Methods and Uses of Microbiota Compositions, Components, or Metabolites for Treating Insulin-Related Disorders

Abstract

Disclosed are methods and compositions (e.g., comprising one or more microbial strains, one or more components, one or more metabolites, or combinations thereof) for treating insulin-related diseases, disorders, and conditions.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 407,592, filed September 16, 2022, the entire contents of which are incorporated herein by reference in their entirety. [Background technology]

[0002] Many insulin-related diseases, disorders, or conditions, including but not limited to diabetes, obesity, cardiovascular disease, nonalcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermadrome, diabetic neuropathy, diabetic foot ulcers, maturity-onset diabetes of the young, pancreatogenic diabetes, polycystic ovary syndrome (PCOS), or Alzheimer's disease (AD), can cause degeneration of various cells (e.g., liver cells, pancreatic cells, etc.) and affect physical and / or mental function. Currently, there are no effective treatments for such diseases, and identifying new drugs or treatment methods is a priority. Summary of the Invention

[0003] The present disclosure provides insight that the compositions (e.g., microbiota compositions) described herein can be used to treat a disease, disorder, or condition (e.g., associated with insulin levels (e.g., an insulin-related disease, disorder, or condition (e.g., diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, AD, etc.)) in a subject (e.g., a mammal (e.g., a human, a mouse, etc.)). Among other things, the present disclosure describes techniques that can be used to treat, prevent, and / or reduce the risk of a disease, disorder, or condition (e.g., associated with insulin levels). In some embodiments, the present disclosure describes compositions and methods for assessing the effect of administering such compositions (e.g., microbiota compositions described herein) to a subject and / or identifying or characterizing the effect and / or modulation of levels of metabolites or metabolome in a subject upon administration of such compositions. In some embodiments, the metabolites that can be modulated can be associated with a particular disease, disorder, or condition. In some embodiments, such techniques may be useful for identifying differences in metabolite levels in a particular subject (e.g., patient) or population (e.g., before and after administration of a disclosed composition). Accordingly, the present disclosure also provides techniques that may be useful for identifying and / or evaluating the properties and effects of the disclosed compositions in specific subjects (e.g., patients) and / or populations, and thus providing subject-specific information regarding how to treat a disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition) in an individual subject or individual population. For example, in some embodiments, the techniques provided herein may be useful for identifying a subject-specific composition based on the metabolome in a subject-specific sample, and treating and / or preventing a disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition) by administering a disclosed composition (e.g., a subject-specific composition) (e.g., to modulate the subject's metabolome).Thus, the technology described herein may be useful as therapeutic agents and tools for reducing the risk of certain diseases, disorders, or conditions (e.g., insulin-related diseases, disorders, or conditions), and for treating and / or preventing such diseases, disorders, or conditions.

[0004] Among other things, the present disclosure provides methods for treating or preventing insulin-related diseases, disorders, or conditions. In some embodiments, the methods include administering to a subject a composition comprising one or more microbial strains or microbial components. In some embodiments, the methods include administering to a subject a composition comprising one or more microbial metabolites. In some embodiments, the methods include administering to a subject a composition comprising (i) one or more microbial strains or microbial components, or (ii) one or more microbial metabolites. In some embodiments, the insulin-related disease, disorder, or condition is or includes diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermadrome, diabetic neuropathy, diabetic foot ulcer, maturity-onset diabetes of the young, pancreatogenic diabetes, or polycystic ovary syndrome (PCOS). In some embodiments, the insulin-related disease, disorder, or condition is or includes diabetes.

[0005] In some embodiments, the subject has been diagnosed with or is at risk of developing an insulin-related disease, disorder, or condition. In some embodiments, the subject is an animal. In some embodiments, the subject is a mammal, e.g., a mammal experiencing or susceptible to a disease, disorder, or condition described herein. In some embodiments, the animal is a vertebrate, e.g., a mammal such as a non-human primate (particularly a higher primate), sheep, dog, rodent (e.g., mouse or rat), guinea pig, goat, pig, cat, rabbit, or cow. In some embodiments, the animal is a non-mammal, such as a chicken, amphibian, reptile, or invertebrate. In some embodiments, the subject is a human.

[0006] In some embodiments, the subject is suffering from or susceptible to one or more insulin-related diseases, disorders, or conditions described herein. In some embodiments, the subject exhibits one or more symptoms of one or more insulin-related diseases, disorders, or conditions described herein. In some embodiments, the subject has been diagnosed with one or more insulin-related diseases, disorders, or conditions described herein. In some embodiments, the subject is undergoing or has undergone a particular therapy to diagnose and / or treat one or more insulin-related diseases, disorders, or conditions.

[0007] In some embodiments, one or more microbial strains are derived from a mammalian microbiota. In some embodiments, one or more microbial strains are derived from a human microbiota. In some embodiments, the human microbiota is a subject's microbiota. In some embodiments, the human microbiota is administered to maintain or regulate the subject's microbiota.

[0008] In some embodiments, the one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3, or Appendix 4. In some embodiments, the metabolites may be derived from one or more microbial strains. In some embodiments, the metabolites may be derived from a source other than a microbial strain, e.g., synthetically produced. In some embodiments, the one or more microbial metabolites are or include bile acids. In some embodiments, the one or more microbial metabolites are or include tauroursodeoxycholic acid. In some embodiments, the one or more microbial components or microbial metabolites are butyrylchamic acid, theobromine, p-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetylcysteine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (Arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro), or any combination thereof.In some embodiments, the one or more microbial components or microbial metabolites are 4-hydroxyphenylpyruvate, ectoine, gramine, N-acetyl-L-phenylalanine, nepsilon-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, transurocanic acid, N-acetyl-L-leucine, sarcosine, isobutyric acid, methyl-L-arginine ... L-carnitine, β-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenaceturic acid, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homocitrulline, mucic acid, homocysteine ​​sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3-indoxyl sulfate, nicotinamide, N-formylglycine, ureidoglycolate, N-methylproline, glucaric acid, butyrylcarnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenaceturic acid, diethanolamine, phosphorylcholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine ​​glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, b-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thiaproline, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0 XA0027, or any combination thereof.

[0009] In some embodiments, the one or more microbial strains are or comprise Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or a combination thereof. In some embodiments, the one or more microbial strains are or comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp., or a combination thereof. In some embodiments, the one or more microbial strains are or include Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or a combination thereof. In some embodiments, the one or more microbial strains are or include Bacillus subtilis.

[0010] In some embodiments, the composition comprises two or more microbial strains, in some embodiments, the composition comprises five or more microbial strains, in some embodiments, the composition comprises ten or more microbial strains.

[0011] In some embodiments, the composition is administered topically, orally, subcutaneously, intravenously, intramuscularly, intracerebrally, intrathecally, intrarectally, ophthalmically, intravitreally, or suprachoroidally. In some embodiments, the composition is administered orally. In some embodiments, the composition is administered intravenously.

[0012] In some embodiments, the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injectable, or eye drops.

[0013] In some embodiments, each microbial strain of the one or more microbial strains is 10 1 ~10 15 In some embodiments, each microbial strain of the one or more microbial strains is present in the composition at a concentration of at least 10 CFU. 6 In some embodiments, each microbial strain of the one or more microbial strains in the composition is present in the composition at a concentration of 10 CFU. 1 Colony forming units (CFU) ~10 20 In some embodiments, each microbial strain of the one or more microbial strains in the composition comprises 10 CFU. 1 Colony forming units (CFU) ~10 15 In some embodiments, each microbial strain of the one or more microbial strains in the composition comprises 10 CFU. 6 CFU~10 15 In some embodiments, each microbial strain of the one or more microbial strains in the composition comprises about 10 CFU. 1 CFU~10 15 CFU, or approximately 10 2 CFU~10 14 CFU, or approximately 10 3 CFU~10 13 CFU, or approximately 10 4 CFU~10 13 CFU, or approximately 10 5 CFU~10 12 CFU, or approximately 10 6 CFU~10 11 CFU, or approximately 10 7 CFU~10 10 CFU, or approximately 10 8 CFU~10 9 CFU, or approximately 10 5 CFU~10 10 CFU, or approximately 10 8 CFU~10 12In some embodiments, each microbial strain of the one or more microbial strains in the composition comprises at least about 10 CFU. 1 , 5×10 1 , 10 2 , 5×10 2 , 10 3 , 5×10 3 , 10 4 , 5×10 4 , 10 5 , 5×10 5 , 10 6 , 5×10 6 , 10 7 , 5×10 7 , 10 8 , 5×10 8 , 10 9 , 5×10 9 , 10 10 , 5×10 10 , 10 11 , 5×10 11 , 10 12 In some embodiments, each of the one or more microbial strains in the composition comprises at most about 10 CFU, or more. 15 , 5×10 14 , 10 14 , 5×10 13 , 10 13 , 5×10 12 , 10 12 , 5×10 11 , 10 11 , 5×10 10 , 10 10 , 5×10 9 , 10 9 , 5×10 8 , 10 8 In some embodiments, each microbial strain of the one or more microbial strains in the composition contains the same number of CFUs. In some embodiments, some microbial strains of the one or more microbial strains in the composition contain different numbers of CFUs.

[0014] The present disclosure includes, inter alia, compositions for treating or for use in treating an insulin-related disease, disorder, or condition, comprising one or more microbial strains, microbial components thereof, or microbial metabolic products thereof. In some embodiments, the compositions described herein comprise one or more microbial metabolic products (e.g., derived from one or more microbial strains, from a source other than the microbial strain (e.g., synthetically derived)), and the compositions are for treating an insulin-related disease, disorder, or condition.

[0015] The present disclosure provides compositions comprising one or more microbial strains selected from Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or combinations thereof. In some embodiments, the compositions comprise one or more microbial strains selected from Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella atypica, Bifidobacterium sp., or combinations thereof. In some embodiments, the compositions comprise a microbial strain. In some embodiments, the microbial strain is Bacillus subtilis. In some embodiments, the composition comprises at least two microbial strains selected from the group consisting of Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or a combination thereof. In some embodiments, the composition comprises at least two microbial strains selected from the group consisting of Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella atypica, Bifidobacterium sp., or a combination thereof.In some embodiments, the composition comprises at least five microbial strains selected from the group consisting of Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or a combination thereof. In some embodiments, the composition comprises at least five microbial strains selected from the group consisting of Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella atypica, Bifidobacterium sp., or a combination thereof. In some embodiments, the composition comprises or consists of Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp. In some embodiments, the composition comprises or consists of Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella atypica, Bifidobacterium sp.

[0016] In some embodiments, the compositions described herein comprise one or more microbial metabolites (e.g., derived from one or more microbial strains, derived from a source other than the microbial strain (e.g., synthetically derived)), and the compositions are for treating an insulin-related disease, disorder, or condition.

[0017] In some embodiments, the composition is for topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal, intraocular, intravitreal, or suprachoroidal administration. In some embodiments, the composition is for oral administration. In some embodiments, the composition is for intravenous administration.

[0018] The present disclosure includes, inter alia, compositions for use in treating insulin-related diseases, disorders, or conditions comprising one or more microbial strains or microbial components. In some embodiments, a composition for use in treating insulin-related diseases, disorders, or conditions comprises one or more microbial metabolites.

[0019] The present disclosure provides that the compositions described herein are for use in regulating one or more microbial metabolites in a subject. In some embodiments, the compositions are for use in regulating one or more characteristics in a subject. In some embodiments, the one or more characteristics are or include (i) the level of cell survival, (ii) the level or activity of a nucleic acid or protein or a form thereof, (iii) weight gain, (iv) fat accumulation in the liver, (v) lipid accumulation in the liver, (vi) blood triglyceride levels, (vii) blood cholesterol levels, (viii) oxidative stress, or (ix) inflammation.

[0020] In some embodiments, the method is for use in characterizing the ability of one or more microbial strains to modulate one or more microbial metabolites in a subject.

[0021] The present disclosure provides for the use of the compositions described herein to treat or ameliorate a disease, disorder, or condition in a subject, wherein the disease, disorder, or condition is an insulin-related disease, disorder, or condition associated with one or more microbial metabolites. In some embodiments, the use of the compositions described herein is to treat or ameliorate diabetes.

[0022] The present disclosure provides a method of screening a microbial strain comprising contacting the microbial strain with a culture comprising pancreatic cells or a pancreatic cell line that models an insulin-related disease, disorder, or condition, and determining whether the microbial strain alters a characteristic of the culture, wherein the characteristic is associated with the insulin-related disease, disorder, or condition.

[0023] In some embodiments, the determining step comprises comparing the characteristic before and after performing the contacting step, hi some embodiments, the determining step comprises comparing the characteristic to an equivalent reference after the contacting step.

[0024] In some embodiments, the comparable reference is a historical reference. In some embodiments, the comparable reference is a negative control reference. In some embodiments, the comparable reference is a positive control reference.

[0025] In some embodiments, the characteristic is a level of cell viability. In some embodiments, the characteristic is a level or activity of a nucleic acid or protein, or a form thereof. In some embodiments, the characteristic is or includes weight gain. In some embodiments, the characteristic is or includes fat accumulation in hepatocytes. In some embodiments, the characteristic is or includes lipid accumulation in hepatocytes. In some embodiments, the characteristic is or includes triglyceride levels. In some embodiments, the characteristic is or includes cholesterol levels. In some embodiments, the characteristic is or includes inflammation.

[0026] In some embodiments, the microbial strain may alter one or more characteristics of the culture. In some embodiments, the one or more characteristics are associated with an insulin-related disease, disorder, or condition. In some embodiments, the one or more characteristics are or include (i) levels of cell survival, (ii) levels or activity of nucleic acids or proteins or forms thereof, (iii) weight gain, (iv) fat accumulation in the liver, (v) lipid accumulation in the liver, (vi) blood triglyceride levels, (vii) blood cholesterol levels, (viii) oxidative stress, or (ix) inflammation.

[0027] The present disclosure provides methods comprising administering to a subject a composition comprising one or more microbial strains or microbial components. In some embodiments, the present disclosure provides methods comprising administering to a subject a composition comprising one or more microbial metabolites. In some embodiments, the microbial metabolites can be derived from one or more microbial strains. In some embodiments, the microbial metabolites can be derived from a source other than a microbial strain, for example, synthetically produced.

[0028] In some embodiments, the microbial strain, microbial component, or microbial metabolite may alter a characteristic of the subject. In some embodiments, the microbial strain, microbial component, or microbial metabolite may alter one or more characteristics of the subject. In some embodiments, the characteristic is the level of cell viability. In some embodiments, the characteristic is the level or activity of a nucleic acid or protein, or a form thereof. In some embodiments, the characteristic is or includes weight gain. In some embodiments, the characteristic is or includes fat accumulation in the liver. In some embodiments, the characteristic is or includes lipid accumulation in the liver. In some embodiments, the characteristic is or includes triglyceride levels. In some embodiments, the characteristic is or includes cholesterol levels. In some embodiments, the characteristic is or includes inflammation.

[0029] In some embodiments, the characteristic is associated with an insulin-related disease, disorder, or condition.

[0030] The present disclosure provides a method for characterizing a microbial strain, comprising adding the microbial strain to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that model an insulin-related disease, disorder, or condition, and determining whether the microbial strain affects the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are associated with the insulin-related disease, disorder, or condition.

[0031] The present disclosure provides a method of manufacturing a pharmaceutical treatment comprising characterizing one or more microbial strains, microbial components, or microbial metabolites, comprising the steps of: adding one or more microbial strains to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that model an insulin-related disease, disorder, or condition; and determining whether the one or more microbial strains affect the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are associated with the insulin-related disease, disorder, or condition.

[0032] The present disclosure provides methods of manufacturing a pharmaceutical treatment comprising formulating one or more microbial strains or microbial components in a syrup, liquid, tablet, troche, gummy candy, capsule, powder, gel, film, injectable, or eye drop.The present disclosure provides methods of manufacturing a pharmaceutical treatment comprising formulating one or more microbial metabolites in a syrup, liquid, tablet, troche, gummy candy, capsule, powder, gel, film, injectable, or eye drop.

[0033] The present disclosure provides methods for evaluating microbial strains for their ability to affect one or more characteristics of the culture, comprising adding the microbial strain to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that model an insulin-related disease, disorder, or condition, and determining whether the microbial strain affects the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are associated with the insulin-related disease, disorder, or condition.

[0034] In some embodiments, the method further comprises determining the level of one or more characteristics of one or more pancreatic cells or pancreatic cell lines in the culture before adding the microbial strain to the culture, determining the level of the same one or more characteristics of the one or more pancreatic cells or pancreatic cell lines in the culture after adding the microbial strain to the culture, and comparing the level of the one or more characteristics determined before adding the microbial strain to the level of the one or more characteristics determined after adding the microbial strain.

[0035] In some embodiments, the one or more characteristics include (i) level of cell survival, (ii) level or activity of a nucleic acid or protein or a form thereof, (iii) weight gain, (iv) fat accumulation in the liver, (v) lipid accumulation in the liver, (vi) blood triglyceride levels, (vii) blood cholesterol levels, (viii) oxidative stress, (ix) inflammation, or (x) a combination thereof.

[0036] The present disclosure provides that the compositions described herein are for use in treating or preventing an insulin-related disease, disorder, or condition, comprising one or more microbial strains or microbial components. The present disclosure provides that the compositions described herein are for use in treating or preventing an insulin-related disease, disorder, or condition, comprising one or more microbial metabolites. In some embodiments, the one or more microbial components or microbial metabolites of the one or more microbial strains are selected from Appendix 1, Appendix 3, or Appendix 4.

[0037] In some embodiments, the compositions described herein are for use in the treatment or prevention of diabetes, where the insulin-related disease, disorder, or condition is diabetes.

[0038] The present disclosure provides an injectable solution comprising the composition described herein.

[0039] The present disclosure provides a dietary supplement comprising the composition described herein.

