IGFBP7 RATIO FOR HFpEF

Abstract

The present invention relates to a method for differentiating between heart failure with preserved ejection fraction (HFpEF) and heart failure with reduced ejection fraction (HFrEF) in a subject suffering from heart failure, said method comprising the steps of determining the amounts of IGFBP7 (Insulin-like growth factor-binding protein 7), of a BNP-type peptide, and optionally of CRP (C-reactive protein) in a sample from the subject, calculating (i) a ratio of the amount of IGFBP7 and the amount of the BNP-peptide or (ii) a ratio of the sum of the amounts of IGFBP7 and CRP and the amount of the BNP-type peptide, comparing the ratio calculated with a reference ratio, and differentiating between heart failure with preserved ejection fraction (HFpEF) and heart failure with reduced ejection fraction (HFrEF). The present invention further concerns a method for the diagnosis of HFpEF.

Claims

1-18. (canceled)

19. A method for measuring a panel of biomarkers in a sample, the method comprising: obtaining the sample from a subject suffering from, or suspected to suffer from, heart failure with preserved ejection fraction (HFpEF) or heart failure with reduced ejection fraction (HFrEF); and determining a measurement for the panel of biomarkers in the sample, wherein the panel comprises the biomarkers IGFBP7 (Insulin-like growth factor-binding protein 7), a BNP-type peptide, and CRP (C-reactive protein), wherein the measurement comprises determining an amount of each of the biomarkers IGFBP7, the BNP-type peptide, and CRP in the panel; and wherein the sample is a blood sample, a plasma sample, or a serum sample.

20. The method of claim 19, wherein the BNP-type peptide is BNP or NT-proBNP.

21. The method of claim 19, wherein determining a measurement for the panel of biomarkers comprises: contacting the sample, or a portion thereof, with a detection agent which specifically binds to IGFBP7, a detection agent which specifically binds to the BNP-type peptide, and a detection agent which specifically binds to CRP; and determining the amount of IGFBP7 bound to the detection agent which specifically binds to IGFBP7, the amount of the BNP-type peptide bound to the detection agent which specifically binds to the BNP-type peptide, and the amount of CRP bound to the detection agent which specifically binds to CRP.

22. The method of claim 21, wherein the detection agent which specifically binds to IGFBP7 is ruthenylated or iridinylated, wherein the detection agent which specifically binds to the BNP-type peptide is ruthenylated or iridinylated, and wherein the detection agent which specifically binds to CRP is ruthenylated or iridinylated.

23. The method of claim 19, wherein the measuring is performed using a quantitative detection method.

24. The method of claim 23, wherein the quantitative detection method is an immunoassay.

25. The method of claim 19, further comprising detecting an elevated ratio of biomarkers, wherein the detecting comprises: calculating a ratio of the sum of the amounts of IGFBP7 and CRP and the amount of the BNP-type peptide, comparing the calculated ratio with a reference ratio, and detecting the calculated ratio is elevated versus the reference ratio.

26. The method of claim 19, further comprising detecting a reduced ratio of biomarkers, wherein the detecting comprises: calculating a ratio of the sum of the amounts of IGFBP7 and CRP and the amount of the BNP-type peptide, comparing the calculated ratio with a reference ratio, and detecting the calculated ratio is reduced versus the reference ratio.

27. The method of claim 19, wherein the subject is a human.

28. A method for measuring an amount of a first biomarker, a second biomarker, and a third biomarker in a sample from a subject, the method comprising: obtaining the sample from the subject; measuring an amount of the first biomarker in the sample, wherein the first biomarker is IGFBP7 (Insulin-like growth factor-binding protein 7); measuring an amount of the second biomarker in the sample, wherein the second biomarker is a BNP-type peptide; and measuring an amount of the third biomarker in the sample, wherein the third biomarker is CRP (C-reactive protein); and contacting the sample, or a portion thereof, with a detection agent which specifically binds to IGFBP7, a detection agent which specifically binds to the BNP-type peptide, and a detection agent which specifically binds to CRP, wherein the sample is a blood sample, a serum sample, or a plasma sample.

