FIELD OF THE INVENTION

This application relates generally to the field of exosomes, and more particularly, to adipocyte-derived exosomes. This application also relates generally to the isolation of exosomes, such as adipocyte-derived exosomes, from biological samples including fluids, such as, for example, urine and blood. Exosomes are actively-shed endocytic vesicles that are capable of transporting functional mRNAs, microRNAs, and proteins between cells over large distances; exosomes therefore may contribute to mechanisms by which adipocytes (and adipose tissue) in obese individuals can induce dysfunction in other organ systems. See Kishida et al. (Ann. Med.,2012, 44(3): 233-41, the entirety of which is incorporated by reference herein; Camussi et al. (Am. J. Cancer Res.,2011, 1(1): 98-110, the entirety of which is incorporated by reference herein; and Zernecke et al. (Sci. Signal,2009, 2(100): ra81), the entirety of which is incorporated by reference herein.

BACKGROUND INFORMATION

Adipose tissue is an active endocrine organ, which synthesizes and releases adipokines (such as leptin, resistin and adiponectin) that act as autocrine, paracrine and endocrine factors to modulate metabolism locally and at distant sites. See Sun et al. (J. Clin. Invest.,2011, 121(6): 2094-101, the entirety of which is incorporated by reference herein). Ectopic adipose accumulation can significantly compromise insulin sensitivity by interfering with insulin signaling. See Shulman (N. Engl. J. Med.,2014, 371(12): 1131-41, the entirety of which is incorporated by reference herein).

Exosomes are nanoparticle-sized endocytic vesicles that transport lipids, sugars, nucleic acids, and proteins between cells. See Camussi et al. and Zernecke et al., supra. Adipose exosomal signaling is largely determined by their microRNA content, which can control gene expression in target cells by inhibiting mRNA translation or enhancing mRNA degradation. See Karelis et al. (J. Clin. Endocrinol. Metab.,2004, 89(6): 2569-75, the entirety of which is incorporated by reference herein); and Bartel (Cell,2009, 136(2): 215-33, the entirety of which is incorporated by reference herein).

Insulin resistance in patients with obesity significantly contributes to the pathogenesis of chronic metabolic diseases such as Type II diabetes mellitus (T2DM). Adipose is a key modulator of systemic insulin resistance and metabolic dysregulation, exerting effects via mechanisms such as releasing pro-inflammatory cytokines into the circulation. See Sun et al. supra; Boden et al. (Diabetes,2014, 63(9): 2977-83, the entirety of which is incorporated by reference herein); Kim et al. (Clin. Invest.,2007, 117(9): 2621-37, the entirety of which is incorporated by reference herein); and Tilg et al. (Nat. Rev. Immunol.,2006, 6(10): 772-83, the entirety of which is incorporated by reference herein). Central adiposity is strongly linked to the pathogenesis of obesity-related comorbidities like insulin resistance in peripheral tissues. See Marette et al. (Rev. Endocr. Metab. Disord.,2014, 15(4): 299-305, the entirety of which is incorporated by reference herein); and Sam et al. (Transl. Res.,2014, 164(4): 284-92, the entirety of which is incorporated by reference herein). However, large variations exist in an individual's response to obesity; some patients with high body mass index (BMI) can have normal glucose homeostasis, while others with normal BMI can have frank diabetes. See Gomez-Ambrosi et al. (Int. J. Obes. (Lond), 2012, 36(2): 286-94, the entirety of which is incorporated by reference herein). The volume of adipose tissue alone therefore does not portend systemic disease, and it is contemplated that adipose function must also play a critical role.

