Intervention strategies for the prevention or treatment of diabetes, autoimmune diseases, inflammatory diseases, or cardiovascular diseases

Abstract

The present invention relates to an intervention strategy in the prevention or treatment of subjects with inflammation-related diseases, such as diabetes, autoimmune diseases, inflammatory diseases, or cardiovascular diseases. The intervention strategy involves the administration of preferably chloro-, fluoro-, or bromo-substituted tryptophan, preferably 6-bromotryptophan, and / or mono- or di-fatty acid substituted glycerolphosphocholine (GPC), preferably selected from the group consisting of 1-myristoyl-2-arachidonoyl-glycero-phosphocholine (MA-GPC) and 1-arachidonoyl-glycero-phosphocholine (A-GPC), or any derivative or functional equivalent thereof. Alternatively, the intervention involves administration of Desulfovibrio species, preferably selected from the group consisting of Desulfovibrio pigel, Desulfovibrio fairfieldensis, Desulfovibrio desulfuricans, Desulfovibrio indonensis, Desulfovibrio alaskensis, Desulfovibrio vulgaris, Desulfovibrio vietnamensis, and Desulfovibrio gigas.

Description

[Technical Field]

[0001] The present invention relates to the prevention and / or treatment of inflammation-related diseases selected from the group consisting of diabetes, autoimmune diseases, inflammatory diseases, and cardiovascular diseases, and more specifically to the use of a composition comprising a specific microorganism and / or its metabolic products in said prevention and / or treatment. [Background technology]

[0002] The role of inflammation in both type 1 and type 2 diabetes has led to increased interest in targeting inflammation to improve the prevention and treatment of this disease. Evidence has shown that inflammatory pathways are key mediators in the pathology of diabetes under the stimulation of risk factors, including obesity or overweight.

[0003] Furthermore, the relationship between diabetes and cardiovascular disease is well established, and the risk of cardiovascular disease is significantly higher in diabetic patients. Atherosclerosis represents the most common cause of coronary artery disease, and the characterization of this disease as a chronic, low-grade inflammatory state is now widely accepted.

[0004] Furthermore, inflammation is also a classic symptom of immune disorders. The disease may flare up, i.e., worsen, or may go into remission, i.e., symptoms improve or disappear. Treatment for autoimmune disorders varies depending on the type of disease, but in most cases, one important goal is to reduce inflammation.

[0005] Current therapeutic agents for diabetes, cardiovascular disease, autoimmune diseases, and inflammatory diseases have anti-inflammatory properties in addition to their primary mechanism of action. Non-pharmaceutical treatments, such as lifestyle interventions, reduce inflammatory status, as measured by, for example, circulating C-reactive protein (CRP) and interleukin-6 (IL-6) levels, and improve cardiovascular and all-cause mortality.

[0006] This makes inflammation-targeted therapeutic approaches an attractive area of ​​research.

Prior Technical Literature

Charter Documents

[0007] [Patent Document 1] WO2019168401

Non-licensed literature

[0008]

Non-patent document 1

Non-patent document 2

Non-patent document 3

Non-patent document 4

Non-patented document 5

Non-patent document 6

Non-patent document 7

Non-patent document 8

Non-patent document 9

Non-patent document 10

Non-patent document 11

Non-patent document 12

Non-patent document 13

Non-patent document 14

Non-patent document 15

Non-patent document 16

Non-patent document 17

Non-patent document 18

Non-patent document 19

Non-patent document 20

Non-patent document 21

Non-patent document 22

[0009] Novel treatment strategies are needed to improve the quality of life for patients with inflammation-related disorders, including diabetes, autoimmune diseases, inflammatory diseases, and cardiovascular diseases. There remains a need to develop new or improved prevention and / or treatment strategies. It is an object of the present disclosure to address this need. [Means for solving the problem]

[0010] The present inventors investigated whether administration of fecal transplants from allogeneic (healthy donors) or autologous (autologous) sources would have beneficial effects in patients with autoimmune disease. Confirming previous studies, it was found that administration of autologous fecal transplants can result in immune resetting in patients with autoimmune disease, thereby reducing the severity of the autoimmune disease.

[0011] The inventors have surprisingly found that this beneficial effect may result from specific components in fecal matter. These components with therapeutic use are most particularly: Bacteria of the genus Desulfovibrio, preferably selected from the group consisting of Desulfovibrio piger, Desulfovibrio fairfieldensis, Desulfovibrio desulfuricans, Desulfovibrio indonensis, Desulfovibrio alaskensis, Desulfovibrio vulgaris, Desulfovibrio vietnamensis, and Desulfovibrio gigas, most preferably Desulfovibrio piger, and The metabolites, 6-bromotryptophan, 1-myristoyl-2-arachidonoyl-glycero-phosphocholine (MA-GPC), and 1-arachidonoyl-glycero-phosphocholine (A-GPC), or more generally chloro-, fluoro-, or bromo-substituted tryptophan, or mono- or di-fatty acid substituted glycerol phosphocholines, or any derivative or functional equivalent thereof. It was found to be related to

[0012] Upon further investigation, it was surprisingly found that Desulfovibrio species and / or metabolites according to the present disclosure, particularly 6-bromotryptophan, have anti-inflammatory effects, i.e., reduce inflammatory conditions and inflammatory markers associated with inflammatory diseases such as type 1 and type 2 diabetes, autoimmune diseases, cardiovascular diseases, and systemic inflammatory response syndrome (SIRS) or sepsis.

[0013] Furthermore, an inverse correlation was found between plasma 6-bromotryptophan levels and the presence of type 2 diabetes, suggesting that Desulfovibrio species and / or metabolites according to the present disclosure, particularly 6-bromotryptophan, may contribute to the prevention of type 2 diabetes and the prevention or treatment of cardiovascular complications (microvascular and macrovascular). Furthermore, it was found that Desulfovibrio species and / or metabolites according to the present disclosure, particularly 6-bromotryptophan, may promote insulin secretion by beta cells.

[0014] Mechanistically, the inventors have determined that the biological effects of 6-BT appear to be distinct from those of tryptophan. While 6-BT does not act through activation of the AhR, it inhibits NFkB activation and enhances mitochondrial metabolism, the latter of which is typically used by cells with an anti-inflammatory phenotype. Due to its broad effects on many cell types, its inhibitory effect on NFkB signaling, and the resulting promotion of mitochondrial metabolism and fitness, 6-BT may be a highly useful therapeutic agent not only in the context of type 1 and type 2 diabetes, but also in many other inflammation-related disorders, such as sepsis, systemic inflammatory response syndrome (SIRS), and cardiovascular disease.

[0015] Furthermore, without being bound by any theory, the inventors believe that the Desulfovibrio species and the metabolites according to the present disclosure may modulate the immune system, for example, by resetting B cell clonal function and regulatory T cells, which may inhibit autoimmune responses.

[0016] During early life, the immune system is thought to be trained through continuous crosstalk with the developing gut microbiome composition. In this way, the gut microbiome plays a key role in modulating the development, composition, and function of adaptive immune cells (see, e.g., Agace and McCoy, Immunity 46, April 18, 2017). It is this process, among others, that results in a properly functioning system free of autoimmune factors.

[0017] However, the crosstalk between the immune system and the gut microbiome, or its end result, can be disrupted, which can lead to the production of autoimmune antibodies (by B cells) and the formation of autoreactive T cells. Treatments according to the present disclosure can overcome this disruption by re-establishing the crosstalk between the immune system and the gut microbiome (including its specific bacteria and / or derived products such as metabolites), resulting in the inhibition of the autoimmune response.

[0018] Thus, the use of the above-described Desulfovibrio species and / or metabolites in autoimmune diseases can halt autoimmune destruction of target tissues and re-establish immune tolerance. The present disclosure can achieve this by stimulating the immune system, wherein the Desulfovibrio species and / or metabolites are preferably administered to the duodenum (directly or indirectly, such as via oral administration). The present disclosure preferably does not aim to alter the gut microbiome, i.e., the composition of the gut microbial community.

[0019] WO2019168401 discloses the use of fecal material for the prevention and treatment of autoimmune diseases, wherein the fecal material is autologous to the subject and is preferably administered to the small intestine, preferably the duodenum, where it can initiate immune resetting and thereby reduce the severity of the autoimmune disease. However, the therapeutic efficacy of the method of WO2019168401 leaves room for improvement, and the procedure is tedious and difficult to scale up.

[0020] Thus, the present disclosure: - type 1 diabetes, - type 2 diabetes, cardiovascular diseases, in particular coronary artery disease (also known as coronary heart disease and ischemic heart disease), peripheral artery disease, cerebrovascular disease (e.g. stroke or TIA, i.e. transient ischemic attack), atherosclerosis, stenosis, renal artery stenosis, aortic disease, aortic aneurysm, cardiomyopathy, hypertensive heart disease, high blood pressure, heart failure, pulmonary heart disease, arrhythmias, cardiovascular inflammation, inflammatory heart disease, endocarditis, inflammatory cardiac hypertrophy, myocarditis, eosinophilic myocarditis, valvular heart disease, congenital heart disease or rheumatic heart disease, inflammatory diseases, in particular cardiovascular inflammation, such as carditis, endocarditis, myocarditis, pericarditis, vasculitis, arteritis, phlebitis or capillitis, or inflammation of the gastrointestinal tract, such as esophagitis, gastritis, gastroenteritis, enteritis, colitis, enterocolitis, duodenitis, ileitis, caecitis, appendicitis or proctitis (inflammatory diseases further include liver inflammation, lung inflammation, skeletal inflammation, systemic immune response syndrome (SIRS), sepsis), and - autoimmune diseases, especially (endocrine) autoimmune diseases (e.g. Hashimoto's hypothyroidism, Graves' hyperthyroidism, rheumatoid arthritis, celiac disease, asthma / COPD, Addison's disease, IBD (Crohn's disease and ulcerative colitis), systemic lupus erythematosus, vasculitis, Guillain-Pare and CIDP, multiple sclerosis, psoriasis (arthritis), vitiligo, and Bechtelew's disease The present invention is intended to prevent or treat an inflammation-related disease selected from the group consisting of:

[0021] Additionally or alternatively, uses according to the present disclosure may be for improving overall health and / or reducing inflammatory conditions.

[0022] That is, the present disclosure also encompasses the prevention of the cited diseases, i.e., type 1 or type 2 diabetes, cardiovascular disease, inflammatory disease, or autoimmune disease. Thus, for example, to avoid the onset of any of the diseases in a subject in whom risk markers associated with a pre-stage or early stage of the respective disease have been detected (prior to diagnosis of the respective disease), the Desulfovibrio species and / or metabolites according to the present disclosure can be administered to the subject. Such primary or secondary prevention strategies may prevent the progression of the disease.

[0023] Some of the autoimmune diseases mentioned herein are currently treated by immunotherapy, for example, by using antibodies against TNFα. However, these expensive immunotherapies are likely to be successful only in a small subset of patients, which has been attributed to deviations in the gut microbiome (Kolho et al., 2015 Am J Gastroenterol. 110(6):921-30). The inventors anticipate that treatment with TNFα antagonists or anti-TNFα may be synergistic with treatments according to the present invention, such as the administration of Desulfovibrio species and / or metabolites according to the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0024] The present disclosure provides: - Desulfovibrio species preferably selected from the group consisting of Desulfovibrio pigel, Desulfovibrio fairfieldensis, Desulfovibrio desulfuricans, Desulfovibrio indonensis, Desulfovibrio alaskensis, Desulfovibrio vulgaris, Desulfovibrio vietnamensis, and Desulfovibrio gigas, most preferably Desulfovibrio pigel, and - the compounds (metabolites), 6-bromotryptophan, 1-myristoyl-2-arachidonoyl-glycero-phosphocholine (MA-GPC) and 1-arachidonoyl-glycero-phosphocholine (A-GPC), or more generally chloro-, fluoro- or bromo-substituted tryptophans, or mono- or di-fatty acid substituted glycerol phosphocholines, or any derivative or functional equivalent thereof; by using one or more agents selected from the group consisting of: The present invention relates to the prevention or treatment of inflammation-related diseases selected from the group consisting of type 1 or type 2 diabetes, autoimmune diseases, cardiovascular diseases, and inflammatory diseases.

[0025] Thus, the present disclosure provides methods for the prevention or treatment of a subject in need thereof, particularly a subject having an inflammation-related disease, such as type 1 or type 2 diabetes, an autoimmune disease (such as an endocrine autoimmune disease), a cardiovascular disease, or an inflammatory disease, comprising administering one or more of the above-mentioned agents.

[0026] Compared to administration of autologous feces as described in WO2019168401, the method according to the present disclosure has been found to have improved therapeutic efficacy in autoimmune diseases, is less cumbersome, simple to administer, easy to manufacture, e.g., under certified Quality Management Systems (QMS) or Good Manufacturing Practice (GMP), and / or is easily scalable.

[0027] The present disclosure, at least a priori, is preferably not aimed at altering the gut microbiome, i.e. the microbial community composition of the gut or in particular the ileum.

[0028] In the context of the present disclosure, the autoimmune disease may be any autoimmune disease, including systemic and local (organ-specific) autoimmune diseases, in particular an autoimmune disease selected from the group consisting of endocrine autoimmune diseases (e.g., type 1 diabetes, Hashimoto's disease, Graves' disease, or Addison's disease), skin autoimmune diseases (e.g., psoriasis or vitiligo), rheumatic autoimmune diseases (e.g., rheumatoid arthritis, systemic lupus erythematosus, vasculitis, or Bechtelew's disease), and gastrointestinal autoimmune diseases (e.g., celiac disease, inflammatory bowel disease), neurological diseases (Guillan-Paré, CIDP, and multiple sclerosis), and pulmonary diseases (COPD / asthma).

[0029] In the context of the present disclosure, a cardiovascular disease may be any cardiovascular disease, such as coronary artery disease (also known as coronary heart disease and ischemic heart disease), peripheral vascular disease, cerebrovascular disease (e.g., stroke or TIA, i.e., transient ischemic attack), atherosclerosis, stenosis, renal artery stenosis, aortic disease, aortic aneurysm, cardiomyopathy, hypertensive heart disease, high blood pressure, heart failure, pulmonary heart disease, arrhythmias, cardiovascular inflammation, inflammatory heart disease, endocarditis, inflammatory cardiac hypertrophy, myocarditis, eosinophilic myocarditis, valvular heart disease, congenital heart disease, or rheumatic heart disease (optionally, any of these diseases may be excluded from the present application).

[0030] Furthermore, in the context of the present disclosure, an inflammatory disease may be any inflammatory disease, particularly cardiovascular inflammation, such as carditis, endocarditis, myocarditis, pericarditis, vasculitis, arteritis, phlebitis, or capillitis, or, for example, inflammation of the gastrointestinal tract, such as esophagitis, gastritis, gastroenteritis, enteritis, colitis, enterocolitis, duodenitis, ileitis, caecitis, appendicitis, or proctitis. Inflammatory diseases according to the present disclosure may further refer to liver inflammation, lung inflammation, or skeletal inflammation. Alternatively, inflammatory diseases may further refer to systemic immune response syndrome (SIRS) or sepsis (optionally, any of these diseases may be excluded from the present application).

[0031] diabetes type 1 diabetes Type 1 diabetes is a chronic endocrine autoimmune disease in which the pancreas produces little or no insulin. It is generally considered to be associated with progressive beta cell destruction and is associated with increased risk of morbidity and mortality compared to healthy subjects. Because beta cell function may also be impaired in type 2 diabetes, the present disclosure may also relate to the prevention and / or treatment of type 2 diabetes.

[0032] It has been found that the medicaments according to the present disclosure can be used to prevent and / or treat type 1 diabetes. Such treatment can also extend the honeymoon period of type 1 diabetes, i.e., the period after diagnosis during which the body's own pancreas still produces insulin in significant amounts sufficient to limit the need for exogenous insulin and maintain blood glucose control. Extending this period can dramatically improve the patient's quality of life. Application of treatment can also reduce the severity of symptoms of type 1 diabetes, such as symptoms or complications associated with impaired function of the eyes, kidneys, nerves, and / or brain.

[0033] More specifically, treatment may inhibit the decline of beta cell function and / or inhibit the production of autoantibodies associated with type 1 diabetes, such as pancreatic islet (beta) cell autoantibodies, autoantibodies against insulin, autoantibodies against GAD (GAD65), autoantibodies against tyrosine phosphatases IA-2 and IA-2β, and / or autoantibodies against zinc transporter 8 (ZnT8).

[0034] Symptoms of type 1 diabetes may include polyuria, polydipsia, polyphagia, weight loss, fatigue, nausea, and blurred vision. The onset of symptomatic disease may be rapid. In this context, it is not uncommon for patients with type 1 diabetes to suffer from diabetic ketoacidosis (DKA). The following diagnostic criteria can be applied to type 1 and type 2 diabetes (American Diabetes Association, ADA): - Fasting plasma glucose (FPG) level ≥ 126 mg / dL (7.0 mmol / L), or - A 2-hour plasma glucose level of 200 mg / dL (11.1 mmol / L) or greater during a 75 g oral glucose tolerance test (OGTT), or - Random plasma glucose ≥ 200 mg / dL (11.1 mmol / L) in patients with classic symptoms of hyperglycemia or hyperglycemic crisis.

[0035] Additionally and / or alternatively, C-peptide responses can be assessed after a mixed meal test as described in the Examples and / or as described in Lachin et al. (2011 PLoS ONE Vol. 6(11) e26471).

[0036] Type 1 diabetes and / or its precursors can be confirmed by the presence of one or more autoimmune markers, including pancreatic islet (beta) cell autoantibodies, autoantibodies to insulin, autoantibodies to GAD (GAD65), autoantibodies to tyrosine phosphatases IA-2 and IA-2β, and / or autoantibodies to zinc transporter 8 (ZnT8), as well as increased HbA1c and altered glucose tolerance.

[0037] type 2 diabetes Type 2 diabetes is a common metabolic condition that develops when the body does not produce enough insulin or when insulin does not work properly (called insulin resistance). Insulin is a hormone that stimulates cells to take up glucose from the blood to use for energy. In the case of diabetes, cells are not instructed by insulin to take up glucose from the blood, meaning blood sugar levels rise (called hyperglycemia).

[0038] People usually develop type 2 diabetes after the age of 40, but people of South Asian origin are at increased risk for the condition, which can develop after the age of 25. The condition is also becoming more common among children and young people across all population groups. Type 2 diabetes often develops as a result of overweight, obesity, and lack of physical activity, and as these problems become more widespread, the prevalence of diabetes is increasing worldwide. Type 2 diabetes accounts for nearly 90% of all diabetes cases (the other form is type 1 diabetes), and treatment approaches include lifestyle changes and the use of medications.