29. The method of claim 28, wherein the BNP-type peptide is BNP or NT-proBNP.

30. The method of claim 28, wherein the subject is a human.

31. The method of claim 28, wherein the measuring is performed using a quantitative detection method.

32. The method of claim 31, wherein the quantitative detection method is an immunoassay.

33. A method of detecting IGFBP7 (Insulin-like growth factor-binding protein 7), a BNP-type peptide, and CRP (C-reactive protein) in a subject, the method comprising: obtaining a blood sample, a serum sample, or a plasma sample from the subject, and detecting whether IGFBP7, the BNP-type peptide, and CRP are present in the sample by contacting the sample with a detection agent which specifically binds to IGFBP7, a detection agent which specifically binds to the BNP-type peptide, and a detection agent which specifically binds to CRP and detecting the binding between IGFBP7, the BNP-type peptide, and CRP and the detection agent which specifically binds to IGFBP7, the detection agent which specifically binds to the BNP-type peptide, and the detection agent which specifically binds to CRP.

34. The method of claim 33, wherein the subject suffers from, or is suspected to suffer from, heart failure with preserved ejection fraction (HFpEF) or heart failure with reduced ejection fraction (HFrEF)

35. The method of claim 33, wherein the BNP-type peptide is BNP or NT-proBNP.

36. The method of claim 33, wherein the subject is a human.

37. The method of claim 33, wherein the detecting is performed using a quantitative detection method.

38. The method of claim 37, wherein the quantitative detection method is an immunoassay.

Description

[0177] The Figures show:

[0178] FIGS. 1A-1D show: Biomarker Measurements for the Diagnosis of HFpEF

[0179] FIG. 1A: Measurement of NT-proBNP in 499 samples of patients with HFrEF and in 123 samples of patients with HFpEF for the Diagnosis of HFpEF

[0180] FIG. 1B: Measurement of CRP in 411 samples of patients with HFrEF and in 107 samples of patients with HFpEF for the Diagnosis of HFpEF

[0181] FIG. 1C: Measurement of GDF15 in 451 samples of patients with HFrEF and in 110 samples of patients with HFpEF for the Diagnosis of HFpEF

[0182] FIG. 1D: Measurement of IGFBP-7 in 366 samples of patients with heart failure with HFrEF and in 96 samples of patients with HFpEF for the Diagnosis of HFpEF

[0183] FIGS. 2A and 2B show: Biomarker Ratio with GDF15 for the Diagnosis of HFpEF

[0184] FIG. 2A: Diagnosis of HFpEF versus HFrEF in patients with heart failure with the Ratio CRP+GDF15 (not elevated in HFrEF versus HFpEF)/NTproBNP (elevated in HFrEF versus HFpEF) in 410 samples of patients suffering from HFrEF versus 107 patients suffering from HFpEF

[0185] FIG. 2B: Diagnosis of HFpEF versus HFrEF in patients with heart failure with the Ratio CRP+GDF15+ST2 (not elevated in HFrEF versus HFpEF)/NTproBNP (elevated in HFrEF versus HFpEF) in 411 samples of patients suffering from HFrEF versus 107 patients suffering from HFpEF

[0186] FIGS. 3A-3C show: Biomarker Ratio with IGFBP-7 for the Diagnosis of HFpEF

[0187] FIG. 3A: Diagnosis of HFpEF versus HFrEF in patients with heart failure with the Ratio CRP+IGFBP-7 (not elevated in HFrEF versus HFpEF)/NTproBNP (elevated in HFrEF versus HFpEF) in 359 samples of patients suffering from HFrEF versus 96 patients suffering from HFpEF

[0188] FIG. 3B: Diagnosis of HFpEF versus HFrEF in patients with heart failure with the Ratio CRP+IGFBP-7+ST2 (not elevated in HFrEF versus HFpEF)/NTproBNP (elevated in HFrEF versus HFpEF) in 359 samples of patients suffering from HFrEF versus 96 patients suffering from HFpEF

[0189] FIG. 3C: Diagnosis of HFpEF versus HFrEF in patients with heart failure with the Ratio GDF-15+IGFBP-7 (not elevated in HFrEF versus HFpEF)/NTproBNP (elevated in HFrEF versus HFpEF)

[0190] All references cited herein are herewith incorporated by reference with respect to their disclosure content in general or with respect to the specific disclosure contents indicated above.

[0191] The invention will now be illustrated by the following Examples which are not intended to restrict or limit the scope of this invention.

EXAMPLES

Example 1: Assessment of HFpEF With Circulating Biomarkers

[0192] NTproBNP, GDF-15, ST2, CRP (hs CRP) and IGFBP-7 levels were determined in HF patients with and without reduced ejection fraction.

