Amuc-1100 polypeptide variants for effecting immune signalling and/or affecting intestinal barrier function and/or modulating metabolic status

A composition comprising an isolated polypeptide and a pharmaceutically or alimentary acceptable carrier, wherein the isolated polypeptide

• a) has at least 30% sequence identity with SEQ ID NO:9;
• and
• b) comprises at least one of the following sets of amino acid residues
• i. R, S, I, S, A, and P or conservative substitutions thereof at positions that correspond to positions 1, 2, 8, 20, 23, and 27, respectively, in SEQ ID NO:9;
• ii. C, K, K, I, and T or conservative substitutions thereof at positions that correspond to positions 92, 93, 95, 97, and 100, respectively, in SEQ ID NO:9;
• iii. W, L, G, and F or conservative substitutions thereof at positions that correspond to positions 105, 106, 107, and 108, respectively, in SEQ ID NO:9;
• iv. F and E or conservative substitutions thereof at positions that correspond to positions 126 and 127, respectively, in SEQ ID NO:9;
• v. V, Y, and R or conservative substitutions thereof at positions that correspond to positions 149, 150, and 151, respectively, in SEQ ID NO:9;
• vi. P, E, I, F, Q, R, S, and V or conservative substitutions thereof at positions that correspond to positions 179, 181, 182, 184, 185, 188, 190, and 191, respectively, in SEQ ID NO:9;
• and
• vii. P, P, P, A, A, P, G, T, A, E, A, P, Q, K, G, and E or conservative substitutions thereof at positions that correspond to positions 220, 222, 229, 230, 231, 234, 248, 258, 260, 262, 264, 172, 175, 279, 283, and 285, respectively, in SEQ ID NO:9,
• wherein the polypeptide affects immune signaling, affects intestinal barrier function, affects glucose homeostasis, cholesterol homeostasis, and/or triglyceride homeostasis.

A genetically modified host cell comprising a nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide

• a) having at least 30% sequence identity with SEQ ID NO:9;
• and
• b) comprising at least one of the following sets of amino acid residues
• i. R, S, I, S, A, and P or conservative substitutions thereof at positions that correspond to positions 1, 2, 8, 20, 23, and 27, respectively, in SEQ ID NO:9;
• ii. C, K, K, I, and T or conservative substitutions thereof at positions that correspond to positions 92, 93, 95, 97, and 100, respectively, in SEQ ID NO:9;
• iii. W, L, G, and F or conservative substitutions thereof at positions that correspond to positions 105, 106, 107, and 108, respectively, in SEQ ID NO:9;
• iv. F and E or conservative substitutions thereof at positions that correspond to positions 126 and 127, respectively, in SEQ ID NO:9;
• v. V, Y, and R or conservative substitutions thereof at positions that correspond to positions 149, 150, and 151, respectively, in SEQ ID NO:9;
• vi. P, E, I, F, Q, R, S, and V or conservative substitutions thereof at positions that correspond to positions 179, 181, 182, 184, 185, 188, 190, and 191, respectively, in SEQ ID NO:9;
• and
• vii. P, P, P, A, A, P, G, T, A, E, A, P, Q, K, G, and E or conservative substitutions thereof at positions that correspond to positions 220, 222, 229, 230, 231, 234, 248, 258, 260, 262, 264, 172, 175, 279, 283, and 285, respectively, in SEQ ID NO:9,
• wherein the polypeptide affects immune signaling, affects intestinal barrier function, affects glucose homeostasis, cholesterol homeostasis, and/or triglyceride homeostasis.

