Lipids That Increase Insulin Sensitivity And Methods Of Using The Same

Abstract

The invention provides, inter alia, fatty acyl hydroxy fatty acid (FAHFA; a novel class of estolide-related molecules) and diagnostic and treatment methods for a variety of disorders—including diabetes-related disorders, Metabolic Syndrome, polycyctic ovarian syndrome, cancer, and inflammatory disorders using them; as well as methods of screening for additional compounds that are useful in treating these disorders and/or that modulate FAHFA levels, FAHFA-mediated signaling, and FAHFA-mediated biological effects.

1 . An isolated fatty acyl hydroxy fatty acid (FAHFA) of formula (I): 
       
         
           
           
               
               
           
         
         Wherein: m is an integer from 0 to 21; 
         n is an integer from 0 to 21; 
         the sum of m and n is an integer from 11 to 21; 
         R 1  is an alkyl group; 
         or a salt thereof. 
       
     
     
         2 . The FAHFA of  claim 1 , wherein R 1  is a (C 15 -C 17 )alkyl group. 
     
     
         3 . The FAHFA of  claim 1 , wherein:
 m is 8;   n is 7; and   R 1  is C 15 H 31  or C 17 H 33 .   
     
     
         4 . The FAHFA of  claim 1 , wherein:
 m is 12;   n is 3; and   R 1  is a C 15 H 31 .   
     
     
         5 . A FAHFA of  claim 1 , wherein the FAHFA is detectably labeled. 
     
     
         6 . A FAHFA of  claim 5 , wherein the FAHFA is isotopically labeled and/or ester- or amide-bound to a detectable moiety, selected from biotin, streptavidin, GST, a fluorous affinity tag, an alkyne suitable for click chemistry, an epitope tag such as FLAG, 6× His, or another affinity tag. 
     
     
         7 . A method of assessing the disease state and/or treatment response of a mammalian subject for a disease or disorder selected from obesity, type 2 diabetes (T2D), impaired glucose tolerance, maturity onset diabetes of the young (MODY), impaired fasting glucose, metabolic syndrome, insulin resistance, polycystic ovarian syndrome, gestational diabetes, cardiovascular disease, inflammatory disorders, and cancer comprising determining the level of one or more fatty acyl-hydroxy fatty acids (FAHFAs), or a precursor or derivative thereof, in an isolated biological sample obtained from the subject, wherein the level of the one or more FAHFAs is indicative of the subject's disease state and/or treatment response for the disease or disorder. 
     
     
         8 . The method of  claim 7 , wherein the inflammatory disorder is selected from sepsis, rheumatoid arthritis (RA), ulcerative colitis, inflammatory bowel disease, Crohn's disease, systemic lupus erythematosus, celiac disease, uveitis, pancreatitis, adult respiratory distress syndrome, asthma, multiple sclerosis, graft-versus host disease, atopic dermatitis, and ankylosing spondylitis. 
     
     
         9 . The method of  claim 7 , wherein the isolated biological sample is a blood fraction, bile salt, pancreas secretions, or a tissue biopsy. 
     
     
         10 . The method of  claim 9 , wherein the blood fraction is plasma or serum. 
     
     
         11 . The method of  claim 9 , wherein the tissue biopsy comprises adipose tissue. 
     
     
         12 . The method of  claim 9 , wherein the tissue biopsy comprises pancreas, liver, kidney, or tumor tissue. 
     
     
         13 . The method  claim 7 , wherein the FAHFA, or precursor or derivative thereof, is detected by a method comprising MS/MS or an immunoassay. 
     
     
         14 . The method of  claim 7 , wherein the disorder is one or more of MODY 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 and, in some embodiments, is determined to be heterozygous or homozygous for a germline mutation in one or more genes selected from HNF4A, GCK, HNF1A, PDX1, TCF2, NEUROD1, KLF11, CEL, PAX4, INS, or BLK. 
     
     
         15 . A method of decreasing pro-inflammatory signaling or increasing glucose uptake, glucose tolerance, insulin secretion, or insulin sensitivity in a cell, comprising contacting the cell with an agent that increases the level of one or more FAHFAs. 
     
     
         16 . A method of decreasing pro-inflammatory signaling or increasing glucose uptake, glucose tolerance, insulin secretion, or insulin sensitivity in a mammalian subject in need thereof, comprising administering to the subject a therapeutically effective amount of an agent that increases the level of one or more FAHFAs. 
     
     
         17 . The method of  claim 15 , wherein the agent that increases the level of one or more FAHFAs is an exogenous FAHFA, or a substrate of carboxyl ester lipase (CEL), an inhibitor of CEL or the agent is a CHREBP expression product. 
     