[0193] The results are shown in FIGS. 1A-1D. NTproBNP was significantly elevated in patients with heart failure with reduced ejection fraction (HFrEF) in comparison to patients with heart failure with preserved ejection fraction (HFpEF) (AUC 0.659), see FIG. 1A. FIG. 1B shows that CRP was not significantly elevated in patients with heart failure with reduced ejection fraction (HFrEF) in comparison to patients with heart failure with preserved ejection fraction (HFpEF) (AUC 0.575). FIG. 1C shows that GDF15 is not significantly elevated in patients with heart failure with reduced ejection fraction (HFrEF) in comparison to patients with heart failure with preserved ejection fraction (HFpEF) (AUC 0.554). FIG. 1D shows that IGFBP-7 is not significantly elevated in patients with heart failure with reduced ejection fraction (HFrEF) in comparison to patients with heart failure with preserved ejection fraction (HFpEF) (AUC 0.494).

[0194] Table 1 shows the titers of circulating biomarkers in the subgroups of heart failure patients with reduced ejection fraction (HFrEF) and in the subgroup of heart failure patients with preserved ejection fraction (HFpEF). Biomarker data of the complete set of biomarkers (NTproBNP, GDF15, ST2, CRPhs and IGFBP7) were available for 366 samples of patients with HFrEF and for 96 patient samples with HFpEF.

TABLE-US-00001 TABLE 1 Titers of circulating biomarkers in HFpEF versus HFrEF patients. Biomarkers HFrEF HFpEF Median population population values n = 366 n = 96 p-value NT-proBNP (pg/mL) 4410 2189 <0.001 GDF-15 (pg/mL) 3939 4304 >0.001 ST2 (ng/mL) 35.7 37.6 >0.001 CRP hs (ng/mL) 6.66 8.54 >0.001 IGFBP7 (ng/mL) 242.1 237.1 >0.001

[0195] Table 1 clearly shows that NTproBNP was detected with significant different marker concentrations in both subgroups (HFrEF population in comparison to HFrEF population) in univariate analyses.

[0196] HFrEF patients exhibited higher NT-proBNP titers in comparison to HFpEF patients [4410 vs. 2189 pg/mL median values, p<0.001]. In contrast for GDF15, ST2, CRP and IGFBP7 only slightly elevated marker concentrations were detected in the subgroup of HFpEF patients in comparison to the subgroup of HFrEF patients. However, the observed differences in marker concentrations between HFpEF and HFrEF were not significant for GDF15, ST2, CRPhs, IGFBP-7 in univariate analyses. HFpEF patients exhibited slightly higher GDF15, ST2, CRPhs, IGFBP-7 titers in comparison to HFrEF patients [4304 vs. 3939 pg/mL median values, 37.6 vs. 35.7 pg/mL; 8.54 vs. 6.66 ng/ml; 237.1 vs. 242.1 ng/ml; p>0.001].

[0197] It follows from the above that differential diagnosis and identification of patients with HFpEF is challenging. Only NTproBNP was observed with significant elevated circulating titers in HFrEF patients with HFpEF patients. In contrast no significant differences between marker levels in HFpEF and HFrEF patients were detected for CRP, GDF15, ST2 or IGFBP-7. Thus there was no obvious hint for an improved diagnosis of HFpEF with a marker combination in comparison to measurement of NTproBNP.

Example 2: Ratio With GDF15 for the Diagnosis of HFpEF

[0198] As shown in Table 1, increased levels of circulating NTproBNP levels could be detected from patients with HFrEF in comparison to patients with HFpEF. In contrast, no significant difference of circulating levels between HF patients with or without reduced ejection fraction could be detected for CRP and GDF15.

[0199] A Ratio CRP+GDF15 (not elevated in HFrEF versus HFpEF)/NTproBNP (elevated in HFrEF versus HFpEF) was assessed for an improved diagnosis of HFpEF.

[0200] As shown in FIG. 2A the observed AUC of the Ratio (CRP+GDF15)/NTproBNP for the detection of HFpEF was 0.671. As shown in FIG. 2B, no further improvement could be achieved with the Ratio (CRP+GDF15+ST2)/NTproBNP in comparison to the Ratio (CRP+GDF15)/NTproBNP. The observed AUC of the Ratio (CRP+GDF15+ST2)/NTproBNP for the detection of HFpEF was 0.671.

