WO2006083798A2 - Procedes de traitement de troubles associes a l'obesite par inhibition de myd88, et procedes d'identification d'inhibiteurs de myd88 - Google Patents

Procedes de traitement de troubles associes a l'obesite par inhibition de myd88, et procedes d'identification d'inhibiteurs de myd88 Download PDF

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WO2006083798A2
WO2006083798A2 PCT/US2006/003296 US2006003296W WO2006083798A2 WO 2006083798 A2 WO2006083798 A2 WO 2006083798A2 US 2006003296 W US2006003296 W US 2006003296W WO 2006083798 A2 WO2006083798 A2 WO 2006083798A2
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myd88
subject
activity
inhibitor
receptor
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PCT/US2006/003296
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WO2006083798A3 (fr
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Anthony W. Ferrante
Stuart P. Weisberg
Rudolph L. Leibel
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The Trustees Of Columbia University In The City Of New York
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/027—New or modified breeds of vertebrates
    • A01K67/0275—Genetically modified vertebrates, e.g. transgenic
    • A01K67/0276—Knock-out vertebrates
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/027—New or modified breeds of vertebrates
    • A01K67/0271—Chimeric vertebrates, e.g. comprising exogenous cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/07—Animals genetically altered by homologous recombination
    • A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A01K2267/035—Animal model for multifactorial diseases

Definitions

  • This invention relates to methods for treating obesity-related disorders, methods for losing weight, and methods for identifying inhibitory compositions.
  • inflammatory response is characterized by the overproduction of inflammatory mediators including iNOS, IL-6, MCP-I 5 and TNF ⁇ ; the overproduction of acute phase proteins such as C-reactive protein, and haptoglobin and increases in white blood cell and monocyte counts.
  • inflammatory mediators including iNOS, IL-6, MCP-I 5 and TNF ⁇ ; the overproduction of acute phase proteins such as C-reactive protein, and haptoglobin and increases in white blood cell and monocyte counts.
  • Overproduction of these inflammatory molecules is associated with activation of inflammatory signaling pathways in leukocytes, liver, muscle and fat tissue.
  • These inflammatory responses include increased activity of NF- KB, and JNK, and overproduction of suppressors of cytokine signaling 1 and 3 (SOCSl and S0CS3). AU of these responses can impair insulin signaling at the cellular level.
  • T2DM type II diabetes mellitus
  • the invention provides for a method for treating an obesity-associated disorder in a subject, the method comprising administering to the subject an effective amount of an inhibitor of Myd88.
  • the invention also provides for a method for preventing an obesity-associated disorder in a human subject, the method comprising administering to the subject an effective amount of an inhibitor of human Myd88.
  • the invention provides for a method for treating insulin resistance in a subject, the method comprising administering to the subject an effective amount of an inhibitor of Myd88.
  • the invention also provides for a method for treating diabetes in a subject, the method comprising administering to the subject an effective amount of an inhibitor of Myd88.
  • the inhibitor of Myd88 may be an inhibitor that inhibits Myd88 gene expression, Myd88 protein production, or function of the Myd88 protein.
  • the inhibitor may comprise a polypeptide, a peptidomimetic, an organic molecule, a carbohydrate, a lipid, an antibody or a nucleic acid.
  • the subject on which the method is employed may be any mammal, e.g. a human, mouse, cow, pig, dog, cat, or monkey.
  • the mammalian members of the receptor superfamily for which Myd88 acts as an adapter include Toll-like receptor/Interleukin-1 receptor superfamily (mammalian): Subgroup 1 (Interleukin 1 receptors) - IL-IRl, IL-IRE, B15R, IL-IRAcP, IL-18R, IL-18RAcP, T1/ST2, IL-lRrp2, IL- IRAPL, TIGGIR, SIGGIR, and Subgroup 2 (Toll-like receptors) - TLR-I through TLR-10.