[0040] The present disclosure provides kits comprising the compositions described herein for use in treating or preventing insulin-related diseases, disorders, or conditions. In some embodiments, the kits include a monitoring device. In some embodiments, the monitoring device is a blood glucose monitor.

[0041] These and other aspects encompassed by the present disclosure are described in more detail below and in the claims.

[0042] definition The scope of the present invention is defined by the claims appended hereto, and is not limited by any particular embodiments described herein. Those skilled in the art will recognize, upon reading this specification, various modifications that are equivalent to such described embodiments or that may otherwise fall within the scope of the claims. Generally, terms used herein follow their understood meanings in the art unless expressly indicated otherwise. Explicit definitions of certain terms are provided below, and the meaning of these and other terms in specific instances throughout this specification will be clear to those skilled in the art from the context.

[0043] The use of ordinal terms such as "first," "second," "third," etc. to modify claim elements in the claims does not, in itself, imply any priority, precedence, or ordering of one claim element over another claim element, or the temporal order in which acts of a method are performed, but rather is merely used as a label to distinguish one claim element having a particular name from another element having the same name (except for the use of ordinal terms) to distinguish between claim elements.

[0044] As used herein, the articles "a" and "an" should be understood to include plural referents unless a clear indication to the contrary is present. A claim or description including "or" between one or more members of a group is deemed applicable if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process, unless a contrary indication is present or otherwise clear from the context. In some embodiments, exactly one member of a group is present in, employed in, or otherwise relevant to a given product or process. In some embodiments, more than one or all group members are present in, employed in, or otherwise relevant to a given product or process. It is to be understood that the present invention encompasses all variations, combinations, and permutations of one or more limitations, elements, clauses, descriptive language, etc. from one or more of the enumerated claims, introduced into another claim dependent on the same base claim (or any other claim, if relevant), unless otherwise indicated or unless a contradiction or inconsistency would be apparent to one skilled in the art. When elements are presented as lists (e.g., in a Markush group or similar format), it is to be understood that each subgroup of elements is also disclosed, and that any element(s) can be removed from the group. In general, when an embodiment or aspect is referred to as "comprising" certain elements, features, etc., it should be understood that a particular embodiment or aspect "consists" or "consists essentially of" such elements, features, etc. For purposes of brevity, those embodiments have not in all instances been specifically described in numerous terms herein. It should also be understood that any embodiment or aspect may be explicitly excluded from the claims, regardless of whether that specific exclusion is recited in the specification.

[0045] Administration: As used herein, the term "administration" typically refers to the administration of a composition to a subject or system to achieve delivery of an agent to the subject or system. In some embodiments, the agent is or is included in a composition, and in some embodiments, the agent is produced through metabolism of the composition or one or more components thereof. Those of skill in the art will recognize various routes that may be utilized for administration to a subject, e.g., a human, in appropriate circumstances. For example, in some embodiments, administration may be ocular, oral, parenteral, topical, etc. In some specific embodiments, administration may be bronchial (e.g., by bronchial instillation), buccal, cutaneous (which may be or include, e.g., one or more of topical to the dermis, intradermal, interdermal, transdermal, etc.), enteral, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, intraspecific organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreous, etc. In many embodiments provided by the present disclosure, administration is oral. In some embodiments, administration may involve only a single dose. In some embodiments, administration may involve the application of a fixed number of doses. In some embodiments, administration may involve administration that is intermittent (e.g., multiple doses separated in time) and / or periodic (e.g., individual doses separated by a common period of time) administration. In some embodiments, administration can involve continuous administration (e.g., perfusion) over at least a selected period of time. Administration of cells can be by any suitable route that results in delivery to a desired location in a subject, where at least a portion of the delivered cells or cellular components remain viable. The period of cell survival after administration to a subject can be as short as a few hours, e.g., 24 hours, to as long as several days or years, i.e., long-term engraftment. In some embodiments, administration involves delivery of a bacterial extract or preparation that contains one or more bacterial metabolic products and / or by-products, but is completely devoid of viable bacterial cells.

[0046] Analog: As used herein, the term "analog" refers to a substance that shares one or more particular structural features, elements, components, or moieties with a reference substance. Typically, an "analog" exhibits significant structural similarity to the reference substance, e.g., sharing a core or consensus structure, but differs in certain individual respects. In some embodiments, an analog is a substance that can be produced from a reference substance, e.g., by chemical manipulation of the reference substance. In some embodiments, an analog is a substance that can be produced through the implementation of a synthetic process that is substantially similar (e.g., shares multiple steps) to that which produces the reference substance. In some embodiments, an analog is produced, or can be produced, through the implementation of a synthetic process that is different from that used to produce the reference substance.

[0047] Approximately: When applied to one or more values ​​of interest, includes values ​​that are similar to a stated reference value. In certain embodiments, the term "approximately" or "about" refers to a range of values ​​that are within ±10% (more or less than) of a stated reference value, unless otherwise stated or otherwise clear from the context (except when such number exceeds 100% of the possible values).

[0048] Equivalent: As used herein, the term "equivalent" refers to two or more agents, entities, circumstances, sets of conditions, subjects, etc. that may not be identical to one another, but are sufficiently similar to permit comparisons between them, so that one of skill in the art will understand that conclusions can be reasonably drawn based on observed differences or similarities. In some embodiments, equivalent sets of conditions, circumstances, individuals, or populations are characterized by multiple substantially identical characteristics and one or a few diverse characteristics. One of skill in the art will understand what degree of identity is required in any given situation for two or more such agents, entities, circumstances, sets of conditions, etc. to be considered equivalent in context. For example, one of skill in the art will understand that sets of circumstances, individuals, or populations are equivalent to one another when they are characterized by a sufficient number and type of substantially identical characteristics to warrant a reasonable conclusion that differences in results obtained under or with different sets of circumstances, individuals, or populations, or in observed phenomena, are caused by or indicate variations in the diverse characteristics.

[0049] Conservative: As used herein, refers to the case when describing a conservative amino acid substitution, which involves replacing an amino acid residue with another amino acid residue having a side chain R group with similar chemical properties (e.g., charge or hydrophobicity). Generally, a conservative amino acid substitution does not substantially alter the desired functional property of a protein, e.g., the ability of a receptor to bind a ligand. Examples of groups of amino acids having side chains with similar chemical properties include aliphatic side chains such as glycine (Gly, G), alanine (Ala, A), valine (Val, V), leucine (Leu, L), and isoleucine (Ile, I); aliphatic hydroxyl side chains such as serine (Ser, S) and threonine (Thr, T); amide-containing side chains such as asparagine (Asn, N) and glutamine (Gln, Q); aromatic side chains such as phenylalanine (Phe, F), tyrosine (Tyr, Y), and tryptophan (Trp, W); basic side chains such as lysine (Lys, K), arginine (Arg, R), and histidine (His, H); acidic side chains such as aspartic acid (Asp, D) and glutamic acid (Glu, E); and sulfate-containing side chains such as cysteine ​​(Cys, C) and methionine (Met, M). Conservative amino acid substitutions include, for example, valine / leucine / isoleucine (Val / Leu / Ile, V / L / I), phenylalanine / tyrosine (Phe / Tyr, F / Y), lysine / arginine (Lys / Arg, K / R), alanine / valine (Ala / Val, A / V), glutamic acid / aspartic acid (Glu / Asp, E / D), and asparagine / glutamine (Asn / Gln, N / Q). In some embodiments, a conservative amino acid substitution can be the substitution of any naturally occurring residue in a protein with alanine, for example, as used in alanine scanning mutagenesis. In some embodiments, conservative substitutions are made that have a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet, GH et al., 1992, Science 256:1443-1445, which is incorporated herein by reference in its entirety. In some embodiments, the substitution is a moderately conservative substitution, and the substitution has a non-negative value in the PAM250 log-likelihood matrix. [Table 1]

[0050] Control: As used herein, refers to the art-understood meaning of "control," which is a standard against which results are compared. Typically, controls are used to enhance the integrity of an experiment by isolating a variable in order to draw conclusions about such a variable. In some embodiments, a control is a reaction or assay performed simultaneously with a test reaction or assay to provide a point of comparison. "Control" also includes "control animals." A "control animal" can have a modification described herein, a different modification described herein, or no modification (i.e., a wild-type animal). In one experiment, the "test" (i.e., the variable being tested) is administered. In a second experiment, the "control," the variable being tested, is not administered. In some embodiments, a control is a historical control (i.e., a control from a previously performed test or assay, or a previously known amount or result). In some embodiments, a control is or includes a printed or otherwise kept record. A control can be a positive or negative control.

[0051] Determining, Measuring, Evaluating, Assessing, Assaying, and Analyzing: Determining, measuring, evaluating, assessing, assaying, and analyzing are used interchangeably herein to refer to any form of measurement, including determining whether an element is present. These terms include both quantitative and / or qualitative determinations. Assaying can be relative or absolute. "Assaying for the presence of" can be determining the amount of something present and / or determining whether it is present or absent.

[0052] Dosage form: Those of skill in the art will understand that the term "dosage form" can be used to refer to a physically discrete unit of an agent (e.g., a therapeutic agent) for administration to a subject. Typically, each such unit contains a predetermined amount of agent. In some embodiments, such amount is a unit dosage (or a whole fraction thereof) appropriate for administration according to a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., a therapeutic regimen). Those of skill in the art will understand that the total amount of a therapeutic composition or agent to be administered to a particular subject is determined by one or more attending physicians and may involve the administration of multiple dosage forms.

[0053] Dosage regimen: Those skilled in the art will understand that the term "dosage regimen" can be used to refer to a set of unit doses (typically more than one) administered individually to a subject, typically separated by a period of time. In some embodiments, a given medication has a recommended dosing regimen, which can involve one or more doses. In some embodiments, a dosing regimen includes multiple doses, each of which is separated in time from the other doses. In some embodiments, the individual doses are separated from each other by periods of equal length, and in some embodiments, a dosing regimen includes multiple doses and at least two different periods separating the individual doses. In some embodiments, all doses within a dosing regimen are the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen includes a first dose in a first dose amount, followed by one or more additional doses in a second dose amount that is different from the first dose amount. In some embodiments, a dosing regimen includes a first dose in a first dose amount, followed by one or more additional doses in a second dose amount that is the same as the first dose amount. In some embodiments, the dosing regimen correlates with a desired or beneficial outcome when administered across a relevant population.

[0054] Engineered: Generally, the term "engineered" refers to the aspect of being manipulated by the hand of man. For example, a cell or organism is considered "engineered" if it has been manipulated so that its genetic information is changed (e.g., new genetic material not previously present is introduced, e.g., by transformation, mating, somatic cell hybridization, transfection, transduction, or other mechanisms, or previously present genetic material is altered or removed, e.g., by substitution or deletion mutations or by mating protocols). As is common practice and understood by those skilled in the art, the progeny of an engineered polynucleotide or cell are typically still referred to as "engineered," despite the actual manipulation performed on the previous entity.

[0055] Excipient: As used herein, refers to an inert (e.g., non-therapeutic) agent that may be included in a pharmaceutical composition to, for example, provide or contribute to a desired consistency or stabilizing effect. In some embodiments, suitable pharmaceutical excipients may include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, nonfat dry milk, glycerol, propylene, glycol, water, ethanol, and the like.

[0056] Functional: As used herein, a "functional" biomolecule is a biomolecule in a form in which it exhibits the property and / or activity for which it is characterized. A biomolecule can have two functions (i.e., bifunctional) or many functions (i.e., multifunctional).

[0057] Gene: As used herein, refers to a DNA sequence in a chromosome that encodes a product (e.g., an RNA product and / or a polypeptide product). In some embodiments, a gene comprises coding sequence (i.e., a sequence that encodes a particular product). In some embodiments, a gene comprises non-coding sequence. In some specific embodiments, a gene may include both coding (e.g., exon) and non-coding (e.g., intron) sequence. In some embodiments, a gene may include one or more regulatory sequences (e.g., promoters, enhancers, etc.) and / or intron sequences that, for example, can control or influence one or more aspects of gene expression (e.g., cell-type specific expression, inducible expression, etc.). For clarity, it is noted that the term "gene," as used in this disclosure, generally refers to a portion of a nucleic acid that encodes a polypeptide or fragment thereof. As will be clear from the context to one of skill in the art, the term may optionally encompass regulatory sequences. This definition is not intended to exclude the application of the term "gene" to non-protein-coding expression units, but is intended to clarify that the term, as used herein, most often refers to a polypeptide-encoding nucleic acid.

[0058] Improve, increase, enhance, inhibit, or reduce: As used herein, the terms “improve,” “increase,” “enhance,” “inhibit,” “reduce,” or their grammatical equivalents refer to a value that is relative to a baseline or other reference measurement. In some embodiments, the value is statistically significantly different from the baseline or other reference measurement. In some embodiments, a suitable reference measurement may be or include a measurement in a particular system (e.g., in a single individual) under otherwise comparable conditions in the absence (e.g., before and / or after) of a particular agent or treatment, or in the presence of an appropriate comparable reference agent. In some embodiments, a suitable reference measurement may be or include a measurement in an equivalent system known or expected to respond in a particular way in the presence of the relevant agent or treatment. In some embodiments, a suitable reference is a negative reference; in some embodiments, a suitable reference is a positive reference.

[0059] Isolated: As used herein, refers to a substance and / or entity that is (1) separated from at least some of the components with which it was associated when originally produced (whether in nature and / or in an experimental setting) and / or (2) designed, produced, prepared, and / or manufactured by the hand of man. In some embodiments, an isolated substance or entity may be concentrated, and in some embodiments, an isolated substance or entity may be pure. In some embodiments, isolated substances and / or entities may be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% of the other components with which they were originally associated. In some embodiments, an isolated agent is about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or greater than about 99% pure. As used herein, a substance is "pure" if it is substantially free of other components. In some embodiments, as will be understood by those of skill in the art, a substance may still be considered "enriched," "isolated," or "pure" after being combined with certain other components, such as, for example, one or more carriers or excipients (e.g., buffers, solvents, water, etc.), and in such embodiments, the percent isolation or purity of the substance is calculated without including such carriers or excipients. Those of skill in the art are aware of various techniques for isolating (e.g., concentrating or purifying) a substance or agent (e.g., using one or more of fractionation, extraction, precipitation, or other separation).

[0060] Level: As used herein, the term "level" refers to a scale of the amount or quantity of a substance (e.g., a metabolite). In some embodiments, the level may simply be the presence or absence of a substance. The level of a substance may be expressed in multiple ways or formats. For example, in some embodiments, the level is expressed as a percentage (%), a measure of weight (e.g., mg, μg, ng, etc.), a measure of concentration (e.g., mg / mL, μg / mL, ng / mL, etc.), a measure of volume (e.g., mL, μL, nL, etc.), a % change, etc.

[0061] Metabolite: As used herein, the term "metabolite" refers to a substance (e.g., a small molecule, a macromolecule, an organic compound, or an inorganic compound) made or used during metabolism. Metabolism is generally understood as the process by which a substance (e.g., a food, a drug, a chemical, a cell, or a tissue) is chemically broken down. In some embodiments, a metabolite is an end product. In some embodiments, a metabolite is an intermediate. Exemplary metabolites are provided herein, for example, in Appendices 1-1, 1-3, and 3. Exemplary metabolic pathways are provided herein, for example, in Appendices 1-2. In some embodiments, a metabolite can be produced or made by an organism. In some embodiments, a metabolite can be produced or made by a microorganism (e.g., a microbial strain). In some embodiments, a microbial metabolite produced or made by a microbial strain. In some embodiments, a metabolite can be produced or made naturally (e.g., by an organism (e.g., a microorganism (e.g., a microbial strain))). In some embodiments, a metabolite can be produced or made synthetically (e.g., from a source that is not a microbial strain (e.g., synthetically produced)).

[0062] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to a composition in which an active agent is formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose amount suitable for administration in a treatment regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition may be specially formulated for administration in solid or liquid form, including those adapted for intraocular, intravitreal, suprachoroidal, oral, subcutaneous, intravenous, intramuscular, intracerebral, or intrathecal administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., tablets targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue, capsules, powders, etc. In some embodiments, the active agent is or may include a cell or cell population (e.g., a culture of an ellagitanin-enzyme-synthesizing (EES) microorganism). In some embodiments, the active agent may be or comprise an extract or component of a cell or cell population (e.g., a culture). In some embodiments, the active agent may be or comprise an isolated, purified, or pure compound. In some embodiments, the active agent may be synthesized in vitro (e.g., via chemical and / or enzymatic synthesis). In some embodiments, the active agent may be or comprise a natural product (whether isolated from its natural source or synthesized in vitro).

[0063] Pharmaceutically acceptable: As used herein, the term "pharmaceutically acceptable," which may be used, for example, in reference to a carrier, diluent, or excipient used to formulate a pharmaceutical composition disclosed herein, means the carrier, diluent, or excipient must be compatible with the other ingredients of the composition and not deleterious to the recipient thereof.

[0064] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, that is involved in carrying or transporting the subject compound from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the subject (e.g., patient). Some examples of materials which can function as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; excipients such as powdered tragacanth, malt, gelatin, talc, cocoa butter, and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar, buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, pH buffer solutions, polyesters, polycarbonates, and / or polyanhydrides, and other non-toxic compatible substances employed in pharmaceutical formulations.

[0065] Prebiotic: As used herein, "prebiotic" refers to an ingredient that enables or promotes specific changes in both composition and / or activity in the gastrointestinal microbiota that may (or may not) confer a benefit to the host. In some embodiments, the prebiotic can include one or more of the following: the prebiotic includes pome fruit extract, berry extract, and walnut extract.

[0066] Prevention: As used herein, the term "prevention" refers to a delay in the onset and / or a reduction in the frequency and / or severity of one or more symptoms of a particular disease, disorder, or condition. In some embodiments, prevention is assessed on a population basis, such that an agent is considered to "prevent" a particular disease, disorder, or condition if a statistically significant reduction in the onset, frequency, and / or intensity of one or more symptoms of the disease, disorder, or condition is observed in a population susceptible to the disease, disorder, or condition. In some embodiments, prevention may be considered complete, for example, when the onset of the disease, disorder, or condition has been delayed for a predetermined period of time.