One potential mechanism linking adipose accumulation to overall glucose regulation is inter-organ signaling from adipose tissue via exosomes. Deng et al. (Diabetes,2009. 58(11): 2498-505, the entirety of which is incorporated by reference herein) suggested that adipocyte-derived exosomes might affect insulin resistance, finding that obesity-related microRNAs targeted key biological pathways that impact insulin signaling using an animal model. Previous studies have described obesity-related differences in the microRNA content of exosomes derived from human obese visceral adipose tissue, as compared to those from lean subjects, and identified adipocyte-derived exosomal signaling as a mediator linking adiposity and insulin resistance in peripheral tissues. See Ferrante et al., supra; and Koeck et al. (J. Surg. Res.,2014, 192(2): 268-75, the entirety of which is incorporated by reference herein). However, previous studies have relied upon isolating adipocyte-derived exosomes from adipose tissue, due to the absence of any technique for effectively isolating adipocyte-derived exosomes from other sources.

There is a strong epidemiological link between obesity and atherosclerosis (Bastien, M., et al., (Prog Cardiovasc Dis,2014. 56(4): p. 369-81, the entirety of which is incorporated by reference herein); and Gupta, N., et al., (Endocr Rev,2012. 33(1): p. 48-70, the entirety of which is incorporated by reference herein). Atherosclerosis, which is one of the hallmarks of cardiovascular disease, is characterized by macrophage cholesterol efflux impairment leading to intracellular accumulation of modified low-density lipoprotein (LDL) and subsequent formation of plaque-forming lipid-rich foam cells. Michael, D. R., et al. (Cytokine,2013. 64(1): p. 357-61, the entirety of which is incorporated by reference herein); and Rohatgi, A., et al., (N Engl J Med,2014. 371(25): p. 2383-93, the entirety of which is incorporated by reference herein). Macrophage cholesterol homeostasis is a delicate balance among influx, endogenous synthesis, esterification/hydrolysis and efflux. See Zhang, M., et al., (Atherosclerosis,2014. 234(1): p. 54-64, the entirety of which is incorporated by reference herein). Large variations exist in individual responses to obesity. For example, some patients with high adiposity have normal cardiovascular health, while others with low adiposity have frank atherosclerosis. See Bradshaw, P. T., K. L. Monda, and J. Stevens. (Obesity(Silver Spring), 2013. 21(1): p. 203-9, the entirety of which is incorporated by reference herein). It is contemplated that adipose health or function, not adipose mass per se, is a determinant of obesity-related comorbidities. However, tools to assess adipose health/function in relation to the comorbidities are lacking.

SUMMARY OF THE INVENTION

The present disclosure relates to techniques for the isolation, quantification, and/or characterization of adipocyte-derived exosomes. In some embodiments of the invention, for example, adipocyte-derived exosomes may be isolated and/or quantified from the circulation and/or the urine, and additionally, their contents may subsequently be characterized. This facile isolation procedure has implications for the generation of convenient and rapid diagnostic tests, and for the development, testing, and monitoring of therapeutics. In some aspects, the subject from which the exosomes are isolated may be a subject in need of weight loss, or may be in need of a modification to their adipocyte-derived exosome microRNA profile. This exosome microRNA profile may include, for example, the presence or absence of particular microRNAs, and/or the abundance of particular microRNAs, including the relative abundance of a microRNA with respect to the abundance of one or more other microRNAs.

According to one embodiment, a process of the present invention involves isolating adipocyte-derived exosomes from a subject. A biological sample is obtained (from a subject) that contains adipocyte-derived exosomes, and the adipocyte-derived exosomes are specifically isolated from the sample using a binding agent which binds to at least one marker specific for adipocyte-derived exosomes. This marker may be, for example, selected from the group consisting of Fatty Acid Binding Protein 4 (FABP4), Pref-1, amino acid transporter Asc-1, proton amino acid transporter PAT-2, purinergic receptor 2X, ligand-gated ion channel 5 (P2RX5), leptin, EBF3, FBX031, EVA1, CD137, Shox2, Hoxc8, Hoxc9, Tbx1, Zic1, Lhx8, Tnsfrsf9, Tmem26, UCP-1, Cidea, Prdm16, N-acetyltransferase 8-like (Nat8L), prolactin receptor (PRLR), neuregulin-4 (Nrg4), transmembrane protein 120B (Tmem120B), adrenergic β3 receptor (Adrb3), aquaporin-7 (Aqp7), G protein-coupled receptor 81 (Grpr81), G protein-coupled receptor 119 (Gpr119), fatty acid transporter (SLC27a1/FATP1), solute carrier family 7 member 10 (SLC7a10/Asc-1 neutral amino acid transporter), mitochondrial protein uncoupling protein-1 (UCP-1), CD300LG, tetraspanin 18 (tspan18), and frizzled-4 (Fzd4).