[0039] It has been found that the medicaments according to the present disclosure can be used to prevent and / or treat type 2 diabetes. Treatment can be administered to reduce the severity of symptoms or complications associated with type 2 diabetes, such as polyuria and polydipsia. More specifically, treatment may reduce the need for exogenous hormone replacement.

[0040] cardiovascular disease Coronary artery disease Coronary artery disease is the most common form of cardiovascular disease. It involves reduced blood flow to the heart muscle due to the buildup of plaque in the arteries of the heart (atherosclerosis). Common symptoms are chest pain or discomfort that may extend to the shoulders, arms, back, neck, or jaw. Often, the first sign is a heart attack. Other complications include heart failure or abnormal heart rhythms.

[0041] Risk factors include high blood pressure, smoking, diabetes, lack of exercise, obesity, high blood cholesterol, poor diet, depression, and excessive alcohol consumption. Several tests can aid in diagnosis, including an electrocardiogram, cardiac stress test, coronary computed tomography angiography, and coronary angiography, among others.

[0042] Inflammatory disorders Subjects suffering from or at risk of developing an inflammatory disorder can be identified by methods known in the art, such as by visual examination of tissue or detection of inflammation associated with tissue or blood. Symptoms of inflammation include pain, redness, and swelling of the affected tissue.

[0043] Systemic inflammatory response syndrome (SIRS) and sepsis Systemic inflammatory response syndrome (SIRS) is the body's excessive defensive response to, for example, infection, trauma, surgery, acute inflammation, ischemia or reperfusion, or malignant tumors. It involves the release of acute phase reactants, which are direct mediators of a wide range of autonomic, endocrine, hematologic, and immune system changes in the subject. Although defensive in nature, an unregulated cytokine storm can trigger a widespread inflammatory cascade that can result in reversible or irreversible end-organ dysfunction and even death. SIRS with a suspected infectious source is termed sepsis. Sepsis with one or more end-organ failure is called severe sepsis, and sepsis with hemodynamic instability despite intravascular volume repletion is called septic shock.

[0044] SIRS can be diagnosed by meeting any two of the following criteria: - a body temperature above 38°C or below 36°C; - a heart rate greater than 90 beats per minute, - respiratory rate greater than 20 breaths / min or CO2 partial pressure less than 32 mmHg; - More than 12000 / μl or less than 4000 / μl white blood cells or more than 10% immature forms or bands.

[0045] Additionally or alternatively, the use of the present disclosure may be for improving overall health and / or reducing an inflammatory condition, preferably as measured by a higher erythrocyte sedimentation rate (ESR) compared to a healthy individual (e.g., an ESR value of at least 35, 40, 45, 50, 55, or 60 mm / hour) and / or a reduced level of C-reactive protein in the blood (plasma) compared to when the composition of the present disclosure is not administered. C-reactive protein may be measured, for example, 1 to 12, 1 to 4, 2 to 8, 4 to 12 weeks, or 1 to 12 months or 1 to 12 years after administration of the composition of the present disclosure. C-reactive protein (CRP) is a protein produced by the liver. The presence of an inflammatory condition anywhere in the body increases CRP levels in the blood. A CRP test measures the amount of CRP in the blood to detect an inflammatory condition.

[0046] Additionally or alternatively, the use according to the present disclosure is for inducing weight loss or reducing body mass index (BMI) in a subject.

[0047] autoimmune disease Autoimmune diseases are a class of diseases in which the immune system responds inappropriately to the subject's own cells, tissues, and / or organs. This can lead to inflammation, damage, and loss of function. Common autoimmune diseases include Hashimoto's hypothyroidism, Graves' hyperthyroidism, rheumatoid arthritis, celiac disease, asthma / COPD, Addison's disease, IBD (Crohn's disease and ulcerative colitis), systemic lupus erythematosus, vasculitis, Guillan-Paré and chronic inflammatory demyelinating polyneuropathy (CIDP), multiple sclerosis, psoriasis (arthritis), vitiligo, type 1 diabetes, and Bechtelew's disease.

[0048] The causes of autoimmune diseases are unknown. However, factors such as infection and genetic predisposition may play a role in triggering autoimmune diseases. Autoimmune diseases are usually diagnosed using a combination of medical history and blood tests (which detect autoantibodies or markers of inflammation or organ function, among other things).

[0049] Although a wide range of treatment options exist depending on the stage and type of autoimmune disease, there is no definitive cure for autoimmune diseases.

[0050] Treatment strategies are generally directed at alleviating symptoms, minimizing organ or tissue damage, and preserving organ function. For example, treatment strategies may include replacing organ function (e.g., insulin for type 1 diabetes or thyroxine for Hashimoto's hypothyroidism), nonsteroidal anti-inflammatory drugs (NSAIDS), corticosteroid anti-inflammatory drugs (e.g., prednisolone), TNFα inhibitors, immunosuppressants, or immunoglobulin replacement therapy.

[0051] endocrine autoimmune diseases Among various autoimmune diseases, autoimmune endocrine disorders are the most common. The endocrine system includes glands that produce hormones and release them directly into the circulatory system, as well as feedback loops that achieve homeostasis. Organs of the endocrine system can be affected by several autoimmune diseases, characterized by varying degrees of impact and severity. Multiple organs may be involved, such as in polyglandular autoimmune syndromes.

[0052] Among the various autoimmune endocrine diseases, type 1 diabetes, Hashimoto's disease, Graves' disease, and Addison's disease are particularly frequent in clinical practice.

[0053] Hashimoto's disease Hashimoto's thyroiditis, also known as Hashimoto's hypothyroidism or chronic lymphocytic thyroiditis, is the most common organ-specific autoimmune disorder. It is also known as Hashimoto's hypothyroidism or chronic lymphocytic thyroiditis and is considered an autoimmune disease that gradually destroys the thyroid gland. The cause of Hashimoto's thyroiditis remains unclear, but it is generally believed to involve an inappropriate cell-mediated immune response and the production of autoantibodies against the thyroid gland. Within the mononuclear lymphoid infiltrate, both B (CD20+ and CD79alpha+) cells (which destroy thyroid follicles and thyroid cells) and overstimulated CD4+ T cells are found. (T helper type 2, or Th2, cells lead to overstimulation and the production of B cells that produce antibodies against thyroid antigens, subsequently promoting thyroiditis in the thyroid gland.) (Marazuela et al., J Clin Endocrinol Metab., September 2006;91(9):3639-46).

[0054] An enlarged thyroid gland is typically the only symptom until hypothyroidism becomes apparent, but the disease can progress to hypothyroidism, which often results in symptoms including edema, weight gain, and easy fatigue (fatality), cold sensitivity, and diarrhea, as well as physical findings such as dry skin, hoarseness, bradycardia, and / or a prolonged relaxation phase of the Achilles tendon reflex.

[0055] Hashimoto's disease can be confirmed by the presence of anti-thyroid peroxidase (TPO) and anti-thyroglobulin (Tg) antibodies in the patient's serum. Furthermore, elevated thyroid-stimulating hormone (TSH) levels and decreased free T4 (FT4) levels, decreased free T3 levels, and / or elevated anti-microsomal antibody levels compared to the average in healthy individuals can aid in obtaining a positive diagnosis.

[0056] Hashimoto's disease is currently treated with thyroid hormone replacement medications, such as levothyroxine (FT4 replacement), triiodothyronine (T3 replacement), or desiccated thyroid extract. The inventors have discovered that agents according to the present disclosure, optionally in combination with the above thyroid hormone replacement medications, can be used to prevent and / or treat Hashimoto's disease. Treatment according to the present disclosure can also be applied to reduce the severity of symptoms of Hashimoto's disease, such as one or more of the symptoms or complications described above.

[0057] Graves' disease Graves' disease is an autoimmune disorder affecting the thyroid gland and is the most common cause of hyperthyroidism. The disease can be characterized by the presence of autoantibodies in the serum that bind to the thyrotropin receptor, i.e., the thyroid-stimulating hormone (TSH) receptor. These anti-TSH receptor antibodies (TBII) can overstimulate the thyroid gland, resulting in goiter and signs of hyperthyroidism, as well as involvement of the eye muscles (Graves' eye disease) in a small group of patients.

[0058] Symptoms include hyperthyroidism, goiter, and orbital disease. Other major symptoms include weight loss (with increased appetite), fatigue, shortness of breath, hyperhidrosis, hand tremors, diarrhea, periodic paralysis (in men), and muscle weakness. With Graves' eye disease, patients may suffer from bulging eyes, blurred vision, and dry / red eyes (which may lead to blindness in rare cases). Two signs, exophthalmos and pretibial myxedema, are truly specific for Graves' disease and are not seen in other hyperthyroid states.

[0059] Graves' disease can be identified by low (sometimes undetectable) serum TSH levels and / or elevated free T3 and free T4 levels compared to healthy individuals. Patients may typically test positive for anti-TSH receptor antibodies (TBII) in their serum.

[0060] Current treatments for Graves' disease may include the administration of antithyroid drugs (block and replacement therapy), the administration of radioactive iodine (radioactive iodine I-131), and / or thyroidectomy (surgical removal of the thyroid gland). Strumazole and methimazole (PTU) are usually prescribed, followed by thyroid hormone replacement medications such as levothyroxine (FT4 replacement), triiodothyronine (T3 replacement), or desiccated thyroid extract.

[0061] Alternatively, or in combination with the above treatments, the inventors have found that medicaments according to the present disclosure can be used for the prevention and / or treatment of Graves' disease, including eye diseases. Treatments according to the present disclosure can also be applied to reduce the severity of symptoms of Graves' disease, such as one or more of the symptoms or complications described above.

[0062] Addison's disease Addison's disease is a chronic endocrine autoimmune disorder in which the adrenal glands fail to produce sufficient steroid hormones. The disease is caused by destruction of the adrenal glands (hormones produced in both the cortex and medulla). The disease may be a manifestation of a polyglandular autoimmune syndrome that includes complications from other organ-specific autoimmune disorders (e.g., type 1 diabetes, Hashimoto's disease, vitiligo).

[0063] Increased pigmentation due to increased secretion of ACTH is the characteristic clinical sign of Graves' disease. Other symptoms include abdominal pain in the stomach region, orthostatic hypotension, and weight loss.

[0064] Medical testing will typically determine whether orthostasis, hypoglycemia, hyponatremia, hyperkalemia, and peripheral blood eosinophilia are present. To confirm Addison's disease, demonstration of low adrenal hormone levels even after stimulation with the synthetic pituitary ACTH hormone tetracosactide (called the ACTH stimulation test or synacthen test) is commonly performed for diagnosis.

[0065] Treatment generally involves replacement therapy with oral hydrocortisone and / or an electrocorticoid such as fludrocortisone (if the adrenal medulla is also involved). The inventors have discovered that medicaments according to the present disclosure can be used to prevent and / or treat Addison's disease, optionally in addition to treatment with hydrocortisone. Treatment according to the present disclosure can also be applied to reduce the severity of symptoms of Addison's disease, such as one or more of the symptoms or complications described above.

[0066] skin autoimmune diseases Psoriasis (arthritis) Psoriasis is a chronic autoimmune disease that causes the rapid production of skin cells. The underlying etiology is that T cells attack healthy skin cells, which overdrives the skin cell production process. The new cells are pushed to the surface of the skin and accumulate there. This results in skin plaques and red, inflamed areas, which are most commonly associated with psoriasis. Subtypes of psoriasis include: (1) Plaque psoriasis, the most common type of psoriasis. It is characterized by inflamed, red patches covering areas of skin, typically the elbows, knees, and scalp. These patches are often covered by silvery-white scales or plaques. (2) Guttate psoriasis, a form of psoriasis common in children, typically causes small pink spots on the trunk, arms, and legs. (3) Pustular psoriasis, which is a more common form of psoriasis in adults and causes white pustules and areas of inflamed red skin, typically on the hands or feet. (4) Inverse psoriasis, which causes light areas of inflamed, red, shiny skin. The patches of inverse psoriasis typically develop in the armpits or chest, groin, or around skin folds. (5) Erythrodermic psoriasis: This is a severe and rare form of psoriasis. This form often covers large areas of the body, and the skin may appear sunburned. People with this form of psoriasis may develop a fever or become very ill, and this form of psoriasis can be fatal. (6) Psoriatic arthritis involving the joints.

[0067] Symptoms of psoriasis vary among patients. Common symptoms include red patches of skin covered with thick, silvery scales, small scale dots (commonly seen in children), dry, cracked skin that may bleed, itching, burning or soreness, thickened, pitted, or raised nails, and / or swollen and stiff joints. Most types of psoriasis may go through cycles of flare-ups lasting weeks or even months, followed by periods of remission or even remission. Psoriatic arthritis (also known as psoriatic arthritis) is a condition in which swollen, sore joints due to arthritis occur together with psoriasis.

[0068] For mild disease involving only small areas of the body, topical treatments (applied to the skin) such as creams, lotions, and sprays are commonly prescribed. Sometimes, local injections of steroids directly into stubborn or resistant, isolated psoriatic plaques can be helpful.

[0069] Tumor necrosis factor (TNF) antagonists (or anti-TNFα therapy) have become first-line agents in the treatment of mild to severe psoriasis or psoriatic arthritis. Examples include infliximab, etanercept, and adalimumab. Anti-TNFα therapy has been found to be effective in treating both psoriasis and psoriatic arthritis and may also reduce the risk of cardiovascular events. The inventors have discovered that agents according to the present disclosure can also or alternatively be used to prevent and / or treat psoriasis and / or psoriatic arthritis. Furthermore, treatment according to the present disclosure can also be applied to reduce the severity of symptoms of psoriasis and psoriatic arthritis, such as one or more of the symptoms or complications described above. In particular, combined treatment of a TNF antagonist or anti-TNFα with a treatment according to the present disclosure may be synergistic.

[0070] vitiligo Vitiligo is a condition in which white patches of skin appear on various parts of the body. It is generally thought to be caused by an autoimmune process that destroys melanocytes, the cells that produce pigment (color) in the skin. Vitiligo can also occur on mucous membranes (such as the inside of the mouth and nose) and in the eyes.

[0071] Recent studies have revealed dysbiosis in the diversity of microbial community structure in the skin microbiome of subjects with vitiligo. While distinct, specific microbiome signatures predominate across vitiligo-specific microbial groups, a clear decrease in taxonomic richness and evenness can be noted in lesional vitiligo (Ganju et al., Sci Rep., 2016 Jan;13;6:18761).

[0072] The white patches of vitiligo are more common in areas of the skin that are regularly exposed to sunlight. The patches can be on the hands, feet, arms, face, and lips, but sometimes also in the armpits and groin, around the mouth, eyes, nostrils, navel, genitals, and rectum. In addition, people with vitiligo often experience early graying of their hair (e.g., before age 35).

[0073] Ultraviolet (UV) light can be used for diagnosis and to assess the effectiveness of UV treatment, especially in the early stages of vitiligo. Skin with vitiligo typically emits blue light when exposed to UV light. In contrast, healthy skin shows no reaction.

[0074] Vitiligo can be classified into segmental vitiligo (SV) and non-segmental vitiligo (NSV), with NSV being the most common type of vitiligo.

[0075] In non-segmental vitiligo (NSV), the depigmented patches are typically symmetrical in location. In extreme cases, almost no pigmented skin remains, which is called generalized vitiligo. NSV can begin at any age, but segmental vitiligo is much more prevalent in the teenage years.

[0076] Segmental vitiligo (SV) tends to affect areas of skin attached to the dorsal roots of the spinal cord and is most often unilateral. It is stable / static throughout its course. SV typically does not improve with UV phototherapy, but surgical treatments such as cell transplants can be effective.

[0077] While there is no definitive cure for vitiligo, several treatment options are available, including ultraviolet light and / or creams. Topical preparations (i.e., creams) of immunosuppressive medications, including corticosteroids or glucocorticoids (such as clobetasol and / or betamethasone) and calcineurin inhibitors (such as tacrolimus and / or pimecrolimus), are considered first-line vitiligo treatments, while UV(B) therapy is considered second-line treatment for vitiligo.

[0078] The inventors have found that, in addition to or instead of the above treatments, the medicaments according to the present disclosure can be used to prevent and / or treat vitiligo. Furthermore, the treatments according to the present disclosure can also be applied to reduce the severity of vitiligo symptoms, such as one or more of the symptoms or complications described above.

[0079] Rheumatic disorders Rheumatoid arthritis Rheumatoid arthritis (RA) can be seen as an autoimmune disease in which the immune system attacks the joints, resulting in inflammation that causes thickening of the tissue lining the joints (the synovium), resulting in joint pain.

[0080] If left untreated, RA can damage cartilage, the elastic tissue that covers the ends of bones in joints, and even the bones themselves. Eventually, cartilage loss occurs, and the joints become loose, unstable, painful, lose their mobility, and can even become deformed. Unfortunately, joint damage is generally irreversible, so early diagnosis and treatment are recommended to control RA.

[0081] RA most commonly affects the joints of the hands, feet, wrists, elbows, knees, and ankles. RA can also affect body systems such as the cardiovascular or respiratory systems, in which case it is called systemic RA. In the early stages, people with RA may experience tenderness and pain in the joints.

[0082] Symptoms of RA typically include six weeks or more of stiffness and joint pain, particularly in the small joints (wrists, hands, and certain joints of the feet). Along with the pain, many people may also experience fatigue, loss of appetite, and a mild fever.

[0083] While no single test can definitively confirm RA, blood tests can be performed to measure inflammation levels and look for biomarkers, such as antibodies, associated with RA.

[0084] High erythrocyte sedimentation rate and high C-reactive protein (CRP) levels compared to healthy individuals are biomarkers of inflammation. Although high ESR or high CRP are not specific for RA, when combined with the presence of RA-associated antibodies, they can confirm the diagnosis of RA.

[0085] Rheumatoid factor (RF) is an antibody found in the majority of people with RA. RF can also occur in other inflammatory diseases, so it is not a definitive sign of having RA. However, a different antibody, anti-cyclic citrullinated peptide (anti-CCP), occurs primarily in RA patients. This makes a positive anti-CCP test a stronger indicator of RA. Additionally, x-rays, ultrasound, or magnetic resonance imaging scans can be performed to look for joint damage, such as erosions and joint space narrowing.

[0086] For treatment, nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly prescribed, which can relieve pain and inflammation caused by arthritis. Examples of NSAIDs include ibuprofen, ketoprofen, and naproxen sodium. Additionally, corticosteroids, including prednisone, prednisolone, and methiprednisolone, can be administered as anti-inflammatory medications.

[0087] DMARDs, or disease-modifying antirheumatic drugs, may be used to slow disease progression. DMARDs include methotrexate, hydroxychloroquine, sulfasalazine, leflunomide, cyclophosphamide, and azathioprine. A subcategory of DMARDs is known as "JAK inhibitors," which block the Janus kinase pathway, or JAK pathway. An example is tofacitinib.