[0067] Reference: As used herein, describes a standard or control against which a comparison is made. For example, in some embodiments, an agent, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control agent, animal, individual, population, sample, sequence, or value. In some embodiments, the reference or control is tested and / or determined substantially contemporaneously with the test or determination of interest. In some embodiments, the reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as understood by those of skill in the art, a reference or control is determined or characterized under conditions or circumstances comparable to those being evaluated. Those of skill in the art will understand when sufficient similarity exists to justify reliance on and / or comparison to a particular potential reference or control. In some embodiments, the reference is a negative control reference, and in some embodiments, the reference is a positive control reference.

[0068] Risk: As understood from the context, "risk" of a disease, disorder, and / or condition refers to the likelihood that a particular individual will develop the disease, disorder, and / or condition. In some embodiments, risk is expressed as a percentage. In some embodiments, risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or up to 100%. In some embodiments, risk is expressed relative to the risk associated with a reference sample or group of reference samples. In some embodiments, the reference sample or group of reference samples has a known risk of the disease, disorder, condition, and / or event. In some embodiments, the reference sample or group of reference samples is derived from individuals comparable to the particular individual. In some embodiments, the relative risk is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more.

[0069] Sample: As used herein, the term "sample" typically refers to an aliquot of material obtained from or derived from a source of interest. In some embodiments, the source of interest is a biological or environmental source. In some embodiments, the source of interest may be or include a cell, or an organism such as a microorganism, a plant, or an animal (e.g., a human). In some embodiments, the source of interest is or includes a biological tissue or fluid. In some embodiments, the biological tissue or fluid may be or include amniotic fluid, aqueous humor, peritoneal fluid, bile, bone marrow, blood, breast milk, cerebrospinal fluid, earwax, chyle, chyme, ejaculate, endolymph, exudate, feces, gastric acid, gastric juice, lymph, mucus, pericardial fluid, perilymph, peritoneal fluid, pleural fluid, pus, catarrhal secretions, saliva, sebum, semen, serum, smegma, sputum, synovial fluid, sweat, tears, urine, vaginal secretions, vitreous humor, vomit, plasma, mucus, digestive fluid, stool, and / or combinations or component(s) thereof. In some embodiments, the biological fluid may be or include intracellular fluid, extracellular fluid, intravascular fluid (plasma), interstitial fluid, lymph, and / or transcellular fluid. In some embodiments, the biological fluid may be or include plant exudates. In some embodiments, the biological tissue or sample may be obtained, for example, by aspirate, biopsy (e.g., fine needle or tissue biopsy), swab (e.g., oral, nasal, skin, or vaginal swab), scraping, surgery, washing or lavage (e.g., bronchoalveolar epithelial, ductal, nasal, ocular, oral, uterine, vaginal, or other washing or lavage). In some embodiments, the biological sample is or comprises cells obtained from an individual. In some embodiments, the sample is a "primary sample" obtained directly from the source of interest by any suitable means. In some embodiments, as will be clear from the context, the term "sample" refers to a preparation obtained by processing the primary sample (e.g., by removing one or more components of the primary sample and / or by adding one or more agents to the primary sample), for example, by filtration using a semi-permeable membrane.Such a "processed sample" may include, for example, nucleic acids or proteins extracted from a sample or obtained by subjecting the primary sample to one or more techniques, such as amplification or reverse transcription of nucleic acids, isolation and / or purification of certain components, etc.

[0070] Small molecule: As used herein, the term "small molecule" refers to a small organic or inorganic molecule with a molecular weight below about 3,000 Daltons. Generally, a small molecule can have a molecular weight less than 3,000 Daltons (Da). The small molecule can be, for example, at least about 100 Da to about 3,000 Da (e.g., about 100 to about 3,000 Da, about 100 to about 2,500 Da, about 100 to about 2,000 Da, about 100 to about 1,750 Da, about 100 to about 1,500 Da, about 100 to about 1,250 Da, about 100 to about 1,000 Da, about 100 to about 750 Da, about 100 to about 500 Da, about 200 to about 1,500, about 500 to about 1,000, about 300 to about 1,000 Da, or about 100 to about 250 Da).

[0071] Subject: As used herein, the term "subject" refers to an individual to whom a provided treatment is administered. In some embodiments, the subject is an animal. In some embodiments, the subject is a mammal, e.g., a mammal experiencing or susceptible to a disease, disorder, or condition described herein. In some embodiments, the animal is a vertebrate, e.g., a mammal such as a non-human primate (particularly a higher primate), sheep, dog, rodent (e.g., a mouse or rat), guinea pig, goat, pig, cat, rabbit, or cow. In some embodiments, the animal is a non-mammal, such as a chicken, amphibian, reptile, or the invertebrate model C. elegans. In some embodiments, the subject is a human. In some embodiments, the subject is suffering from or susceptible to one or more diseases, disorders, or conditions described herein. In some embodiments, the subject exhibits one or more symptoms of one or more diseases, disorders, or conditions described herein. In some embodiments, the subject has been diagnosed with one or more diseases, disorders, or conditions described herein. In some embodiments, the subject is undergoing or has undergone a particular treatment to diagnose and / or treat a disease, disorder, or condition, hi other embodiments, the subject is an experimental animal or surrogate animal used as a disease model.

[0072] Substantially: As used herein, refers to the qualitative state of exhibiting the full or nearly full extent or degree of a desired characteristic or property. Those skilled in the biological arts will understand that biological and chemical phenomena rarely, if ever, proceed to completion and / or perfection or achieve or avoid absolute results. Thus, the term "substantially" is used herein to capture the potential lack of perfection inherent in many biological and chemical phenomena.

[0073] Treatment regimen: A "treatment regimen," as that term is used herein, refers to a dosing regimen, the administration of which across a relevant population can be correlated with a desired or beneficial therapeutic outcome.

[0074] Therapeutically effective amount: As used herein, it refers to the amount at which it is administered that produces a desired effect. In some embodiments, the term refers to an amount that, when administered to a population suffering from or susceptible to a disease, disorder, and / or condition according to a therapeutic regimen, is sufficient to treat the disease, disorder, and / or condition. In some embodiments, a therapeutically effective amount is an amount that reduces the incidence and / or severity of and / or delays the onset of one or more symptoms of the disease, disorder, and / or condition. Those skilled in the art will understand that the term "therapeutically effective amount" does not, in fact, require that successful treatment be achieved in a particular individual. Rather, a therapeutically effective amount may be an amount that, when administered to subjects (e.g., patients) in need of such treatment, provides a specific, desired pharmacological response in a significant number of subjects. In some embodiments, reference to a therapeutically effective amount may be a reference to the amount measured in one or more specific tissues (e.g., tissues affected by a disease, disorder, or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). Those skilled in the art will appreciate that in some embodiments, a therapeutically effective amount of a particular agent or therapy may be formulated and / or administered in a single dose, while in some embodiments, a therapeutically effective agent may be formulated and / or administered in multiple doses, e.g., as part of a dosing regimen.

[0075] Treatment: As used herein, the term "treatment" (also "treat" or "treating") refers to any administration of a therapy that partially or completely alleviates, ameliorates, relieves, inhibits, delays the onset of, reduces the severity of, and / or reduces the incidence of one or more symptoms, characteristics, and / or causes of a particular disease, disorder, and / or condition. In some embodiments, such treatment may be treatment of a subject who does not exhibit signs of the relevant disease, disorder, and / or condition and / or who exhibits only early signs of the disease, disorder, and / or condition. Alternatively, or in addition, such treatment may be treatment of a subject who exhibits one or more established signs of the relevant disease, disorder, and / or condition. In some embodiments, treatment may be treatment of a subject who has been diagnosed with the relevant disease, disorder, and / or condition. In some embodiments, treatment may be treatment of a subject who is known to have one or more susceptibility factors that statistically correlate with an increased risk of developing the relevant disease, disorder, and / or condition. [Brief explanation of the drawings]

[0076] [Figure 1] Plots of IDE relative to actin are shown for each of the five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice. Groups are: G1 = untreated wild-type, G2 = untreated transgenic, G3 = transgenic CT10, G4 = transgenic CT10x, and G5 = transgenic CT10m. Plots show ANOVA comparisons between group G2 (left) and group G1 (right). [Figure 2] Plots of the relative values ​​of NSE to actin are shown for each of the five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice: G1 = untreated wild-type, G2 = untreated transgenic, G3 = transgenic CT10, G4 = transgenic CT10x, and G5 = transgenic CT10m. [Figure 3]Plots of p-Akt Ser473 relative to actin (left) and Akt1 relative to actin (right) are shown for each of the five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice: G1 = untreated wild-type, G2 = untreated transgenic, G3 = transgenic CT10, G4 = transgenic CT10x, and G5 = transgenic CT10m. [Figure 4] Plots of insulin receptor β relative to actin are shown for each of the five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice: G1 = untreated wild-type, G2 = untreated transgenic, G3 = transgenic CT10, G4 = transgenic CT10x, and G5 = transgenic CT10m. [Figure 5] Plots of relative Glut3 to actin levels are shown for each of the five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice: G1 = untreated wild-type, G2 = untreated transgenic, G3 = transgenic CT10, G4 = transgenic CT10x, and G5 = transgenic CT10m. [Figure 6] Plots of RBAP48 relative to actin are shown for each of the five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice: G1 = untreated wild-type, G2 = untreated transgenic, G3 = transgenic CT10, G4 = transgenic CT10x, and G5 = transgenic CT10m. [Figure 7] Plots of p-4EBP1 relative to actin are shown for each of the five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice: G1 = untreated wild-type, G2 = untreated transgenic, G3 = transgenic CT10, G4 = transgenic CT10x, and G5 = transgenic CT10m. [Figure 8]Plots of relative NRF2 levels to actin levels are shown for each of the five study groups (G1, G2, G3, G4, and G5) in TG2576 AD mice: G1 = untreated wild-type, G2 = untreated transgenic, G3 = transgenic CT10, G4 = transgenic CT10x, and G5 = transgenic CT10m. DETAILED DESCRIPTION OF THE INVENTION

[0077] Insulin-Related Diseases, Disorders, and Conditions Insulin-related diseases, disorders, and conditions are a comprehensive term for a range of diseases, disorders, and conditions that primarily result from fluctuating or unregulated insulin levels in the human body. Insulin is a hormone that helps the body utilize glucose as an energy source from food. Several pathological conditions exist that are associated with impaired insulin secretion and utilization by the body. Many insulin-related diseases, disorders, or conditions, including, but not limited to, diabetes, obesity, cardiovascular disease, nonalcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermadrome, diabetic neuropathy, diabetic foot ulcers, maturity-onset diabetes of the young, pancreatogenic diabetes, polycystic ovary syndrome (PCOS), or Alzheimer's disease (AD), can cause degeneration of various cells (e.g., liver cells, pancreatic cells, etc.), affecting physical and / or mental function. Some of these are temporary, but others are untreatable (e.g., chronic) and debilitating conditions that result in progressive degeneration and / or death of cells (e.g., liver cells, pancreatic cells, etc.), which manifest as an observed decline in physical and / or mental function.

[0078] Unregulated insulin levels, or insulin resistance, also known as decreased insulin sensitivity, occurs when cells in muscle, fat, and the liver fail to respond to insulin as they should. Muscle, fat, and liver cells may respond inappropriately to insulin, resulting in inefficient uptake or storage of glucose from the blood. As a result, the pancreas produces more insulin to overcome increasing blood glucose levels. This is called hyperinsulinemia. As long as the pancreas can produce enough insulin to overcome the weak response to insulin, blood glucose levels remain within a healthy range. When cells become overly resistant to insulin, it leads to elevated blood glucose levels (hyperglycemia), which can eventually lead to prediabetes and type 2 diabetes. In addition to type 2 diabetes, insulin resistance is associated with several other conditions, including obesity, cardiovascular disease, nonalcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, polycystic ovary syndrome (PCOS), and / or other diseases, disorders, or conditions disclosed herein.

[0079] Elevated insulin levels can also cause weight gain, which in turn worsens insulin resistance, which is also associated with higher triglyceride levels, hardening of the arteries (atherosclerosis), high blood pressure (hypertension), etc.

[0080] Insulin resistance is also a key feature of metabolic syndrome, a set of features that link excess fat around the waist and insulin resistance to an increased risk of cardiovascular disease, stroke, and type 2 diabetes. Features of metabolic syndrome include elevated blood glucose levels, elevated triglyceride levels, low levels of high-density lipoprotein (HDL) cholesterol, and / or high blood pressure.

[0081] Several factors and conditions can cause various degrees of insulin resistance.In particular, excess body fat around the abdomen and physical inactivity are considered to be two major contributing factors to insulin resistance.Apart from these, insulin resistance can also be caused by diet, certain drugs, hormonal disorders (e.g., Cushing's syndrome, acromegaly, hypothyroidism, etc.), genetic conditions (e.g., type A insulin resistance syndrome, Rabson-Mendenhall syndrome, Donahue syndrome, etc.), and other hereditary conditions (e.g., myotonic dystrophy, Alström syndrome, Werner syndrome, hereditary lipodystrophy, etc.).

[0082] The present disclosure provides compositions (e.g., microbiota compositions) and methods that inhibit one or more events or processes that occur in insulin-related diseases, disorders, or conditions. The present disclosure is based, in part, on the discovery that one or more microbial strains or compositions comprising one or more microbial strains are particularly suitable as therapeutic agents for insulin-related diseases, disorders, or conditions.

[0083] Microbial preparation(s) and / or component(s) The present disclosure provides systems and methods for evaluating, characterizing, and identifying one or more microbial strains of a microbiome. For example, the present disclosure provides systems and methods for evaluating, characterizing, and identifying one or more microbial strains of a microbiome having one or more capabilities. Such systems and methods can be useful for evaluating, characterizing, and identifying one or more microbial strains that affect the health of humans, livestock, and / or pets. In some embodiments, the one or more microbial strains affect the health of humans, livestock, and / or pets by modulating their respective metabolomes, cell viability, ATP levels, one or more other parameters or characteristics (e.g., of an organ of interest), or a combination thereof, to prevent, treat, or reduce the risk of a disease, disorder, or condition. For example, the technology described herein may result in modulation of a subject's metabolome, improved cell survival, increased ATP levels, modulation of one or more other parameters or characteristics (e.g., levels or activity of nucleic acids or proteins or forms thereof, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.), or combinations thereof, which results in a decrease in the production of toxic components and / or components that are indicative of or are markers of cellular damage in the subject (e.g., in the subject's blood) (e.g., liver cell damage, pancreatic cell damage, neuronal cell damage (e.g., increased blood levels of neurofilament light chain protein (NF-L)).

[0084] The present disclosure also provides systems and methods for producing pharmaceutical compositions that include evaluating, characterizing, and identifying one or more microbial strains of a microbiota.

[0085] In some embodiments, the method includes evaluating, characterizing, and identifying one or more microbial strains from the microbiota of a snake, lizard, fish, or bird. In some embodiments, the method includes evaluating, characterizing, and identifying one or more microbial strains from the microbiota of a mammal. The mammalian microbiota can be the microbiota of a dog, cat, horse, cow, sheep, goat, or pig. In some embodiments, the microbiota used in the systems or methods described herein can prevent or treat a disease or condition.

[0086] The microbiota can be isolated from any system or tissue of an organism that supports microbial growth. For example, the microbiota can be skin microbiota, oral microbiota, nasal microbiota, gastrointestinal microbiota, brain microbiota, lung microbiota, or urogenital microbiota. A list of exemplary microbial strains found in gastrointestinal microbiota is included in Table 1 below. Those skilled in the art will understand that microbiota samples can be obtained by various methods known in the art. For example, skin, oral, nasal, lung, or urogenital microbiota samples can be obtained using swabs or tissue scrapings. In some embodiments, gastrointestinal microbiota can be sampled from feces. Skin microbiota, oral microbiota, nasal microbiota, gastrointestinal microbiota, brain microbiota, lung microbiota, or urogenital microbiota samples can be obtained via biopsy.

[0087] In some embodiments, the microbiota is that of a healthy individual or an individual who does not suffer from or is not at risk of developing a particular disease or disorder. In some embodiments, the microbiota is that of an individual who suffers from or is at risk of developing a particular disease, disorder, or condition. In some embodiments, the microbiota is that of an individual who is known to suffer from a particular disease, disorder, or condition. In some embodiments, the human microbiota is that of a human who has an unknown risk of one or more diseases, disorders, or conditions.

[0088] In some embodiments, the microbiota is a reference microbiota. The reference microbiota can be the microbiota of a healthy individual or an individual who is not suffering from or at risk of developing a particular disease, disorder, or condition. In some cases, the reference microbiota can be derived from the same individual as the microbiota to be evaluated or characterized, but obtained at a different time. In some cases, the reference microbiota can be derived from the same individual as the microbiota to be evaluated or characterized, but obtained from a different system or tissue.

[0089] In some embodiments, individual microbial strains or combinations of microbial strains may be evaluated, characterized, or identified in relative abundances that differ from those found in the microbiota when such one or more strains are present. For example, the effect of modulating a cell or organism in response to a single strain may be evaluated, characterized, or identified using in vitro methods (e.g., mammalian cells) or in vivo methods using a mammal (e.g., mouse, human, etc.) described herein. In some embodiments, the effect of modulating a cell or organism to treat, prevent, or reduce the risk of a disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition described herein) may be evaluated, characterized, or identified using in vitro methods (e.g., mammalian cells) or in vivo methods using a mammal (e.g., mouse, human, etc.) described herein. In some embodiments, the effect of modulating a cell or organism to treat, prevent, or reduce the risk of a disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition described herein) by, for example, modulating one or more metabolites of the cell or organism, one or a characteristic or parameter of the cell or organism (e.g., cell survival, the level or activity of a nucleic acid or protein or form thereof, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.), or a combination thereof, can be evaluated, characterized, or identified using in vitro methods (e.g., mammalian cells) or in vivo methods using a mammal (e.g., mouse, human, etc.) described herein. As another example, the effect of modulating a cell or organism (e.g., the level of one or more metabolites) to treat, prevent, or reduce the risk of a disease, disorder, or condition described herein in response to two microbial strains can be evaluated, characterized, or identified together using the methods described herein.