  • Toll-like receptor/Interleukin-1 receptor superfamily (mammalian): Subgroup 1 (Interleukin 1 receptors) - IL-IRl, IL-IRE, B15R, IL-IRAcP, IL-18R, IL-18RAcP, T1/ST2, IL-lRrp2, IL- IRAPL, TIGGIR, SIGGIR, and Subgroup 2 (Toll-like receptors
  • the administration of the agent may be effected by intralesional, intraperitoneal, intramuscular or intravenous injection; by infusion; or may involve liposome- mediated delivery; or topical, nasal, oral, anal, ocular or otic delivery.
  • administration of the inhibitor may comprise daily, weekly, monthly or hourly administration, the precise frequency being subject to various variables such as age and condition of the subject, amount to be administered, half- life of the agent in the subject, area of the subject to which administration is desired and the like.
  • a therapeutically effective amount of the inhibitor may include dosages which take into account the size and weight of the subject, the age of the subject, the severity of the obesity-related symptoms, the method of delivery of the agent and the history of the symptoms in the subject.
  • FIGs 1A-1B Graphical representations showing the genotyping of blood cells and weights of mice transplanted with Myd88 -/- bone marrow. Mice lethally irradiated and reconstituted with Myd88 -/- bone marrow are engrafted normally and they grow at the same rate on a high fat diet as control animals that were lethally irradiated and reconstituted with wild type bone marrow cells.
  • Figure IA Four weeks after bone marrow transplantation, blood cells from the recipient mice were genotyped using qPCR.
  • Figure IB Three weeks after bone marrow transplantation, blood cells from the recipient mice were genotyped using qPCR.
  • Circulating levels of MCP-I (28.5 ⁇ 15.5 vs. 70.2 ⁇ 28.1 vs. 24 ⁇ 7 pg/ml) were measured ( Figure 2A), IL-6 (6.5 ⁇ 1.5 vs. 13.5 ⁇ 4.6 vs. 4.6 ⁇ 0.84 pg/ml) ( Figure 2B), and PAI-I (4600 ⁇ 2200 vs. 7000 ⁇ 1100 vs. 2000 ⁇ 800 pg/ml) ( Figure 2C).
  • the circulating monocyte fraction of peripheral blood leukocytes was determined by staining peripheral blood leukocytes for CdI Ib and using flow cytometry to quantify the monocyte population (9 ⁇ 1.7 vs. 11 ⁇ 1.5 vs. 6.9 ⁇ 0.3%) (Figure 2D).
  • FIGs 3A-3D Obese animals reconstituted with Myd88 -/- bone marrow have improved insulin sensitivity.
  • Figure 3A After 20 weeks on high fat diet, obese mice that had been transplanted with Myd88 -/- (white circles) and Myd88 +/+ (black squares) bone marrow were fasted for six hours and given an intraperitoneal injection of insulin (1.5 u/kg).
  • Figure 3B Blood glucose was measured 0, 15, 30, 45 and 60 minutes after injection. At each time point, mice with Myd88 -/- hematopoietic cells had significantly lower absolute blood glucose levels and a greater percentage reduction in glucose levels compared to mice with Myd88 +/+ hematopoietic cells.
  • Figure 3C Figure 3C.
  • mice with Myd88 -/- hematopoietic cells had significantly lower blood glucose levels at time points 45, 60, 90 and 120 minutes after injection.
  • FIGs 4A-4D Obese animals reconstituted with Myd88 -/- bone marrow have decreased hepatomegaly, hepatic steatosis and decreased hepatic SOCSl expression. After 24 weeks on high fat diet whole livers were removed and weighed.
  • Figure 4A Liver weights were normalized to body mass (0.0456 ⁇ 0.00880 vs. 0.0686 ⁇ 0.00727; p value ⁇ 0.01).
  • Figure 4B Hepatic steatosis was assessed by measuring total triglyceride content per gram of liver tissue (52.4 ⁇ 17.9 vs. 80.4 ⁇ 27.3 mg/g; p value ⁇ 0.05).