[0088] Biologics act more rapidly than traditional DMARDs and are given by injection or infusion. In many people with RA, biologics can slow, modify, or even halt the disease. Particularly preferred are tumor necrosis factor (TNF) antagonists (anti-TNFα therapy).

[0089] The inventors have found that in addition to or instead of the above treatments, agents according to the present disclosure can be used to prevent and / or treat rheumatoid arthritis and / or one or more of its symptoms as described above. Combination treatments according to the present disclosure with TNF antagonists or anti-TNFα may be synergistic.

[0090] Bechtelew's disease Bechtelew's disease (or ankylosing spondylitis) is a chronic autoimmune rheumatic disorder that specifically involves the axial skeleton. It typically presents in male adults between the ages of 20 and 30.

[0091] The most serious symptom is pain in the neck and lower back. Typical symptoms are nighttime pain and inflammation of the sacroiliac joints. In some patients, bone deformities of the spine can occur, which leads to limited movement. Apart from these spinal pains, inflammation of the peripheral joints is common.

[0092] To diagnose Bechtelew's disease, a spinal examination is performed to evaluate the mobility of the cervical and lumbar spine. A Schober's test can help estimate the amount of restriction of lumbar forward bending. The diagnosis can be confirmed by the detection of the HLA-B27 antigen in the patient's blood.

[0093] Treatment options include the administration of NSAIDs, sulfasalazine, methotrexate, leflunomide, corticosteroids, and TNFα inhibitors. The inventors have found that, in addition to or instead of the above treatments, medicaments according to the present disclosure can be used to prevent and / or treat Bechtelew's disease and / or one or more of its symptoms, as described above. In particular, the combination of a TNF antagonist or anti-TNFα with a treatment according to the present disclosure may be synergistic.

[0094] Systemic lupus erythematosus Systemic lupus erythematosus (SLE), also known simply as lupus, is an autoimmune disease in which the body's immune system mistakenly attacks healthy tissue throughout the body. Symptoms vary from person to person and can range from mild to severe. SLE significantly increases the risk of cardiovascular disease, making it the most common cause of death. With modern treatment, approximately 80% of affected individuals survive more than 15 years after diagnosis. Common symptoms include joint pain and swelling, fever, chest pain, hair loss, mouth ulcers, swollen lymph nodes, fatigue, and a red rash, most commonly on the face. Periods of illness called flares and periods of remission with few symptoms are common. There is no cure for SLE. Treatments may include NSAIDs, corticosteroids, immunosuppressants, hydroxychloroquine, and methotrexate. Corticosteroids are effective quickly, but long-term use can cause side effects. The inventors have found that in addition to or instead of the above treatments, medicaments according to the present disclosure can be used to prevent and / or treat SLE disease and / or one or more of its symptoms as described above.

[0095] vasculitis Vasculitides range from inflamed large vessels to small vessels. Large-vessel vasculitic diseases include giant cell arteritis (or temporal arteritis) and Takayasu's disease (Takayasu's arteritis). Medium- to large-vessel vasculitic diseases include polyarteritis nodosa (PAN) and Kawasaki disease. Small-vessel vasculitic diseases include microscopic polyangiitis, GPA (granulomatosis with polyangiitis, also known as Wegener's disease), EGPA (eosinophilic granulomatosis with polyangiitis, also known as Churg-Strauss syndrome), Henoch-Schönlein syndrome, anti-GBM (Goodpasture's syndrome), and cryoglobulinemia-associated vasculitis. The inventors have discovered that the agents disclosed herein can be used to prevent and / or treat vasculitides and / or one or more of their symptoms, as described above.

[0096] Gastrointestinal tract autoimmune diseases Celiac disease Celiac disease (also known as coeliac disease) is an autoimmune disorder in which gluten ingestion leads to damage to small intestinal epithelial cells. It typically occurs in combination with type 1 diabetes in genetically predisposed people. Celiac disease and type 1 diabetes have similar etiologies, and inherited genetic factors as well as dietary and microbial exposures may play a role, particularly in early life (see, e.g., Verdu and Danska, Nature Immunology, Vol. 19, July 2018, pp. 685-695).

[0097] When people with celiac disease eat gluten (a protein found in wheat, rye, and barley), their bodies mount an immune response that attacks the small intestine, resulting in damage to the villi (tiny finger-like protrusions that line the small intestine). When the villi are damaged, nutrients cannot be properly absorbed by the intestine. Symptoms include abdominal cramps, malnutrition, and osteoporosis.

[0098] There are several serum (blood) tests available to screen for celiac disease antibodies, but the most commonly used is the tTG-IgA test. For this test to be successful, the patient must be consuming gluten. Furthermore, a diagnosis of celiac disease can be reached by endoscopic biopsy. The biopsy involves obtaining a sample of the small intestine, which can then be analyzed to see if there is any damage consistent with celiac disease. If there is improvement on a gluten-free diet, the diagnosis can be confirmed.

[0099] Currently, the only treatment for celiac disease is a strictly gluten-free diet. People who live without gluten must avoid foods containing wheat, rye, and barley, such as bread and beer. Even ingesting small amounts of gluten can cause damage to the small intestine. The inventors have found that in addition to or instead of the above treatments, the medicament according to the present disclosure can be used to prevent and / or treat celiac disease and / or one or more of its symptoms as described above.

[0100] inflammatory bowel disease Inflammatory bowel disease (IBD) is a term for two conditions (Crohn's disease and ulcerative colitis) characterized by chronic inflammation of the gastrointestinal (GI) tract. IBD is thought to be caused by a dysregulated immune response. Symptoms of IBD include persistent diarrhea, abdominal pain, rectal bleeding / bloody stools, weight loss, and fatigue. In IBD, the immune system responds improperly to environmental triggers, which causes inflammation of the GI tract. There also appears to be a genetic component, and individuals with a family history of IBD are more likely to develop this inappropriate immune response.

[0101] IBD is diagnosed using a combination of endoscopy (for Crohn's disease) or colonoscopy (for ulcerative colitis) and imaging studies such as contrast-enhanced X-ray, magnetic resonance imaging (MRI), or computed tomography (CT).

[0102] Several types of medications can be used to treat IBD, including aminosalicylates, corticosteroids (e.g., prednisone), immunomodulators, and the newest class of "biologics" approved for IBD, such as anti-TNF alpha. Several vaccinations are recommended for IBD patients to prevent infection. Severe IBD can require surgery to remove damaged sections of the gastrointestinal tract, but advances in pharmaceutical treatments mean that surgery is less common than it was decades ago. The inventors have discovered that, in addition to or instead of the above treatments, medicaments according to the present disclosure can be used to prevent and / or treat IBD and / or reduce the severity of one or more of its symptoms, as described above.

[0103] Neurological disorders Guillain-Barré Guillain-Barré syndrome (GBS) is a rapid-onset muscle weakness (acute polyneuropathy) caused by the immune system damaging the peripheral nervous system. Initial symptoms are typically altered sensation or pain accompanied by muscle weakness, beginning in the feet and hands and often spreading to the arms and upper body. Both sides are affected. Symptoms may develop over several hours and may last for several weeks. During the acute phase, the disorder can be fatal, and approximately 15% of people may develop respiratory muscle weakness and therefore require mechanical ventilation.

[0104] Although the cause is unknown, the underlying mechanism involves an autoimmune disorder in which the body's immune system mistakenly attacks peripheral nerves, damaging their myelin insulation. This immune dysfunction can be triggered by infection or, less commonly, by surgery or, in rare cases, by vaccination. Diagnosis is usually based on signs and symptoms, ruling out alternative causes, and is supported by tests such as nerve conduction studies and cerebrospinal fluid tests. Several subtypes exist based on the area of ​​weakness, the results of nerve conduction studies, and the presence of certain antibodies. Treatment with intravenous immunoglobulin or plasmapheresis, along with supportive care, results in a good recovery in the majority of people. Recovery can take weeks to years, and approximately one-third experience some permanent weakness. The inventors have discovered that, in addition to or instead of the above-mentioned treatments, the medicaments disclosed herein can be used to prevent and / or treat GBS and / or reduce the severity of one or more of its symptoms, as described above.

[0105] CDIP Chronic inflammatory demyelinating polyneuropathy (CIDP) is an acquired immune-mediated inflammatory disorder of the peripheral nervous system. This disorder is sometimes referred to as chronic polyneuropathy (CRP) or chronic inflammatory demyelinating polyradiculoneuropathy (because it involves the nerve roots). CIDP is closely related to Guillain-Barré syndrome and is considered the chronic counterpart of this acute disease. The inventors have discovered that the medicaments of the present disclosure can be used to prevent and / or treat CDIP and / or reduce the severity of one or more of its symptoms, as described above.

[0106] Multiple sclerosis Multiple sclerosis (MS) is a demyelinating disease in which the insulating covering of nerve cells in the brain and spinal cord is damaged. This damage disrupts the ability of parts of the nervous system to transmit signals, resulting in a variety of signs and symptoms, including physical, psychological, and sometimes spiritual problems. Specific symptoms can include double vision, blindness in one eye, muscle weakness, and impaired sensation or coordination. MS comes in several forms, and new symptoms occur in isolated attacks (relapsing forms) or accumulate over time (progressive forms). Symptoms may disappear completely between attacks, but permanent neurological damage often remains, especially with disease progression. The cause is unclear, but the underlying mechanism is thought to be immune system destruction or a failure of myelin-producing cells. Proposed causes include genetic factors and environmental factors, such as triggers by viral infections. MS is usually diagnosed based on the presenting signs and symptoms and the results of supporting medical tests. There is no known cure for multiple sclerosis. Treatment attempts to improve function after an attack and prevent new attacks. The inventors have found that medicaments according to the present disclosure can be used to prevent and / or treat MS as described above and / or reduce the severity of one or more of its symptoms.

[0107] Asthma and COPD In the context of the present disclosure, the prevention and / or treatment of asthma is also envisaged, in view of the autoimmune mechanisms that are also thought to be at play in asthma.

[0108] Asthma is a common chronic inflammatory disease of the lung airways. It can be characterized by reversible airway obstruction and bronchospasm. Symptoms include coughing, wheezing, chest tightness, and shortness of breath.

[0109] Currently, there is no definitive diagnostic test for asthma, and diagnosis is typically based on the pattern of symptoms and response to treatment over time. A diagnosis of asthma can be made if there is a history of recurrent wheezing, cough, or difficulty breathing, and these symptoms are caused or worsened by exercise, viral infections, allergens, and / or air pollution. FEV1 testing with bronchodilators is also performed to examine the effect on lung function.

[0110] Effective treatment for asthma involves identifying triggers of the disease, such as cigarette smoke, pets, or aspirin, and eliminating exposure to these triggers. In addition, bronchodilators are often recommended. In cases of mild but persistent disease, low-dose inhaled corticosteroids, or leukotriene antagonists or mast cell stabilizers can be used. For patients with severe asthma, i.e., daily attacks, higher doses of inhaled corticosteroids can be used.

[0111] The inventors have found that in addition to or instead of the above treatments, medicaments according to the present disclosure can be used to prevent and / or treat asthma and / or one or more of its symptoms as described above.

[0112] The efficacy of treatment according to the present disclosure confirms the link between the composition of the gut microbiome and the risk of developing asthma asserted by Korpela et al. (Nat Commun 2016 Jan 26, 7:10410).

[0113] Emphysema is one of the diseases that comprise COPD (chronic obstructive pulmonary disease). Emphysema involves the progressive damage of lung tissue, specifically the thinning and destruction of alveoli or air sacs. The medicament according to the present disclosure can be used to prevent and / or treat COPD, specifically emphysema, and / or one or more of its symptoms, as described above.

[0114] Other conditions The present disclosure may also be used in connection with the prevention and / or treatment of other autoimmune diseases, including, inter alia, autoimmune hepatitis, type 1a and / or type 1b diabetes, autoimmune polyglandular syndrome, myasthenia gravis, pernicious anemia, primary biliary cirrhosis, sclerosing cholangitis, antiphospholipid syndrome, dermatomyositis, mixed connective tissue disease, polymyalgia rheumatica, polymyositis, scleroderma, and Sjogren's syndrome, although it is anticipated that any of the above diseases may be excluded from the present disclosure.

[0115] Additionally, medicaments according to the present disclosure may be used to prevent and / or treat allergies, also known as allergic diseases, which are conditions caused by hypersensitivity of the immune system to typically harmless substances in the environment. Common allergies include hay fever (plant pollen allergy) and food allergies (e.g., associated with milk, soy, egg, wheat, peanuts, tree nuts, fish, and / or shellfish).

[0116] The present disclosure also enables the prevention and / or treatment of the following diseases, although these diseases are optionally excluded from the scope of the present disclosure: gastrointestinal disorders, Clostridium difficile infection, Crohn's disease, ulcerative colitis or inflammatory bowel disease (IBD), and / or irritable bowel syndrome (IBS). Alternatively and / or in addition, any of the following diseases may be excluded from the present disclosure: systemic and local (organ-specific) autoimmune diseases, endocrine autoimmune diseases, type 1 diabetes, type 2 diabetes, Hashimoto's disease, Graves' disease, or Addison's disease, skin autoimmune diseases, psoriasis or vitiligo, rheumatoid autoimmune diseases, rheumatoid arthritis, Bechteleu's disease and gastrointestinal autoimmune diseases, celiac disease, vasculitis, COPD, CIDP, MS, SLE, Guillain-Pare's disease. Diseases according to the present invention do not have to be inflammation-related.

[0117] Treatment according to the present disclosure The agent for use in the prevention or treatment of an autoimmune disease as described herein may be a Desulfovibrio species, preferably Desulfovibrio pigelii (ATCC 29098), Desulfovibrio fairfieldensis (ATCC 700045), Desulfovibrio desulfuricans (Essex 6 ATCC 29577), D. desulfuricans (MB ATCC 27774), Desulfovibrio indonensis (NCIMB 13468), Desulfovibrio alaskensis (NCIMB 13491), Desulfovibrio vietnamensis (DSM 10520), Desulfovibrio gigas (DSM 1382), Desulfovibrio intestinalis (DSM 10520), Desulfovibrio spp. (ATCC 10520), Desulfovibrio spp. (ATCC 10520), Desulfovibrio gigas (ATCC 10520), Desulfovibrio spp. ... 11275), Desulfovibrio longreachensis (ACM 3958), Desulfovibrio termitidis (DSM 5308), Desulfovibrio vulgaris subsp. vulgaris (DSM 644), and Desulfovibrio vulgaris subsp. oxamicus (DSM 1925). Additionally or alternatively, the agent may be a Bacteroides species, preferably Bacteroides stercoris, or a relative thereof, such as a relative having at least 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.5, 99.9% sequence identity with the 16S rDNA sequence of a type strain of Bacteroides stercoris.

[0118] Most preferably, the Desulfovibrio species is Desulfovibrio pigel or a relative thereof having at least 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.5, or 99.9% sequence identity with the 16S rDNA sequence of Desulfovibrio pigel (e.g., SEQ ID NO: 1). Such cutoff values ​​based on 16S rDNA similarity can define species with similar characteristics and / or functionality.

[0119] Preferably, in a composition comprising, for example, Desulfovibrio species, at least 10 per ml or g of said composition. 4 , 10 5 , 10 6 , 10 7 , 10 8 Alternatively or additionally, a total of 10 Desulfovibrio cells may be used in a composition containing Desulfovibrio species, for example, per ml or g of the composition. 4 ~10 16 , 10 4 ~10 15 , 10 4 ~10 14 , 10 4 ~10 12 , 10 6 ~10 12 , preferably 10 8 ~10 10 Desulfovibrio cells can be preferably used.

[0120] Alternatively or additionally, the Desulfovibrio cells may be viable, but have been obtained, for example, after pasteurization, or after incubation at, for example, 50-100, 60-80, 65-75, or 70°C, preferably for at least 5, 10, 15, 20, 25, 30, 40, 50 minutes, or preferably for at least 1, 5, 10, 20, 30 seconds, or 1, 5, 10, 15, 20, 25, 30, 40, 50 minutes of UV or It is also envisioned to use attenuated or killed cells (only) obtained after exposure to gamma irradiation or after incubation with oxygen, e.g., a gas containing at least 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 99, 100% by volume of oxygen, preferably for at least 1, 5, 10, 20, 30 seconds, or 1, 5, 10, 15, 20, 25, 30, 40, 50 minutes. Preferably, the Desulfovibrio species is the first, second, third, fourth, or fifth most abundant bacterial species in the composition, i.e., has the highest cell count or is at least in the top five compared to other bacterial species contained in the composition.

[0121] The Desulfovibrio species according to the present disclosure are preferably not present in the feces, or if present in the feces (e.g., as an alternative to the compositions described above), they are enriched, i.e., the number of Desulfovibrio cells is higher than in feces of the prior art, e.g., Desulfovibrio cells have been added to the feces or the feces have been exposed to conditions favorable for the growth of said Desulfovibrio species. If Desulfovibrio species according to the present disclosure are present in the feces, e.g., preferably at least 10 per ml or per g of feces. 4 , 10 5 , 2 × 10 5 , 3×10 5 , 4×10 5 , 5×10 5 , 6×10 5 , 7×10 5 , 8×10 5 , 9×10 5 , 10 6 , 2 × 10 6 , 3×10 6 , 4×10 6 , 5×10 6 , 6×106 , 7×10 6 , 8×10 6 , 9×10 6 , 10 7 , 2 × 10 7 , 3×10 7 , 4×10 7 , 5×10 7 , 6×10 7 , 7×10 7 , 8×10 7 , 9×10 7 , 10 8 , 10 9 , 10 10 , 10 11 , 10 12 , 10 13 Desulfovibrio cells are contained in the feces. Preferably, the Desulfovibrio species is the first, second, third, fourth, or fifth most abundant bacterial species in the feces, i.e., has the greatest cell count or is at least in the top five compared to other bacterial species contained in the feces.