[0090] Extracts, components, or compounds of microbial strains can also be evaluated, characterized, or identified together using the methods described herein. In some cases, extracts, components, or compounds of microbial strains that are determined to treat, prevent, or reduce the risk of diseases, disorders, or conditions in organisms (e.g., mammals) described herein can be evaluated, characterized, or identified. Evaluating, characterizing, or identifying extracts, components, or compounds of microbial strains that treat, prevent, or reduce the risk of diseases, disorders, or conditions in organisms (e.g., mammals) can provide additional information about potential biomarkers, targets, or protective agents in the microbiome.

[0091] A variety of techniques are known in the art that can be used to prepare extracts of microbial strains and / or to isolate or process extracts, components, or compounds therefrom (e.g., to isolate and / or purify one or more components or compounds therefrom). Such techniques may include, for example, one or more of organic extraction, vacuum concentration, chromatography, etc., to name just a few.

[0092] Biological effects assessment The present disclosure provides insight that the compositions (e.g., microbiota compositions) described herein can be used to treat, prevent, and / or reduce the risk of a disease, disorder, or condition in an organism (e.g., a mammal (e.g., a human)) by contacting the composition(s) with the organism (e.g., feeding the composition to the organism, administering the composition to the organism). In some embodiments, the organism can be suffering from or at risk of suffering from a disease, disorder, or condition (e.g., a mammalian disease, disorder, or condition). To determine whether one or more compositions treat, prevent, or reduce the risk of a disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition), the level of one or more metabolites can be observed, measured, or assessed in a sample contacted with one or more compositions. For example, the level of one or more metabolites can be observed, measured, or assessed in a sample at different times (e.g., before administration of the composition, after administration of the composition, during administration of the composition, etc.). To determine whether one or more compositions treat, prevent, or reduce the risk of a disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition), one or more characteristics or parameters can be observed, measured, or evaluated in a sample contacted with one or more compositions. For example, one or more characteristics or parameters can be observed, measured, or evaluated in a sample at different times (e.g., before administration of the composition, after administration of the composition, during administration of the composition, etc.).

[0093] In some embodiments, the methods described herein utilize a first sample and a second sample. In some embodiments, the first sample is a reference sample. In some embodiments, the reference sample can be a sample obtained from a subject contacted with (e.g., administered or provided with) a composition, e.g., a CT10 composition, a CT10m composition, a CT10x composition, a CT6 composition, or a CT6m composition. In some embodiments, the reference sample can be a sample obtained from a subject contacted with (e.g., administered or provided with) a composition, e.g., a CT10 composition, a CT10m composition, a CT10x composition, a CT6 composition, or a CT6m composition, at a first time point. In some embodiments, the reference sample can be a sample obtained from a subject before contacting (e.g., administered or provided with) a composition, e.g., a CT10 composition, a CT10m composition, a CT10x composition, a CT6 composition, or a CT6m composition ... healthy individual. In some embodiments, the reference sample may be a sample obtained from an individual who may be suffering from or at risk for a disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition). In some embodiments, the reference sample is a control sample. In some embodiments, the reference sample is a negative control sample. In some embodiments, the reference sample is a positive control sample. In some embodiments, the reference sample may be a historical reference (e.g., a value across a control sample). In some embodiments, the reference sample may be derived from a printed publication (e.g., a textbook, a journal, etc.).

[0094] In some embodiments, the second sample can be a test sample. In some embodiments, the test sample can be a sample obtained from a subject contacted with (e.g., administered or provided with) a composition, such as a CT10 composition, a CT10x composition, a CT10m composition, a CT6 composition, or a CT6m composition. In some cases, the subject (e.g., a patient or a population) can be suffering from or at risk of a disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition). In some cases, the subject (e.g., a patient or a population) can have an unknown risk of one or more diseases, disorders, or conditions described herein. In some embodiments, the test can be a sample obtained from a subject contacted with (e.g., administered or provided with) a composition, such as a CT10 composition, a CT10m composition, a CT10x composition, a CT6 composition, or a CT6m composition, at a second time point.

[0095] In some embodiments, the methods described herein involve comparing one or more metabolite levels (e.g., metabolome) or one or more parameters or characteristics (e.g., cell survival, levels or activity of nucleic acids or proteins or forms thereof, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.) obtained from a test sample with one or more metabolite levels (e.g., metabolome) or one or more parameters or characteristics (e.g., cell survival, levels or activity of nucleic acids or proteins or forms thereof, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.) obtained from a reference sample. In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from the test sample with one or more metabolite levels, parameters, or characteristics obtained from the reference sample, a composition described herein can be evaluated, characterized, or identified as useful for treating, preventing, or reducing the risk of a disease, disorder, or condition described herein (e.g., an insulin-related disease, disorder, or condition). In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from a test sample with one or more metabolite levels, parameters, or characteristics obtained from a reference sample, it can be determined that a composition disclosed herein increases the severity or incidence of a disease, disorder, or condition phenotype. In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from a test sample with one or more metabolite levels, parameters, or characteristics obtained from a reference sample, it can be determined that a composition disclosed herein reduces the severity or incidence of a disease, disorder, or condition phenotype.In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from a test sample with one or more metabolite levels, parameters, or characteristics obtained from a reference sample, it can be determined that the compositions disclosed herein do not affect the severity or incidence of a disease, disorder, or condition phenotype. In some embodiments, by comparing one or more metabolite levels, parameters, or characteristics obtained from a test sample with one or more metabolite levels, parameters, or characteristics obtained from a reference sample, it can be determined that the compositions disclosed herein prevent a disease, disorder, or condition phenotype.

[0096] The present disclosure also recognizes that the compositions and methods provided herein can be used to monitor the progression of a disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition) in an individual. For example, a decrease in relative amount of a metabolite level, parameter, or characteristic (e.g., cell survival, level or activity of a nucleic acid or protein or form thereof, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.) determined to increase the severity of the disease, disorder, or condition can indicate that the disease, disorder, or condition is being attenuated, for example, by a treatment or immune response.

[0097] The present disclosure also provides insight that the compositions and methods provided herein can be used to tailor treatments (e.g., therapeutics, nutraceuticals, and / or probiotics) to individual patients. In some embodiments, the compositions and methods provided herein can provide "personalized" therapy. In some cases, metabolite levels, characteristics, or parameters within an individual (e.g., cell viability, levels or activity of nucleic acids or proteins or forms thereof, weight gain, fat accumulation in the liver, lipid accumulation in the liver, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.) can be assessed, characterized, or identified to determine whether the individual has a disease, disorder, or condition. Based on the results, the individual can be treated with one or more compositions to adjust metabolite levels (i.e., their metabolome), characteristics, or parameters. In some cases, this will affect the disease, disorder, or condition the individual is suffering from or at risk of developing. For example, if an individual is determined to have relatively low levels of one or more metabolites determined to reduce the severity of a disease, disorder, or condition, administering to the individual one or more compositions (or extracts, components, or compounds thereof) determined to reduce the severity of the disease, disorder, or condition may attenuate the severity of the individual's disease or condition.

[0098] The present disclosure provides insight that the compositions and methods provided herein can be used recursively to treat, prevent, or ameliorate a disease, disorder, or condition. In some embodiments, for example, one or more compositions disclosed herein can be administered (e.g., fed, injected, etc.) to a subject after determining the effect of the one or more compositions on the subject's metabolite levels or on a characteristic or parameter (e.g., cell survival, level or activity of a nucleic acid or protein or form thereof, weight gain, liver fat accumulation, liver lipid accumulation, blood triglyceride levels, blood cholesterol levels, oxidative stress, etc.) of the subject. In some embodiments, the composition can be administered once. In some embodiments, the composition can be administered more than once. In some embodiments, the composition can be administered daily, weekly, biweekly, monthly, bimonthly, etc. In each of these cases, changes in the level of one or more metabolites or characteristics or parameters can be monitored. In some embodiments, changes in the level of one or more metabolites (e.g., the metabolome), or characteristics or parameters can be monitored prior to administration of the composition. In some embodiments, changes in the levels of one or more metabolites (eg, the metabolome), or characteristics or parameters, may be monitored following administration of the composition.

[0099] Pharmaceutical Compositions Compositions are provided herein that include individual microbial strains, or combinations of microbial strains, their metabolites, extracts, or components thereof. In some embodiments, the compositions include individual microbial strains or combinations of microbial strains from a mammalian microbiota, its metabolites, extracts, and / or components thereof that have been assessed, identified, characterized, or assayed using the methods described herein. In some embodiments, the compositions provided herein include one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more microbial strains from a mammalian microbiota, its extracts, its metabolites, and / or components thereof that have been assessed, identified, characterized, or assayed using the methods described herein.

[0100] Also provided herein are compositions comprising one or more components or metabolites.In some embodiments, the components or metabolites in the compositions herein are derived from a source other than a microbial strain, for example, are synthetically produced.In some embodiments, the components or metabolites in the compositions may be identified from a microbial strain, but are independent of the microbial strain and are not produced by the microbial strain, for example, they can be synthetically produced.

[0101] In some embodiments, the compositions provided herein comprise two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more microbial strains listed in Table 1 below. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7]

[0102] In some embodiments, the compositions provided herein comprise Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or a combination thereof. In some embodiments, the compositions comprise at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or all of Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, and Acidaminococcus sp. In some embodiments, for example, the composition includes all of Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, and Acidaminococcus sp., and may be referred to by different names, including, but not limited to, CT10 composition, CT10 cocktail, etc.

[0103] In some embodiments, the compositions provided herein comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp., or a combination thereof. In some embodiments, the compositions comprise at least two, at least three, at least four, at least five, or all of Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., and Bifidobacterium sp. In some embodiments, for example, a composition comprises all of Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., and Bifidobacterium sp., and may be referred to by different names, including, but not limited to, a CT6 composition, a CT6 cocktail, etc. In some embodiments, the compositions provided herein comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or a combination thereof. In some embodiments, the compositions comprise at least two, at least three, at least four, at least five, or all of Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, and Bifidobacterium breve.In some embodiments, for example, the composition includes all of Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, and Bifidobacterium breve, and may be referred to by different names, including, but not limited to, CT6 composition, CT6 cocktail, etc.

[0104] Exemplary microbiota compositions described herein (e.g., CT10, CT10m, CT10x, CT6, CT6m) and their compositional details are listed in Tables 2-6 below. [Table 3] [Table 4] [Table 5] [Table 6] [Table 7]

[0105] In some embodiments, the compositions provided herein comprise one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more metabolites. Metabolites that may be assessed, identified, characterized, or assayed for and / or included in the compositions disclosed herein include, for example, those listed in the appendices filed herewith (e.g., Appendix 1-1, 1-2, 1-3, 2, 3, or 4).

[0106] In some embodiments, the metabolite can be butyrylcamitine, theobromine, p-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro), or any combination thereof.

[0107] In some embodiments, the metabolite is 4-hydroxyphenylpyruvate, ectoine, gramine, N-acetyl-L-phenylalanine, nepsilon-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, transurocanic acid, N-acetyl-L-leucine, sarcosine, isobutyrylcarnitine, b-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenaceturic acid, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homocitrulline, mucic acid, homocysteine ​​sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3-indoxyl sulfate, nicotinamide, N-formylglycine, ureidoglycolate, N-methylproline, glucaric acid, butyrylcarnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenaceturic acid, diethanolamine, phosphorylcholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine ​​glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, b-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thiaproline, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0 XA0027, or any combination thereof.

[0108] In some embodiments, the individual microbial strains or combinations of microbial strains from the mammalian microbiota are killed (e.g., heat-killed). Alternatively, in some embodiments, the individual microbial strains or combinations of microbial strains from the mammalian microbiota may comprise viable or living cells.

[0109] In some embodiments, the one or more microbial strains include viable or living individual microbial strains or combinations of microbial strains, for example, from the mammalian microbiota.

[0110] In some embodiments, the one or more microbial strains include, for example, viable or living individual microbial strains or combinations of microbial strains derived from a mammalian microbiota, and include and / or are formulated through the use of one or more cell cultures and / or supernatants or pellets thereof, and / or powders formed therefrom, as described herein.

[0111] In some embodiments, a composition for use according to the present disclosure is, for example, a pharmaceutical composition for administration to a mammal (e.g., a human) (e.g., topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal (e.g., rectal intubation), intraocular, intravitreal, or suprachoroidal administration). Pharmaceutical compositions typically include an active agent (e.g., an individual microbial strain or combination of microbial strains derived from a mammalian microbiota, an extract thereof, and / or components thereof) and a pharmaceutically acceptable carrier. Certain exemplary pharmaceutically acceptable carriers include, for example, saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration.

[0112] In some embodiments, pharmaceutical compositions for use according to the present disclosure may comprise and / or be administered in conjunction with one or more supplementary active compounds. In certain embodiments, such supplemental active agents include ginger, curcumin, probiotics (e.g., probiotic strains of one or more of the following bacterial genera: Bifidobacterium, Saccharomyces, Enterococcus, Streptococcus, Pediococcus, Leuconostoc, Bacillus, and / or Escherichia coli) (see Fijan, Int J Environ Res Public Health. 2014 May;11(5):4745-4767, incorporated herein by reference in its entirety), prebiotics (non-digestible food ingredients that help support the growth of probiotic bacteria, e.g., fructans such as fructooligosaccharides (FOS) and inulin, galactans such as galactooligosaccharides (GOS), dietary fiber such as resistant starch, pectin, beta-glucan, and xylooligosaccharides (see Hutkins et al., Curr Opin Biotechnol. 2016, incorporated herein by reference in its entirety). Feb;37:1-7), as well as combinations thereof.

[0113] In some embodiments, the prebiotic comprises fructooligosaccharides, inulin, isomaltooligosaccharides, lactylol, lactosucrose, lactolose, soybean oligosaccharides, transgalactooligosaccharides, xylooligosaccharides, seaweed, or a combination thereof. In some embodiments, the prebiotic comprises seaweed. In some embodiments, the prebiotic comprises pome fruit extract, berry extract, and walnut extract.

[0114] In some embodiments, the probiotic composition may be formulated for oral administration. In some embodiments, the probiotic composition may be a food, beverage, feed composition, or dietary supplement. In some embodiments, the ellagitanin composition, the enzyme composition, or both may be a liquid, syrup, tablet, lozenge, gummy, capsule, powder, gel, or film. In some embodiments, the probiotic composition is an enteric-coated formulation.

[0115] In some embodiments, the probiotic comprises a prebiotic, which in some embodiments comprises fructooligosaccharides, inulin, isomaltooligosaccharides, lactylol, lactosucrose, lactolose, soybean oligosaccharides, transgalactooligosaccharides, xylooligosaccharides, seaweed, pome fruit extract, berry extract, and walnut extract, or a combination thereof.

[0116] Pharmaceutical compositions are typically formulated to be compatible with their intended administration route. Examples of administration routes include topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal (e.g., rectal intubation), intraocular, intravitreal, or suprachoroidal administration. Methods for formulating suitable pharmaceutical compositions are known in the art, see, for example, Remington: The Science and Practice of Pharmacy, 21st ed., 2005, and the books in the series Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs (Dekker, NY), which are incorporated herein by reference in their entirety. Oral compositions generally contain an inert diluent or an edible carrier (e.g., a pharmaceutically acceptable diluent, a pharmaceutically acceptable carrier). In some embodiments, oral formulations can be or include syrups, liquids, tablets, lozenges, gummies, capsules (e.g., gelatin capsules), powders, gels, films, etc., to name just a few examples. Similarly, ophthalmic compositions (e.g., for intraocular, intravitreal, or suprachoroidal administration) can contain various additives, such as inert diluents or carriers (e.g., pharmaceutically acceptable diluents, pharmaceutically acceptable carriers), viscosity enhancers, permeation enhancers, cyclodextrins, etc. Examples of viscosity enhancers include hydroxymethylcellulose, hydroxyethylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, and polyalcohols. Examples of permeation enhancers include chelating agents, preservatives, surfactants, bile salts, benzalkonium chloride, polyoxyethylene glycol ethers (lauryl, stearyl, and oleyl), ethylenediaminetetraacetic acid sodium salt, sodium taurocholate, saponin, and Cremophor EL. For example, in some embodiments, the ophthalmic formulation can be or include a suspension, emulsion (e.g., water-in-oil or oil-in-water), nanocarrier (e.g., nanoparticles, nanosuspensions, liposomes, nanomicelles, dendrimers, etc.), ointment, gel, eye drops, etc.Brain compositions (e.g., for intracerebral or intrathecal administration) may include inert diluents or excipients and / or additives. In some embodiments, the brain compositions do not include preservatives. In some embodiments, the brain compositions are sterile.

[0117] In some embodiments, pharmaceutically compatible binders and / or adjuvant materials can be included as part of the pharmaceutical composition. In some specific embodiments, the pharmaceutical composition can contain, for example, any one or more of the following inactive ingredients or compounds of a similar nature: binders such as microcrystalline cellulose, gum tragacanth, or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate or sterotes; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylate, or orange flavoring. In some embodiments, the composition can be consumed directly or sprinkled on or mixed into food or liquids (such as water). In some embodiments, compositions that can be administered to a mammal described herein can be or can include ingestible items (e.g., foods or beverages) that contain (e.g., are supplemented with) individual or combinations of microbial strains from the mammal's microbiota, extracts thereof, and / or components thereof.