  • Figure 4C Figure 4C.
  • Figures 5A-5D Myd88 -/- (open circles), heterozygous (black triangles) and wild type littermates (black squares) were placed on a high fat diet at five weeks of age and their growth was monitored for two months.
  • Figure 5D Myd88 -/- (open circles), heterozygous (black triangles) and wild type littermates (black squares) were placed on a high fat diet at five weeks of age and their growth was monitored for two months.
  • Figure 5A At five weeks of age Myd88 -/- mice weighed significantly less than heterozygous (19.8 +/- 1.8 v
  • FIGS 6A - 6B cDNA sequence for human Myd88 (SEQ ID NO: 1) as reported in Hardiman, G., Rock, F.L., Balasubramanian, S., Kastelein, R.A. and Bazan, J.F, Molecular characterization and modular analysis of human MyD88, Oncogene 13 (11), 2467- 2475 (1996); Accession No. NM_002468.
  • Figure 7 The translation, or protein sequence for human Myd88 (SEQ ID NO:2) based on the nucleotide sequence of SEQ ID NO:1.
  • US20030148986A1 Methods for treating vascular disease by inhibiting myeloid differentiation factor 88; European Publication No. EP1474444A1, A Novel Splice Variant Of Myd88 And Uses Thereof; and European Publication No. EP1465905A2, Antisense Modulation of Myd88 Expression.
  • Myd88 myeloid differentiation primary response gene
  • TIR Toll/Interleukin-1 receptor
  • TLRs toll-like receptors
  • Many signaling events downstream of these receptors depend on Myd88 because it is required to recruit signaling mediators such as Interleukin 1 receptor associated kinase (IRAK) to the signaling complex.
  • IRAK Interleukin 1 receptor associated kinase
  • This invention provides for the discovery that Myd88 can be used as a target in a drug screening assay to identify drugs that are capable of inhibiting Myd88 activity or function, thereby decreasing insulin resistance.
  • the present invention provides that Myd88 plays a role in the overproduction of cytokines during obesity and thereby influences the development of T2DM.
  • Myd88 deficiency plays a role in the overproduction of cytokines during obesity and thereby influences the development of T2DM.
  • the levels of inflammatory markers in their blood were examined as well as their metabolic phenotype.
  • Myd88 in hematopoietic cells plays an important role in obesity induced inflammation and insulin resistance.
  • Obesity is associated with a proinflammatory state characterized by the increased accumulation in adipose tissue of macrophages, the increased production of cytokines such as TNF-alpha, IL-6 and MCP-I.
  • cytokines such as TNF-alpha, IL-6 and MCP-I.
  • the protein Myd88 plays a central role in activation of the innate immune response by recruiting downstream signaling mediators in response to engagement of pattern recognition receptors. Animals with genetic deficiency of Myd88 exhibit impaired innate immune responses to exogenous pathogens as well as resistance to atherosclerosis.
  • Inhibitors of Myd88 include nucleic acid compounds that inhibit Myd88, such as an antisense Myd88 RNA, a ribozyme designed to cleave Myd88 RNA and double- stranded RNA that is sufficiently homologous to the Myd88 gene product to inhibit the encoding function of Myd88 mRNA (See, U.S. Serial No. 2003 0148986 Al, U.S. Serial No. 2003 0186903 Al).
  • Another inhibitor of Myd88 is a Myd88-specific small interfering RNA (siRNA) which was shown to knockdown expression of Myd88 in a macrophage cell line (Yang et al., J Virol 78(20): 11152-11160 (2004)).
  • Myd88-specific antibodies are available from commercial vendors, for example, Abeam, eBiosciences and ProSci, Inc.
  • Inhibitors such as peptides or peptidomimetics can be designed based on structural studies of Myd88, TLRs and IL-IRs (Khan et al., J Biol Chem 279(30):31664- 31670 (2004); Dunne et al, J Biol Chem 278(42): 41443-41451 (2003)).