[0122] The agent according to the present disclosure may also or instead be an amino acid substituted with one or more halogens, preferably one halogen, preferably a chloro-, fluoro-, or bromo-substituted amino acid. Aromatic amino acids optionally substituted with one or more halogens, preferably one halogen, for example, at the 6-position, are preferred, for example, aromatic amino acids preferably chloro-, fluoro-, or bromo-substituted at the 6-position. Tryptophan, tyrosine, or phenylalanine optionally substituted with one or more halogens, preferably one halogen (for example, at the 6-position), for example, tryptophan, tyrosine, or phenylalanine preferably chloro-, fluoro-, or bromo-substituted at the 6-position, are more preferred, for example, chloro-, fluoro-, or bromo-substituted tryptophan. For example, halogenated tryptophan at the 6-position, preferably chlorotryptophan, fluorotryptophan, or bromotryptophan, is even more preferred. 6-Bromotryptophan or any derivative or functional equivalent thereof is most preferred. The drug may be used in an amount of, for example, at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, or 900 μg (micrograms) per ml or g of the composition it is contained in. Alternatively or additionally, for example, a total of 0.1 to 10, 0.5 to 15, 1 to 20, 1 to 100, 5 to 100, 1 to 500, or 50 to 750 μg (micrograms) per ml or g of the composition containing the drug may be preferably used. Alternatively, the drug may be used in an amount of at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mg per ml or g of the composition it is contained in. Alternatively or additionally, a total of 0.1 to 10, 0.5 to 15, 1 to 20, 1 to 100, 5 to 100, 1 to 500, or 50 to 750 μg (micrograms) per ml or g of the composition may be preferably used.Alternatively or additionally, for example, a total amount of 0.1 to 10, 0.5 to 15, 1 to 20, 1 to 100, 5 to 100, 1 to 500, or 50 to 750 mg per ml or gram of drug-containing composition may be preferably used. Administration may be oral, subcutaneous, or intravenous. The total amount to be administered can be determined by a specialist based on the body weight of the subject being treated. For example, a single dose may contain 10 micrograms to 100 g, or 10 mg to 50 g, or 50 mg to 10 g, or 100 mg to 5 g. Doses may be administered periodically as described elsewhere herein. Additionally or alternatively, the drug is preferably not present in the feces, or, if present in the feces (e.g., as an alternative to the compositions described above), it is enriched, i.e., the amount of drug is greater than in prior art feces, i.e., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10% by weight greater, compared to any feces or fecal microbiome explant to which no drug has been added. According to the present invention, the agent can be added to feces. When the agent is contained in feces, for example, the agent is preferably added in an amount of at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900 ng per ml or g of feces, or at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900 μg (micrograms), or at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100 mg of the drug is contained in the feces. Preferably, the drug is the first, second, third, fourth or fifth most abundant metabolite in the feces, i.e., has the largest mass or is at least in the top ten or top five relative to other metabolites contained in the feces.

[0123] Additionally or alternatively, an agent according to the present disclosure may be a mono- or di-fatty acid substituted glycerolphosphocholine (GPC), preferably wherein the fatty acids are (independently) saturated or (mono- or poly-) substituted fatty acids.

[0124] Unsaturated fatty acids such as myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, α-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, and docosahexaenoic acid are preferred. Substituted glycerol phosphocholines (GPCs) containing one or more of myristoleic acid and arachidonic acid are more preferred. 1-Myristoyl-2-arachidonoyl-glycerophosphocholine (MA-GPC) and 1-arachidonoyl-glycerophosphocholine (A-GPC), or any derivatives or functional equivalents thereof, are more preferred and have shown good results. The drug may be used in an amount of at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, or 900 μg (micrograms) per ml or g of the composition it is contained in. Alternatively or additionally, a total of 0.1 to 10, 0.5 to 15, 1 to 20, 1 to 100, 5 to 100, 1 to 500, or 50 to 750 μg (micrograms) per ml or g of the composition it is contained in may be preferably used. Instead, the drug may be used in an amount of, for example, at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mg per ml or g of the composition it is contained in. Alternatively or additionally, for example, a total of 0.1 to 10, 0.5 to 15, 1 to 20, 1 to 100, 5 to 100, 1 to 500, or 50 to 750 μg (micrograms) per ml or g of the composition containing the drug may be preferably used. Alternatively or additionally, for example, a total of 0.1 to 10, 0.5 to 15, 1 to 20, 1 to 100, 5 to 100, 1 to 500, or 50 to 750 mg per ml or g of the composition containing the drug may be preferably used. Administration may be oral, subcutaneous, or intravenous. The total amount to be administered can be determined by a specialist depending on the body weight of the subject to be treated.For example, a single dose may comprise 10 micrograms to 100 g, or 10 mg to 50 g, or 50 mg to 10 g, or 100 mg to 5 g. Alternatively or additionally, administration may be such that a plasma concentration of 0.1 to 100, 0.2 to 50, 0.5 to 25, 0.5 to 20, 0.5 to 3, 1 to 15, 2 to 10, or 2 to 5 nmol / ml or 0.1 to 100, 0.2 to 50, 0.5 to 3, 0.5 to 25, 0.5 to 20, 1 to 15, 2 to 10, or 2 to 5 μmol / ml, or 50% thereof in the case of pediatric use, may be achieved in the treated subject. Doses may be administered periodically as described elsewhere herein. Additionally or alternatively, the agent is preferably not present in the feces, or if present in the feces (e.g., as an alternative to the compositions described above), it is enriched, i.e., the amount of agent is greater than in prior art feces, i.e., at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10% by weight greater, compared to any feces or fecal microbiome transplant without added agent. According to the present invention, the agent can be added to the feces. If the drug is contained in feces, for example, it is preferably at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900 ng per ml or g of feces, or at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900 μg (micrograms), or at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100 mg of the drug is contained in the feces. Preferably, the drug is the first, second, third, fourth or fifth most abundant metabolite in the feces, i.e., has the largest mass or is at least in the top ten or top five relative to other metabolites contained in the feces.

[0125] The agents disclosed herein may be used in any combination in the prevention or treatment of autoimmune diseases as described herein. For example, Desulfovibrio species may be combined with chloro-, fluoro-, or bromo-substituted tryptophan, such as 6-BT, and / or mono- or di-fatty acid-substituted glycerolphosphocholine (GPC), such as MA-GPA or A-GPC. Alternatively, chloro-, fluoro-, or bromo-substituted tryptophan, such as 6-BT, may be combined with mono- or di-fatty acid-substituted glycerolphosphocholine (GPC), such as MA-GPC or A-GPC. Alternatively, MA-GPC may be combined with A-GPC.

[0126] Agents according to the present disclosure can modulate the immune system by resetting the function of B cell clones and regulatory T cells that inhibit autoimmune responses.

[0127] Preferably, an agent according to the present disclosure may be derived from, but not contained in, or associated with, feces. Additionally or alternatively, the agent may be contained in a composition comprising no more than 100, 90, 80, 70, 60, 50, 40, 30, 20, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 bacterial species.

[0128] Preferably, the agent according to the present disclosure is included in the (pharmaceutical) composition in an amount of at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, or 0.1-10, 0.5-15, 1-20, 1-100 mg, 5-50 mg, or 1-25 mg, for example, per gram or ml of composition or carrier (e.g., an aqueous solution comprising 0.5-1.5% by weight NaCl, e.g., 0.9% by weight NaCl, particularly for intravenous administration).

[0129] Prevention and / or treatment according to the present disclosure may involve administering the agent to the subject orally or to the small intestine, preferably the duodenum. In this regard, feces may be administered to the intestine, preferably by oral, nasal, or rectal administration, and / or by duodenal administration, such as by a (naso)duodenal tube. In particular for any of the substituted amino acid agents according to the present disclosure, and in particular for GPCs according to the present disclosure, such as MA-GPC and A-GPC, intravenous administration and subcutaneous administration (e.g., by a rod-sized (4 x 44 mm) implantable device (which is then replaced again) subcutaneous delivery system suitable for delivery for 3-6 months of treatment and effective, sustained long-term dosing) are also envisioned.

[0130] The medicaments of the present disclosure are administered to the gastrointestinal tract of a subject, preferably to the small intestine of a subject, most preferably to the duodenum. The duodenum is the first portion of the small intestine in most higher vertebrates, including mammals. The duodenum is located before the jejunum and ileum and is the smallest portion of the small intestine. In humans, the duodenum is a 25-38 cm hollow tube connecting the stomach and the distal duodenum. It begins at the duodenal bulb and ends at the suspensory muscle of the duodenum. While it is also possible to administer the medicament to the colon (or cecum) of a subject, administration to the colon (or cecum) of a subject is preferably not encompassed by the present disclosure.

[0131] The drug according to the present disclosure has the following structure: - Firmicutes, for example Eubacterium, Intestinimonas, Faecalibacterium, Christensenella, Anaerostipes, Agathobacter, Roseburia, Coprococcus, Clostridium, Subdoligranulum, Anaerostipes, those belonging to the genera Anaerotruncus, Flavinobacter, Ruminococcus, Butyricicoccus, Butyrovibrio, Sporobacter, Papilibacter, Oscillobacter, Oscillospora, Veilonella, Lactobacillus, and Streptococcus; Proteobacteria, for example those belonging to the genera Escherichia or Enterobacter, Actinobacteria, for example those belonging to the genus Bifidobacterium or Colinsella, Bacteroidetes, for example those belonging to the genera Bacteroides, Prevotella or Alistipes, and / or - Verrucomicrobia, for example those belonging to the genus Akkermansia The bacteria may be combined with a microorganism or enteric microbial cell, which may be one (or a combination) selected from the group consisting of:

[0132] The agent may be combined with a microorganism or gut microbial cells selected from the group consisting of eukaryotic bacteria, archaea, and bacteria, preferably selected from the group of 1057 species disclosed by Rajilic-Stojanovic and de Vos (2014 FEMS Microbiol Rev. 38(5):996-1047).

[0133] Preferably, for example, 10 per ml or g of carrier 4 ~10 16 , 10 4 ~10 15 , 10 4 ~10 14 , 10 6 ~10 12 , preferably 10 8 ~10 10 Any of the above microbial cells may be used.

[0134] The drug may be applied in an effective amount, i.e., an amount sufficient to achieve the desired therapeutic and / or prophylactic effect, for example, an amount that results in the treatment and / or prevention of the respective condition. For therapeutic or prophylactic applications, the amount to be administered to a subject may depend on the type and severity of the disease or condition, as well as the characteristics of the subject, such as general health, age, sex, weight, and tolerance to the drug. This may also depend on the degree, severity, and type of the disease or condition. A specialist can determine the appropriate dosage depending on these and other factors.

[0135] In preferred embodiments, prevention and / or treatment according to the present disclosure involves at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and / or at most 10, 20, 30, 40, 50 separate administrations of the agent, with intervals between said separate administrations preferably of at least 1, 2, 3, 4, 5, 6, 7, 8, 10, and / or at most 10, 20, 30, 40, 50 weeks. Prevention and / or treatment may involve daily, weekly, monthly administrations, e.g., one or two administrations every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 days / weeks / months, and / or for a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50 weeks (or even months or years).

[0136] The agent may be contained in a liquid vehicle and / or is preferably not combined with a solid (e.g., a composition comprising a solid) having a diameter greater than 1, 2, 3, 4, 5, 10, 15, 20, 25, 50, 75, 100, 200, 400, 600, 800, or 1000 μm. The liquid vehicle may be an aqueous solution containing 0.5-1.5% NaCl by weight, e.g., 0.9% NaCl by weight. The term "solid" refers to discrete particles containing at most 30, 20, 10, 5, or 1% water by weight.

[0137] It is further envisioned that the agents according to the present disclosure are contained in a composition, preferably a pharmaceutical composition, more preferably a liquid or solid dosage form, most preferably a capsule, tablet, or powder.

[0138] For oral administration, the drug may be administered in solid dosage forms, such as capsules, tablets, and powders, or in liquid dosage forms, such as elixirs, syrups, and suspensions. Carriers such as activated charcoal may also be employed.

[0139] The agent may be used as a pharmaceutical and / or may be accompanied by a physiologically acceptable carrier, which may be any inert carrier. For example, non-limiting examples of suitable physiologically or pharmaceutically acceptable carriers include any well-known physiological or pharmaceutical carriers, buffers, diluents, and excipients. It will be recognized that the selection of a suitable physiological carrier depends on the intended mode of administration of the compositions taught herein (e.g., oral) and the intended form of the composition (e.g., beverage, yogurt, powder, capsule, etc.). A person skilled in the art will be familiar with how to select a physiologically acceptable carrier that is suitable for or compatible with the compositions for use as taught herein.

[0140] It is particularly preferred that the drug be contained within and / or encapsulated by an (enteric) coating, preferably one that does not dissolve and / or disintegrate in the subject's stomach environment. Such a coating can help the drug reach the intended site for delivery, e.g., the duodenum, without being degraded by the acidic environment of the stomach. A preferred (enteric) coating works by presenting a stable surface at the highly acidic pH found in the stomach, but disintegrating more rapidly at lower pH. For example, it will not dissolve in the gastric acid of the stomach (pH approximately 3), but will dissolve in the alkaline environment (pH 7-9) present in the small intestine or duodenum.

[0141] In embodiments, an agent according to the present disclosure may be combined with or contained in a composition comprising a mucosal binding agent. As used herein, the term "mucosal binding agent" or "mucosal binding polypeptide" refers to an agent or polypeptide capable of binding itself to the intestinal mucosal surface of the intestinal mucosal barrier of a mammal (e.g., a human). Various mucosal binding polypeptides have been disclosed in the art. Non-limiting examples of mucosal binding polypeptides include membrane-binding subunits of bacterial toxins, including the B subunit of cholera toxin, the B subunit of Escherichia coli heat-labile enterotoxin, subunits S2, S3, S4, and / or S5 of Bordetella pertussis toxin, the B fragment of Diphtheria toxin, and the membrane-binding subunit of Shiga toxin or Shiga-like toxin. Other suitable mucosa-binding polypeptides include bacterial pilus proteins, including E. coli fimbriae K88, K99, 987P, F41, FAIL, CFAIII ICES1, CS2 and / or CS3, CFA1IV ICS4, CS5 and / or CS6), P. fimbriae, etc. Non-limiting examples of other fimbriae include Bordetella pertussis fimbriae hemagglutinin, Vibrio cholerae toxin coregulatory pilus (TCP), mannose-sensitive hemagglutinin (MSHA), fucose-sensitive hemagglutinin (PSHA), etc. Still other mucosa-binding agents include viral attachment proteins, including influenza and Sendai virus hemagglutinins, and animal lectins or lectin-like molecules, including immunoglobulin molecules or fragments thereof, calcium-dependent (C-type) lectins, selectins, collectins, or plant lectins with mucosa-binding subunits, including helix pomatia hemagglutinin, concanavalin A, wheat germ agglutinin, phytohemagglutinin, abrin, ricin, and the like.

[0142] In embodiments, compositions containing agents for use as taught herein may be in liquid form, e.g., a stabilized suspension containing one or more of the agents, or in solid form, e.g., a lyophilized powder of an agent as taught herein. For example, cryoprotectants such as lactose, trehalose, or glycogen may be employed.

[0143] Optionally, the agents according to the present disclosure may be encapsulated in capsules, such as gelatin capsules, optionally with inactive ingredients and powdered carriers, such as glucose, lactose, sucrose, mannitol, starch, cellulose or cellulose derivatives, magnesium stearate, stearic acid, sodium saccharin, talcum, magnesium carbonate, and the like.

[0144] In embodiments, a medicament according to the present disclosure may comprise one or more ingredients suitable for promoting the survival and / or viability and / or maintenance and / or integrity of the medicament, for example, during storage and / or exposure to bile and / or passage through the gastrointestinal tract of a mammal (e.g., a human). Non-limiting examples of such ingredients include enteric coatings as previously described herein and / or controlled-release agents that allow passage through the stomach. A person skilled in the art will be familiar with how to select suitable ingredients that ensure that the feces reach the intended destination where they will exert their action.

[0145] In embodiments, compositions comprising agents for uses taught herein may further comprise an ingredient selected from the group consisting of prebiotics, probiotics, carbohydrates, polypeptides, lipids, vitamins, minerals, pharmaceuticals, preservatives, antibiotics, or any combination thereof.

[0146] In particularly preferred embodiments, the agent according to the present disclosure is preferably selected from the group consisting of Bifidobacterium animalis subsp. lactis or Bifidobacterium breve, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Eubacterium hallii, Intestinimonas butyriciproducens, and / or Akkermansia muciniphila. muciniphila, Eubacterium, Intestinimonas, Bifidobacteria, Lactobacillales, and / or Ackermansia. 4 ~10 14 , 10 6 ~10 12 , preferably 10 8 ~10 10 (per ml or per gram) of such bacterial cells may be used. The above combinations may produce synergistic effects. The drug and bacteria may be contained in different compositions or may be contained together in a single composition (e.g., in a capsule or other dosage form described herein).

[0147] The medicaments of the present disclosure may also or instead be combined with hormone replacement drugs (such as thyroid, hydrocortisone, or insulin), tumor necrosis factor alpha (TNFα) inhibitors, and / or DMARDs (for rheumatoid arthritis), preferably selected from the group consisting of infliximab, adalimumab, certolizumab pegol, and golimumab. The inventors believe that treatment with a TNFα inhibitor will enhance the response to treatment with the medicaments of the present disclosure, and / or conversely, treatment with a medicament of the present disclosure will enhance the response to treatment with a TNFα inhibitor. Preferably, the TNFα inhibitor is administered in a separate or the same composition as the medicament (e.g., a capsule or other dosage form described herein). The TNFα inhibitor may be administered weekly or daily at least (or at most) 1, 2, 3, 4 times, and / or intravenously / or orally, e.g., at a dose of 1-10, 2-8, 3-7, 4-6, or 5 mg / kg.

[0148] It is further envisioned that the agent for use according to the present disclosure, particularly Desulfovibrio species, is present in lyophilized and / or microencapsulated form, e.g., in a capsule containing the agent. Preferably, the agent, e.g., Desulfovibrio species, is present in a solid, lyophilized or dried form (i.e., containing less than 20, 10, 5, 2, or 1% water by weight), e.g., in the form of a powder or granules. For example, it may be present in a microencapsulated form. Those skilled in the art can lyophilize or microencapsulate the agent based on well-known techniques in which oxygen-free conditions are applied to preserve the viability of any bacteria contained in feces.

[0149] The technique of microencapsulation is well known in the art for preserving bacteria (e.g., as reviewed by Serna-Cock and Vallejo-Castillo, 2013, Afr J of Microbiol Res, 7(40): 4743-4753). For example, any of the preservation techniques and systems taught by Serna-Cock and Vallejo-Castillo may be employed in the present disclosure.

[0150] Freeze-drying methods include, but are not limited to, slow, gradual freezing to -40°C followed by drying, rapid freezing by placing at -80°C followed by drying, or ultra-rapid freezing by dripping cells into liquid nitrogen with a cryoprotectant followed by drying. A cryoprotectant is often employed to protect the composition during freeze-drying and extend shelf life. The cryoprotectant may be selected from the group consisting of, but not limited to, sucrose, maltose, maltodextrin, trehalose, mannitol, sorbitol, inulin, glycerol, DMSO, ethylene glycol, propylene glycol, 2-methyl-2,4-pentanediol, polyethylene glycol, polyvinylpyrrolidone, polyvinyl alcohol, polyglycerol, skim milk powder, milk protein, whey protein, UHT milk, betaine, adonitol, sucrose, glucose, lactose, or any combination thereof.

[0151] Prebiotics such as starch and bran may also be added to the formulation of the present disclosure to enhance its efficiency, for example, prior to freeze-drying. The addition of antioxidants such as riboflavin, riboflavin phosphate, or physiologically acceptable salts thereof, glutathione, ascorbate, glutathione, and cysteine ​​to the freeze-dried mixture can enhance the viability of any included bacteria.