[0118] In some embodiments, the food product can be or include one or more of bars, candy, baked goods, cereals, savory snacks, pasta, chocolate, and other solid foods, liquid or semi-solid foods including yogurt, soups, and stews, and beverages such as smoothies, shakes, juices, and other carbonated or non-carbonated beverages. In some embodiments, the food product is prepared by the subject by mixing individual or combinations of microbial strains, extracts thereof, and / or components thereof from the mammalian microbiota.

[0119] The compositions can be included in a kit, container, pack, or dispenser together with instructions for administration or use in the methods described herein.

[0120] Those skilled in the art will understand, upon reading this disclosure, that in some embodiments, the compositions (e.g., pharmaceutical compositions) described herein can be or include one or more cells, tissues, or organisms (e.g., plant or microbial cells, tissues, or organisms) that produce (e.g., have produced and / or are producing) the relevant compounds.

[0121] Those skilled in the art will understand that in some embodiments, techniques for preparing and / or for preparing compositions and / or preparations (particularly for preparing pharmaceutical compositions) may include one or more steps of evaluating or characterizing the compound, preparation, or composition, e.g., as part of quality control. In some embodiments, if assayed material does not meet predetermined specifications for relevant evaluation, it is discarded. In some embodiments, if such assayed material meets predetermined specifications, it continues to be processed as described herein.

[0122] In some embodiments, the pharmaceutical compositions provided herein are capable of promoting colonization of individual microbial strains or combinations of microbial strains from a mammal's microbiota, particularly microbial strain(s) that have been identified, characterized, or evaluated as reducing the severity or incidence of a mammalian disease, disorder, or condition in a mammal suffering from or at risk for the disease, disorder, or condition. In some embodiments, the pharmaceutical compositions provided herein are capable of attenuating colonization of individual microbial strains or combinations of microbial strains from a mammal's microbiota, particularly microbial strain(s) that have been identified, characterized, or evaluated as increasing the severity or incidence of a mammalian disease, disorder, or condition (e.g., an insulin-related disease, disorder, or condition) in a mammal suffering from or at risk for the disease, disorder, or condition. In some embodiments, the pharmaceutical compositions provided herein are capable of promoting colonization of individual microbial strains or combinations of microbial strains from the microbiota of a mammal, particularly microbial strain(s) that have been identified, characterized, or assessed as not affecting the severity or incidence of the mammalian disease, disorder, or condition, but that have been identified, characterized, or assessed as being capable of outcompeting one or more microbial strains that have been identified, characterized, or assessed as increasing the severity or incidence of the mammalian disease, disorder, or condition, in a mammal suffering from or at risk for the mammalian disease, disorder, or condition.

[0123] In some embodiments, each of the one or more microbial strains in the composition is 10 1 Colony forming units (CFU) ~10 20 In some embodiments, each of the one or more microbial strains in the composition comprises 10 CFU. 1 Colony forming units (CFU) ~10 15 In some embodiments, each of the one or more microbial strains in the composition comprises 10 CFU. 6 CFU~10 15In some embodiments, each of the one or more microbial strains in the composition comprises about 10 CFU. 1 CFU~10 15 CFU, or approximately 10 2 CFU~10 14 CFU, or approximately 10 3 CFU~10 13 CFU, or approximately 10 4 CFU~10 13 CFU, or approximately 10 5 CFU~10 12 CFU, or approximately 10 6 CFU~10 11 CFU, or approximately 10 7 CFU~10 10 CFU, or approximately 10 8 CFU~10 9 CFU, or approximately 10 5 CFU~10 10 CFU, or approximately 10 8 CFU~10 12 In some embodiments, each of the one or more microbial strains in the composition comprises at least about 10 CFU. 1 , 5×10 1 , 10 2 , 5×10 2 , 10 3 , 5×10 3 , 10 4 , 5×10 4 , 10 5 , 5×10 5 , 10 6 , 5×10 6 , 10 7 , 5×10 7 , 10 8 , 5×10 8 , 10 9 , 5×10 9 , 10 10 , 5×10 10 , 10 11 , 5×10 11 , 10 12 In some embodiments, each of the one or more microbial strains in the composition comprises at most about 10 CFU, or more. 15 , 5×10 14 , 10 14 , 5×10 13, 10 13 , 5×10 12 , 10 12 , 5×10 11 , 10 11 , 5×10 10 , 10 10 , 5×10 9 , 10 9 , 5×10 8 , 10 8 In some embodiments, each of the one or more microbial strains in the composition contains the same number of CFUs, or fewer. In some embodiments, some of the one or more microbial strains in the composition contain different numbers of CFUs.

[0124] In some embodiments, the composition comprises a total of 10 1 CFU~10 20 In some embodiments, the composition comprises a total of 10 CFU. 6 CFU~10 15 In some embodiments, the composition comprises about 10 CFU. 1 CFU~10 20 CFU, or approximately 10 5 CFU~10 15 CFU, or approximately 10 5 CFU~10 12 CFU, approximately 10 5 CFU~10 10 CFU, or approximately 10 8 CFU~10 12 In some embodiments, the composition may comprise about 10 CFU of one or more microbial strains. 1 CFU~10 15 CFU, or approximately 10 2 CFU~10 14 CFU, or approximately 10 3 CFU~10 13 CFU, or approximately 10 4 CFU~10 13 CFU, or approximately 10 5 CFU~10 12 CFU, or approximately 10 6 CFU~10 11 CFU, or approximately 10 7 CFU~10 10CFU, or approximately 10 8 CFU~10 9 CFU, or approximately 10 5 CFU~10 10 CFU, or approximately 10 8 CFU~10 12 In some embodiments, the composition may comprise at least about 10 CFU of one or more microbial strains. 1 , 5×10 1 , 10 2 , 5×10 2 , 10 3 , 5×10 3 , 10 4 , 5×10 4 , 10 5 , 5×10 5 , 10 6 , 5×10 6 , 10 7 , 5×10 7 , 10 8 , 5×10 8 , 10 9 , 5×10 9 , 10 10 , 5×10 10 , 10 11 , 5×10 11 , 10 12 In some embodiments, the composition may contain at most about 10 CFU of one or more microbial strains. 15 , 5×10 14 , 10 14 , 5×10 13 , 10 13 , 5×10 12 , 10 12 , 5×10 11 , 10 11 , 5×10 10 , 10 10 , 5×10 9 , 10 9 , 5×10 8 , 10 8 The culture medium may contain one or more microbial strains at or below CFU.

[0125] In some embodiments, a pharmaceutical composition is tailored to a particular mammal (e.g., a particular human, e.g., a patient) based on the microbiota of that mammal (e.g., a human). In some embodiments, a pharmaceutical composition is specific to the microbiota of an individual mammal (e.g., a human). In some embodiments, a pharmaceutical composition is specific to the microbiota of a population of mammals (e.g., humans). A population of mammals can include, but is not limited to, a family, mammals in the same geographic location (e.g., neighborhood, city, state, or country), mammals with the same disease or condition, mammals of a particular age or age range, or mammals consuming a particular diet (e.g., food, food source, or caloric intake).

[0126] Treatment methods The present disclosure recognizes that the compositions described herein may be useful for treating a subject. The methods provided by the present disclosure include methods for treating certain diseases, disorders, and conditions. In some embodiments, the relevant disease, disorder, or condition may be or include an insulin-related disease, disorder, or condition. In some embodiments, the insulin-related disease, disorder, or condition may be ALS, AD, PD, or HD.

[0127] Generally, the methods of treatment provided by the present disclosure involve administering to a subject in need of, or determined to be in need of, such treatment a therapeutically effective amount of a composition described herein, alone or in combination with other compositions and / or treatments.

[0128] In some embodiments, the methods of treatment provided herein are prophylactic or preventative, e.g., may be administered to a subject prior to the onset of noticeable symptoms and / or prior to exposure to a particular anticipated trigger associated with an insulin-related disease, disorder, or condition described herein. In some embodiments, the methods of treatment provided herein are therapeutic, e.g., may be administered to a subject after the onset of noticeable symptoms associated with an insulin-related disease, disorder, or condition.

[0129] In some embodiments, the methods of treatment provided are administered to a subject that is a mammal, e.g., a mammal experiencing a disease, disorder, or condition described herein; in some embodiments, the subject is a human or non-human animal subject, e.g., an ape, cat, monkey, or pig.

[0130] In many embodiments, treatment involves ameliorating at least one symptom of a disease, disorder, or condition associated with an insulin-related disease, disorder, or condition. In some embodiments, the method of treatment can be prophylactic.

[0131] In some embodiments, the methods can include administration of a therapeutically effective amount of a composition disclosed herein before, during (e.g., simultaneously with), or after administration of a treatment expected to be associated with an insulin-related disease, disorder, or condition.

[0132] In some embodiments, a subject receiving a treatment described herein may be undergoing and / or have undergone other treatments (e.g., pharmacological treatments / therapies, surgery, etc.) that may be intended to treat, for example, one or more symptoms of a disease, disorder, or condition described herein (e.g., an insulin-related disease, disorder, or condition), and thus provided compositions are administered in combination with such other treatments (i.e., treatments) to treat the relevant disease, disorder, or condition.

[0133] In some embodiments, the compositions described herein may be administered in a form containing one or more pharmaceutically acceptable carriers. Suitable carriers have been described previously and vary depending on the desired form and mode of administration of the composition. For example, pharmaceutically acceptable carriers may include diluents or excipients such as fillers, binders, wetting agents, disintegrants, surfactants, glidants, and lubricants. Typically, carriers may be solid (including powders), liquids, or any combination thereof. Each carrier is preferably "acceptable" in the sense that it is compatible with the other ingredients in the composition and not harmful to the subject. Carriers may be biologically acceptable and inert (e.g., allowing the composition to maintain the viability of biological materials until delivered to the appropriate site).

[0134] The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth, or gelatin, an excipient such as starch or lactose, a disintegrating agent such as alginic acid, primogel, or corn starch, a lubricant such as magnesium stearate or sterols, a glidant such as colloidal silicon dioxide, a sweetener such as sucrose or saccharin, or a flavoring agent such as peppermint, methyl salicylate, orange flavor, or other suitable flavor, which are for purposes of example only and are not intended to be limiting.

[0135] Oral compositions can include an inert diluent or an edible carrier. For oral therapeutic administration, the active compound can be incorporated with an excipient and used in the form of tablets, lozenges, drops, troches, or capsules, such as gelatin capsules. Oral compositions can also be prepared by combining the compositions of the present disclosure with food. In some embodiments, the microorganisms (e.g., one or more microbial strains) can be formulated into food. Some non-limiting examples of food products that can be used with the methods and compositions described herein include popsicles, cheese, cream, chocolate, milk, meat, beverages, pickles, kefir, miso, sauerkraut, etc. In other embodiments, the food product may be juice, soft drinks, tea-based drinks, drink preparations, jelly drinks, and functional drinks; alcoholic beverages such as beer; carbohydrate-containing foods such as processed rice foods, noodles, bread, and pasta; fish, ham, sausage, and fish paste products; retort pouch products such as curry, foods with thick sauces, and Chinese soup; dairy products such as soup, milk, dairy drinks, ice cream, and yogurt; fermented products such as miso, fermented drinks, and pickles; soy products; various confectionery products including biscuits, cookies, candy, chewing gum, and gummies; cold desserts including jellies, custard puddings, and frozen desserts; and instant foods such as instant soups and instant miso soups. Preferably, the food preparation does not require cooking after mixing with the microbial strain(s) to avoid killing any microorganisms. In one embodiment, the food used for administration is chilled, e.g., ice-cold, flavored water. In certain embodiments, the food product is not a potentially allergenic food (e.g., not soy, wheat, peanut, tree nut, dairy, egg, shellfish, or fish). Pharmaceutically compatible binding agents and / or adjuvant materials can be included as part of the composition.

[0136] Ophthalmic formulations (e.g., for intraocular, intravitreal, or suprachoroidal administration) can include an inert diluent or carrier. For the purpose of ophthalmic therapeutic administration, the active compound can be incorporated with an excipient and used in the form of a suspension, emulsion (e.g., water-in-oil or oil-in-water), nanocarrier (e.g., nanoparticles, nanosuspensions, liposomes, nanocells, dendrimers, etc.), ointment, gel, eye drops, etc. In some embodiments, administration of such formulations is topical (e.g., eye drops). In some embodiments, administration of such formulations is via injection (e.g., intravitreal, suprachoroidal, etc.).

[0137] Brain formulations (e.g., for intracerebral or intrathecal administration) can contain an inert diluent or carrier. For the purpose of brain therapeutic administration, the active compound can be incorporated with an excipient and used in the form of a suspension, emulsion (e.g., water-in-oil or oil-in-water), nanocarrier (e.g., nanoparticles, nanosuspensions, liposomes, nanocells, dendrimers, etc.), ointment, gel, etc. In some embodiments, such formulations are administered topically (e.g., ointment). In some embodiments, such formulations are administered via injection (e.g., intracerebral, intrathecal, etc.).

[0138] In some such embodiments, the compositions described herein are administered to a subject according to a dosage regimen that achieves the introduction of administered cells into the subject's microbiota.In some embodiments, the compositions are administered to a subject in a single dose.In some embodiments, the compositions are administered to a subject in multiple doses.In some embodiments, a single dose of the composition is administered to a subject twice a day, daily, weekly, or monthly.

[0139] In some embodiments, each of the one or more microbial strains in a dose is 10 1 ~10 15 In some embodiments, each of the one or more microbial strains in a dose comprises 10 colony forming units (CFU). 6 ~10 15In some embodiments, each of the one or more microbial strains in a single dose contains the same number of CFU. In some embodiments, some of the one or more microbial strains in a single dose contain different numbers of CFU.

[0140] In some embodiments, a single dose of one or more microbial strains comprises a total of 10 6 ~10 15 In some embodiments, a single dose of one or more microbial strains comprises a total of 10 CFU. 7 ~10 15 In some embodiments, a single dose of the one or more microbial strains comprises between 5 and 200 billion CFUs. In some embodiments, a single dose of the one or more microbial strains comprises between 5 and 50 billion CFUs. In some embodiments, a single dose of the one or more microbial strains comprises between 5 and 20 billion CFUs. In some embodiments, a single dose of the one or more microbial strains comprises between 50 and 100 billion CFUs. In some embodiments, a single dose of the one or more microbial strains comprises between 100 and 200 billion CFUs.

[0141] In some embodiments, efficacy can be assessed by measuring the level of cellular oxidative stress in biological samples before and after administration of the compositions described herein. The level of cellular oxidative stress can be assessed, for example, by measuring the expression of oxidative stress biomarkers, such as reactive oxygen species (ROS) levels, or lipid, protein, and nucleic acid damage levels, or by determining the ratio of oxidized to reduced forms of one or more biomarkers. Because high levels of oxidative stress can be cytotoxic, the level of oxidative stress can be measured by assessing the concentration of intracellular proteins present in the systemic circulation from inflamed or lysed cells (e.g., nerve cells). [Example]

[0142] Example 1: Evaluation of the efficacy of microbiota compositions in affecting insulin signaling and its upstream molecule, IDE, in an AD mouse model This example provides an evaluation of the efficacy of microbiota compositions, specifically CT10, CT10x, and CT10m compositions, in affecting insulin signaling and its upstream molecule IDE, which is associated with Aβ clearance and neuroprotection, in an in vivo mouse model of AD.

[0143] Background: The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeletal assembly, synaptogenesis, neurotransmitter and memory function, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). Several Aβ-degrading enzymes, including neprilysin (NEP), insulin-degrading enzyme (IDE), and endothelin-converting enzyme (endothelin-converting enzyme), reduce Aβ levels and protect against cognitive impairment in mouse models of AD (Miners et al., 2011). Insulin-degrading enzyme (IDE) is a thiol metalloprotease that degrades several peptide hormones, including insulin, glucagon, atrial natriuretic peptide (ANP), and IGF-II (Duckworth et al., 1998). IDE has also been identified as the primary protease involved in the degradation of Aβ peptides (Kurochkin and Goto, 1994; Vekrellis et al., 2000; Farris et al., 2003, 2004). Decreased IDE protein and mRNA levels have been observed in the hippocampus and cortex of AD patients carrying the apolipoprotein E-e4 (ApoE4) allele (Cook et al., 2003; Zhao et al., 2004).

[0144] Mouse model: Nine-month-old Tg2576 mice were used in these experiments. This mouse model is one of the most popular transgenic mouse models, overexpressing a mutant form of the amyloid precursor protein (APP) (isoform 695) with the Swedish mutation (KM670 / 671NL), leading to increased levels of amyloid beta (Aβ) and ultimately amyloid plaques. Wild-type mice were used as controls in all experiments.

[0145] Study: Tg2576 mice were divided into five groups of 15 mice per group and provided with sham (DPBS) or microbiota compositions (CT10, CT10m, or CT10x, composition details listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS, (ii) G2: Tg2576 transgenic mice treated with DPBS, (iii) G3: Tg2576 transgenic mice treated with the CT10 composition, (iv) G4: Tg2576 transgenic mice treated with the CT10x composition, and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, after which histological analysis was performed.

[0146] Methods: To determine whether CT10, CT10x, or CT10m treatment affected insulin signaling and its upstream molecules related to Aβ clearance and neuroprotection, insulin-degrading enzyme (IDE) protein levels were examined by Western blot in brain samples from TG2576, an AD mouse model. Brain lysates were obtained from the cortex, hippocampus, and thalamus by immersing the tissue in PhosphoSafe buffer (EMD Millipore) and lysing in tubes containing Lysing Matrix D (MP Biomedicals). After centrifugation of the samples, the protein concentration of the supernatant was determined by the Bradford method using BioRad Protein Assay Reagent (Cat. No. 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using sodium dodecyl sulfate (SDS) sample buffer, boiled at 70°C for 10 min. Samples were run on custom-made SDS-polyacrylamide Bis-Tris gels (4-12%, Invitrogen) using MES running buffer and subsequently transferred to PVDF membranes (catalog no. IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membranes were blocked for 1 hour using Intercept® (TBS) protein-free blocking buffer (catalog no. 927-80001, LI-COR) or 5% nonfat dry milk solution, and then incubated with IDE-specific antibody (catalog no. AB9210, Millipore) at a dilution of 1:1000 in Intercept® T20 (TBS) protein-free antibody diluent (catalog no. 927-85001, LI-COR) overnight at 4°C with gentle shaking. The next day, membranes were thoroughly washed in TBST (0.1% Tween 20) and incubated in horseradish peroxidase (HRP)-conjugated secondary anti-rabbit antibody (Cat. No. 7074, Cell Signaling) at a dilution of 1:2000 for 1 hour at room temperature. After visualization using a G:Box Mini (Syngene), densitometric quantification of immunoblots was performed using GeneTools (Syngene). Target bands were normalized using the respective β-actin loading controls.