  • TIR Toll/interleukin-1 receptor
  • the domain is conserved among the TLRs, IL-IRs and Myd88.
  • Peptide inhibitors or peptidomimetics that share homology with this domain could be used to block the recruitment of Myd88 to the receptors, thus creating a lesion in the signaling pathway and inhibiting downstream events.
  • U.S. Patent No. 6,222,019 discloses fragments of Myd88 that function as dominant negative versions of full-length Myd88. For example, a Myd88 fragment comprising amino acid residues 106-296 inhibits IL-lR-induced NF- ⁇ B activity.
  • the invention provides for a method for treating an obesity-associated disorder in a subject, the method comprising administering to the subject an effective amount of an inhibitor of Myd88.
  • the invention also provides for a method for preventing an obesity-associated disorder in a human subject, the method comprising administering to the subject an effective amount of an inhibitor of human Myd88.
  • the invention provides for a method for treating insulin resistance in a subject, the method comprising administering to the subject an effective amount of an inhibitor of Myd88.
  • the invention also provides for a method for treating diabetes in a subject, the method comprising administering to the subject an effective amount of an inhibitor of Myd88.
  • the inhibitor comprises an antagonist of Myd88, a Myd88-specific antibody, an antisense oligonucleotide that specifically binds to a region of a nucleic acid that encodes Myd88, an organic molecule, a peptide, or a peptidomimetic.
  • the inhibitor comprises a small interfering RNA (siRNA) that specifically binds to a region of a nucleic acid that encodes Myd88, wherein expression of Myd88 is inhibited.
  • siRNA small interfering RNA
  • the inhibitor comprises a peptide that is a fragment of Myd88, a fragment of a Toll-like receptor, or a fragment of an interleukin 1 receptor.
  • the inhibitor comprises a peptidomimetic of a peptide fragment of Myd88, a peptide fragment of a Toll-like receptor, or a peptide fragment of an interleukin 1 receptor.
  • the inhibitor comprises a peptide fragment, which is of a Toll/interleukin 1 receptor (TIR) domain.
  • TIR Toll/interleukin 1 receptor
  • the obesity-associated disorder comprises diabetes, insulin resistance, hyperinsulinemia, decreased glucose clearance, dyslipidemia, non-alcoholic fatty liver disease, hypertension, inflammation, hepatomegaly, hepatic steatosis, myocardial infarction, asthma, stroke, or any combination thereof.
  • the invention provides for embodiments where the administering comprises contacting hematopoietically-derived cells or myeloid cells with the inhibitor.
  • the invention provides for embodiments where the administering comprises contacting bone marrow of the subject with the inhibitor.
  • the invention provides for embodiments where the administering comprises ex vivo treatment of cells or tissue taken from the subject with the inhibitor, and returning the cells or tissues to the subject.
  • the invention provides for embodiments where the administering comprises intralesional, intraperitoneal, intramuscular or intravenous injection; infusion; liposome-mediated delivery; or topical, nasal, oral, ocular, otic delivery, or any combination thereof.
  • the invention provides for embodiments where the subject is a human, mouse, rabbit, monkey, rat, bovine, pig or dog.
  • the invention provides for methods for treating an obesity-associated disorder in a subject, the method comprising transplanting Myd88-deflcient bone marrow into the subject.
  • the invention also provides methods for reducing or preventing obesity in a subject, the method comprising administering to the subject an effective amount of an inhibitor of Myd88.
  • the invention provides for methods for lowering body weight in a mammal comprising administering to the mammal in need of losing weight an inhibitor of Myd88 in an effective amount to reduce the weight of the subject, thereby lowering the body weight.
  • the invention provides for embodiments where the administration is systemic administration.
  • the invention provides for embodiments where the subject is suffering from diabetes.
  • the invention provides for embodiments where the mammal is a dog, a mouse, a human, a horse, a monkey, a primate, or a feline.
  • the invention also provides for drug screening methods.
