foetidus J115 T ( Figure 1A ).

[0204] This first set of data convincingly shows that a non-invasive method using, for example, Bacillus foetidus J115 T can be associated with a significant delay in cancer development, and surprisingly, this is not due to the anti-obesity effect of Bacillus foetidus J115 T .

[0205] In a second set of independent experiments (Figure 2), the effect of Bacillus foetidus J115 was fully reproduced TThe same beneficial effect on reducing tumor growth ( Figure 2B and 2C ), and even when selecting a volume of 400 mm 3 ( Figure 2D and 2E ) instead of 300 mm 3 as the threshold, this effect still exists significantly.

[0206] Finally, a third set of independent experiments (Figure 3) confirmed the beneficial effect of Bacillus foetidus J115 T on reducing tumor growth in the case of obesity ( Figure 3B ).

[0207] In the present invention, the ability of Bacillus foetidus J115 T to reduce or delay cancer development in the non-obese case was also tested in vivo.

[0208] Cells were subcutaneously injected into the fifth mammary fat pad of mice ( Figure 4A ). Then the mice were treated daily with placebo or Bacillus foetidus J115 T . On the 16th day, compared with the untreated mice, the mice treated with Bacillus foetidus J115 T showed significantly reduced tumor growth ( Figure 4B ). These experiments showed that Bacillus foetidus J115 T delayed cancer development independent of obesity.

[0209] Next, an in vitro test was performed using the fermentation medium of Bacillus foetidus J115 T to study the effect of potential metabolites produced by Bacillus foetidus J115 T (Figure 5). The fermentation medium of Bacillus foetidus J115 T significantly reduced the cell density of two different triple-negative breast cancer models (E0771 and PY8119 cells) in a dose-dependent manner after 24 hours of treatment ( Figure 5A and 5C ). In contrast, the non-fermented bacterial medium did not affect this parameter.

[0210] Since Bacillus foetidus J115 T produces short-chain fatty acids SCFA (butyrate, acetate), the non-fermented bacterial medium was tested, its pH value was adjusted and SCFA was supplemented (so it was content-matched (for the fermentation medium)). Neither the non-fermented medium nor the non-fermented medium rich in SCFA affected the cell density (Figure 5, SCFA-matched).

[0211] Bacillus foetidus J115 TThe fermentation medium reduced the cell proliferation of E0771 and PY8119 cells after 24 hours of treatment, while the non-fermented medium had no effect on this parameter. However, cell proliferation was reduced in the SCFA-matched medium but did not reach the same level as that obtained from T Bacteroides caccae J115. T The fermentation medium completely inhibited cell proliferation at a concentration of 20%, so its potency was 10 to 20 times higher than that of SCFA ( Figure 5B and 5D , SCFA-matched compared to fermented).

[0212] These data suggest that the antitumor effect of the bacteria is mediated by the action of specific metabolites produced by T Bacteroides caccae J115 that are different from SCFA.

Claims

1. A composition for preventing and / or treating breast cancer in a subject in need thereof, wherein the composition comprises at least: (a) bacteria from the genus Dysosmobacter, and / or variants thereof, and / or extracts and / or fragments thereof, and / or (b) culture supernatants of bacteria from the genus Dysosmobacter and / or variants thereof.

2. The composition for use according to claim 1, wherein the bacteria belong to the species Dysosmobacter welbionis.

3. The composition used according to claim 1 or 2, wherein the bacterium belongs to strain J115 T , which was deposited with BCCM / LMG on March 14, 2018 under the accession number LMG P-30603.

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 consisting of or comprising: progesterone receptor (PR), estrogen receptor (ER), and human epidermal growth factor receptor-2 (HER2).

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

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

7. The composition for use according to any one of claims 1 to 6, wherein the subject also suffers from one or more diseases or conditions that accelerate tumor growth, the diseases or conditions being selected from the group consisting of or comprising: obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver (NAFL), cirrhosis, diabetes, glucose intolerance, hyperglycemia, lipid metabolism disorder, dyslipidemia, hypercholesterolemia, elevated LDL-cholesterol, decreased HDL-cholesterol, elevated triglycerides, and intestinal inflammation, and combinations thereof.

8. The composition used according to any one of claims 1 to 7, wherein the composition comprises a therapeutically effective amount of said bacteria, preferably from 1×10 2 to about 1×10 15 CFU.

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 the composition further comprises at least one other 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 which further comprises a pharmaceutically acceptable carrier.

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

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

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