[0152] Agents, particularly Desulfovibrio species, can be stored for extended periods (e.g., at least 10, 20, 40, 52 weeks, or at least 1, 2, 3 years) after addition of a cryoprotectant, such as glycerol, as disclosed herein and / or freezing at -80°C. Additionally or alternatively, lyophilization can render the agent stable over such periods. Finally, Desulfovibrio species can also be inoculated as described by de Vos (2013 Microb Biotechnol. July 2013, 6(4):316-25).

[0153] In embodiments, the agents for use as taught herein may be or may be included in a food or supplement composition, which may include a dairy product, more preferably a fermented dairy product, preferably a yogurt or drinking yogurt.

[0154] In embodiments, the medicament for the uses taught herein or the composition comprising same may further comprise one or more ingredients that further enhance the nutritional and / or therapeutic value of the feces taught herein. For example, it may be advantageous to add one or more ingredients (e.g., nutritional ingredients, veterinary drugs, or pharmaceuticals) selected from proteins, amino acids, enzymes, mineral salts, vitamins (e.g., thiamine HCl, riboflavin, pyridoxine HCl, niacin, inositol, choline chloride, calcium pantothenate, biotin, folic acid, ascorbic acid, vitamin B12, p-aminobenzoic acid, vitamin A acetate, vitamin K, vitamin D, vitamin E, etc.), sugars and complex carbohydrates (e.g., water-soluble and water-insoluble monosaccharides, disaccharides, and polysaccharides), pharmaceutical compounds (e.g., antibiotics), antioxidants, trace element ingredients (e.g., cobalt, copper, manganese, iron, zinc, tin, nickel, chromium, molybdenum, iodine, chlorine, silicon, vanadium, selenium, calcium, magnesium, sodium, and potassium, etc.). Those skilled in the art are familiar with methods and ingredients suitable for enhancing nutritional value and / or therapeutic / pharmaceutical value.

[0155] The present disclosure also provides a method for predicting the response of an autoimmune disease patient to treatment with an agent (or with autologous feces) according to the present disclosure, the method comprising: - determining the level of abundance in the patient's fecal microbiome of at least one bacterium selected from the group consisting of Bacteroides caccae and Coprococcus catus; A measured level higher than the reference level indicates that the autoimmune disease patient is responding to treatment. The reference level may be, for example, 50 to 150%, preferably 75 to 125%, more preferably 90 to 120, 95 to 110, or 98 to 105% of the abundance level of the at least one bacterium selected from the group consisting of Bacteroidetes cacae and Coprococcus catus in the fecal microbiota of a healthy control subject.

[0156] In the context of the present disclosure, the subject to be treated is preferably an animal, more preferably a mammal, and most preferably a human. As will be apparent, the treatment is preferably not conducted as a control or placebo treatment and / or a clinical trial, i.e., a study in which participants are assigned to receive one or more interventions / treatments, one or more control or placebo interventions / treatments, or no intervention, so that researchers can evaluate the effect of the intervention on a biomedical or health-related outcome.

[0157] In this specification and the claims, the verb "comprise" and its conjugations are used in their open-ended sense to mean that the items following the word are included, but items not specifically mentioned are not excluded. Furthermore, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that more than one element is present, unless the context clearly requires that only one element should be present. That is, the indefinite article "a" or "an" normally means "at least one."

[0158] "Sequence identity" can be determined by aligning two peptide or two nucleotide sequences using an alignment algorithm (e.g., when optimally aligned using the programs GAP or BESTFIT with default parameters). GAP uses the Needleman and Wunsch global alignment algorithm to align two sequences over their entire length, maximizing the number of matches and minimizing the number of gaps. Generally, the GAP default parameters are used, with a gap insertion penalty of 50 (nucleotides) / 8 (proteins) and a gap extension penalty of 3 (nucleotides) / 2 (proteins). For nucleotides, the default scoring matrix used is nwsgapdna, and for proteins, the default scoring matrix is ​​Blosum62 (Henikoff and Henikoff, 1992, PNAS 89, 915-919). Sequence alignment and scoring for percentage sequence identity may be determined using computer programs such as the GCG Wisconsin Package, version 10.3, available from Accelrys Inc., 9685 Scranton Road, San Diego, CA 92121-3752 USA, or EmbossWin version 2.10.0 (using the program "needle"). Alternatively, the percentage of similarity or identity may be determined by searching a database using algorithms such as FASTA, BLAST, etc. By way of illustration, by a polynucleotide having a nucleotide sequence having at least, e.g., 95% "identity" with a reference nucleotide sequence encoding a polypeptide of a particular sequence, it is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence, except that the polynucleotide sequence may contain up to 5 point mutations per each 100 nucleotides of the reference polypeptide sequence, which may be (conservative) substitutions, deletions, and / or insertions.In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted and / or substituted with alternative nucleotides, and / or up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These mutations in the reference sequence may occur at the 5' or 3' terminal positions of the reference nucleotide sequence, or anywhere between these terminal positions, either individually among nucleotides in the reference sequence or interspersed in one or more contiguous groups within the reference sequence. Similarly, by a polypeptide having an amino acid sequence having at least, e.g., 95% "identity" with a reference amino acid sequence of SEQ ID NO: 1, it is intended that the amino acid sequence of the polypeptide is identical to the reference sequence, except that the polypeptide sequence may contain up to 5 amino acid changes per 100 amino acids of each of the reference amino acids of SEQ ID NO: 1. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a reference amino acid sequence, up to 5% of the amino acid residues in the reference sequence may be deleted or substituted with alternative amino acids, or up to 5% of the total amino acid residues in the reference sequence may be inserted into the reference sequence. These variations in the reference sequence can occur at the amino or carboxy terminal positions of the reference amino acid sequence, or anywhere between these terminal positions, either individually among residues in the reference sequence or interspersed in one or more contiguous groups within the reference sequence. Sequence identity can be determined over the entire length of the sequence under consideration.

[0159] Sequence Listing

[0160] 16S rDNA sequence of Desulfovibrio pigeli (SEQ ID NO: 1)

[0161] [ka] [Brief explanation of the drawings]

[0162] [Figure 1]The top 10 small intestinal microbial groups with relative importance that best predicted treatment group assignment (XGBoost predictive modeling algorithm). Percentages are scaled with a maximum of 100%. The top four microbial groups stand out with their high relative importance. [Figure 2A] Top 10 metabolites that best predicted treatment group assignment (XGBoost predictive modeling algorithm). Percentages are scaled with a maximum of 100%. The top three metabolites stand out as having high relative importance in the analysis. [Figure 2B] Relative abundance of the top three metabolites plotted against time for each treatment group (in each figure, the predominantly upper line represents the autologous FMT group, and the predominantly lower line represents the allogeneic FMT group). Median ± IQR is reported. P values ​​were calculated using the Mann-Whitney U test between groups at 12 months. 1-Myristoyl-2-arachidonoyl-GPC differed between groups at 12 months, p=0.020. 1-Arachidonoyl-GPC differed between groups at 12 months, p=0.020. [Figure 2C] Relative abundance of the top three metabolites plotted against time for each treatment group (in each figure, the predominantly upper line represents the autologous FMT group, and the predominantly lower line represents the allogeneic FMT group). Median ± IQR is reported. P values ​​were calculated using the Mann-Whitney U test between groups at 12 months. 1-Myristoyl-2-arachidonoyl-GPC differed between groups at 12 months, p=0.020. 1-Arachidonoyl-GPC differed between groups at 12 months, p=0.020. [Figure 2D] Relative abundance of the top three metabolites plotted against time for each treatment group (in each figure, the predominantly upper line represents the autologous FMT group, and the predominantly lower line represents the allogeneic FMT group). Median ± IQR is reported. P values ​​were calculated using the Mann-Whitney U test between groups at 12 months. 1-Myristoyl-2-arachidonoyl-GPC differed between groups at 12 months, p=0.020. 1-Arachidonoyl-GPC differed between groups at 12 months, p=0.020. [Figure 2E] Spearman correlation between change in fasting C-peptide and change in 1-myristoyl-2-arachidonoyl-GPC. [Figure 2F] Abundance of D. pigelii in feces over time. P values ​​were calculated using the Mann-Whitney U test. At 6 months, p = 0.024; at 12 months, p = 0.023. [Figure 2G] Fold change in D. pigelii between groups (mainly upper line represents autologous FMT group). Delta p-values ​​were calculated by performing a Mann-Whitney U test on the delta from 0 to 12 months for each group. p-value = 0.006. [Figure 2H] Spearman correlation plot of Delta (0–12 months) fecal D. Pigel and fasting C-peptide Delta (0–12 months). [Figure 2I] Correlation plot between fecal D. pigel and 1-arachidonoyl-GPC. [Figure 2J] Correlation plot between fecal D. pigel and small intestinal Prevotella 1. [Figure 2K] Correlation plot between fecal D. pigel and small intestinal Prevotella 2. [Figure 2L] Figure 2L. [Figure 3] Predictive modeling output showing the top 30 species causing differential changes in fecal microbial communities between treatment groups. [Figure 4A] The number of responders at 6 and 12 months and the number of subjects in each treatment group are shown. Response was defined as a decrease in the AUC of C-peptide to less than 10% compared to baseline. 12-month responders were used for all analyses. [Figure 4B] Individual subject lines for C-peptide AUC over time are shown. [Figure 4C] The abundance of B. cacae and C. catus over time is shown, respectively. In both figures, the upper line represents responders. P values ​​were calculated using the Mann-Whitney U test between groups at each time point. p value = 0.0099 for B. cacae at baseline and p value = 0.00049 for C. catus at baseline. [Figure 4D] The abundance of B. cacae and C. catus over time is shown, respectively. In both figures, the upper line represents responders. P values ​​were calculated using the Mann-Whitney U test between groups at each time point. p value = 0.0099 for B. cacae at baseline and p value = 0.00049 for C. catus at baseline. [Figure 4E] Correlation between Delta C. catus (0–12 months) and AUC of Delta C peptide (0–12 months) is shown. Spearman's rho (r) is shown and p-values ​​were calculated using Spearman's rank. [Figure 5] Predictive modeling output showing the top 30 species causing differential changes in fecal microbial communities between treatment groups. [Figure 6-1] Figure 6. Abundance over time of five fecal microbial groups from the top 10 species (see Figure 3) that best discriminated between responders and non-responders at baseline (in Figures 6A, 6B, and 6D, the top lines represent responders, and the other lines represent non-responders. In Figures 6C and 6E, the top lines represent non-responders, and the other lines represent responders). Strains that differed between responders and non-responders at baseline or over the course of the study were selected for presentation. P values ​​were calculated using the Mann-Whitney U test at each time point. Figure 6A: Paraprevotella spp., p = 0.019. Figure 6B: Eubacterium ramulus, p = 0.043. Figure 6C: Collinsella aerofaciens, p = 0.043. Figure 6D: Bacteroides eggerthii, p=0.006. Figure 6E: Ruminococcus callidus, p=0.026. Faecalibacterium prausnitzii (10th from the top 10) was not significantly different at baseline (p=0.063). [Figure 6-2] Continued from Figure 6. [Figure 7] Effects of various doses of 6-bromotryptophan (6-BT), 1-arachidonoyl-glycero-phosphocholine (20:0) (A-GPC), 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphoethanolamine (16:0–18:2 PE), and 1-myristoyl-2-arachidonoyl-glycero-phosphocholine (MA-GPC) on activation of the NFκB pathway. [Figure 8] Effects of 6-BT, MA-GPC, and A-GPC on myeloid cells. Mouse monocytes activated with LPS (10 ng / ml) for 24 hours were CD11b+ - TLR4 stimulated. [Figure 9] Effects of 6-BT, MA-GPC, and A-GPC on myeloid cells. Mouse monocytes activated with the dsDNA analog poly I:C for 24 hours were stimulated with CD11b+ TLR3. [Figure 10] Effect of 6-BT on human monocytes upon LPS stimulation. [Figure 11] Effects of 6-BT, A-GPC, and MA-GPC on T lymphocytes. Mouse CD4+ T cells activated with anti-CD3 and anti-CD28 mAbs. [Figure 12] INS1e beta cells treated with 6-BT for 24 hours. Gene expression of beta cell differentiation markers. [Figure 13] INS1e beta cells treated with 6-BT for 24 hours. Gene expression of beta cell differentiation markers. [Figure 14] Monocytes isolated from mouse bone marrow (Christ A., Cell 2018) or bone marrow-derived macrophages (Swansen, JEM 2017) were exposed to the indicated concentrations (10-100 μM) of 6-BT in the presence or absence of 10 ng / ml LPS, 10 μg / ml P3C, or 10 μg / ml poly(I:C) for 24 hours. Using ELISA assays, we found that 6-BT inhibited the secretion of the pro-inflammatory cytokine TNFα via TLR4 and TLR2 activation and IFN-beta via TLR3 activation. [Figure 15]We investigated the effects of 6-BT on murine DCs differentiated with bone marrow cells and GM-CSF (40 ng / ml). As for monocytes / macrophages, 6-BT inhibited the secretion of the proinflammatory cytokines TNFα and IFNβ by DCs after activation of TLR4 (by 100 ng / ml LPS) or TLR3 (by 10 μg / ml poly(I:C)), respectively. [Figure 16] We further investigated the effects of 6-BT on CD4 T cells. To mimic antigen presentation, mouse CD4 T cells (isolated from the pancreas, Uchimura T, Immunity 2019) were activated with monoclonal antibodies against CD3 and CD28 (2.5 and 1 μg / ml, respectively). Consistent with findings in myeloid cells, 6-BT significantly reduced production of the Th1 cytokine IFN-gamma. [Figure 17] We investigated whether 6-BT could exert a direct effect on beta cells. Indeed, we found that 6-BT induced gene expression of the transcription factors PDX1 and MAFA in IS1E beta cells, which is important for beta cell maturation and functionality. Consistently, 6-BT also promoted insulin secretion during steady-state and glucose-stimulated insulin secretion (data presented as the difference between insulin release in starvation [1 mM glucose] and hyperglycemic [22 mM] conditions) (Paula S, FASEB J 2015). [Figure 18] We investigated the effect of 6-BT on the activation of the NF-kB pathway, a central pathway in all inflammatory diseases (not just autoimmune diseases). To this end, we quantified the expression of the phosphorylated form of the p65 subunit, which is considered a marker of NF-kB activation. Upon T cell activation with PMA (50 ng / ml) and ionomycin (1 μg / ml), 6-BT was able to inhibit NF-kB signaling at a very early time point (5–10 min after activation). This effect was observed in both mouse and human (Jurkat) CD4 T cells. [Figure 19]Using the RAW264.7 murine macrophage cell line stably expressing an NFkB luciferase reporter (Groeneweg M, J Lipid Res 2006), we demonstrated that overnight exposure of macrophages to 6-BT (10-200 μM) dose-dependently inhibited the transcriptional activity of the NFkB complex following 2-hour stimulation with LPS (10 ng / ml). [Figure 20] In mouse CD4 T lymphocytes (isolated from mouse pancreas, Uchimura T, Immunity 2019), 6-BT exerted an inhibitory effect on CD3 / CD28-mediated IFN-gamma production, whereas tryptophan did not, indicating that 6-BT and tryptophan induce distinct biological activities. [Figure 21] Exposure of monocytes (isolated from mouse bone marrow, Christ A., Cell 2018) to 6-BT or tryptophan shows that the anti-inflammatory effect is specific to the 6-bromotryptophan molecule, but not to tryptophan. [Figure 22] We found that 6-BT (100 μM) promoted mitochondrial metabolism in mouse and human (Jurkat) CD4 T cells. OCR, the oxygen consumption rate (OCR), was used as a surrogate for cellular utilization of mitochondrial oxidative phosphorylation. OCR was measured using a Seahorse XF Analyzer (Uchimura T, Immunity 2019; Chou, Nature 2021). [Figure 23] Exposure to 6-BT can enhance mitochondrial metabolism in pro-inflammatory M1 macrophages (differentiated in the presence of LPS and IFN-gamma (Cheng N, JCI Insight 2018)) without affecting glycolytic flux. Intracellular metabolism was measured using a Seahorse XF Analyzer (Uchimura T, Immunity 2019; Chou, Nature 2021). [Figure 24]We investigated whether 6-BT could affect mitochondrial metabolism in beta cells. This depends on mitochondrial metabolic production for the exocytosis of ATP and insulin. 6-BT increased mitochondrial metabolism in beta cells (INS1E beta cells) both in the steady state and under high glucose conditions (25 mM glucose). Furthermore, we examined the effect of tryptophan on intracellular metabolism and found that tryptophan exerted different effects than 6-BT on inflammatory markers. Intracellular metabolism was measured using a Seahorse XF Analyzer (Uchimura T, Immunity 2019; Chou, Nature 2021). [Figure 25] Relative abundance of fecal Desulfovibrio in diabetic and nondiabetic subjects. [Figure 26] Diabetes: Effect of Desulfovibrio. Odds ratio for diabetes. [Figure 27] Plasma 6BT levels and fecal 6BT in (1) non-diabetic subjects (top line) and (2) diabetic patients (bottom line). Example 1

[0163] Relationship with the relative abundance of Desulfovibrio spp. Patients with the conditions indicated below were treated as follows: 1. Empty enteric-coated capsules administered orally daily for 2 years. 2. Approximately 1×10 8 Enteric-coated capsules containing cells of one Desulfovibrio species (Desulfovibrio pigel, Desulfovibrio desulfuricans, or Desulfovibrio fairfieldensis) were administered orally daily for two years. 3. Enteric-coated capsules containing 50 mg of chloro-, fluoro-, or bromo-substituted tryptophan (6-bromotryptophan (6-BT) or 6-fluorotryptophan (6-FT)) administered orally daily for 2 years. 4. Daily oral administration of 50 mg of enteric-coated capsules containing mono- or di-fatty acid substituted glycerolphosphocholine (GPC) (1-myristoyl-2-arachidonoyl-glycero-phosphocholine (MA-GPC) or 1-arachidonoyl-glycero-phosphocholine (A-GPC)) for 2 years.

[0164] [Table 1A]

[0165] [Table 1B]

[0166] [Table 1C]

[0167] [Table 1D]

[0168] [Table 1E]

[0169] [Table 1F]

[0170] [Table 1G]

[0171] It is anticipated that results similar to the predicted effects shown in Table 1 above will be obtained in larger patient cohorts. Example 2

[0172] Patients with recent-onset (less than 6 weeks) T1D were randomized into two groups to receive three autologous or allogeneic (healthy donor) fecal microbiota transplants (FMTs) over a four-month period.

[0173] Several plasma metabolites (derived from the microbiome) and (small) intestinal bacterial strains were found to be associated with improved residual beta cell function in type 1 diabetes.