In some embodiments, the process may involve using Fatty Acid Binding Protein 4 (FABP4) as a marker. Additionally, the biological sample obtained may be, but is not limited to, milk, blood, serum, plasma, ascites, cyst fluid, pleural fluid, peritoneal fluid, cerebral spinal fluid, tears, urine, saliva, sputum, or combinations thereof. In particular embodiments, the biological sample obtained is blood, serum, or urine.

In some embodiments, the process may involve using a binding agent such as an anti-FABP4 antibody, or an antigen-binding fragment thereof.

In some embodiments, the process may involve isolating the adipocyte-derived exosomes using magnetic beads. The isolated adipocyte-derived exosomes may be substantially free of other non-adipocyte-derived exosomes, or completely free of other non-adipocyte-derived exosomes, if desired.

The sample may be collected from a lean subject, an overweight subject, or an obese subject. In some embodiments, the sample may be collected from an obese subject prior to initiation of a weight-loss treatment, surgery, or regimen. Alternatively, the sample may be collected from an obese subject after initiation or completion of a weight-loss treatment, surgery, or regimen.

In a second aspect of the invention, a kit for isolating adipocyte-derived exosomes in a biological sample is provided. The kit may contain a binding agent which specifically binds to at least one marker specific for adipocyte-derived exosomes, a bioparticle for isolation of the adipocyte-derived exosomes, and instructions for performing the adipocyte-derived exosome isolation.

The kit may include at least one marker, as a control for example. The marker may be, but is not limited to, Fatty Acid Binding Protein 4 (FABP4), Pref-1, amino acid transporter Asc-1, proton amino acid transporter PAT-2, purinergic receptor 2X, ligand-gated ion channel 5 (P2RX5), leptin, EBF3, FBX031, EVA1, CD137, Shox2, Hoxc8, Hoxc9, Tbx1, Zic1, Lhx8, Tnsfrsf9, Tmem26, UCP-1, Cidea, Prdm16, N-acetyltransferase 8-like (Nat8L), prolactin receptor (PRLR), neuregulin-4 (Nrg4), transmembrane protein 120B (Tmem120B), adrenergic β3 receptor (Adrb3), aquaporin-7 (Aqp7), G protein-coupled receptor 81 (Grpr81), G protein-coupled receptor 119 (Gpr119), fatty acid transporter (SLC27a1/FATP1), solute carrier family 7 member 10 (SLC7a10/Asc-1 neutral amino acid transporter), mitochondrial protein uncoupling protein-1 (UCP-1), CD300LG, tetraspanin 18 (tspan18), and/or frizzled-4 (Fzd4).

The bioparticle for isolation of the adipocyte-derived exosomes may comprise, for example, a magnetic bead or an agarose bead. In particular embodiments, the bioparticle is at least one magnetic bead selected from the group consisting of a sepharose magnetic bead, a streptavidin magnetic bead, a Protein A magnetic bead, a Protein G magnetic bead, a ferromagnetic bead, a magnetic nanosphere, a magnetic microsphere, and a polyvinyl alcohol magnetic particle (M-PVA Magnetic Bead)

In some embodiments, the kit provided for performing the process of isolating adipocyte-derived exosomes in a biological sample includes a binding agent that specifically binds to Fatty Acid Binding Protein 4 (FABP4), at least one magnetic bead, and instructions for performing the adipocyte-derived exosome isolation.

In a third aspect of the invention, a method of detecting an obesity-related disorder in a subject is provided. The method may involve, for example, obtaining a biological sample from a subject which contains adipocyte-derived exosomes, isolating adipocyte-derived exosomes from the biological sample using a binding agent which binds to at least one marker specific for adipocyte-derived exosomes, and detecting that the subject has an obesity-related disorder by a measurable change in a profile of the isolated adipocyte-derived exosomes from the subject. The measurable change may be relative to, for example, a profile of adipocyte-derived exosomes from one or more control subjects.