[0147] Results: Figure 1 shows the results of this experiment. Each dot in Groups 1, 4, and 5 represents pooled brain lysates from two animals. IDE protein levels were significantly increased in brain lysates from the TG2576 (G2), CT10 (G3), and CT10m (G5) groups compared with the control group (G1), and the mean IDE levels were higher in CT10m (G5) than in TG2576 (G2). *p<0.05, **p<0.01. Data are expressed as mean ± SEM. Statistical analysis was performed using GraphPad Prism. Data sets were analyzed by one-way ANOVA followed by Dunnett's test. The results suggest that CT10 and CT10m treatment increased IDE levels and its clearance effect on Aβ peptides, indicating neuroprotection.

[0148] Example 2: Evaluation of the efficacy of microbiota compositions in affecting insulin signaling and its upstream molecule NSE in an AD mouse model This example provides an evaluation of the efficacy of microbiota compositions, specifically CT10, CT10x, and CT10m compositions, in affecting insulin signaling and its upstream molecule NSE, which is associated with Akt activation and neuroprotection, in an in vivo mouse model of AD.

[0149] Background: The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeletal assembly, synaptogenesis, neurotransmitter and memory function, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). Neuron-specific enolase (NSE) is a glycolytic isoenzyme that is a highly specific marker of central and peripheral neurons and neuroendocrine cells. NSE is also expressed in microglia (Hafner et al., 2013; Pislar et al., 2017) and astrocytes, particularly reactive astrocytes (Vinores et al., 1985). NSE levels in serum and CSF have been used as a biomarker in injury, cancer, and neurodegenerative diseases (Schmidt et al., 2014; Isgro et al., 2015). Elevated NSE promotes glycolysis, cell proliferation, activation, and migration via the PI3K / AKT and MAPK / ERK pathways. NSE-mediated activation of PI3K also regulates RhoA kinase, which affects actin cytoskeleton reorganization and induction of neurite outgrowth (Haque et al., 2018).

[0150] Mouse model: The mouse model described in Example 1 was used in this study.

[0151] Study: Tg2576 mice were divided into five groups of 15 mice per group and provided with sham (DPBS) or microbiota compositions (CT10, CT10m, or CT10x, composition details listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS, (ii) G2: Tg2576 transgenic mice treated with DPBS, (iii) G3: Tg2576 transgenic mice treated with the CT10 composition, (iv) G4: Tg2576 transgenic mice treated with the CT10x composition, and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, after which histological analysis was performed.

[0152] Methods: To determine whether CT10, CT10x, or CT10m treatment affected insulin signaling and its upstream molecule, NSE, which is involved in Akt activation and neuroprotection, NSE protein levels were examined by Western blot in brain samples from TG2576, an AD mouse model. Brain lysates were obtained from the cortex, hippocampus, and thalamus by immersing the tissue in PhosphoSafe buffer (EMD Millipore) and lysing in tubes containing Lysing Matrix D (MP Biomedicals). After centrifugation of the samples, the protein concentration of the supernatant was determined by the Bradford method using BioRad Protein Assay Reagent (Cat. No. 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer boiled at 70°C for 10 min. Samples were run on custom-made SDS-polyacrylamide Bis-Tris gels (4-12%, Invitrogen) using MES running buffer and subsequently transferred to PVDF membranes (catalog no. IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membranes were blocked for 1 hour using Intercept® (TBS) protein-free blocking buffer (catalog no. 927-80001, LI-COR) or 5% nonfat dry milk solution, and then incubated with NSE-specific antibody (catalog no. sc-21738, Santa Cruz Biotechnology) at a dilution of 1:1000 in Intercept® T20 (TBS) protein-free antibody diluent (catalog no. 927-85001, LI-COR) overnight at 4°C under gentle shaking. The next day, membranes were thoroughly washed in TBST (0.1% Tween 20) and incubated in HRP-conjugated secondary anti-rabbit antibody (Cat. No. 7074, Cell Signaling) at a dilution of 1:2000 for 1 hour at room temperature. After visualization using a G:Box Mini (Syngene), densitometric quantification of immunoblots was performed using GeneTools (Syngene). Target bands were normalized using the respective β-actin loading controls.

[0153] Results: Figure 2 shows the results of this experiment. Each dot in groups 1, 4, and 5 represents pooled brain lysates from two animals. NSE protein levels were significantly increased in brain lysates from the CT10 (G3) and CT10m (G5) groups compared with TG2576 (G2). **p<0.01, ***p<0.001. Data are expressed as mean ± SEM. Statistical analysis was performed using GraphPad Prism. Data sets were analyzed by one-way ANOVA followed by Dunnett's test. These results suggest that CT10 and CT10m treatment increased NSE levels, which may induce insulin-Akt signaling activation and neuroprotection.

[0154] Example 3: Evaluation of the efficacy of microbiota compositions in affecting Akt insulin signaling in an AD mouse model This example provides an evaluation of the efficacy of microbiota compositions, specifically CT10, CT10x, and CT10m compositions, in affecting Akt insulin signaling in an in vivo mouse model of AD.

[0155] Background: The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeletal assembly, synaptogenesis, neurotransmitter function, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). AD is associated with a relative state of insulin resistance in the brain and reduced insulin / IGF-1 expression and Akt signaling. There are fewer insulin receptors and reduced tyrosine kinase activity (IR desensitization) (Steen E & de la Monte 2005; Frohlich L & Hoyer S. Ann. 1999). AD is associated with decreased insulin-mediated glucose uptake. Insulin levels are reduced in the AD brain and CSF (Frohlich 1998; Craft 1998).

[0156] Mouse model: The mouse model described in Example 1 was used in this study.

[0157] Study: Tg2576 mice were divided into five groups of 15 mice per group and provided with sham (DPBS) or microbiota compositions (CT10, CT10m, or CT10x, composition details listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS, (ii) G2: Tg2576 transgenic mice treated with DPBS, (iii) G3: Tg2576 transgenic mice treated with the CT10 composition, (iv) G4: Tg2576 transgenic mice treated with the CT10x composition, and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, after which histological analysis was performed.

[0158] Methods: To determine whether CT10, CT10x, or CT10m treatment affected Akt signaling, the active form of Akt, p-Akt (Ser473), and total Akt1 enzyme protein levels were measured by Western blot in brain samples from TG2576 mice. Brain lysates were obtained from the cortex, hippocampus, and thalamus by immersing the tissue in PhosphoSafe buffer (EMD Millipore) and lysing in tubes containing Lysing Matrix D (MP Biomedicals). After centrifugation of the samples, the protein concentration of the supernatant was determined by the Bradford method using BioRad Protein Assay Reagent (Cat. No. 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer boiled at 70°C for 10 min. Samples were run on custom-made SDS-polyacrylamide Bis-Tris gels (4-12%, Invitrogen) using MES running buffer and subsequently transferred to PVDF membranes (catalog no. IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membranes were blocked for 1 hour using Intercept® (TBS) protein-free blocking buffer (catalog no. 927-80001, LI-COR) or 5% nonfat dry milk solution, and then incubated with p-Akt (Ser473), Akt1-specific antibody (catalog no. 4060, 4691, Cell Signaling Tech.) at a dilution of 1:1000 in Intercept® T20 (TBS) protein-free antibody diluent (catalog no. 927-85001, LI-COR) overnight at 4°C under gentle shaking. The next day, membranes were thoroughly washed in TBST (0.1% Tween 20) and incubated in HRP-conjugated secondary anti-rabbit antibody (Cat. No. 7074, Cell Signaling) at a dilution of 1:2000 for 1 hour at room temperature. After visualization using a G:Box Mini (Syngene), densitometric quantification of immunoblots was performed using GeneTools (Syngene). Target bands were normalized using the respective β-actin loading controls.

[0159] Results: Figure 3 shows the results of this experiment. Each dot in Groups 1, 4, and 5 represents pooled brain lysates from two animals. p-Akt (Ser473) protein levels were significantly reduced in brain lysates from TG2576 mice compared with non-Tg mice. CT10 (G3), CT10x (G4), and CT10m (G5)-treated mice exhibited significantly restored p-Akt (Ser473) protein levels compared with vehicle-treated TG2576 mice (G2). Total Akt1 protein kinase levels in brain lysates from CT10 (G3), CT10x (G4), or CT10m (G5)-treated TG2576 mice were elevated by approximately 120% relative to the control group (non-Tg, G1). *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001. Data are expressed as mean ± SEM. Statistical analysis was performed using GraphPad Prism. Data sets were analyzed by one-way analysis of variance followed by Dunnett's test. The results suggest that CT10, CT10x, and CT10m treatments increase and restore the insulin / Akt signaling pathway, which benefits neuronal survival and memory as well as other brain functions.

[0160] Example 4: Evaluation of the efficacy of microbiota compositions in affecting insulin receptor β levels in an AD mouse model This example provides an evaluation of the efficacy of microbiota compositions, specifically CT10, CT10x, and CT10m compositions, in affecting insulin receptor β levels in an in vivo mouse model of AD.

[0161] Mouse model: The mouse model described in Example 1 was used in this study.

[0162] Study: Tg2576 mice were divided into five groups of 15 mice per group and provided with sham (DPBS) or microbiota compositions (CT10, CT10m, or CT10x, composition details listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS, (ii) G2: Tg2576 transgenic mice treated with DPBS, (iii) G3: Tg2576 transgenic mice treated with the CT10 composition, (iv) G4: Tg2576 transgenic mice treated with the CT10x composition, and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, after which histological analysis was performed.

[0163] Methods: To determine whether CT10, CT10x, or CT10m treatment affected insulin signaling, insulin receptor β protein levels were measured by Western blot in brain samples from TG2576 mice. Brain lysates were obtained from the cortex, hippocampus, and thalamus by immersing the tissue in PhosphoSafe buffer (EMD Millipore) and lysing in tubes containing Lysing Matrix D (MP Biomedicals). After centrifugation of the samples, the protein concentration of the supernatant was determined by the Bradford method using BioRad Protein Assay Reagent (Cat. No. 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer boiled at 70°C for 10 min. Samples were run on a custom-made SDS-polyacrylamide Bis-Tris gel (4-12%, Invitrogen) using MES running buffer and subsequently transferred to a PVDF membrane (catalog no. IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using Intercept® (TBS) protein-free blocking buffer (catalog no. 927-80001, LI-COR) or 5% nonfat dry milk solution, and then incubated with an insulin receptor β-specific antibody (catalog no. 3025, Cell Signaling Tech.) at a dilution of 1:1000 overnight at 4°C under gentle shaking. The next day, the membrane was thoroughly washed in TBST (0.1% Tween 20) and incubated for 1 hour at room temperature in an HRP-conjugated secondary anti-rabbit antibody (catalog no. 7074, Cell Signaling) at a dilution of 1:2000. After visualization using G:Box Mini (Syngene), densitometric quantification of immunoblots was performed using GeneTools (Syngene). Target bands were normalized using the respective β-actin loading controls.

[0164] Results: Figure 4 shows the results of this experiment. Each dot in Groups 1, 4, and 5 represents pooled brain lysates from two animals. Compared with the control group (non-Tg), the mean level of insulin receptor β was decreased in TG2576, although not significantly. Insulin receptor β protein levels were significantly increased in brain lysates from the CT10m (G5) group compared with TG2576 (G2). **p<0.01. Data are expressed as mean ± SEM. Statistical analysis was performed using GraphPad Prism. Data sets were analyzed by one-way ANOVA followed by Dunnett's test. The results suggest that CT10m treatment increases and restores the insulin / Akt signaling pathway by increasing insulin receptor β levels.

[0165] Example 5: Evaluation of the efficacy of microbiota compositions in affecting glucose transport 3 levels in an AD mouse model This example provides an evaluation of the efficacy of microbiota compositions, specifically CT10, CT10x, and CT10m compositions, in affecting glucose transport 3 levels in an in vivo mouse model of AD.

[0166] Mouse model: The mouse model described in Example 1 was used in this study.

[0167] Study: Tg2576 mice were divided into five groups of 15 mice per group and provided with sham (DPBS) or microbiota compositions (CT10, CT10m, or CT10x, composition details listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS, (ii) G2: Tg2576 transgenic mice treated with DPBS, (iii) G3: Tg2576 transgenic mice treated with the CT10 composition, (iv) G4: Tg2576 transgenic mice treated with the CT10x composition, and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, after which histological analysis was performed.

[0168] Methods: To determine whether CT10, CT10x, or CT10m treatment affected insulin signaling and its downstream targets, glucose transporter 3 (Glut3) protein levels were measured by Western blot in brain samples from TG2576 mice. Brain lysates were obtained from the cortex, hippocampus, and thalamus by immersing the tissue in PhosphoSafe buffer (EMD Millipore) and lysing in tubes containing Lysing Matrix D (MP Biomedicals). After centrifugation of the samples, the protein concentration of the supernatant was determined by the Bradford method using BioRad Protein Assay Reagent (Cat. No. 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer boiled at 70°C for 10 min. Samples were run on a custom-made SDS-polyacrylamide Bis-Tris gel (4-12%, Invitrogen) using MES running buffer and subsequently transferred to a PVDF membrane (catalog no. IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using Intercept® (TBS) protein-free blocking buffer (catalog no. 927-80001, LI-COR) or 5% nonfat dry milk solution, and then incubated with a glucose transporter 3 (Glut3)-specific antibody (catalog no. ab191071, Abcam) at a dilution of 1:1000 overnight at 4°C under gentle shaking. The next day, the membrane was thoroughly washed in TBST (0.1% Tween 20) and incubated for 1 hour at room temperature in an HRP-conjugated secondary anti-rabbit antibody (catalog no. 7074, Cell Signaling) at a dilution of 1:2000. After visualization using G:Box Mini (Syngene), densitometric quantification of immunoblots was performed using GeneTools (Syngene). Target bands were normalized using the respective β-actin loading controls.

[0169] Results: Figure 5 shows the results of this experiment. Each dot in Groups 1, 4, and 5 represents pooled brain lysates from two animals. Compared with the control group (non-Tg), the mean level of Glut3 was not significantly decreased in TG2576 (G2). Glut3 protein levels were significantly increased in brain lysates from the CT10m (G5) group compared with TG2576 (G2). *p<0.05. Data are expressed as mean ± SEM. Statistical analysis was performed using GraphPad Prism. Data sets were analyzed by one-way ANOVA followed by Dunnett's test. The results suggest that CT10m treatment increases and restores the insulin / Akt signaling pathway and energy metabolism by increasing glucose transporter 3 (Glut3) protein levels.

[0170] Example 6: Evaluation of the efficacy of microbiota compositions in affecting protein RBAP48 levels in an AD mouse model This example provides an evaluation of the efficacy of microbiota compositions, specifically CT10, CT10x, and CT10m compositions, in affecting memory function-related protein RBAP48 levels in an in vivo mouse model of AD.

[0171] Background: The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeletal assembly, synaptogenesis, neurotransmitter and memory function, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). RBAP48 interacts with histones and modifies histone acetylation, which is essential for memory consolidation. Loss of RBAP48 is key to age-related memory decline (Pavlopoulos et al., 2013). RBAP48 interacts with CREB-binding protein and phosphorylated CREB complexes, which are involved in learning and memory (Zhang et al., 2000).

[0172] Mouse model: The mouse model described in Example 1 was used in this study.

[0173] Study: Tg2576 mice were divided into five groups of 15 mice per group and provided with sham (DPBS) or microbiota compositions (CT10, CT10m, or CT10x, composition details listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS, (ii) G2: Tg2576 transgenic mice treated with DPBS, (iii) G3: Tg2576 transgenic mice treated with the CT10 composition, (iv) G4: Tg2576 transgenic mice treated with the CT10x composition, and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, after which histological analysis was performed.

[0174] Methods: To determine whether CT10, CT10x, or CT10m treatment affected insulin signaling and its downstream target molecules related to memory function, RBAP48 protein levels were measured by Western blot in brain samples from TG2576, an AD mouse model. Brain lysates were obtained from the cortex, hippocampus, and thalamus by immersing the tissue in PhosphoSafe buffer (EMD Millipore) and lysing in tubes containing Lysing Matrix D (MP Biomedicals). After centrifugation of the samples, the protein concentration of the supernatant was determined by the Bradford method using BioRad Protein Assay Reagent (Cat. No. 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer boiled at 70°C for 10 min. Samples were run on a custom-made SDS-polyacrylamide Bis-Tris gel (4-12%, Invitrogen) using MES running buffer and subsequently transferred to a PVDF membrane (catalog no. IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membrane was blocked for 1 hour using Intercept® (TBS) protein-free blocking buffer (catalog no. 927-80001, LI-COR) or 5% nonfat dry milk solution, and then incubated with RBAP48-specific antibody (catalog no. GTX70232, GeneTex) at a dilution of 1:1000 overnight at 4°C under gentle shaking. The next day, the membrane was thoroughly washed in TBST (0.1% Tween 20) and incubated for 1 hour at room temperature in HRP-conjugated secondary anti-mouse antibody (catalog no. 7076, Cell Signaling) at a dilution of 1:2000. After visualization using G:Box Mini (Syngene), densitometric quantification of immunoblots was performed using GeneTools (Syngene). Target bands were normalized using the respective β-actin loading controls.