  • the invention provides for methods for identifying whether a test compound is capable of inhibiting Myd88 activity, the method comprising: (a) contacting a test compound with a solution containing (i) Myd88, and (ii) either a Toll-like receptor, an interleukin 1 receptor, or both, (b) measuring Myd88 activity in the solution of step (a), and (c) comparing the Myd88 activity in step (b) with Myd88 activity in the absence of the test compound, so as to determine whether or not the test compound is capable of inhibiting Myd88 activity.
  • the invention provides for methods for identifying a compound capable of inhibiting Myd88, the method comprising: (a) administering a test compound to a subject, and (b) measuring inhibition of Myd88 activity in the subject as compared to another subject that was not administered the test compound, so as to determine whether or not the test compound inhibits Myd88 activity.
  • the invention also provides for a method for identifying a compound capable of inhibiting Myd88 activity, the method comprising: (a) contacting a test compound with a cell, (b) contacting the cell with a toll-like receptor agonist or interleukin 1 receptor agonist, (c) measuring Myd88 activity in the cell, and (d) comparing the activity measured in (c) with Myd88 activity in a cell in the absence of the test compound, so as to determine whether the test compound is capable of inhibiting Myd88.
  • the invention provides for embodiments where the method is carried out in a high-throughput way.
  • the invention provides for embodiments where the activity measured comprises Myd88 gene expression, Myd88 protein production, Myd88 translocation, Myd88 protein function, Myd88 binding activity, cytokine production, immune response, blood glucose concentration, insulin resistance, insulin concentration, NF- ⁇ B activity, SOCSl expression, MCP-I production, PAI-I production, IL-6 production, blood monocyte fraction, or any combination thereof.
  • Myd88 activity is detected as a complex formed with Myd88 and either Toll-like receptor, interleukin 1 receptor, or both.
  • the invention provides for embodiments where the solution comprises an extract from a cell contacted with a Toll-like receptor agonist or an interleukin 1 receptor agonist.
  • the invention provides for embodiments where the inhibition of Myd88 activity is measured by detecting decreased levels of an inflammatory marker in blood of the subject.
  • the invention provides for embodiments where the inflammatory marker comprises MCP-I, IL-6, PAI-I, monocytes, C-reactive protein or any combination thereof.
  • the invention provides for embodiments where the inhibition of Myd88 activity determined by detecting increased insulin sensitivity, decreased circulating glucose levels, decreased hepatomegaly, decreased hepatic steatosis, decreased hepatic suppressor of cytokine signaling (SOCSl) expression, or any combination thereof.
  • SOCSl cytokine signaling
  • the invention provides for embodiments where glucose levels are measured by a glucose tolerance test (GTT) or measuring fasting glucose levels, or a combination thereof.
  • GTT glucose tolerance test
  • ITT insulin tolerance test
  • HOMA-IR homeostatic model assessment of insulin resistance
  • euglycemic hyperinsulinemic clamp or any combination thereof.
  • the cell is a hematopoietic cell, a myeloid cell, a macrophage, or any combination thereof.
  • the invention provides for embodiments where the agonist comprises lipopolysaccharide, interleukin 1, interferon gamma, saturated fatty acids, dsRNA, bacterial cellular components, viral protein, carbohydrate, nucleic acid components, or any combination thereof.
  • the invention provides for embodiments where the inhibition of Myd88 activity is measured as decreased NFKB activity, IRAK activation, IRAK recruitment, decreased production of cytokines, or any combination thereof.
  • the cytokine comprises MCP-I 5 EL-6, PAI-I, TNF-a, MCP-3, adiponection, resistin, or any combination thereof.
  • the inhibitor of Myd88 can be combined with a carrier.
  • carrier is used herein to refer to a pharmaceutically acceptable vehicle for a pharmacologically active agent.
  • the carrier facilitates delivery of the active agent to the target site without terminating the function of the agent.