In a fourth aspect of the invention, a method of detecting an increased risk of a subject for having an obesity-related disorder is provided. The method may involve, for example, obtaining a biological sample from a subject which contains adipocyte-derived exosomes, isolating adipocyte-derived exosomes from the biological sample using a binding agent which binds to at least one marker specific for adipocyte-derived exosomes, and detecting the subject has an increased risk of having an obesity-related disorder by a measurable change in a profile of the isolated adipocyte-derived exosomes from the subject. The measurable change may be relative to, for example, a profile of adipocyte-derived exosomes from a control subject.

The detection methods of the present invention may further include applying the profile of the isolated adipocyte-derived exosomes from the subject against the profile of adipocyte-derived exosomes from one or more control subjects.

The detection methods of the present invention may further include diagnosing the subject as having an obesity-related disorder, or an increased risk of having an obesity-related disorder.

The detection methods may further include determining the profile of the isolated adipocyte-derived exosomes.

The detection methods may further include administering at least one therapeutic agent to the subject to treat the obesity-related disorder.

The detection methods may further include recommending to the subject, a therapy comprising a therapeutic regimen, a surgical intervention, a behavioral modification, or a combination thereof.

In a fifth aspect of the invention, a method for identifying a therapy for an obesity-related disorder in a subject is provided. The method involves obtaining a biological sample from the subject which contains adipocyte-derived exosomes, isolating adipocyte-derived exosomes from the biological sample using a binding agent which binds to at least one marker specific for adipocyte-derived exosomes, and detecting the subject has an obesity-related disorder, or has an increased risk of having an obesity-related disorder, by a measurable change in a profile of the isolated adipocyte-derived exosomes from the subject. The measurable change may be relative to, for example, a profile of adipocyte-derived exosomes from a control subject. Based on this detected measurable change, a therapy for the obesity-related disorder is then identified. The therapy may include, for example, a therapeutic regimen, a surgical intervention, a behavioral modification, or a combination thereof.

In particular embodiments, the surgical intervention may be bariatric surgery or bypass surgery.

The therapeutic regimen may include administering at least one therapeutic agent, a dietary intervention, exercise, or a combination thereof.

The method for identifying a therapy for an obesity-related disorder in a subject may further include determining the profile of the isolated adipocyte-derived exosomes from the subject.

The method for identifying a therapy for an obesity-related disorder in a subject may further include applying the profile of the isolated adipocyte-derived exosomes from the subject against the profile of adipocyte-derived exosomes from the control subject.

The method for identifying a therapy for an obesity-related disorder in a subject may further include administering the identified therapy to the subject.

The method for identifying a therapy for an obesity-related disorder in a subject may further include recommending the identified therapy to the subject.

In a sixth aspect of the invention, a method for identifying at least one therapeutic agent for an obesity-related disorder in a subject is provided. The method involves obtaining a biological sample from a subject which contains adipocyte-derived exosomes, isolating adipocyte-derived exosomes from the biological sample using a binding agent which binds to at least one marker specific for adipocyte-derived exosomes, detecting the subject has an obesity-related disorder, or an increased risk of having an obesity-related disorder, by a measurable change in a profile of the isolated adipocyte-derived exosomes from the subject relative to a profile of adipocyte-derived exosomes from a control subject, and identifying the at least one therapeutic agent for treatment or prevention of the obesity-related disorder.

The therapeutic agent identified includes a therapeutic agent for treatment or prevention of a glucose homeostasis disorder, an agent for treatment or prevention of a respiratory disorder, an agent for treatment or prevention of a cardiovascular disorder, an agent for treatment or prevention of a neoplastic disease, an agent for treatment or prevention of a gallbladder disease, an agent for treatment or prevention of arthritis, an agent for dyslipidemia, an agent for mental illness, an agent for pain syndromes, or a combination thereof.