[0175] Results: Figure 6 shows the results of this experiment. Each dot in Groups 1, 4, and 5 represents pooled brain lysates from two animals. RBAP48 protein levels were significantly increased in brain lysates from the CT10 (G3), CT10x (G4), and CT10m (G5) groups compared with TG2576 (G2). *p<0.05, ****p<0.0001. Data are expressed as mean ± SEM. Statistical analysis was performed using GraphPad Prism. Data sets were analyzed by one-way ANOVA followed by Dunnett's test. These results suggest that CT10, CT10x, and CT10m treatments increase and restore the insulin / Akt signaling pathway and its downstream targets for memory function by increasing RBAP48 protein levels.

[0176] Example 7: Evaluation of the efficacy of microbiota compositions in affecting p-4EBP1, Akt, and mTOR levels in an AD mouse model This example provides an evaluation of the efficacy of microbiota compositions, specifically CT10, CT10x, and CT10m compositions, in affecting p-4EBP1, Akt, and mTOR levels in an in vivo mouse model of AD.

[0177] Background: The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeletal assembly, synaptogenesis, neurotransmitter and memory function, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). Hyperphosphorylation of 4EBP1 leads to activation of cap-dependent translation, a process responsible for protein synthesis (Pause et al., 1994). Both the PI3 kinase / Akt pathway and mTOR kinase regulate 4EBP1 activity (Brunn et al., 1997; Gingras et al., 1998). Tg2576 mice showed decreased levels of phospho-4EBP1 (Thr37 / 46) in the brain hippocampus. Inhibition of mTOR signaling correlates with reduced synaptic plasticity in an AD mouse model (Ma et al., 2010).

[0178] Mouse model: The mouse model described in Example 1 was used in this study.

[0179] Study: Tg2576 mice were divided into five groups of 15 mice per group and provided with sham (DPBS) or microbiota compositions (CT10, CT10m, or CT10x, composition details listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS, (ii) G2: Tg2576 transgenic mice treated with DPBS, (iii) G3: Tg2576 transgenic mice treated with the CT10 composition, (iv) G4: Tg2576 transgenic mice treated with the CT10x composition, and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, after which histological analysis was performed.

[0180] Methods: To determine whether CT10, CT10x, or CT10m treatment affected insulin signaling and its downstream target molecules involved in protein synthesis and translation, phospho-4EBP1 (Thr37 / 46) protein levels were measured by Western blot in brain samples from TG2576, an AD mouse model. Brain lysates were obtained from the cortex, hippocampus, and thalamus by immersing the tissue in PhosphoSafe buffer (EMD Millipore) and lysing in tubes containing Lysing Matrix D (MP Biomedicals). After centrifugation of the samples, the protein concentration of the supernatant was determined by the Bradford method using BioRad Protein Assay Reagent (Cat. No. 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer boiled at 70°C for 10 min. Samples were run on custom-made SDS-polyacrylamide Bis-Tris gels (4-12%, Invitrogen) using MES running buffer and subsequently transferred to PVDF membranes (catalog no. IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membranes were blocked for 1 hour using Intercept® (TBS) protein-free blocking buffer (catalog no. 927-80001, LI-COR) or 5% nonfat dry milk solution, and then incubated with phospho-4EBP1 (Thr37 / 46)-specific antibody (catalog no. 2855, Cell Signaling) at a dilution of 1:1000 in Intercept® T20 (TBS) protein-free antibody diluent (catalog no. 927-85001, LI-COR) overnight at 4°C under gentle shaking. The next day, membranes were thoroughly washed in TBST (0.1% Tween 20) and incubated in HRP-conjugated secondary anti-rabbit antibody (Cat. No. 7074, Cell Signaling) at a dilution of 1:2000 for 1 hour at room temperature. After visualization using a G:Box Mini (Syngene), densitometric quantification of immunoblots was performed using GeneTools (Syngene). Target bands were normalized using the respective β-actin loading controls.

[0181] Results: Figure 7 shows the results of this experiment. Each dot in groups 1, 4, and 5 represents pooled brain lysates from two animals. Phospho-4EBP1 (Thr37 / 46) protein levels were significantly increased in brain lysates from the CT10x (G4) and CT10m (G5) groups compared with TG2576 (G2). **p<0.01, ****p<0.0001. Data are expressed as mean ± SEM. Statistical analysis was performed using GraphPad Prism. Data sets were analyzed by one-way ANOVA followed by Dunnett's test. These results suggest that CT10x and CT10m treatments increase the insulin / Akt signaling pathway and its downstream targets of protein synthesis by increasing phospho-4EBP1 (Thr37 / 46) protein levels.

[0182] Example 8: Evaluation of the efficacy of microbiota compositions in affecting antioxidant response regulator NRF2 levels in AD mouse models This example provides an evaluation of the efficacy of microbiota compositions, specifically CT10, CT10x, and CT10m compositions, in affecting antioxidant response regulator NRF2 levels in an in vivo mouse model of AD.

[0183] Background: The insulin / IGF pathway supports neuronal growth, survival, differentiation, migration, energy metabolism, gene expression, protein synthesis, cytoskeletal assembly, synaptogenesis, neurotransmitter and memory function, and plasticity (Chesik et al., 2008; de la Monte and Wands, 2005; Gong et al., 2008; Liang et al., 2007). NRF2 (nuclear factor erythroid 2-related factor 2) is a master regulator of the cellular antioxidant response, regulating the expression of over 200 genes containing antioxidant response elements (AREs). The dietary herb Bungeanum ameliorates cognitive dysfunction and neurological deficits in aging mouse models through activation of the PI3K / Akt / NRF2 signaling pathway (Zhao et al., 2020). NRF2 activation through the PI3K / GSK-3 axis protects neurons from Aβ-mediated oxidative and metabolic damage (Sotolongo et al., 2020). Activation of the NRF2 / ARE pathway attenuates cognitive impairment in a Tg mouse model of AD through regulation of oxidative stress (Tian et al., 2018).

[0184] Mouse model: The mouse model described in Example 1 was used in this study.

[0185] Study: Tg2576 mice were divided into five groups of 15 mice per group and provided with sham (DPBS) or microbiota compositions (CT10, CT10m, or CT10x, composition details listed in Tables 2, 3, and 4) by daily oral gavage for 6 months. The groups were: (i) G1: wild-type mice treated with DPBS, (ii) G2: Tg2576 transgenic mice treated with DPBS, (iii) G3: Tg2576 transgenic mice treated with the CT10 composition, (iv) G4: Tg2576 transgenic mice treated with the CT10x composition, and (v) G5: Tg2576 transgenic mice treated with the CT10m composition. Animals were sacrificed at the end of the study, after which histological analysis was performed.

[0186] Methods: To determine whether CT10, CT10x, or CT10m treatment affected insulin signaling and its downstream target molecules related to antioxidant responses and neuroprotection, NRF2 protein levels were measured by Western blot in brain samples from TG2576, an AD mouse model. Brain lysates were obtained from the cortex, hippocampus, and thalamus by immersing the tissue in PhosphoSafe buffer (EMD Millipore) and lysing in tubes containing Lysing Matrix D (MP Biomedicals). After centrifugation of the samples, the protein concentration of the supernatant was determined by the Bradford method using BioRad Protein Assay Reagent (Cat. No. 5000002, Bio-Rad). Total protein lysates (30 μg) were prepared using SDS sample buffer boiled at 70°C for 10 min. Samples were run on custom-made SDS-polyacrylamide Bis-Tris gels (4-12%, Invitrogen) using MES running buffer and subsequently transferred to PVDF membranes (catalog no. IB24001, Invitrogen) using the iBlot2 (Invitrogen) system. The membranes were blocked for 1 hour using Intercept® (TBS) protein-free blocking buffer (catalog no. 927-80001, LI-COR) or 5% nonfat dry milk solution, and then incubated with NRF2-specific antibody (catalog no. 12721, Cell Signaling) at a dilution of 1:1000 in Intercept® T20 (TBS) protein-free antibody diluent (catalog no. 927-85001, LI-COR) overnight at 4°C under gentle shaking. The next day, membranes were thoroughly washed in TBST (0.1% Tween 20) and incubated in HRP-conjugated secondary anti-rabbit antibody (Cat. No. 7074, Cell Signaling) at a dilution of 1:2000 for 1 hour at room temperature. After visualization using a G:Box Mini (Syngene), densitometric quantification of immunoblots was performed using GeneTools (Syngene). Target bands were normalized using the respective β-actin loading controls.

[0187] Results: Figure 8 shows the results of this experiment. Each dot in Groups 1, 4, and 5 represents pooled brain lysates from two animals. NRF2 protein levels were significantly increased in brain lysates from the CT10 (G3), CT10x (G4), and CT10m (G5) groups compared with TG2576 (G2). *p<0.05, **p<0.01. Data are expressed as mean ± SEM. Statistical analysis was performed using GraphPad Prism. Data sets were analyzed by one-way ANOVA followed by Dunnett's test. The results suggest that CT10, CT10x, and CT10m treatments increase the insulin / Akt signaling pathway and its downstream targets for antioxidant response and neuroprotection by increasing NRF2 protein levels.

[0188] Other embodiments Those skilled in the art will appreciate that various changes, modifications, and improvements to this disclosure will readily occur to those skilled in the art. Such changes, modifications, and improvements are intended to be part of this disclosure and are intended to be within the spirit and scope of the invention. Accordingly, the foregoing description and drawings are by way of example only, and any inventions described in this disclosure as more particularly set forth in the following claims.

[0189] Those of ordinary skill in the art will understand the typical standard deviation or error attributed to values ​​obtained from the assays or other processes described herein. Publications, websites, and other reference materials referred to herein to describe the background of the invention and to provide additional details regarding its practice are hereby incorporated by reference in their entirety.

[0190] While embodiments of the invention have been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate, but not limit, the scope of the invention, which is defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

[0191] equivalent Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. The scope of the present invention is not intended to be limited to the above Description, but rather is as set forth in the following claims. [Table 8-1] [Table 8-2] [Table 8-3] [Table 8-4] [Table 9-1] [Table 9-2] [Table 9-3] [Table 9-4] [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4] [Table 11-1] [Table 11-2] [Table 11-3]

Table 11-4

Table 12-1

Table 12-2

Table 13-1

Table 13-2

Table 13-3

Table 13-4

Table 14-1

Table 14-2

Table 14-3

Table 14-4

Table 15-1

Table 15-2

Table 15-3

Table 15-4

Table 16-1

Table 16-2

Table 21-1

Table 21-2

Table 22-1

Table 22-2

Table 24-1

Table 24-2

Table 24-3

Table 24-4

Table 25-1

Table 25-2

Table 25-3

Table 25-4

Table 26-1

Table 26-2

Table 26-3

Table 26-4

Table 28-1

Table 28-2

Table 28-3

Table 28-4

Table 29-1

Table 29-2

Table 29-3

Table 30-1

Table 30-2

Table 30-3

Table 30-4

Table 30-5

Table 30-6

Table 30-7

Table 30-8

Table 30-9

Table 30-10

Claims

1. 1. A method comprising: The method comprises administering to a subject a composition comprising one or more microbial strains or microbial components, wherein the subject has been diagnosed with or is at high risk for developing an insulin-related disease, disorder, or condition.

2. 1. A method comprising: The method comprises administering to a subject a composition comprising one or more microbial metabolites, wherein the subject has been diagnosed with or is at high risk of developing an insulin-related disease, disorder, or condition.

3. 1. A method comprising: Target, (i) one or more microbial strains or microbial components; or (ii) administering a composition comprising one or more microbial metabolites; The method, wherein the subject has been diagnosed with or is at high risk of developing an insulin-related disease, disorder, or condition.

4. 10. The method of any one of the preceding claims, wherein the insulin-related disease, disorder, or condition is or comprises diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermadrome, diabetic neuropathy, diabetic foot ulcer, maturity-onset diabetes of the young, pancreatogenic diabetes, or polycystic ovary syndrome (PCOS).

5. 10. The method of any one of the preceding claims, wherein the subject is a mammal.

6. 10. The method of any one of the preceding claims, wherein the subject is a human.

7. 7. The method of any one of claims 1 or 3 to 6, wherein the one or more microbial strains are derived from the microbiota of a mammal.

8. 8. The method of any one of claims 1 or 3 to 7, wherein the one or more microbial strains are derived from the human microbiome.

9. 9. The method of claim 8, wherein the human microbiota is the microbiota of the subject.

10. 10. The method of claim 9, wherein the human microbiota is administered to maintain or modulate the microbiota of the subject.

11. 10. The method of any one of the preceding claims, wherein the one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3, or Appendix 4.

12. 10. The method of any one of the preceding claims, wherein the one or more microbial metabolic products are or comprise bile acids.

13. 10. The method of any one of the preceding claims, wherein the one or more microbial metabolites is or comprises tauroursodeoxycholic acid.

14. 10. The method of any one of the preceding claims, wherein the one or more microbial components or microbial metabolites are butyrylcamitin, theobromine, p-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro), or any combination thereof.

15. One or more microbial components or microbial metabolites may be 4-hydroxyphenylpyruvate, ectoine, gramine, N-acetyl-L-phenylalanine, nepsilon-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, trans-urocanic acid, N-acetyl-L-leucine, sarcosine, isobutyrylcarnitine , b-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenaceturic acid, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homocitrulline, mucic acid, homocysteine ​​sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3-indoxyl sulfate, nicotinamide, N-formylglycine, ureidoglycolate, N-methylproline, glucaric acid, butyrylcarnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenaceturic acid, diethanolamine, phosphorylcholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine ​​glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, b-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thiaproline, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0 XA0027, or any combination thereof.

16. 16. The method of any one of claims 1 or 3-15, wherein the one or more microbial strains are or comprise Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or a combination thereof.

17. 17. The method of any one of claims 1 or 3-16, wherein the one or more microbial strains are or comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp., or a combination thereof.

18. 18. The method of any one of claims 1 or 3-17, wherein the one or more microbial strains are or comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or a combination thereof.

19. 19. The method of any one of claims 1 or 3 to 18, wherein the one or more microbial strains is or comprises Bacillus subtilis.

20. 20. The method of any one of claims 1 or 3-19, wherein the composition comprises two or more microbial strains.

21. 21. The method of any one of claims 1 or 3 to 20, wherein the composition comprises five or more microbial strains.

22. 22. The method of any one of claims 1 or 3 to 21, wherein the composition comprises 10 or more microbial strains.

23. 10. The method of any one of the preceding claims, wherein the composition is administered topically, orally, subcutaneously, intravenously, intramuscularly, intracerebrally, intrathecally, intrarectally, ophthalmically, intravitreally, or suprachoroidally.

24. 24. The method of claim 23, wherein the composition is administered orally.

25. 24. The method of claim 23, wherein the composition is administered intravenously.

26. 10. The method of any one of the preceding claims, wherein the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injectable, or eye drops.

27. Each microbial strain of the one or more microbial strains is １ ~10 １５ 27. The method of any one of claims 1 or 3 to 26, wherein the microbial composition is present in the composition at a concentration of CFU.

28. Each microbial strain of the one or more microbial strains is ６ 28. The method of any one of claims 1 or 3 to 27, wherein the microbial composition is present in the composition at a concentration of CFU.

29. A composition for use in treating an insulin-related disease, disorder, or condition, comprising one or more microbial strains or microbial components.

30. A composition for use in treating an insulin-related disease, disorder, or condition, comprising one or more microbial metabolites.

31. 30. The composition of claim 29, wherein the one or more microbial strains are derived from a mammalian microbiota.

32. 32. The composition of claim 29 or 31, wherein the one or more microbial strains are derived from the human microbiome.

33. 33. The composition of claim 32, wherein the human microbiota is the microbiota of a subject.

34. 34. The composition of claim 33, wherein the human microbiota is administered to maintain or modulate the microbiota of the subject.

35. 35. The composition of any one of claims 29 to 34, wherein the one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3, or Appendix 4.

36. 36. The composition of any one of claims 29 to 35, wherein the one or more microbial metabolic products is or comprises a bile acid.

37. 37. The composition of any one of claims 29 to 36, wherein the one or more microbial metabolites is or comprises tauroursodeoxycholic acid.

38. 36. The composition of any one of claims 29-35, wherein the one or more microbial components or microbial metabolites are butyrylcamitin, theobromine, p-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro), or any combination thereof.

39. One or more microbial components or microbial metabolites may be 4-hydroxyphenylpyruvate, ectoine, gramine, N-acetyl-L-phenylalanine, nepsilon-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, trans-urocanic acid, N-acetyl-L-leucine, sarcosine, isobutyrylcarnitine , b-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenaceturic acid, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homocitrulline, mucic acid, homocysteine ​​sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3-indoxyl sulfate, nicotinamide, N-formylglycine, ureidoglycolate, N-methylproline, glucaric acid, butyrylcarnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenaceturic acid, diethanolamine, phosphorylcholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine ​​glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, b-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thiaproline, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0 XA0027, or any combination thereof.

40. 40. The composition of any one of claims 29 or 31-39, wherein the one or more microbial strains are or comprise Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or a combination thereof.

41. 41. The composition of any one of claims 29 or 31-40, wherein the one or more microbial strains are or comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp., or a combination thereof.

42. 42. The composition of any one of claims 29 or 31-41, wherein the one or more microbial strains are or comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or a combination thereof.

43. 43. The composition of any one of claims 29 or 31 to 42, wherein the one or more microbial strains are or comprise Bacillus subtilis.

44. 44. The composition of any one of claims 29 or 31-43, wherein the composition comprises two or more microbial strains.

45. 45. The composition of any one of claims 29 or 31-44, wherein the composition comprises five or more microbial strains.

46. 46. ​​The composition of any one of claims 29 or 31-45, wherein the composition comprises 10 or more microbial strains.

47. 47. The composition of any one of claims 29 to 46, wherein the composition is for topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal, intraocular, intravitreal, or suprachoroidal administration.

48. 48. The composition of claim 47, wherein the composition is for oral administration.