  • suitable forms of the carrier include solutions, creams, gels, gel emulsions, jellies, pastes, lotions, salves, sprays, ointments, powders, solid admixtures, aerosols, emulsions (e.g., water in oil or oil in water), gel aqueous solutions, aqueous solutions, suspensions, liniments, tinctures, and patches suitable for topical administration.
  • the term "effective" is used herein to indicate that the inhibitor is administered in an amount and at an interval that results in the desired treatment or improvement in the disorder or condition being treated (e.g., an amount effective to reduce body weight of a subject, or to reduce insulin resistance).
  • the subject is a mammal.
  • mammals include: human, primate, mouse, otter, rat, and dog.
  • compositions include those suitable for oral or parenteral (including intramuscular, subcutaneous and intravenous) administration.
  • forms suitable for parenteral administration also include forms suitable for administration by inhalation or insufflation or for nasal, or topical (including buccal, rectal, vaginal and sublingual) administration.
  • the formulations may, where appropriate, be conveniently presented in discrete unit dosage forms and may be prepared by any of the methods well known in the art of pharmacy. Such methods include the step of bringing into association the active compound with liquid carriers, solid matrices, semi-solid carriers, finely divided solid carriers or combinations thereof, and then, if necessary, shaping the product into the desired delivery system.
  • the following examples illustrate the present invention, and are set forth to aid in the understanding of the invention, and should not be construed to limit in any way the scope of the invention as defined in the claims which follow thereafter.
  • Example 1 Body Mass of Myd88 -/- Mice and Bone Marrow Chimeras Is Lower Than Myd88 +/+ Mice
  • the mammal such as a mouse, a primate, or a human
  • Systemic inhibitors include siRNA or antisense nucleic acids specifically targeted to affect the expression of Myd88.
  • Other inhibitors include antibodies that specifically bind to Myd88.
  • MCP-I has been linked to the development of atherosclerosis.
  • IL-6 is an important stimulator of c-reactive protein production and both have been linked to progression of coronary artery disease.
  • obesity induced insulin resistance has been linked to overproduction of iNOS, TNF- ⁇ , MCP-I and IL-6.
  • Example 2 Inhibition of Myd88 in Hematopoietic Cells Results in Decreased Insulin Resistance - A treatment for obesity-induced inflammation
  • mice that had been transplanted with Myd88 +/+ bone marrow cells had elevated plasma concentrations of MCP-I (70.2 ⁇ 28.1 vs. 24 ⁇ 7 pg/mL; p value ⁇ 0.01), IL-6 (13.5 ⁇ 4.6 vs. 4.6 ⁇ 0.84 pg/mL; p value ⁇ 0.01), and PAI-I (7000 ⁇ 1100 vs. 2000 ⁇ 800 pg/mL; p value ⁇ 0.05) compared to lean mice.
  • Myd88 deficiency in hematopoietic cells completely prevented the obesity induced increase in plasma MCP-I levels (28.5 ⁇ 15.5 vs.
  • mice with Myd88 -/- hematopoietic cells became as obese as wild types but had an attenuated inflammatory response, the development of insulin resistance in mice with Myd88 -/- hematopoietic cells might be similarly attenuated.
  • mice with Myd88 -/- bone marrow cells had lower blood glucose concentrations than wild type controls (166 ⁇ 26.5 vs. 242 ⁇ 41.8 mg/dL; p value ⁇ 0.01) and had a much greater hypoglycemic response to an injected bolus of insulin.
  • mice with Myd88 -/- hematopoietic cells had significantly lower blood glucose concentrations at every time point and the percentage decrease in blood glucose concentration was significantly greater at every time point ( Figures 3A-3D).
  • mice with Myd88 -/- hematopoietic cells had significantly lower blood glucose levels (100.4 ⁇ 20.5 vs. 127 ⁇ 23; p value ⁇ 0.05) and blood insulin levels that were half that of mice with Myd88 +/+ hematopoietic cells (0.75 ⁇ 0.31 vs. 1.6 ⁇ 0.65; p value ⁇ 0.05).