At least one therapeutic agent for a glucose homeostasis disorder includes orlistat, lorcaserin, phentermine, liraglutide, insulin, amylin, leptin, glucagon, glucagon-like peptide (GLP-1), glucagon-like peptide (GLP-1) receptor agonist, a sulfonylurea, a biguanide, somatostatin, diazoxide, a sodium-glucose co-transporter 2 (SGLT-2) inhibitor, acarbose, miglitol, a dopamine agonist, a dipeptidyl peptidase 4 (DPP-4) inhibitor, a meglitinide, an α-glucosidase inhibitor, a thiazolidinedione, or a combination thereof.

At least one therapeutic agent for a cardiovascular disorder includes an aldosterone receptor antagonist, an angiotensin II receptor antagonist, an angiotensin converting enzyme inhibitor, a β-adrenergic receptor antagonist, a calcium channel blocker, an α2-adrenergic receptor agonist, an α1-adrenergic receptor antagonist, a vasodilator, a cardiac glycoside, a diuretic, an inotropic agent, a phosphodiesterase inhibitor, an antiarrhythmic agent, potassium, a proprotein convertase subtilisin kexin type 9 (PCSK9) inhibitor, a nitrate, warfarin, a cholesterol lowering agent, or a combination thereof. At least one therapeutic agent for a respiratory disorder includes a β-adrenergic receptor agonist, an antihistamine, a leukotriene modifier, a mast cell stabilizer, theophylline, an immunomodulator, an anticholinergic, and a corticosteroid, or a combination thereof.

The method for identifying at least one therapeutic agent for an obesity-related disorder in a subject may further include applying the profile of the isolated adipocyte-derived exosomes from the subject against the profile of adipocyte-derived exosomes from the control subject.

The method for identifying at least one therapeutic agent for an obesity-related disorder in a subject may further include determining the profile of the isolated adipocyte-derived exosomes from the subject.

The method for identifying at least one therapeutic agent for an obesity-related disorder in a subject may further include administering the at least one therapeutic agent to the subject.

The method for identifying at least one therapeutic agent for an obesity-related disorder in a subject may further comprise recommending the at least one therapeutic agent to the subject.

In a seventh aspect of the invention, a method of treating or preventing an obesity-related disorder in a subject is provided, which method may comprise obtaining a biological sample from a subject which contains adipocyte-derived exosomes, isolating adipocyte-derived exosomes from the biological sample using a binding agent which binds to at least one marker specific for adipocyte-derived exosomes, detecting the subject has an obesity-related disorder or an increased risk of having an obesity-related disorder by a measurable change in a profile of the isolated adipocyte-derived exosomes from the subject relative to a profile of adipocyte-derived exosomes from a control subject, and administering a therapy to the subject to treat or prevent the obesity-related disorder. The therapy may include, for example, a therapeutic regimen, surgical intervention, behavioral modification, or a combination thereof. The surgical intervention includes bariatric surgery or bypass surgery. The therapeutic regimen includes administering at least one therapeutic agent.

The obesity-related disorder may include, for example, a glucose homeostasis disorder, a respiratory disorder, a cardiovascular disorder, a neoplastic disease, gallbladder disease, arthritis, or a combination thereof.

The glucose homeostasis disorder may include, for example, diabetes, insulin resistance, glycosuria, hyperglycemia, hyperinsulinism, and/or hypoglycemia. Diabetes may be diabetes mellitus type I, diabetes mellitus type II and gestational diabetes.

The cardiovascular disorder may include, for example, atherosclerosis, coronary artery disease, stroke, peripheral artery disease, angina, myocardial infarction, hypertension, and hypertensive heart disease.

The respiratory disorder may include, for example, asthma, sleep apnea, obesity hypoventilation syndrome and chronic obstructive pulmonary disease.

The profile of the isolated adipocyte-derived exosomes may be, for example, a microRNA expression profile, a protein expression profile, an mRNA expression profile, a lipid profile, a sugar profile, and/or a nucleic acid profile.