[0174] Materials and Methods We conducted a double-blind, randomized, controlled clinical trial using computer-generated randomization in subjects with new-onset T1D to examine the effects of infusion of allogeneic (healthy donor) gut microbiota compared with autologous (self) gut microbiota on residual beta-cell function and autoimmune T-cell responses in relation to changes in the (small)gut microbiota 1 year after treatment.

[0175] Patient recruitment Patients with newly diagnosed T1D were recruited from outpatient clinics in the Amsterdam area. Patient inclusion criteria were male / female with residual beta-cell function (as indicated by plasma C-peptide >0.2 mmol / L and / or >1.2 ng / mL after MMT), age 18-35 years, and normal BMI (18.5-25 kg / m). 2 ), and a diagnosis of T1D within a maximum of 6 weeks prior to inclusion. Exclusion criteria were a diagnosis or symptoms of another autoimmune disease (e.g., hypo- or hyperthyroidism, celiac disease, rheumatoid arthritis, or inflammatory bowel disease), (expected) long-term immune impairment (due to recent cytotoxic chemotherapy or HIV infection with a CD4 count below 240), and use of antibiotics in the last 3 months, use of proton pump inhibitors, and any other type of systemic medication that interferes with insulin.

[0176] Donor recruitment Lost weight (BMI 25kg / m 2Healthy, omnivorous, Caucasian men and women (<10 years of age) were recruited to serve as fecal donors. Donors completed a complete questionnaire regarding dietary and bowel habits, travel history, comorbidities including family history of diabetes, and medication use. Donors were screened for the presence of infectious diseases as previously described (van Nood et al., 2013). Blood was screened for human immunodeficiency virus, human T-lymphotropic virus, hepatitis A, B, and C viruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), Strongyloides, amebiasis, and syphilis. Active infections were excluded, but previous and inactive infections with EBV and CMV were allowed. Donors were also excluded if screening of their feces revealed the presence of pathogenic parasites (e.g., Blastocystis hominis, Dientamoeba fragilis, Giardia lamblia), multidrug-resistant bacteria (Shigella, Campylobacter, Yersinia, MRSA, ESBL, Salmonella, enteropathogenic E. coli, and Clostridium difficile), or viruses (noro-, rota-, astro-, adeno-(40 / 41 / 52)-, entero-, parecho-, and sapo-viruses), as previously recommended.

[0177] Clinical trial visit Participants were required to complete an online nutrition diary for one week prior to each study visit to monitor calorie intake, including dietary carbohydrates, fat, protein, and fiber. Blood pressure, length, weight, and daily insulin use were recorded during study visits. Fasting blood samples were collected at each visit, centrifuged, and stored at -80°C for later analysis. Whole blood sodium heparin tubes were kept at room temperature and processed for immunological analysis within 24 hours.

[0178] Three fecal transplants using freshly produced feces were performed at 0, 2, and 4 months. A mixed meal test (for residual beta cell function) and gut microbiome analysis were performed at 0, 2, 6, 9, and 12 months. Plasma metabolites were measured at 0, 6, and 12 months. Biometric measurements and fasting plasma samples were performed at all time points to monitor safety parameters.

[0179] Explanation of clinical trial visits All visits were conducted after an overnight fast, with subjects not taking long-acting insulin the previous night. Blood, fecal, and urine samples, as well as biometric measurements, were collected at each visit. At baseline / 0 months, a nasoduodenal tube was placed. After tube placement, while patients were adequately awake, a standardized 2-hour mixed meal test (Nestlé sustacal boost®) was performed to assess residual beta-cell function, as previously described (Moran et al., 2013). At 2, 9, and 12 months, patients again underwent a mixed meal test for residual beta-cell C-peptide secretion. At that time, a duodenal tube was placed via CORTRAK intestinal access, and the fecal transplant procedure was repeated. At 6 months, a mixed meal test was performed.

[0180] Fecal transplant procedure Subjects were randomly assigned to receive three autologous or allogeneic fecal transplants. All patients and investigators were blinded to treatment assignments. After admission, a duodenal tube was placed via gastroscopy or the CORTRAK intestinal access system. Each patient then underwent complete bowel cleansing with 2-4 L of Klean Prep® (macrogol) through the duodenal tube for approximately 3 hours until the investigator determined the bowel was adequately cleansed (i.e., no solid waste, only clear fluid). 200-300 grams of donor feces were then diluted with 500 mL of 0.9% saline solution and filtered through unfolded cotton gauze. Two hours after the last dose of Klean Prep®, the filtrate was passed through the duodenal tube using a 50 cc syringe in approximately 30 minutes and used for transplantation. After a brief observation period, patients were discharged home.

[0181] Mixed meal test The night before each mixed meal test, T1D patients discontinued their long-acting insulin injections. After an overnight fast, the mixed meal test was performed without morning short-acting insulin administration, with Boost High Protein (Nestlé Nutrition, Vervey, Switzerland) at 6 ml / kg body weight, up to a maximum of 360 ml per subject. Blood samples were subsequently collected at -10, 0, 15, 30, 45, 60, 90, and 120 minutes for stimulated C-peptide. AUC (area under the curve) values ​​were derived according to the trapezoidal rule.

[0182] Plasma metabolites Fasting plasma metabolite measurements were performed by Metabolon (Durham, NC) using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS / MS). Raw data were normalized to account for interday variability. Levels of each metabolite were then rescaled so that the median across all samples equaled 1. Missing values, generally due to sample measurements below the detection limit, were considered the minimum observed value for each metabolite.

[0183] biochemistry Glucose and C-reactive protein (CRP, Roche, Switzerland) were determined in fasting plasma samples. C-peptide was measured by radioimmunoassay (Millipore). Total cholesterol, high-density lipoprotein cholesterol (HDLc), and triglycerides (TG) were determined in EDTA-containing plasma using commercially available enzymatic assays (Randox, Antrim, UK and DiaSys, Germany). All analyses were performed using a Selectra autoanalyzer (Sopachem, The Netherlands). Low-density lipoprotein cholesterol (LDLc) was calculated using the Friedewald formula. Fecal calprotein was determined using a commercially available ELISA (Buhlmann, Switzerland).

[0184] Shotgun sequencing and metagenomic pipeline for fecal samples Fecal microbial communities were analyzed using shotgun sequencing for donor fecal samples and those collected at 0, 6, and 12 months after the start of the clinical trial. For shotgun metagenomics, DNA was extracted from fecal samples and subsequently subjected to shotgun metagenomic sequencing (Clinical Microbiomics, Copenhagen, Denmark). Prior to sequencing, DNA sample quality was assessed using agarose gel electrophoresis, a NanoDrop 2000 spectrophotometer, and quantification using a Qubit 2.0 fluorometer. Genomic DNA was randomly distributed into approximately 350-bp fragments. Fragmented DNA was used for library construction using the NEBNext Ultra Library Prep Kit for Illumina (New England Biolabs). The prepared DNA libraries were evaluated using a Qubit 2.0 fluorometer for quantification and an Agilent 2100 Bioanalyzer for fragment size distribution. Quantitative real-time PCR (qPCR) was used to determine the concentration of the final library before sequencing. Libraries were sequenced on an Illumina HiSeq platform, generating 2 × 150 bp paired-end reads. Raw reads were quality-filtered using Trimmomatic (v0.38), adapter removal, and trimming the first 5 bp. Reads were then quality-trimmed using a 4 bp sliding window and a minimum Q-score of 15. After trimming, reads shorter than 70 bp were discarded. To remove human reads, the remaining paired reads were mapped to the human genome (GRCh37_hg19) using Bowtie2 (v2.3.4.3). Finally, the remaining quality-filtered non-human reads were subsampled to 20 million reads per sample and processed using Metaphlan2 (v2.7.7) to infer metagenomic microbial species composition and Humann2 (v0.11.2) to extract gene counts and functional pathways.Briefly, reads were mapped using Bowtie2 against the microbial pangenome, and unmapped reads were translated and mapped against the full Uniref90 protein database using diamond (v0.8.38). Pathway collection was performed using the MetaCyc database.

[0185] Analysis of small intestinal microbial communities The biopsy sample was placed in a bead-beating tube with 300 μl of fecal transfer and collection (STAR) buffer and 0.25 g of sterile zirconia beads (0.1 mm). Six μl of proteinase K (20 mg / ml; QIAGEN, Venlo, The Netherlands) was added and incubated at 55°C for 1 h. The biopsy sample was then homogenized three times by bead-beating (60 s × 5.5 ms) and then incubated at 1000 rpm for 15 min at 95°C. The sample was then centrifuged at 14,000 g for 5 min at 4°C, and the supernatant was transferred to a sterile tube. The pellet was reprocessed with 200 μl of STAR buffer, and both supernatants were pooled. DNA purification was performed using a custom-made kit (AS1220; Promega) using 250 μl of the final supernatant pool. DNA was eluted in 50 μl of DNAse- and RNAse-free water, and the DNA concentration was measured using a DS-11 FX+ Spectrophotometer / Fluorometer (DeNovix Inc., Wilmington, USA) and a Qubit™ dsDNA BR Assay kit (Thermo Scientific, Landsmeer, The Netherlands). The V5-V6 region of the 16S ribosomal RNA (rRNA) gene was amplified for each sample in a duplex PCR reaction with a total reaction volume of 50 μl. A first-step PCR using the 27F and 1369R primers was used for primary enrichment. The mixture consisted of 1 μl of 10 μM primers, 1 μl of dNTP mix, 0.5 μl of Phusion Green Hot Start II High-Fidelity DNA Polymerase (2 U / μl; Thermo Scientific, Landsmeer, The Netherlands), 10 μl of 5× Phusion Green HF Buffer, and 36.5 μl of DNAse- and RNAse-free water. The amplification program included an initial denaturation step at 98°C for 30 s, followed by five cycles of denaturation at 98°C for 30 s, annealing at 52°C for 40 s, extension at 72°C for 90 s, and a final extension step at 72°C for 7 min.Nested PCR was performed on the PCR products using a master mix containing 1 μl of uniquely barcoded primers, 784F-n and 1064R-n (10 μM each per reaction), 1 μl of dNTP mixture, 0.5 μl of Phusion Green Hot Start II High-Fidelity DNA Polymerase (2 U / μl; Thermo Scientific, Landsmeer, The Netherlands), 10 μl of 5× Phusion Green HF Buffer, and 36.5 μl of DNAse- and RNAse-free water. The amplification program included an initial denaturation step at 98°C for 30 seconds, followed by five cycles of denaturation at 98°C for 10 seconds, annealing at 42°C for 10 seconds, extension at 72°C for 10 seconds, and a final extension step at 72°C for 7 minutes. PCR products (approximately 280 bp) were visualized on a 1% agarose gel and purified using a Clean PCR kit (CleanNA, Alphen aan den Rijn, The Netherlands). The concentration of the purified PCR products was measured using a Qubit dsDNA BR Assay Kit (Invitrogen, California, USA). 200 ng of microbial DNA from each sample was pooled to create a final amplicon library, which was sequenced (150 bp, paired-end) on an Illumina HiSeq. 2500 platform (GATC Biotech, Constance, Germany).

[0186] Raw reads were demultiplexed using the Je software suite (v2.0), which does not allow mismatches in barcodes. After removing barcodes, linkers, and primers, reads were mapped to the human genome using Bowtie 2 to remove human reads. The remaining microbial forward and reverse reads were pipelined separately using DADA2 (Callahan et al., 2016) (v1.12.1). Amplicon sequence variants (AVS) inferred from the reverse reads were converted to their reverse complements and matched against ASVs inferred from the forward reads. Only nonchimeric forward-read ASVs that matched the reverse-complemented reverse-read ASVs were retained. ASV sample counts were inferred from the forward reads. ASV taxonomy was assigned using DADA2 and the SILVA (v132) database. The resulting ASV tables and taxonomy assignments were integrated using the PhyloSec R package (v1.28.0) and diluted to 60,000 counts per sample.

[0187] Power calculations and statistics Using a two-sided test with an α of 0.05 and a 10% dropout rate, a sample size of 17 patients per group (34 patients total) was required to provide 80% power to detect a 50% difference in C-peptide AUC between treatment groups at 12 months (360 mmol / L / min vs. 180 mmol / L / min with a standard deviation of 170). All analyses were based on prespecified intention-to-treat cohorts with known measurements (complete case analysis), and missing values ​​were assumed to be missing at random. The primary endpoint of the trial was the preservation of residual (MMT-stimulated) beta-cell function at 6 and 12 months compared with baseline (month 0). Other secondary endpoints included changes in whole blood leukocyte subsets, parameters of glycemic control, and fasting plasma metabolites for immune markers of autoimmunity over these 12 months. Finally, changes in small intestinal epithelial genes between baseline and 6 months after the initiation of FMT were determined. Analyses were performed by intention-to-treat analysis.

[0188] Baseline differences between groups were evaluated using unpaired Student's t-tests or Mann-Whitney U tests, depending on the distribution of the data. Data are presented as mean ± standard deviation or median and interquartile range. Postprandial results (e.g., C-peptide) are reported as the area under the curve (AUC) for the 2-hour postprandial follow-up, calculated using the trapezoidal method. For correlation analyses, Spearman's rank test was used (because all parameters were nonparametric). For comparison of primary endpoints, a linear mixed model (LMM) was used (lme4 package in R), with "allocation" and "time point" as fixed effects and "patient entry number" as a random effect. p-values ​​for the interaction between "allocation" and "time point" were reported. Additionally, parameters were compared between groups at various time points using the Mann-Whitney U test. A p-value of less than 0.05 was considered statistically significant. The clinical trial was conducted at the Academic Medical Center (Amsterdam) in accordance with the Declaration of Helsinki (updated version of 2013). All participants provided written informed consent, and all study procedures were approved by the Institutional Review Board (IRB) of the Academic Medical Center. The study was prospectively registered in the Dutch Trial Registry (NTR3697).

[0189] Machine learning and follow-up statistical analysis The Extreme Gradient Boosting (XGBoost) machine learning classification algorithm was applied in combination with a stability selection procedure to identify which parameters (as baseline values ​​or relative change) best predicted treatment group and responder / non-responder. This approach was used for duodenal microbial composition (16S rRNA sequencing of biopsy samples), fecal microbial community composition and metabolic pathway abundance, and plasma metabolite levels. The relative change (delta) of each parameter between 0 and 12 months was used to predict treatment group. For duodenal microbiota, the delta between 0 and 6 months was used. To predict responder / non-responder, the baseline value, delta between 0 and 6 months, and delta between 0 and 12 months were used. Each analysis yielded a ranked list of the top 30 species with the most distinctive features. Top parameters were selected from each analysis that accurately (i.e., ROC AUC ≥ 0.8) or moderately (ROC AUC > 0.7) predicted group assignment in more thorough studies, using arbitrary but reasonable cutoff values. This cutoff value generally had a relative importance of approximately 30% or greater. Changes in the selected parameters over time were then visualized (Wilcoxon signed-rank test), and differences between groups were finally examined (Mann-Whitney U test) using Spearman's rank test at each time point used. These parameters were correlated with the primary endpoint and other important parameters thus identified.

[0190] result Patients were randomly assigned to donor FMT (n = 11 subjects) or autologous FMT (n = 10 subjects). One participant withdrew consent after the first study visit. Due to lack of funding, the trial was discontinued after 20 subjects enrolled and completed the study. Seven healthy, lean donors (three of whom were used twice) provided allogeneic gut microbiota transfers to 10 new-onset DM1 patients, and the same donors were used for three consecutive FMTs in each DM1 patient. There were no differences between the two groups at baseline or throughout the follow-up period. There were no serious adverse events or adverse changes in plasma biochemistry in either treatment group.

[0191] Autologous FMT sustains (stimulated) C-peptide levels better than allogeneic FMT. Mean fasting plasma C-peptide at baseline was similar between groups (327 pmol / l ± 89 in the allogeneic group vs. 319 ± 118 in the autologous group, p = 0.86, Student's T-test) but decreased in the allogeneic FMT group compared with the autologous FMT group at 12 months (348 pmol / l ± 115 vs. 202 ± 85, Student's T-test, p value = 0.0049, LMM p = 0.00019). A similar effect was seen in the stimulated C-peptide response AUC, which was similar between groups at baseline (361 ± 154 mmol / l·min in the allogeneic group vs. 355 ± 97 in the autologous group, p = 0.92, Student's T-test), but residual beta-cell function was significantly more preserved at 12 months after autologous FMT (392 ± 124 vs. 248 ± 153 mmol / l·min, Student's T-test, p = 0.033, LMM p = 0.000067). As expected, exogenous insulin treatment reduced HbA1c levels at 12 months in both groups. Despite similar daily exogenous insulin requirements between the allogeneic (0.45 ± SD IU / kg / day) and autologous FMT groups (0.47 IU / kg / day), non-significantly improved glycemic control was observed in the autologous FMT compared with the allogeneic FMT group (HbA1c 53.5 mmol / mol vs. 46, MWU p = 0.19, LMM p = 0.12). Glucose metabolism parameters were determined at 0, 6, and 12 months. Finally, BMI, fecal calprotein, microalbuminuria, lipid profile, and dietary intake (separate assessments of total calories, fat, saturated fat, protein, carbohydrates, and fiber) did not differ at baseline or over the course of the study.

[0192] The therapeutic success of autologous FMT can be predicted by changes in plasma metabolites and microbial community composition. Differences in small intestinal microbial communities between FMT treatment groups The alpha diversity of the small intestinal microbial community was not significantly different between treatment groups at baseline, but was significantly different between the autologous and allogeneic FMT groups at 6 months (p = 0.054), consistent with a significant increase in diversity in the allogeneic FMT group (p = 0.009). When plotted along the vertical axis in a redundancy analysis (RDA plot), the small intestinal microbial community composition clustered differentially between groups at baseline and changed differentially between treatment groups. FMT treatment group assignment was reliably predicted by changes in specific small intestinal microbial strains (AUC ROC 0.89 ± 0.18), including two species, Prevotella and Streptococcus oralis (Figure 1). However, changes at the phylum, family, genus, and species levels did not significantly alter the small intestinal microbial community composition. The relative abundance of all these species decreased after autologous fecal transplantation but increased after allogeneic fecal transplantation. Of note, the relative abundance of Prevotella 1 showed baseline differences between groups (p = 0.033). Delta differed significantly between groups for Prevotella 2 (p = 0.048), but not for Prevotella 1 (p = 0.069) or S. oralis. Furthermore, a significant inverse correlation was observed between the relative abundance of Prevotella 1 and the AUC of stimulated C-peptide (Spearman p = 0.015, rho = -0.55).