     
         18 . The method of  claim 16 , wherein the agent that increases the level of one or more FAHFA's is an exogenous FAHFA, or a substrate of carboxyl ester lipase (CEL), an inhibitor of CEL, or the agent is a CHREBP expression product. 
     
     
         19 . The method of  claim 16 , wherein the subject has Type-2 Diabetes. 
     
     
         20 . The method of  claim 16 , wherein the pro-inflammatory signaling is dendritic cell maturation, activation, or proliferation; or macrophage cell maturation, activation, or proliferation. 
     
     
         21 . The method of  claim 20 , wherein the dendritic cell maturation, activation, or proliferation comprises an increase in CD40+, CD80+, CD86+, MHCII+ cells, or combinations thereof. And/or cytokine production. 
     
     
         22 . The method of  claim 21 , wherein the increased number of CD40+, CD80+, CD86+, or MHCII+ cells are also CD11c+. 
     
     
         23 . The method of  claim 16 , wherein the pro-inflammatory signaling is release of a proinflammatory cytokine selected from TNF-α, IL-1β, IL-12p70, IL-6, or combinations thereof. 
     
     
         24 . A method of identifying an agent that modulates the level of one or more FAHFAs, comprising contacting a cell with a candidate agent and measuring the level of one or more FAHFAs in the cell, wherein a change in the level of one or more FAHFAs in the cell, relative to a control cell not contacted with the agent, indicates that the agent modulates the level of one or more FAHFAs. 
     
     
         25 . The methods of  claim 24 , wherein the modulation results in an increase in the level of one or more FAHFAs. 
     
     
         26 . The method of  claim 24 , wherein the modulation results in a decrease in the level of one or more FAHFAs. 
     
     
         27 . The method of  claim 24 , wherein the agent modulates the level of a FAHFA synthase. 
     
     
         28 . The method of  claim 24 , wherein the agent modulates the level of a FAHFA esterase. 
     
     
         29 . The method of  claim 24 , wherein the cell is an isolated, cultured animal cell. 
     
     
         30 . The method of  claim 29 , wherein the cell is a macrophage (such as a RAW cell), an islet cell (such as Ins1 cell line), or a hepatic cell (such as HepG2). 
     
     
         31 . The method of  claim 29 , wherein the cell is cultured in the presence of low glucose. 
     
     
         32 . The method of  claim 29 , wherein the cell is cultured in the presence of a modulator of PPAR α, γ, or β. 
     
     
         33 . The method of  claim 29 , wherein the cell is located in a non-human animal. 
     
     
         34 . The method of  claim 33 , wherein the non-human animal is a mammal. 
     
     
         35 . The method of  claim 34 , wherein the mammal is murine. 
     
     
         36 . The method of  claim 35 , wherein the murine is an AG4OX mouse, an AG4KO mouse, a ChREBPKO mouse, or a ChREBPOX mouse. 
     
     
         37 . A detectable biotinylated amino-FAHFA of formula (II): 
       
         
           
           
               
               
           
         
         Wherein: m is an integer from 0 to 21; 
         n is an integer from 0 to 21; 
         the sum of m and n is an integer from 11 to 21; 
         X is selected from (OCH 2 CH 2 ) p  or (CH 2 ) p , wherein p is an integer from 2 to 20; 
         or a salt thereof. 
       
     
     
         38 . A method of identifying a modulator of FAHFA-mediated signaling, and/or FAHFA-mediated biological effects, comprising forming a mixture comprising an isolated mammalian cell that expresses a G-protein coupled receptor (GCPR), one or more FAHFAs, and a test compound and monitoring the FAHFA-mediated signaling, wherein a change in the FAHFA-mediated signaling indicates that the test compound is a modulator of FAHFA-mediated signaling. 
     
     
         39 . The method of  claim 38 , wherein the GCPR is GPR120 or GPR40. 
     
     
         40 . The method of  claim 38 , wherein the one or more FAHFAs comprise 9-PAHSA, 5-PAHSA, 9-OAHSA, or a combination thereof. 
     
     
         41 . The method  claim 38 , wherein FAHFA-mediated signaling and/or FAHFA-mediated biological effect is selected from decreasing pro-inflammatory signaling, stimulating insulin secretion, stimulating GLP1 secretion, stimulating calcium flux including calcium entry into cells or calcium exit from intracellular organelles into the cytoplasm, G-protein activation, and combinations thereof. 
     
     
         42 . The method of  claim 41 , wherein the FAHFA-mediated signaling is stimulating insulin secretion and the isolated mammalian cell is a pancreatic islet cell, an intestinal enteroendocrine cell, a macrophage, a T lymphocyte, or dendritic cell. 
     
     
         43 . The method of  claim 38 , wherein the mammalian cell is a human cell.