A method for treating a subject to treat and/or preventing a disorder selected from the group consisting of obesity, metabolic syndrome, insulin-deficiency or insulin-resistance related disorders, type 2 diabetes, type 1 diabetes, gestational diabetes, preeclampsia, inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), glucose intolerance, abnormal lipid metabolism, atherosclerosis, hypertension, cardiac pathology, stroke, non-alcoholic fatty liver disease, alcoholic fatty liver disease, hyperglycemia, hepatic steatosis, dyslipidemias, dysfunction of the immune system associated with obesity (weight gain), allergy, asthma, autism, Parkinson's disease, multiple sclerosis, neurodegenerative diseases, depression, other diseases related to compromised barrier function, wound healing, behavioral disorders, alcohol dependence, cardiovascular diseases, high cholesterol, elevated triglycerides, atherosclerosis, sleep apnea, osteoarthritis, gallbladder disease, cancer, and conditions altering the physical integrity of the gut mucosal barrier such as food allergies, immaturity of the gut, the method comprising:

• administering to a mammal in need thereof an effective amount of a polypeptide
• a) having at least 30% sequence identity with SEQ ID NO:9;
• and
• b) comprising at least one of the following sets of amino acid residues
• i. R, S, I, S, A, and P or conservative substitutions thereof at positions that correspond to positions 1, 2, 8, 20, 23, and 27, respectively, in SEQ ID NO:9;
• ii. C, K, K, I, and T or conservative substitutions thereof at positions that correspond to positions 92, 93, 95, 97, and 100, respectively, in SEQ ID NO:9;
• iii. W, L, G, and F or conservative substitutions thereof at positions that correspond to positions 105, 106, 107, and 108, respectively, in SEQ ID NO:9;
• iv. F and E or conservative substitutions thereof at positions that correspond to positions 126 and 127, respectively, in SEQ ID NO:9;
• v. V, Y, and R or conservative substitutions thereof at positions that correspond to positions 149, 150, and 151, respectively, in SEQ ID NO:9;
• vi. P, E, I, F, Q, R, S, and V or conservative substitutions thereof at positions that correspond to positions 179, 181, 182, 184, 185, 188, 190, and 191, respectively, in SEQ ID NO:9;
• and
• vii. P, P, P, A, A, P, G, T, A, E, A, P, Q, K, G, and E or conservative substitutions thereof at positions that correspond to positions 220, 222, 229, 230, 231, 234, 248, 258, 260, 262, 264, 172, 175, 279, 283, and 285, respectively, in SEQ ID NO:9,
• wherein the polypeptide affects immune signaling, affects intestinal barrier function, affects glucose homeostasis, cholesterol homeostasis, and/or triglyceride homeostasis.

A method for producing a polypeptide

• a) having at least 30% sequence identity with SEQ ID NO:9;
• and
• b) comprising at least one of the following sets of amino acid residues
• i. R, S, I, S, A, and P or conservative substitutions thereof at positions that correspond to positions 1, 2, 8, 20, 23, and 27, respectively, in SEQ ID NO:9;
• ii. C, K, K, I, and T or conservative substitutions thereof at positions that correspond to positions 92, 93, 95, 97, and 100, respectively, in SEQ ID NO:9;
• iii. W, L, G, and F or conservative substitutions thereof at positions that correspond to positions 105, 106, 107, and 108, respectively, in SEQ ID NO:9;
• iv. F and E or conservative substitutions thereof at positions that correspond to positions 126 and 127, respectively, in SEQ ID NO:9;
• v. V, Y, and R or conservative substitutions thereof at positions that correspond to positions 149, 150, and 151, respectively, in SEQ ID NO:9;
• vi. P, E, I, F, Q, R, S, and V or conservative substitutions thereof at positions that correspond to positions 179, 181, 182, 184, 185, 188, 190, and 191, respectively, in SEQ ID NO:9;
• vii. P, P, P, A, A, P, G, T, A, E, A, P, Q, K, G, and E or conservative substitutions thereof at positions that correspond to positions 220, 222, 229, 230, 231, 234, 248, 258, 260, 262, 264, 172, 175, 279, 283, and 285, respectively, in SEQ ID NO:9,
• wherein the polypeptide affects immune signaling, affects intestinal barrier function, affects glucose homeostasis, cholesterol homeostasis, and/or triglyceride homeostasis, the method comprising the steps of:
• culturing bacteria of the species Akkermansia muciniphila or Akkermansia glycaniphila in a suitable culture medium;
• and
• optionally, isolating the polypeptide produced in step (a).