49. 48. The composition of claim 47, wherein the composition is for intravenous administration.

50. 50. The composition of any one of claims 29 to 49, wherein the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injectable, or eye drops.

51. Each microbial strain of the one or more microbial strains is １ ~10 １５ 51. The composition of any one of claims 29 or 31-50, wherein the composition is present in a concentration of CFU.

52. Each microbial strain of the one or more microbial strains is ６ 51. The composition of any one of claims 29 or 31-50, wherein the composition is present in a concentration of CFU.

53. 53. Use of a composition according to any one of claims 29 to 52 for modulating one or more microbial metabolites in a subject.

54. 53. Use of a composition according to any one of claims 29 to 52 for modulating one or more characteristics in a subject.

55. The one or more features are (i) the level of cell survival; (ii) the level or activity of a nucleic acid or protein or a form thereof; (iii) weight gain, (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride levels; (vii) blood cholesterol levels; (viii) oxidative stress, or (ix) Use of the composition of claim 54, which is or comprises inflammation.

56. 53. Use of a composition according to any one of claims 29 to 52 for characterising the ability of one or more microbial strains to modulate one or more microbial metabolites in a subject.

57. 53. Use of the composition of any one of claims 29 to 52 for treating or ameliorating a disease, disorder, or condition in a subject, wherein the disease, disorder, or condition is an insulin-related disease, disorder, or condition associated with one or more microbial metabolites.

58. 58. The use of the composition of claim 57, wherein the disease, disorder, or condition is or comprises diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermadrome, diabetic neuropathy, diabetic foot ulcer, maturity-onset diabetes of the young, pancreatogenic diabetes, or polycystic ovary syndrome (PCOS).

59. 59. The use of the composition of claim 58, wherein the disease, disorder, or condition is diabetes.

60. 1. A method for screening a microbial strain, comprising: contacting the microbial strain with a culture comprising pancreatic cells or a pancreatic cell line that models an insulin-related disease, disorder, or condition; and determining whether the microbial strain has altered a characteristic of the culture, wherein the characteristic is associated with the insulin-related disease, disorder, or condition.

61. 61. The method of claim 60, wherein said determining step comprises comparing said characteristics before and after said contacting step is performed.

62. 61. The method of claim 60, wherein said determining step comprises comparing said characteristic to an equivalent reference after said contacting step.

63. 63. The method of claim 62, wherein the equivalent reference is a historical reference.

64. 64. The method of claim 63, wherein the equivalent reference is a negative control reference.

65. 64. The method of claim 63, wherein the equivalent reference is a positive control reference.

66. 66. The method of any one of claims 60 to 65, wherein the characteristic is the level of cell survival.

67. 66. The method of any one of claims 60 to 65, wherein the characteristic is the level or activity of a nucleic acid or protein, or a form thereof.

68. 66. The method of any one of claims 60 to 65, wherein the characteristic is or comprises weight gain.

69. 66. The method of any one of claims 60 to 65, wherein the characteristic is or comprises fat accumulation in liver cells.

70. 66. The method of any one of claims 60 to 65, wherein the characteristic is or comprises lipid accumulation in hepatocytes.

71. 66. The method of any one of claims 60 to 65, wherein the characteristic is or comprises triglyceride level.

72. 66. The method of any one of claims 60 to 65, wherein the characteristic is or comprises a cholesterol level.

73. 66. The method of any one of claims 60 to 65, wherein the characteristic is or comprises inflammation.

74. the microbial strain alters one or more characteristics of the culture, the one or more characteristics being associated with the insulin-related disease, disorder, or condition, and the one or more characteristics being: (i) the level of cell survival; (ii) the level or activity of a nucleic acid or protein or a form thereof; (iii) weight gain, (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride levels; (vii) blood cholesterol levels; (viii) oxidative stress, or (ix) The method of any one of claims 60 to 65, which is or comprises inflammation.

75. 1. A method comprising: The method comprises administering to the subject a composition comprising one or more microbial strains or microbial components.

76. 1. A method comprising: The method comprises administering to a subject a composition comprising one or more microbial metabolites.

77. 77. The method of claim 75 or 76, wherein the subject is a mammal.

78. 78. The method of any one of claims 75 to 77, wherein the subject is a human.

79. 79. The method of any one of claims 75 or 77-78, wherein the one or more microbial strains are derived from the microbiota of a mammal.

80. 80. The method of any one of claims 75 or 77 to 79, wherein the one or more microbial strains are derived from the human microbiome.

81. 81. The method of claim 80, wherein the human microbiota is the microbiota of the subject.

82. 82. The method of claim 81, wherein the human microbiota is administered to maintain or modulate the microbiota of the subject.

83. 83. The method of any one of claims 75 to 82, wherein the one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3, or Appendix 4.

84. 84. The method of any one of claims 75 to 83, wherein the one or more microbial metabolic products is or comprises a bile acid.

85. 85. The method of any one of claims 75 to 84, wherein the one or more microbial metabolites is or comprises tauroursodeoxycholic acid.

86. 84. The method of any one of claims 75-83, wherein the one or more microbial components or microbial metabolites are butyrylcamitin, theobromine, p-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro), or any combination thereof.

87. One or more microbial components or microbial metabolites may be 4-hydroxyphenylpyruvate, ectoine, gramine, N-acetyl-L-phenylalanine, nepsilon-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, trans-urocanic acid, N-acetyl-L-leucine, sarcosine, isobutyrylcarnitine , b-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenaceturic acid, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homocitrulline, mucic acid, homocysteine ​​sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3-indoxyl sulfate, nicotinamide, N-formylglycine, ureidoglycolate, N-methylproline, glucaric acid, butyrylcarnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenaceturic acid, diethanolamine, phosphorylcholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine ​​glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, b-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thiaproline, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0 XA0027, or any combination thereof.

88. 88. The method of any one of claims 75 or 77-87, wherein the one or more microbial strains are or comprise Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or a combination thereof.

89. 89. The method of any one of claims 75 or 77-88, wherein the one or more microbial strains are or comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp., or a combination thereof.

90. 90. The method of any one of claims 75 or 77-89, wherein the one or more microbial strains are or comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or a combination thereof.

91. 91. The method of any one of claims 75 or 77-90, wherein the one or more microbial strains is or comprises Bacillus subtilis.

92. 92. The method of any one of claims 75 or 77-91, wherein the composition comprises two or more microbial strains.

93. 93. The method of any one of claims 75 or 77-92, wherein the composition comprises five or more microbial strains.

94. 94. The method of any one of claims 75 or 77-93, wherein the composition comprises 10 or more microbial strains.

95. 95. The method of any one of claims 75-94, wherein the composition is administered topically, orally, subcutaneously, intravenously, intramuscularly, intracerebrally, intrathecally, intrarectally, ophthalmically, intravitreally, or suprachoroidally.

96. 96. The method of claim 95, wherein the composition is administered orally.

97. 96. The method of claim 95, wherein the composition is administered intravenously.

98. 98. The method of any one of claims 75 to 97, wherein the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injectable, or eye drops.

99. Each microbial strain of the one or more microbial strains is １ ~10 １５ 99. The method of any one of claims 75 or 77-98, wherein the microbial composition is present in the composition at a concentration of CFU.

100. Each microbial strain of the one or more microbial strains is ６ 100. The method of any one of claims 75 or 77-99, wherein the microbial composition is present in the composition at a concentration of CFU.

101. 101. The method of any one of claims 75 to 100, wherein the microbial strain or the microbial metabolite alters a characteristic of the subject.

102. 102. The method of claim 101, wherein the characteristic is the level of cell viability.

103. 102. The method of claim 101, wherein the characteristic is the level or activity of a nucleic acid or protein, or a form thereof.

104. 102. The method of claim 101, wherein the characteristic is or comprises weight gain.

105. 102. The method of claim 101, wherein the characteristic is or comprises fat accumulation in the liver.

106. 102. The method of claim 101, wherein the characteristic is or comprises lipid accumulation in the liver.

107. 102. The method of claim 101, wherein the characteristic is or comprises triglyceride level.

108. 102. The method of claim 101, wherein the characteristic is or comprises a cholesterol level.

109. 102. The method of claim 101, wherein the characteristic is or comprises inflammation.

110. The microbial strain alters one or more characteristics of the subject, and the one or more characteristics are: (i) the level of cell survival; (ii) the level or activity of a nucleic acid or protein or a form thereof; (iii) weight gain, (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride levels; (vii) blood cholesterol levels; (viii) oxidative stress, or (ix) The method of claim 101, which is or comprises inflammation.

111. 111. The method of any one of claims 101-110, wherein the characteristic is associated with the insulin-related disease, disorder, or condition.

112. 1. A method for characterizing a microbial strain, comprising: adding the microbial strain to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that model an insulin-related disease, disorder, or condition; and determining whether the microbial strain affects the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are associated with the insulin-related disease, disorder, or condition.

113. 1. A method of manufacturing a pharmaceutical treatment comprising characterizing one or more microbial strains, microbial components, or microbial metabolites, comprising: adding the one or more microbial strains to a culture comprising one or more pancreatic cells or one or more pancreatic cell lines that model an insulin-related disease, disorder, or condition; and determining whether the one or more microbial strains affect the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are associated with the insulin-related disease, disorder, or condition.

114. 1. A method of manufacturing a pharmaceutical treatment, comprising: The method comprises formulating one or more microbial strains or microbial components in a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injectable, or eye drop.

115. 1. A method of manufacturing a pharmaceutical treatment, comprising: The method comprises formulating one or more microbial metabolites in a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injectable, or eye drop.

116. 1. A method for evaluating a microbial strain for its ability to affect one or more characteristics of a culture, comprising: adding the microbial strain to the culture comprising one or more pancreatic cells or one or more pancreatic cell lines that model an insulin-related disease, disorder, or condition; and determining whether the microbial strain affects the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines, wherein the one or more characteristics are associated with the insulin-related disease, disorder, or condition.

117. determining the level of one or more characteristics of the one or more pancreatic cells or pancreatic cell lines in the culture prior to adding the microbial strain to the culture; determining the level of the same one or more characteristics of the one or more pancreatic cells or pancreatic cell lines in the culture after adding the microbial strain to the culture; 117. The method of any one of claims 112-114 or 116, further comprising comparing the level of the one or more characteristics determined before adding the microbial strain with the level of the one or more characteristics determined after adding the microbial strain.

118. The one or more features are (i) the level of cell survival; (ii) the level or activity of a nucleic acid or protein or a form thereof; (iii) weight gain, (iv) fat accumulation in the liver; (v) lipid accumulation in the liver; (vi) blood triglyceride levels; (vii) blood cholesterol levels; (viii) oxidative stress, (ix) inflammation, or (x) The method of any one of claims 112 to 117, including combinations thereof.

119. 119. The method of any one of claims 60-118, wherein the disease, disorder, or condition is or comprises diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermadrome, diabetic neuropathy, diabetic foot ulcer, maturity-onset diabetes of the young, pancreatogenic diabetes, or polycystic ovary syndrome (PCOS).

120. A composition for use in treating or preventing an insulin-related disease, disorder, or condition, comprising one or more microbial strains or microbial components.

121. A composition for use in treating or preventing an insulin-related disease, disorder, or condition, comprising one or more microbial metabolites.

122. 121. The composition for use of claim 120, wherein the one or more microbial strains are derived from the microbiota of a mammal.

123. 123. The composition for use of claim 120 or 122, wherein the one or more microbial strains are derived from the human microbiome.

124. 124. The composition for use of claim 123, wherein the human microbiota is the microbiota of a subject.

125. 125. The composition for use of claim 124, wherein the human microbiota is administered to maintain or regulate the microbiota of the subject.

126. 126. A composition for use according to any one of claims 120 to 125, wherein the one or more microbial components or microbial metabolites are selected from Appendix 1, Appendix 3 or Appendix 4.

127. 127. A composition for use according to any one of claims 120 to 126, wherein the one or more microbial metabolic products is or comprises a bile acid.

128. 128. A composition for use according to any one of claims 120 to 127, wherein the one or more microbial metabolites is or comprises tauroursodeoxycholic acid.

129. 127. The composition for use of any one of claims 120-126, wherein the one or more microbial components or microbial metabolites are butyrylcamitin, theobromine, p-hydroxyphenylpyruvic acid, propionic acid, picolinic acid, 2-hydroxy-4-methylvaleric acid, N6-acetyllysine, urocanic acid, N5-ethylglutamine, trigonelline, stachydrine, ectoine, 5-hydroxylysine, arginine (arg), cholic acid, 2-(4-hydroxyphenyl)propionic acid, N-acetyltryptophan, hydroxyproline, argininosuccinic acid, glutamic acid (Glu), sarcosine, 5-methoxyindoleacetic acid, indole-3-lactic acid, isovalerylalanine, N-acetylleucine, 1-methylhistidine, N-acetylphenylalanine, proline (Pro), or any combination thereof.

130. One or more microbial components or microbial metabolites may be 4-hydroxyphenylpyruvate, ectoine, gramine, N-acetyl-L-phenylalanine, nepsilon-acetyl-L-lysine, stachydrine, trigonelline, 3-ureidopropionic acid, theobromine, hippuric acid, imidazolepropionic acid, NG-methyl-L-arginine, trans-urocanic acid, N-acetyl-L-leucine, sarcosine, isobutyrylcarnitine , b-hydroxyisovaleric acid, L-theanine / N5-ethylglutamine, 5-hydroxylysine, phenaceturic acid, betaine, hydroxyproline, picolinic acid, 2-aminoadipic acid, glycerophosphocholine, carnitine, glycerol 3-phosphate, argininosuccinic acid, creatine, terephthalic acid, homocitrulline, mucic acid, homocysteine ​​sulfinic acid, trimethyllysine, spermidine, glyoxylic acid, XA0013 C6H6O4S, 3-indoxyl sulfate, nicotinamide, N-formylglycine, ureidoglycolate, N-methylproline, glucaric acid, butyrylcarnitine, methionine sulfoxide, carboxymethyllysine, glycolic acid, phenaceturic acid, diethanolamine, phosphorylcholine, guanidinosuccinic acid, N-acetylhistidine, glyceric acid, S-methylmethionine, cysteine ​​glutathione disulfide, kynurenine, N-acetylphenylalanine, threonic acid, malic acid, 7,8-dihydrobiopterin, homovanillic acid, taurocholic acid, 5-methoxyindoleacetic acid, butyrate, b-hydroxyisovaleric acid, 2-oxoglutaric acid, N-acetyltryptophan, thiaproline, hypotaurine, cholic acid, acetoacetic acid, ethanolamine, guanidoacetic acid, S-sulfocysteine, myristic acid C14:0 XA0027, or any combination thereof.

131. 131. The composition for use according to any one of claims 120 or 122-130, wherein the one or more microbial strains are or comprise Gluconacetobacter hansenii, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Clostridium butyricum, Paenibacillus sp., Veillonella sp., Bifidobacterium sp., Bacillus subtilis, Acidaminococcus sp., or a combination thereof.

132. 132. The composition for use according to any one of claims 120 or 122-131, wherein the one or more microbial strains are or comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus sp., Lactobacillus plantarum, Veillonella sp., Bifidobacterium sp., or a combination thereof.

133. 133. The composition for use of any one of claims 120 or 122-132, wherein the one or more microbial strains are or comprise Gluconacetobacter hanseni, Terrisporobacter glycolicus, Coprococcus catus, Lactobacillus plantarum, Veillonella atypica, Bifidobacterium breve, or a combination thereof.

134. 134. A composition for use according to any one of claims 120 or 122 to 133, wherein the one or more microbial strains are or comprise Bacillus subtilis.

135. 135. A composition for use according to any one of claims 120 or 122 to 134, wherein the composition comprises two or more microbial strains.

136. 136. The composition for use according to any one of claims 120 or 122 to 135, wherein the composition comprises five or more microbial strains.

137. 137. A composition for use according to any one of claims 120 or 122 to 136, wherein the composition comprises 10 or more microbial strains.

138. 138. The composition for use according to any one of claims 120 to 137, wherein the composition is for topical, oral, subcutaneous, intravenous, intramuscular, intracerebral, intrathecal, rectal, intraocular, intravitreal, or suprachoroidal administration.

139. 139. The composition for use according to claim 138, wherein the composition is for oral administration.

140. 139. The composition for use of claim 138, wherein the composition is for intravenous administration.

141. 141. The composition for use according to any one of claims 120 to 140, wherein the composition is formulated as a syrup, liquid, tablet, lozenge, gummy, capsule, powder, gel, film, injectable, or eye drops.

142. Each microbial strain of the one or more microbial strains is １ ~10 １５ 142. The composition for use of any one of claims 120 or 122-141, wherein the composition is present in a concentration of CFU.

143. wherein each microbial strain of said one or more microbial strains is at least 10 ６ 142. The composition for use of any one of claims 120 or 122-141, wherein the composition is present in a concentration of CFU.

144. 144. The composition for use of any one of claims 120 to 143, wherein the insulin-related disease, disorder, or condition is or comprises diabetes, obesity, cardiovascular disease, non-alcoholic fatty liver disease, Wolfram syndrome, metabolic syndrome, insulin resistance, diabetic ketoacidosis, hyperosmolar hyperglycemic state, gestational diabetes, diabetic dermadrome, diabetic neuropathy, diabetic foot ulcer, maturity-onset diabetes of the young, pancreatogenic diabetes, or polycystic ovary syndrome (PCOS).

145. 145. The composition for use of claim 144, wherein the insulin-related disease, disorder, or condition is diabetes.

146. An injection comprising the composition of any one of claims 29 to 52.

147. A dietary supplement comprising the composition of any one of claims 29 to 52.

148. 121. A kit comprising the composition of any one of claims 29-52 or 120 for use in treating or preventing an insulin-related disease, disorder, or condition.

149. 149. The kit of claim 148, comprising a monitoring device.

150. 150. The kit of claim 149, wherein the monitoring device is a blood glucose monitor.

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