[0193] Changes in fasting plasma metabolites during FMT Fasting plasma metabolite levels differed between DM1 and donors and were altered by FMT. Treatment group assignment was reliably predicted by changes in fasting plasma metabolites between months 0 and 12 (ROC AUC 0.79 ± 0.23). The relative importance of the 10 most predictive metabolites is shown in Figure 2A. Of the top three metabolites, 1-myristoyl-2-arachidonoyl-GPC (MA-GPC) (p = 0.02) and 1-arachidonoyl-GPC (A-GPC) (p = 0.02, Mann-Whitney U test) showed between-group differences at 12 months, whereas 1-(1-enyl-palmitoyl)-2-linoleoyl-GPE (EPL-GPE) did not (Figures 2B-D). Furthermore, changes in plasma MA-GPC levels were significantly correlated with changes in fasting C-peptide (p=0.012, Mann-Whitney U test, Figure 2E).

[0194] Changes in fecal microbiota during FMT Fecal microbiota composition differed between Dm1 and healthy donors at baseline and showed differential changes between treatment groups. However, alpha diversity did not differ significantly between FMT treatment groups or between donors and recipients at baseline, 6 months, or 12 months. Some variation was observed at the phylum, family, genus, and species levels between groups. Prediction of group assignment based on taxonomic changes in fecal microbiota between 0 and 12 months showed a moderate receiver operating characteristic curve (ROC) AUC of 0.72 ± 0.24. Desulfovibrio pigelii stood out as the most differentially differentiated species among treatment groups (Figure 3). Prediction of treatment group assignment based on metabolic pathways showed a relatively poor ROC AUC of 0.68 ± 0.27. Interestingly, D. pigelii abundance showed differential changes between treatment groups at 6 months (p = 0.024, MWU) and 12 months (p = 0.023) follow-up (Figures 2G-H). Furthermore, changes in D. pigel were positively correlated with changes in fasting C-peptide (p = 0.009, Figure 2I) and plasma l-arachidonoyl-GPC levels (p = 0.004, Figure 2J). Furthermore, changes in the relative abundance of D. pigel were inversely correlated with changes in the relative abundance of Prevotella 1 (Figure 2K) and Prevotella 2 (Figure 2L). Furthermore, changes in D. pigel were positively correlated with changes in fasting C-peptide (p = 0.009, Figure 2I) and plasma l-arachidonoyl-GPC levels (p = 0.004, Figure 2J). Furthermore, changes in the relative abundance of D. pigel were inversely correlated with changes in the relative abundance of Prevotella 1 (Figure 2K) and Prevotella 2 (Figure 2L).

[0195] Baseline fecal microbiota composition predicts FMT response. Because gut microbiota composition differed between healthy and T1D subjects in various age groups, we also reasoned that FMT, which introduces fecal material into the small intestine, is essentially an intervention in autoimmune disease. Therefore, we conducted a post-hoc analysis to examine responders to FMT compared with non-responders, regardless of treatment group. Specifically, we investigated whether baseline characteristics of T1D patients could predict response to FMT treatment at 12-month follow-up, and which bacterial strains and plasma metabolites were associated with this response. Clinical response was defined as a decline of less than 10% in beta cell function compared to baseline at 12-month follow-up, which is significantly less than the expected 20% natural decline in beta cell function over one year. At 6-month follow-up, i.e., 2 months after the final FMT, 12 of 20 subjects were responders. At 12 months of follow-up, clinical responses were sustained in 10 subjects, three of whom underwent allogeneic FMT and seven of whom underwent autologous FMT (Figures 4A-B). Therefore, we selected responders at 12 months for analysis because the primary endpoint (MMT-stimulated C-peptide) was significantly different at 12 months (but not at 6 months), and because there was less interference from the honeymoon phase at 12 months compared with 6 months. We then used predictive modeling to determine which baseline parameters (their baseline values ​​or delta values ​​from 0 to 12 months) predicted clinical response to FMT.

[0196] Baseline fecal microbiota composition best predicts clinical response in FMT. Baseline fecal microbiota composition highly accurately predicted clinical response during FMT (AUC ROC 0.93 ± 0.14). In this regard, gut levels of Bacteroidetes cacae and Coprococcus catus stood out as the most differential microorganisms (Figure 5), and both were significantly more abundant in responders than in non-responders at baseline (Figures 4C–4D). Among the top 10 most differential gut bacterial strains, Paraprevotella species, Collinsella aerofaciens, Bacteroides aegertii, and Ruminococcus callidus also significantly differed between responders and non-responders at baseline (Figures 6A–6E). A significant (negative) correlation was observed between changes in C. catus abundance and stimulated C-peptide AUC (p = 0.053, r = −0.44, Figure 4E). Changes in fecal microbiota composition predicted response less accurately than baseline composition (AUC ROC 0.76 ± 0.23), suggesting that gut microbiota composition can predict the efficacy of gut microbiota-based treatments in diagnosing T1D. The most distinct species were Bacteroidales bacterium ph8, Actinomyces viscosus, Bacteroides thetaoitaomicron, Streptococcus salivarius, Ruminococcus bromii, and Clostridium leptum. Among them, B. bacterium ph8 (p = 0.015, Mann-Whitney U test) and R. bromii (p = 0.013) were less abundant in responders compared to non-responders, S. salivarius (p = 0.045) was more abundant in responders compared to non-responders, and B. thetaiotaomicron was significantly different at baseline and showed a tendency to decrease in responders.

[0197] Integration of multi-omics analysis in FMT Correlations between parameters found to be significantly affected by FMT were explored. Because responders were found in both treatment groups, correlations were explored first in our pooled dataset (n = 20), then within treatment groups individually and among clinical responders to FMT. In the pooled dataset, we found an intertwined cluster of notable parameters positively and negatively associated with markers of glucose regulation (i.e., C-peptide AUC, fasting C-peptide, and HbA1c). On the one hand, the highly correlated plasma metabolites MA-GPC and A-GPC, which accurately predict preservation of insulin distribution, positively correlate with D. pigelii, which positively correlates with fasting C-peptide. On the other hand, Prevotella 1, Prevotella 2, and S. oralis negatively correlate with glucose regulation and the metabolites MC-GPC and A-GPC. When treatment groups were analyzed separately, preserved beta-cell function (high C-peptide) in the autologous group was characterized by high Coprocococcus catus at baseline followed by a decrease in Ruminococcus bromii. In the allogeneic group, preserved beta-cell function was characterized by a decrease in fecal Roseburia intestinalis (which coincidentally correlates positively with Prevotella 1 and 2). Finally, in clinical responders, preserved beta-cell function was characterized by decreases in duodenal Prevotella 1, Prevotella 2, fecal Coprocococcus catus, and the metabolite EPL-GPE, while D. pigerii increased.

[0198] analysis Here, we report for the first time that FMT can have an effect on residual beta cell function in patients with new-onset T1D. This is consistent with recent observational studies supporting the role of the gut microbiome in T1D subjects. Contrary to expectations, autologous FMT performed better than healthy donor FMT, while even in the allogeneic group, residual beta cell function appeared to decline less than expected without treatment. A compelling explanation would be that the beneficial immunological effects of FMT would be more pronounced and durable if the FMT donor's microbiome better matched the host's immunological status. This is to the extent that the beneficial effects of healthy donor feces are attenuated by (immune) incompatibility. Other observations also suggest an immunomodulatory role for specific plasma metabolites derived from the diet and transformed by the gut microbiome. Although the overall clinical effects of FMT were modest and showed wide variation among subjects with new-onset T1D, the intervention was safe and without side effects. We propose that changes in plasma metabolites, primarily fatty acids and tryptophan derivatives, as a result of altered gut microbial community composition may explain the observed beneficial effects of FMT on residual beta cell function in patients with new-onset T1D.

[0199] Preservation of beta cell function is associated with changes in specific gut microbial strains. D. pigel may attenuate autoimmunity in T1D via plasma l-arachidonoyl-GPC. Predictive modeling demonstrated that baseline fecal microbiota taxonomy and metabolic pathways accurately predicted response at 12 months. However, identified microorganisms (e.g., B. cacae and C. catus) did not correlate with any of our relevant immune parameters, small intestinal genes, or plasma metabolites. This suggests that fecal microbiota composition is a consequence, rather than a cause, of response-related host immunological features. The exception to this was the sulfate-reducing bacterial strain D. pigel, whose beneficial effect may be mediated by its production of hydrogen sulfide. Furthermore, we identified D. pigel as a prominent fecal microbiota predictive of FMT treatment group assignment. Interestingly, this small intestinal bacterial strain was also beneficially associated with altered stimulated C-peptide responses during FMT, with its abundance increasing in the autologous group and in all responders. Interestingly, D. pigel positively correlated with plasma levels of 1-arachidonoyl-GPC, another key metabolite associated with improved C-peptide production (Figure 2J). In conclusion, D. pigel may be a strong candidate for attenuating autoimmunity through the production of A-GPC, for example, through uptake by immune cell dendrites projecting into the intestinal lumen. Interestingly, D. pigel was recently cultured from the human intestinal tract, making it possible to test this bacterial strain in human T1D (Chen et al., 2019, Letters in Applied Microbiology 68(6) pp. 553-561). Other bacterial species in the duodenum that best differentiated between treatment groups were two unnamed Prevotella species and Streptococcus oralis. Subsequently, our exploratory integration of multi-omics analyses shows that these Prevotella species and S. oralis are negatively associated with our key beneficial metabolite, the glycerophospholipid MA-GPC. B. stercolis correlated positively with D. pigel and A-GPC, negatively with S. oralis, but not with C-peptide.Finally, although both strains of Ruminococcus bromii and Roseburia intestinalis are generally considered beneficial microorganisms that grow on a high-fiber diet, produce SCFAs, and promote intestinal integrity, changes in Ruminococcus bromii (autologous FMT group) and Roseburia intestinalis (allogeneic FMT group) were negatively associated with changes in C-peptide.

[0200] conclusion Fecal transplantation of gut-derived microbes into the small and large intestines of T1D patients effectively prolongs residual beta cell function and thereby the honeymoon phase. Furthermore, several novel bacterial strains, including D. pigelii and B. stercoli in the feces and Prevotella species and S. oralis in the duodenum, have been identified as having therapeutic potential. Consequently, increases in plasma metabolites such as 1-myristoyl-2-arachidonoyl-GPC, 1-arachidonoyl-GPC, and 6-bromotryptophan during FMT have been associated with beneficial changes. Example 3

[0201] In this example, the effects of the following compounds were evaluated in cell-based assays: - 6-Bromotryptophan (6-BT) - 1-Arachidonoyl-glycero-phosphocholine (20:0) (A-GPC) - 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphoethanolamine (16:0-18:2 PE) - 1-Myristoyl-2-arachidonoyl-glycero-phosphocholine (MA-GPC)

[0202] Materials and Methods Metabolite preparation and cell culture 6-Bromotryptophan (6-BT) (Alichem) was purchased as a powder and dissolved in DMSO at 50 mM.

[0203] 1-Arachidonoyl-glycero-phosphocholine (20:0) (LysoPC(20:0)) (Avanti Polar) was purchased as a powder and dissolved in PBS at 0.9 mM.

[0204] 1-Palmitoyl-2-linoleoyl-sn-glycero-3-phosphoethanolamine (16:0-18:2 PE) (Avanti Polar) was purchased at 10 mg / ml in chloroform. 200 μl (equivalent to 2 mg) was transferred to a glass tube, and the chloroform was evaporated under a stream of nitrogen to give a clear film, which was subsequently dissolved in PBS to 1 mM.

[0205] 1-Myristoyl-2-arachidonoyl-glycero-phosphocholine (MA-GPC) (Syncom, custom synthesis) was purchased as a powder and dissolved in DMSO at 10 mM.

[0206] All cell types were cultured at 37°C in a 5% CO atmosphere and treated with metabolites for no longer than 24 hours. In control conditions, the appropriate vehicle (DMSO or PBS) was added to the culture medium.

[0207] NF-κB reporter macrophage and luciferase assay Activation of NF-κB signaling was assessed by assaying luciferase activity. RAW264.7 cells stably transfected with the 3×-κB-luc plasmid (a DNA construct containing three NF-κB sites from the Ig κ light chain promoter linked to the gene encoding firefly luciferase) were grown in DMEM medium supplemented with 10% heat-inactivated fetal bovine serum, penicillin (100 U / ml), streptomycin (100 μg / ml), and L-glutamine (2 mM). 0.5 × 10 cells were cultured per well in a 96-well F-bottom plate. 5Cells were seeded at a density of 1 / 3 and stimulated with LPS (10 / 100 ng / ml) for 2 hours the following day with or without various concentrations of metabolites (6-BT 0.1–100 μM, LysoPC (20:0) 1–10 μM, 16:0–18:2 PE 1–50 μM, MA-GPC 1–100 μM). Cells were then lysed with 25 μl / well of 1x passive lysis buffer, and firefly luciferase activity was measured using the Luciferase® Assay System (Promega, E1500) on a GloMax® Multi Detection System (Promega).

[0208] In vitro stimulation of primary monocytes Naive bone marrow monocytes were isolated from BM cells. Briefly, hind limbs were isolated from three mice and cleared of surrounding muscle. The femurs and tibias were cut at their distal ends, and the BM contents were flushed with cold PBS using a 10 ml syringe and a 25 G needle and filtered through a 40 μm strainer. Red blood cells (RBCs) were lysed with 1x RBC lysis buffer (Biolegend) for 5 minutes on ice. CD11b+ monocytes were further purified by positive selection using a cocktail of CD11b magnetic beads (Miltenyi Biotec, #130-049-601) and a magnetized MS column (Miltenyi Biotec) according to the manufacturer's instructions. Subsequently, 1 × 10 cells were plated per well in an F-bottom 96-well plate in RPMI 1640 supplemented with 10% heat-inactivated fetal bovine serum, penicillin (100 U / ml), streptomycin (100 μg / ml), L-glutamine (2 mM), and activated with 10 μg / ml poly(I:C). 5 Monocytes were seeded with cells. Monocytes were activated with 10 ng / ml LPS (Sigma-Aldrich) and treated with 6-BT (10 / 100 μM), LysoPC (20:0) (10 / 50 μM), MA-GPC (10 / 50 / 100 μM), or the appropriate vehicle. Cells were maintained in a final volume of 200 μl / well for 24 h, after which the supernatants were harvested and stored at -80°C.

[0209] In vitro stimulation of mouse macrophages / dendritic cells (DCs) Bone marrow-derived macrophages (BMDMs) and DCs (BMDCs) were obtained by differentiation of freshly isolated BM cells (as described above) from the femurs and tibias of C57 / Bl6 mice (n=3 per experiment). For BMDMs, 3 × 10 cells were cultured per 10 cm dish. 6 BM cells were seeded and cultured for 7 days in 12 ml of RPMI 1640 medium containing 20% ​​fetal bovine serum and 30% L-929 cell-conditioned medium as a source of mouse macrophage colony-stimulating factor (M-CSF). BMDCs were cultured at 0.5 × 10 cells per 25 ml in a 10 cm dish. 6 BM cells were seeded at 1 × 10 cells / ml and cultured in 5% FBS-RPMI 1640 medium in the presence of GM-CSF for 7 days. After differentiation, BMDM / DCs were harvested, counted, and plated at 1 × 10 cells / ml per well in an F-bottom 96-well plate. 5 Cells were seeded at a density of 100 μM and allowed to adhere for 20 h before experiments were performed. Macrophages were activated with 10 μg / ml polyinosinic-polycytidylic acid (poly(I:C)) (InvivoGen) in the presence or absence of 6-BT (10 / 100 μM), LysoPC (20:0) (10 / 50 μM), or MA-GPC (10 / 50 / 100 μM) for 24 h in a final volume of 200 μl / well. Supernatants were harvested at the end of the assay and stored at -80°C.

[0210] In vitro CD4+ T cell activation assay Primary CD4+ T cells were freshly isolated from the spleens of C57 / Bl6 mice (n=3 per experiment) by negative selection. Briefly, spleens were crushed in a culture dish and passed twice through strainers (70 μm and 40 μm, respectively) to obtain a single-cell suspension. After lysing red blood cells with 1x RBC lysis buffer (Biolegend) (on ice for 10 min), cells were counted and stained with a cocktail of biotin-conjugated antibodies against CD8a, CD11b, CD11c, CD19, CD45R (B220), CD49b (DX5), CD105, anti-anti-MHC class II, Ter-119, and TCRγ / δ, followed by magnetic labeling with anti-biotin microbeads using negative selection (CD4+ T Cell Isolation Kit, Miltenyi Biotec, #130-104-454). Non-CD4 T cells were depleted by retention on LS magnetic columns (Miltenyi Biotec).

[0211] Isolated CD4 T cells were cultured in 96-well plates (1 × 10 per well). 5 Immediately after seeding for 24 h in 200 μl / well of complete RPMI 1640 medium, cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 U / ml penicillin, 100 μg / ml streptomycin, and 2 mM L-glutamine. Treatment with metabolites 6-BT (1 / 10 μM), LysoPC(20:0) (10 / 50 μM), or MA-GPC (10 / 50 / 100 μM) and activation with 2.5 μg / ml soluble anti-CD3 antibody (145-2C11, eBioscience) and 1 μg / ml soluble anti-CD28 antibody (37.51, eBioscience) were initiated. At the end of the treatment, supernatants were harvested and stored at −80°C.

[0212] In vitro stimulation of human monocytes Mononuclear cell fractions were isolated from the blood of healthy volunteers (Sanquin Bloodbank, Amsterdam, The Netherlands) by density centrifugation using Lymphoprep™ (Axis-Shield) and human CD14 magnetic beads and MACS® cell separation columns (Miltenyi Biotec) according to the manufacturer's instructions. Isolated primary human monocytes were counted and plated at 1 × 10 in 24-well plates with 1 ml of medium supplemented with 10% fetal bovine serum, 100 U / ml penicillin, 100 μg / ml streptomycin, and 2 mM L-glutamine. 6 Cells were seeded at 1000 x g / well. After seeding, cells were stimulated with 10 ng / ml LPS or 25 mM D-glucose (Sigma-Aldrich) for 24 h, with or without 100 μM 6-BT. Cells were then lysed with Tripure isolation reagent (Roche) and stored at -80°C until RNA isolation.

[0213] In vitro stimulation of pancreatic beta cells INS1E cells (rat pancreatic beta cell line) were maintained in complete RPMI 1640 supplemented with 5% fetal bovine serum, 2 mM L-glutamine, 5 μM beta-mercaptoethanol, 1 mM sodium pyruvate, 10 mM HEPES, 100 units / ml penicillin, and 100 μg / ml streptomycin at 1 × 10 per well in a 48-well plate. 5 INSIE cells were seeded at a density of 1000 ng / well and allowed to rest for 1 day. The medium was then replaced with 0.5 ml / well of INSIE complete RPMI 1640 medium containing vehicle or the metabolites 6-BT (1 / 10 / 25 μM), LysoPC (20:0) (5 / 10 / 50 μM), or MA-GPC (10 / 50 / 100 μM). After 24 h, cells and supernatants were harvested and stored at -80°C for further analysis (gene expression and insulin secretion, respectively). Cells were lysed with Tripure Isolation Reagent (Roche) prior to storage.