  • mice with Myd88 +/+ hematopoietic cells were estimated to be 2.5 times more insulin resistant than mice with Myd88 -/- hematopoietic cells (4.9 ⁇ 3.0 vs. 12.9 ⁇ 6.3; p value ⁇ 0.05).
  • mice with Myd88 -/- hematopoietic cells cleared the excess glucose faster than mice with Myd88 +/+ hematopoietic cells ( Figures 3A-3D).
  • the triglyceride content of liver from obese mice with Myd88 -/- hematopoietic cells was 35% less than those from obese mice with Myd88 -/- hematopoietic cells (52.4 ⁇ 17.9 vs. 80.4 ⁇ 27.3; p value ⁇ 0.05) ( Figures 4A-4D).
  • IL-6 is a potent stimulator of SOCSl and SOCS3 expression. Because of the observation that Myd88 in hematopoietic cells is important for stimulating IL-6 production during obesity, Myd88 might thereby regulate hepatic SOCSl and/or SOCS3 expression. Therefore, quantitative RT-PCR was used to measure hepatic expression of SOCSl and SOCS3 in the two groups of mice.
  • Myd88 in hematopoietic cells plays an important role in the development of several inflammatory responses during diet-induced obesity in mice.
  • Mice that had been transplanted with Myd88 deficient bone marrow cells prior to being placed on a high fat diet became equally obese as controls that had been transplanted with wild type bone marrow cells. But they had lower circulating levels of MCP-I, IL-6, and PAI-I. Consistent with having lower circulating levels of MCP-I, mice with Myd88 -/- hematopoietic cells had a lower fraction of circulating monocytes compared to controls. And consistent with having lower circulating levels of IL-6, mice with Myd88 -/- hematopoietic cells had lower SOCSl expression in liver.
  • the data presented here provide direct evidence that Myd88-dependent signaling pathways in hematopoietic cells play a role in causing obesity induced insulin resistance.
  • obese mice with Myd88 -/- hematopoietic cells had decreased fasting blood glucose levels, increased insulin sensitivity as assessed by HOMA-IR modeling and an ITT, improved ability to clear a glucose load during a GTT as well as decreased hepatic steatosis, hepatomegaly and hepatic SOCSl expression.
  • the most parsimonious explanation for these findings is that inflammatory molecules produced downstream of the Myd88 dependent signaling pathways activated by obesity cause insulin resistance.

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Abstract

L'invention concerne des procédés de traitement de troubles associés à l'obésité chez un sujet, consistant à administrer au sujet une quantité efficace d'un inhibiteur de Myd88. L'invention concerne également des procédés permettant de réduire le poids du corps chez un mammifère, consistant à administrer audit mammifère pour lequel une perte de poids est requise, un inhibiteur de Myd88, en une quantité efficace pour réduire le poids du sujet.
PCT/US2006/003296 2005-01-31 2006-01-31 Procedes de traitement de troubles associes a l'obesite par inhibition de myd88, et procedes d'identification d'inhibiteurs de myd88 WO2006083798A2 (fr)

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* Cited by examiner, † Cited by third party
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Publication number Priority date Publication date Assignee Title
WO2009121152A2 (fr) 2008-04-03 2009-10-08 Katholieke Universiteit Leuven Signatures géniques
WO2009121152A3 (fr) * 2008-04-03 2009-12-17 Katholieke Universiteit Leuven Signatures géniques
US8524656B2 (en) 2008-07-08 2013-09-03 Jacques Galipeau GM-CSF and truncated CCL2 conjugates and methods and uses thereof
WO2023174356A1 (fr) * 2022-03-17 2023-09-21 圣诺生物医药技术(苏州)有限公司 Composition pharmaceutique d'interférence d'acide nucléique, et médicament pour le traitement du cancer colorectal, du cancer gastrique et du cancer de la prostate

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