In some embodiments, the profile of the isolated adipocyte-derived exosomes from the subject, and the profile of the adipocyte-derived exosomes from the control subject, are each an microRNA expression profile that includes an expression level of at least one microRNA. The expression level of the at least one microRNA from the subject may be higher than the corresponding expression level of the at least one microRNA from the control subject.

In other embodiments, the profile of the isolated adipocyte-derived exosomes from the subject, and the profile of the adipocyte-derived exosomes from the control subject, are each an microRNA expression profile that includes an expression level of at least one microRNA, and the expression level of the at least one microRNA from the subject may be lower than the corresponding expression level of the at least one microRNA from the control subject.

The microRNA expression profile may include an expression level of at least one microRNA, such as miR-33a, miR-10b, miR-20a, miR-374b, miR-504, miR-101-1, let-7a-5p and miR-16-5p.

The subject may be a mammalian subject such as a human.

The biological sample may or may not include adipose tissue, and in some embodiments, adipose tissue is excluded from the biological sample. In some embodiments, the biological sample may include, for example, milk, blood, serum, plasma, ascites, cyst fluid, pleural fluid, peritoneal fluid, cerebral spinal fluid, tears, urine, saliva, sputum, or combinations thereof. In some embodiments, the biological sample is blood, serum, or urine.

The biological sample may be obtained from a lean subject or an obese subject. In some embodiments, the biological sample may be obtained from an obese mammalian subject prior to initiation of a weight-loss treatment, surgery, or regimen. Alternatively, the biological sample may be obtained from an obese mammalian subject after initiation or completion of a weight-loss treatment, surgery, or regimen.

The at least one marker includes, but is not limited to, Fatty Acid Binding Protein 4 (FABP4), Pref-1, amino acid transporter Asc-1, proton amino acid transporter PAT-2, purinergic receptor 2X, ligand-gated ion channel 5 (P2RX5), leptin, EBF3, FBX031, EVA1, CD137, Shox2, Hoxc8, Hoxc9, Tbx1, Zic1, Lhx8, Tnsfrsf9, Tmem26, UCP-1, Cidea, Prdm16, N-acetyltransferase 8-like (Nat8L), prolactin receptor (PRLR), neuregulin-4 (Nrg4), transmembrane protein 120B (Tmem120B), adrenergic β3 receptor (Adrb3), aquaporin-7 (Aqp7), G protein-coupled receptor 81 (Grpr81), G protein-coupled receptor 119 (Gpr119), fatty acid transporter (SLC27a1/FATP1), solute carrier family 7 member 10 (SLC7a10/Asc-1 neutral amino acid transporter), mitochondrial protein uncoupling protein-1 (UCP-1), CD300LG, tetraspanin 18 (tspan18), and/or frizzled-4 (Fzd4). In a particular embodiment, the marker is Fatty Acid Binding Protein 4 (FABP4).

In a particular embodiment, an anti-FABP4 antibody, or antigen-binding fragment thereof, is the binding agent.

In another embodiment, the methods of the invention include use of magnetic beads to isolate adipocyte-derived exosomes.

The binding agent and the at least one marker specific for adipocyte-derived exosomes form a complex.

In one particular aspect of the invention, a method for isolating FABP4+ adipocyte-derived exosomes from a human subject is provided. The method involves obtaining a biological sample from the human subject which contains FABP4+ adipocyte-derived exosomes, and isolating FABP4+ adipocyte-derived exosomes from the biological sample using a FABP4+ antibody. The biological sample may be, for example, blood, serum or urine, and the FABP4+ adipocyte-derived exosomes are isolated using magnetic beads.

In another aspect of the invention, a method of detecting an obesity-related disorder in a human subject is provided. The method involves obtaining a biological sample from the human subject which contains FABP4+ adipocyte-derived exosomes, isolating FABP4+ adipocyte-derived exosomes from the biological sample using a FABP4+ antibody, and detecting the human subject has the obesity-related disorder by a measurable change in a microRNA expression profile of the FABP4+ isolated adipocyte-derived exosomes from the human subject relative to a corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from a control human subject. The biological sample is blood, serum or urine, and the FABP4+ adipocyte-derived exosomes are isolated using magnetic beads. An expression level of at least one microRNA in the microRNA expression profile of isolated of FABP4+ adipocyte-derived exosomes from the human subject is higher relative to an expression level of at least one microRNA in the corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from the control human subject.

