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W 2011/056572 Al (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date 1 - - 12 May 2011 (12.05.2011) W 2011/056572 Al (51) International Patent Classification: AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, G01N 33/68 (2006.01) CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (21) International Application Number: HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, PCT/US20 10/054096 KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, (22) International Filing Date: ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, 26 October 2010 (26.10.2010) NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, (25) Filing Language: English TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (26) Publication Language: English (84) Designated States (unless otherwise indicated, for every (30) Priority Data: kind of regional protection available): ARIPO (BW, GH, 61/255,381 27 October 2009 (27.10.2009) US GM, KE, LR, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, 61/359,695 29 June 2010 (29.06.2010) US ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, (71) Applicant (for all designated States except US): THE EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, BOARD OF TRUSTEES OF THE UNIVERSITY OF LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, ILLINOIS [US/US]; 506 South Wright Street, 352 A d SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, ministration Building, Urbana, IL 61801 (US). GW, ML, MR, NE, SN, TD, TG). (72) Inventor; and Published: (75) Inventor/Applicant (for US only): DUDLEY, Samuel — with international search report (Art. 21(3)) [US/US]; 25 E. Superior St. #3002, Chicago, IL 6061 1 (US). — before the expiration of the time limit for amending the claims and to be republished in the event of receipt of (74) Agents: HONG, Julie, J. et al; Marshall, Gerstein & amendments (Rule 48.2(h)) Borun LLP, 233 S. Wacker Drive, 6300 Willis Tower, Chicago, IL 60606-6357 (US). — with sequence listing part of description (Rule 5.2(a)) (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, (54) Title: METHODS OF DIAGNOSING DIASTOLIC DYSFUNCTION (57) Abstract: Diagnostic methods relating to a cardiac ventricular dysfunction are provided. In some embodiments, the diagnos o tic method is a method of diagnosing diastolic dysfunction in the absence of systolic dysfunction in a subject. The method com- prises assaying a sample obtained from the subject for evidence of activation of renin-angiontensin system (RAS), evidence of ox- idative stress, a level of adiponectin, or a combination thereof, wherein, when there is a lack of evidence of RAS activation, a lack of evidence of oxidative stress, a reduction in the level of adiponectin, or a combination thereof, as compared to a control subject, the subject is diagnosed with diastolic dysfunction in the absence of systolic dysfunction. A method of diagnosing a type of car- diac ventricular dysfunction, a method of determining a therapeutic regimen for a subject suffering from a cardiac ventricular dys function and methods of treating diastolic dysfunction in the absence of systolic dysfunction are also provided. METHODS OF DIAGNOSING DIASTOLIC DYSFUNCTION CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority to U.S. Provisional Patent Application No. 61/255,381, filed on October 27, 2009, and U.S. Provisional Patent Application No. 61/359,695, filed on June 29, 2010, each of which is incorporated by reference in their entirety. GRANT FUNDING [0002] This invention was made with government support under Grant Nos. R01 HL 085558, R01 HL 085520, R01 HL 073753, and P01 HL 058000, awarded by the National Institutes of Health. The government has certain rights in the invention. BACKGROUND [0003] Patients presenting signs and/or symptoms of heart failure may be suffering from systolic dysfunction, diastolic dysfunction, or both types of dysfunction. Successful treatment of the patient depends on the type of cardiac dysfunction present, since the treatment of diastolic dysfunction without systolic dysfunction is very different from the treatments used for patients presenting with systolic dysfunction (with or without systolic dysfunction). [0004] Methods of diagnosing diastolic dysfunction are known in the art and include, for example, Doppler echocardiography, cardiac catheterization, magnetic resonance imaging, tissue Doppler imaging, and measurement of left ventricular end diastolic pressure and systolic function. However, these methods can be invasive, time-consuming, and costly. Accordingly, there exists a need in the art for non-invasive, time-effective, and cost-effective methods of accurately diagnosing diastolic dysfunction in the absence of systolic dysfunction. Such methods would also facilitate practitioners to choose the appropriate treatment for the patient. SUMMARY [0005] Presented herein for the first time are data which demonstrate that diastolic dysfunction in the absence of systolic dysfunction is associated with neither activation of the renin-angiontensisn system (RAS) nor oxidative stress. Also presented herein for the first time are data which demonstrate that diastolic dysfunction in the absence of systolic dysfunction is associated with reduced levels of adiponectin. Accordingly, provided herein are diagnostic methods relating to a cardiac dysfunction, e.g., diastolic dysfunction, systolic dysfunction. In some embodiments, the diagnostic method is a method of diagnosing diastolic dysfunction in the absence of systolic dysfunction in a subject, e.g., a subject exhibiting a sign or symptom of heart failure. The method comprises assaying a sample obtained from the subject for evidence of activation of renin-angiontensin system (RAS), evidence of oxidative stress, a level of adiponectin, or a combination thereof. In some embodiments, the subject is diagnosed with diastolic dysfunction in the absence of systolic dysfunction, when there is a lack of evidence of RAS activation, a lack of evidence of oxidative stress, a reduction in the level of adiponectin, or a combination thereof, as compared to a control subject, e.g., a control subject exhibiting a sign or symptom of heart failure. [0006] In some embodiments, the diagnostic method of the present disclosures is a method of diagnosing a type of heart failure in a subject suffering from a heart failure. The method comprises assaying a sample obtained from the subject for evidence of activation of renin- angiontensin system (RAS), evidence of oxidative stress, a level of adiponectin, or a combination thereof. In some embodiments, the subject is diagnosed with heart failure with preserved ejection fraction, e.g., diastolic heart failure, when there is a lack of evidence of RAS activation, a lack of evidence of oxidative stress, a reduction in the level of adiponectin, or a combination thereof, as compared to a control subject, e.g., a control subject suffering from heart failure (e.g., systolic heart failure, heart failure with systolic dysfunction). [0007] Further provided herein is a method of determining a therapeutic regimen for a subject exhibiting a sign or symptom of heart failure. The method comprises assaying a sample obtained from the subject for evidence of activation of renin-angiontensin system (RAS), evidence of oxidative stress, a level of adiponectin, or a combination thereof. In some embodiments, the therapeutic regimen is determined to be a therapeutic regimen for treating diastolic dysfunction in the absence of systolic dysfunction, when there is a lack of evidence of RAS activation, a lack of evidence of oxidative stress, a reduction in the level of adiponectin, or a combination thereof, as compared to a control subject, e.g., a control subject exhibiting a sign or symptom of heart failure. [0008] A method of treating a subject for diastolic dysfunction in the absence of systolic dysfunction is furthermore provided herein. The method comprises (a) assaying a sample obtained from the subject for evidence of activation of renin-angiontensin system (RAS), evidence of oxidative stress, a level of adiponectin, or a combination thereof, and (b) administering to the subject a therapeutic agent suitable for treating diastolic dysfunction in the absence of systolic dysfunction in an amount effective to treat the diastolic dysfunction. [0009] Moreover, provided herein is a method of treating diastolic dysfunction in the absence of systolic dysfunction in a subject, comprising administering to the subject an agent which increases the level of adiponectin in the subject. [0010] Further provided herein is a method of treating or preventing heart failure with preserved ejection fraction in a subject. The method comprises administering to the subject an agent which increases the level of adiponectin in the subject. BRIEF DESCRIPTION OF THE DRAWINGS [0011] Figure 1 represents a graph of the multivariate odds ratios for association with early diastolic dysfunction {BMI, Body mass index; E CyS , Redox potential of reduced to oxidized cysteine; E GSH, Redox potential of reduced to oxidized glutathione; DROM , Derivatives of reactive oxygen metabolites; IsoP, Isoprostanes; ACE , Angiotensin converting enzyme levels.} [0012] Figure 2 represents a Western blot demonstrating the protein expression of ecSOD in blood samples from DD and control groups. The results depicted in the graph are presented as mean + SE (DD (n=6) and controls (n=12)). [0013] Figure 3 represents a series of graphs demonstrating a comparison of (Top) total, (Center) high molecular weight, and (Bottom) mid + low molecular weight adiponectin levels between patients with and without diastolic dysfunction (DD).
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