[0214] For glucose-stimulated insulin secretion (GSIS) assays, cells were preincubated in Krebs-Ringer bicarbonate buffer (KRB) [115 mM NaCl, 5 mM KCl, 2.56 mM CaCl, 1 mM MgCl, 10 mM NaHCO, 15 mM HEPES, and 0.3% BSA (pH 7.4)] for 30 min at 37°C, followed by stimulation with 1 mM glucose in KRB (0.5 ml / well) for 1 h and 22 mM glucose in KRB (0.5 ml / well) for an additional 1 h at 37°C. After treatment with 1 mM glucose (Sigma-Aldrich) and 22 mM glucose, supernatants were collected and cells were stored at -80°C. GSIS was performed on beta cells after 24 h of treatment with 10 μM 6-BT.

[0215] ELISA The concentrations of TNFα, IFNβ, and IFNγ were measured in the cell supernatants of mouse monocytes, macrophages, and T cells, respectively, using specific ELISAs (R&D Systems) according to the manufacturer's instructions. The concentration of insulin after 24 h of treatment with metabolites or after GSIS was determined using a rat insulin ELISA (Mercodia) according to the manufacturer's instructions. GSIS was calculated by subtracting the concentration of insulin at 1 mM glucose from the percentage of insulin at 22 mM glucose.

[0216] Gene expression analysis Total RNA was extracted from cell lysates using Tripure Isolation Reagent. RNA was converted to cDNA using the iScript kit (BioRad). Quantitative polymerase chain reaction (qPCR) was performed using SYBR Green-SensiMix (Bioline) on a CFX384 Touch Real-Time PCR Detection System (BioRad). Gene expression was calculated as the fold change compared to the control (unstimulated condition) using the delta-delta Ct method.

[0217] statistics Statistical analysis was performed using Student's t-test for two-group comparisons and one-way ANOVA and Dunnett's test for multiple-group comparisons. Data are expressed as the mean and standard error of the mean (SEM). P<0.05 was considered significant.

[0218] result The results are shown in Figures 7 to 13. - 6-BT, A-GPC, and MA-GPC can inhibit the activation of the NFκB pathway in macrophages at different doses (Figure 7). - 6-BT, A-GPC, and MA-GPC abolish cytokine secretion by monocytes (Figure 8). - 6-BT and MA-GPC impair type 1 IFN secretion (Figure 9). - 6-BT reduces cytokine production by human monocytes (Figure 10). - 6-BT and A-GPC attenuate Th1 responses in CD4 T cells (Figure 11). - 6-BT enhances pancreatic beta cell function (Figures 12 and 13).

[0219] Of particular interest is 6-bromotryptophan, which inhibits activation of the NFκB pathway, disrupting immune responses in monocytes / macrophages and CD4 T cells and improving pancreatic beta cell function. MA-GPC inhibits activation of the NFκB pathway, disrupting immune responses in monocytes / macrophages and CD4 T cells. Example 4

[0220] Plasma 6-bromotryptophan levels are inversely correlated with the presence of type 2 diabetes and glycemic control in a cross-sectional cohort (n=369 subjects). In a cohort of 369 subjects, evidence was found that 6-bromotryptophan may protect against the onset and progression of type 2 diabetes. More specifically, plasma 6-bromotryptophan levels were found to be inversely correlated with the presence of type 2 diabetes and glycemic control. This suggests that 6-BT may contribute to the prevention and treatment of type 2 diabetes and, therefore, the improvement of cardiovascular complications (both microvascular and macrovascular) in type 2 diabetes. 6-BT may also help reduce macrovascular disease (i.e., cardiovascular disease) and microvascular complications in non-type 2 diabetic patients.

[0221] Materials and Methods Targeted metabolite measurements in fasting plasma were performed by Metabolon (Durham, NC) using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS / MS) as previously described [Koh A, Molinaro A, Stahlman M, et al., "Microbially Produced Imidazole Propionate Impairs Insulin Signaling through mTORC1," Cell 2018;175:947-961, e17. doi:10.1016 / j.cell.2018.09.055].

[0222] result The results are shown in Table 2 and Table 3.

[0223] [Table 2]

[0224] [Table 3]

[0225] 6-Bromotryptophan halts the inflammatory response in myeloid cells. 6-BT is a bromoindole derivative of tryptophan and is known to be metabolized by indigenous gut microorganisms in the colon and small intestine. To date, the physiological role of 6-BT remains unknown. Our clinical findings suggest a protective role against inflammation and diabetes.

[0226] Through a series of in vitro / ex vivo experiments, the inventors elucidated some of the functions of 6-BT on immune cells as well as insulin-producing pancreatic beta cells.

[0227] 6-BT can inhibit the secretion of the pro-inflammatory cytokine TNFα through the engagement of TLR4 and TLR2, and the secretion of IFN-beta through the activation of TLR3. Here, monocytes isolated from mouse bone marrow (Christ A, "Western Diet Triggers NLRP3-Dependent Innate Immune Reprogramming," Cell 2018) or bone marrow-derived macrophages (Swanson KV, "A noncanonical function of cGAMP in inflammasome priming and activation," JEM 2017) were exposed to the indicated concentrations (10-100 μM) of 6-BT for 24 hours in the presence or absence of 10 ng / ml LPS, 10 μg / ml P3C, or 10 μg / ml poly(I:C). Using ELISA assays, we found that 6-BT inhibited the secretion of the pro-inflammatory cytokine TNFα via TLR4 and TLR2 engagement and IFN-beta via TLR3 activation. See Figure 14.

[0228] Inhibition of cytokine secretion induced by TLR signaling is particularly important for therapeutic approaches to inflammatory and infectious diseases in which damage is driven by an excessive inflammatory response (e.g., sepsis and systemic inflammatory response syndrome (SIRS)).

[0229] 6-BT inhibits the secretion of the pro-inflammatory cytokines TNFα and IFN-beta. Because dendritic cells (DCs) are the primary antigen-presenting cells important in T cell activation, we next examined the effect of 6-BT on mouse DCs differentiated with bone marrow cells and GM-CSF (40 ng / ml). For monocytes / macrophages, 6-BT inhibited the secretion of the pro-inflammatory cytokines TNFα and IFNβ by DCs after activation of TLR4 (by 100 ng / ml LPS) or TLR3 (by 10 μg / ml poly(I:C)), respectively. See Figure 15.

[0230] 6-BT significantly reduced the production of the Th1 cytokine IFN-gamma. Particularly with regard to autoimmune diabetes, T cell activation drives the onset and progression of the disease. Therefore, we further investigated the effects of 6-BT on CD4 T cells. To mimic antigen presentation, mouse CD4 T cells (isolated from the pancreas, Uchimura T, "The Innate Immune Sensor NLRC3 Acts as a Rheostat that Fine-Tunes T Cell Responses in Infection and Autoimmunity," Immunity 2019) were activated with monoclonal antibodies against CD3 and CD28 (2.5 and 1 μg / ml, respectively). Consistent with findings in myeloid cells, 6-BT significantly reduced the production of the Th1 cytokine IFN-gamma. See Figure 16.

[0231] 6-BT stimulates beta cell differentiation and insulin production. Given the positive correlation between plasma 6-BT levels and C-peptide concentrations (found in clinical studies), we next investigated whether 6-BT could exert a direct effect on beta cells. Indeed, we now find that 6-BT induces gene expression of the transcription factors PDX1 and MAFA in IS1E beta cells, which are important for beta cell maturation and functionality. Consistently, 6-BT also promotes insulin secretion during steady-state and glucose-stimulated insulin secretion (data shown as the difference between insulin release under starvation [1 mM glucose] and hyperglycemic [22 mM] conditions) (Paula S, "Exercise increases pancreatic beta-cell viability in a model of type 1 diabetes through IL-6 signaling," FASEB J 2015). See Figure 17.

[0232] Mechanism of action of 6-bromotryptophan To explore the molecular mechanisms underlying the action of 6-BT, we first examined the effect of 6-BT on the activation of the NF-kB pathway, a central pathway in all inflammatory diseases (not just autoimmune diseases). To this end, we quantified the expression of the phosphorylated form of the p65 subunit, which is considered a marker of NF-kB activation. Upon T cell activation with PMA (50 ng / ml) and ionomycin (1 μg / ml), 6-BT was able to inhibit NF-kB signaling at a very early time point (5–10 min after activation). This effect was observed in both mouse and human (Jurkat) CD4 T cells. See Figure 18.

[0233] 6-BT inhibits the activation of NFkB in macrophages. Similar to what we observed in lymphocytes, 6-BT inhibits NFkB activation in macrophages. Using the RAW264.7 murine macrophage cell line stably expressing an NFkB luciferase reporter (Groeneweg M, "Lipopolysaccharide-induced gene expression in murine macrophages is enhanced by prior exposure to oxLDL," J Lipid Res 2006), we demonstrated that overnight exposure of macrophages to 6-BT (10-200 μM) dose-dependently inhibited the transcriptional activity of the NFkB complex after 2 hours of stimulation with LPS (10 ng / ml). See Figure 19.

[0234] 6-BT and tryptophan induce distinct biological activities. Next, we investigated whether the effects of 6-BT were specific or whether they were also mediated by tryptophan. In mouse CD4 T lymphocytes (isolated from mouse pancreas, Uchimura T, "The Innate Immune Sensor NLRC3 Acts as a Rheostat that Fine-Tunes T Cell Responses in Infection and Autoimmunity," Immunity 2019), 6-BT exerted an inhibitory effect on CD3 / CD28-mediated IFN-gamma production, whereas tryptophan did not. This indicates that 6-BT and tryptophan induce distinct biological activities. (See Figure 20.)

[0235] Consistent with the results in DCs, exposure of monocytes (isolated from mouse bone marrow, Christ A, "Western Diet Triggers NLRP3-Dependent Innate Immune Reprogramming," Cell 2018) to 6-BT or tryptophan indicates that the anti-inflammatory effect is specific to the 6-bromotryptophan molecule, but not to tryptophan. See Figure 21.

[0236] 6-BT affects intracellular metabolism. Finally, we found that 6-BT also affects intracellular metabolism. Specifically, 6-BT (100 μM) was found to promote mitochondrial metabolism in mouse and human (Jurkat) CD4 T cells. See Figure 22.

[0237] OCR is the oxygen consumption rate used as a surrogate for cellular utilization of mitochondrial oxidative phosphorylation. OCR was measured using a Seahorse XF Analyzer. Uchimura T, "The Innate Immune Sensor NLRC3 Acts as a Rheostat that Fine-Tunes T Cell Responses in Infection and Autoimmunity," Immunity 2019; Chou, "AIM2 in regulatory T cells restrains autoimmune diseases," Nature 2021.

[0238] 6-BT enhances mitochondrial metabolism. Similarly, exposure to 6-BT can enhance mitochondrial metabolism in pro-inflammatory M1 macrophages (differentiated in the presence of LPS and IFN-gamma (Cheng et al. JCI Insight. 2018;3(22):e120638)) without affecting glycolytic flux. Intracellular metabolism was measured using a Seahorse XF Analyzer (Uchimura T, "The Innate Immune Sensor NLRC3 Acts as a Rheostat that Fine-Tunes T Cell Responses in Infection and Autoimmunity," Immunity 2019; Chou, "AIM2 in Regulatory T Cells Restrains Autoimmune Diseases," Nature 2021). See Figure 23.

[0239] 6-BT was able to rescue beta cell dysfunction in both type 1 and type 2 diabetes. Finally, we investigated whether 6-BT could affect mitochondrial metabolism in beta cells. This relies on mitochondrial metabolic production for the exocytosis of ATP and insulin. 6-BT increased mitochondrial metabolism in beta cells (INS1E beta cells) both in the steady state and under high glucose conditions (25 mM glucose). Furthermore, we examined the effect of tryptophan on intracellular metabolism and found that tryptophan exerted different effects than 6-BT on inflammatory markers. Intracellular metabolism was measured using a Seahorse XF Analyzer (Uchimura T, "The Innate Immune Sensor NLRC3 Acts as a Rheostat that Fine-Tunes T Cell Responses in Infection and Autoimmunity," Immunity 2019; Chou, "AIM2 in Regulatory T Cells Restrains Autoimmune Diseases," Nature 2021).

[0240] Importantly, defects in mitochondrial and oxidative metabolism have been reported in T2D (Haythorne, Nature Communications, Vol. 10, Paper No.: 2474 (2019)), suggesting that 6-BT may rescue beta cell dysfunction in both type 1 and type 2 diabetes. Furthermore, increased utilization of mitochondrial metabolism may resist ectopic intracellular lipid accumulation, thereby combating obesity.

[0241] in conclusion, 6-BT exerts pleiotropic effects on multiple cell types. 6-BT has anti-inflammatory effects on myeloid and lymphoid cells. 6-BT stimulates insulin secretion by beta cells.

[0242] Mechanistically, the biological effects of 6-BT are distinct from those of tryptophan. 6-BT does not act through activation of the AhR, but inhibits NFkB activation and enhances mitochondrial metabolism, which is typically utilized by cells with an anti-inflammatory phenotype.

[0243] application Due to its broad effects on multiple cell types, its inhibitory action on NFkB signaling, and its promotion and adaptation of mitochondrial metabolism, 6-BT may be a novel therapeutic option not only for type 1 and type 2 diabetes, but also for many other inflammation-related disorders such as sepsis, systemic inflammatory response syndrome (SIRS), and cardiovascular disease. Example 5

[0244] Desulfovibrio levels are inversely associated with the presence of type 2 diabetes and glycemic control in a cross-sectional cohort (n=369 subjects). In the same cohort of 369 subjects previously discussed herein, evidence was found that bacteria belonging to the genus Desulfovibrio (e.g., Desulfovibrio pigel) may confer protection against the onset and progression of type 2 diabetes. More specifically, the relative abundance of fecal bacteria belonging to the genus Desulfovibrio was found to be inversely correlated with the presence of type 2 diabetes and glycemic control. This suggests that administration of Desulfovibrio may contribute to the prevention and treatment of type 2 diabetes and thus to the improvement of cardiovascular complications (both microvascular and macrovascular) in type 2 diabetes. Desulfovibrio may also help reduce macrovascular disease (i.e., cardiovascular disease) and microvascular complications in non-type 2 diabetic patients.

[0245] Materials and Methods Targeted metabolite measurements in fasting plasma were performed by Metabolon (Durham, NC) using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS / MS) as previously described [Koh A, Molinaro A, Stahlman M, et al., "Microbially Produced Imidazole Propionate Impairs Insulin Signaling through mTORC1," Cell 2018;175:947-961, e17. doi:10.1016 / j.cell.2018.09.055].

[0246] result The results are shown in Table 4 and in Figures 25 and 26.

[0247] [Table 4]

[0248] Furthermore, a clear association was observed between the relative abundance of Desulfovibrio in feces and plasma 6BT levels in type 2 diabetic patients. See Figure 27.

Claims

1. A composition comprising 6-bromotryptophan for use in the prevention or treatment of an inflammation-related disease selected from the group consisting of diabetes, autoimmune diseases, inflammatory diseases, and cardiovascular diseases, wherein when the use is for the prevention or treatment of diabetes or autoimmune diseases, 6-bromotryptophan is not present in the feces.

2. 2. The composition of claim 1, wherein the diabetes is selected from type 1 diabetes and type 2 diabetes.

3. 2. The composition of claim 1, wherein the autoimmune disease is selected from the group consisting of type 1 diabetes, Hashimoto's disease, Graves' disease, Addison's disease, psoriasis, vitiligo, rheumatoid arthritis, Bechteleu's disease, celiac disease, inflammatory bowel disease, asthma, chronic obstructive pulmonary disease (COPD), Addison's disease, vasculitis, multiple sclerosis (MS), chronic inflammatory demyelinating polyneuropathy (CDIP), and Guillain-Barré syndrome (GBS).

4. 2. The composition of claim 1, wherein the inflammatory disease is selected from the group consisting of cardiovascular inflammation, carditis, endocarditis, myocarditis, pericarditis, vasculitis, arteritis, phlebitis, capillaritis, gastrointestinal inflammation, esophagitis, gastritis, gastroenteritis, enteritis, colitis, enterocolitis, duodenitis, ileitis, caecitis, appendicitis, proctitis, liver inflammation, lung inflammation, skeletal inflammation, systemic inflammatory response syndrome (SIRS), and sepsis.

5. 2. The composition of claim 1, wherein the cardiovascular disease is selected from coronary artery disease, peripheral artery disease, cerebrovascular disease, atherosclerosis, stenosis, renal artery stenosis, aortic disease, aortic aneurysm, cardiomyopathy, hypertensive heart disease, hypertension, heart failure, pulmonary heart disease, arrhythmia, cardiovascular inflammation, inflammatory heart disease, endocarditis, inflammatory cardiac hypertrophy, myocarditis, eosinophilic myocarditis, valvular heart disease, congenital heart disease, and rheumatic heart disease.

6. 6. The composition of any one of claims 1 to 5, in combination with a tumor necrosis factor alpha (TNFα) inhibitor, preferably selected from the group consisting of infliximab, adalimumab, certolizumab pegol, and golimumab.

7. 7. The composition of any one of claims 1 to 6, in combination with bacteria of the genera Eubacterium, Intestinimonas, Bifidobacteria, Lactobacillales and / or Ackermansia, preferably selected from the group consisting of Bifidobacterium animalis subsp. lactis or Bifidobacterium breve, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Eubacterium hallii, Intestinimonas butyriciproduscens and / or Ackermansia muciniphila.

8. 8. The composition according to any one of claims 1 to 7, which is administered enterally, preferably orally or nasally, or by subcutaneous, intravenous, rectal administration and / or nasoduodenal tube administration.

9. 9. The composition according to any one of claims 1 to 8, wherein the use comprises administering the composition to the small intestine, preferably the duodenum.

10. A composition according to any one of claims 1 to 9, which is preferably a pharmaceutical composition, more preferably a liquid or solid dosage form, most preferably a capsule, tablet or powder.

11. The composition of claim 10, comprising at least 1, 5, 10, 25, 50, or 100 mg of 6-bromotryptophan.

12. 12. A composition according to any one of claims 1 to 11, which is contained within and / or encapsulated by an enteric coating, preferably such that the enteric coating does not dissolve and / or disintegrate in the gastric environment.

13. 13. The composition of any one of claims 1 to 12, wherein the use comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10 separate administrations of the composition, preferably with an interval of at least 1, 2, 3, 4, 5, 6, 7, 8 weeks between said separate administrations.

14. The composition according to any one of claims 1 to 13, wherein the subject to be treated is a mammal, preferably a human.

Images