In yet another aspect of the invention, a method of detecting susceptibility of a subject to an obesity-related disorder is provided. The method involves obtaining a biological sample from a subject which contains FABP4+ adipocyte-derived exosomes, isolating FABP4+ adipocyte-derived exosomes from the biological sample using a FABP4+ antibody, and detecting the human subject has an increased risk of having an obesity-related disorder by a measurable change in a microRNA expression profile of the FABP4+ isolated adipocyte-derived exosomes from the human subject relative to a corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from a control human subject. The biological sample may be, for example, blood, serum or urine, and the FABP4+ adipocyte-derived exosomes may be isolated using magnetic beads. An expression level of at least one microRNA in the microRNA expression profile of isolated of FABP4+ adipocyte-derived exosomes from the human subject is higher relative to an expression level of at least one microRNA in the corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from the control human subject.

In still another aspect of the invention, a method for identifying a therapy for an obesity-related disorder in a human subject is provided. The method involves obtaining a biological sample from the human subject which contains FABP4+ adipocyte-derived exosomes, isolating FABP4+ adipocyte-derived exosomes from the biological sample using a FABP4+ antibody, detecting the human subject has the obesity-related disorder or an increased risk of having an obesity-related disorder, by a measurable change in a microRNA expression profile of the FABP4+ isolated adipocyte-derived exosomes from the human subject relative to a corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from a control human subject, and identifying the therapy for the obesity-related disorder. The biological sample may be, for example, blood, serum or urine, and the FABP4+ adipocyte-derived exosomes may be isolated using magnetic beads. An expression level of at least one microRNA in the microRNA expression profile of isolated of FABP4+ adipocyte-derived exosomes from the human subject is higher relative to an expression level of at least one microRNA in the corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from the control human subject.

In a particular aspect of the invention, a method of identifying at least one therapeutic agent for an obesity-related disorder in a human subject is provided. The method involves obtaining a biological sample from the human subject which contains FABP4+ adipocyte-derived exosomes, isolating FABP4+ adipocyte-derived exosomes from the biological sample using a FABP4+ antibody, detecting the human subject has the obesity-related disorder or an increased risk of having an obesity-related disorder, by a measurable change in a microRNA expression profile of the FABP4+ isolated adipocyte-derived exosomes from the human subject relative to a corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from a control human subject, and identifying the at least one therapeutic agent for the obesity-related disorder. The biological sample may be, for example, blood, serum or urine, and the FABP4+ adipocyte-derived exosomes may be isolated using magnetic beads. An expression level of at least one microRNA in the microRNA expression profile of isolated of FABP4+ adipocyte-derived exosomes from the human subject is higher relative an expression level of at least one microRNA in the corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from the control human subject.

In still another particular aspect of the invention, a method for treating or preventing an obesity-related disorder in a human subject is provided. The method involves obtaining a biological sample from the human subject which contains FABP4+ adipocyte-derived exosomes, isolating FABP4+ adipocyte-derived exosomes from the biological sample using a FABP4+ antibody, detecting the human subject has the obesity-related disorder or an increased risk of having an obesity-related disorder, by a measurable change in a microRNA expression profile of the isolated FABP4+ adipocyte-derived exosomes from the human subject relative to a corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from a control human subject, and administering a therapy to treat or prevent the obesity-related disorder. The biological sample may be, for example, blood, serum or urine, and the FABP4+ adipocyte-derived exosomes may be isolated using magnetic beads. An expression level of at least one microRNA in the microRNA expression profile of isolated of FABP4+ adipocyte-derived exosomes from the human subject is higher relative to an expression level of at least one microRNA in the corresponding microRNA expression profile of FABP4+ adipocyte-derived exosomes from the control human subject.