[0201] Based on the results, it was concluded, that the Ratio (CRP+GDF15)/NTproBNP improves the diagnosis of HFpEF in comparison to single biomarker determinations of NTproBNP or GDF15 or CRP.

Example 3: Biomarker Ratio With IGFBP-7 for the Diagnosis of HFpEF

[0202] As shown in Table 1, increased levels of circulating NTproBNP levels could be detected from patients with HFrEF in comparison to patients with HFpEF. In contrast, no significant difference of circulating levels between HF patients with or without reduced ejection fraction could be detected for CRP and IGFBP-7. The Ratio CRP+IGFBP-7 (not elevated in HFrEF versus HFpEF)/NTproBNP (elevated in HFrEF versus HFpEF) was assessed for an improved diagnosis of HFpEF.

[0203] As shown in FIG. 3A the observed AUC of a calculated index of the sum of the biomarkers (CRP+IGFBP-7)/NTproBNP for the detection of HFpEF was 0.699. In comparison the observed AUC of the ratio IGFBP-7/NTproBNP for detection of HFpEF was 0.680. As shown in FIG. 3C the observed AUC of a calculated index of the sum of the biomarkers (GDF-15+IGFBP-7)/NTproBNP for the detection of HF was 0.693. As shown in FIG. 3B comparable performances were observed with the Ratio (CRP+IGFBP-7+ST2)/NTproBNP in comparison to the Ratio (CRP+IGFBP-7)/NTproBNP. The observed AUC of the Ratio (CRP+IGFBP-7+ST2):NTproBNP for the detection of HFpEF was 0.703. Surprisingly the calculated index of the sum of the biomarkers (CRP +IGFBP-7):NTproBNP even showed superior performance for diagnosis of HFpEF versus the Ratio (CRP+GDF15)/NTproBNP. The Ratio (CRP+IGFBP-7)/NTproBNP even shows superior performance versus the Ratio (CRP+GDF15+ST2)/NTproBNP, which has been described by Sinning et al. (AUC 0.699 vs. AUC 0.671).

[0204] Table 2 shows fold changes of calculated indices in HFpEF versus HFrEF patients.

TABLE-US-00002 Fold change HFpEF vs Algorithms HFrEF (IGFBP-7)/NTproBNP 74.97% (IGFBP-7 + CRP)/NTproBNP 87.81% (GDF15 + CRP)/NTproBNP 66.49% (GDF15 + CRP + ST2)/NTproBNP 65.34%

[0205] Fold changes were calculated ((median HFpEF)/(median HFrEF))-1*100.

[0206] Table 2 clearly shows the improved performance in differential diagnosis of HFpEF versus HFrEF of the calculated index of the sum of (IGFBP-7+CRP)/NTproBNP in comparison to the calculated index of the sum of (GDF15+CRP+ST2)/NTproBNP (Fold change 87.81% versus 65.34%). In comparison for the ratio IGFBP-7/NTproBNP a Fold change of 74.97% was observed. [0207] In conclusion IGFFBP-7 improves the identification of HFpEF in comparison to NTproBNP, GDF15, CRP based scores without IGFBP-7. [0208] The present score with IGFBP-7 shows a better effect size in comparison to the published score from Sinning et al. [0209] It was concluded, that an algorithm including the Ratio (CRP+IGFBP-7)/NTproBNP is useful to aid in diagnosis of patients with HFpEF. The algorithm may be used in diagnosis of HFpEF in patients with heart failure and for identification of HFpEF in individuals suspected of having heart failure.

Example 4: Case Studies

[0210] A 76 year old female obese patient with hypertension presents with shortness of breath. NTproBNP, IGFBP-7 and CRP are determined in a serum sample obtained from the patient (With Elecsys NTproBNP, Elecsys IGFBP-7 and Cobas CRP). Since NTproBNP values might be in the grey zone because of the presence of obesity differential diagnosis of heart failure is performed. The NTproBNP value is 476 pg/mL, the IGFBP-7 value 190.8 ng/mL and the CRP value is 5.8 ng/mL. The calculated index (CRP+IGFBP-7): NTproBNP is 0.41which is elevated versus a calculated reference ratio (0.09). The reference ratio was calculated as median ratio in a reference cohort. The obtained ratio (0.41) is indicative of heart failure with preserved ejection fraction and the patient has been identified to benefit more from a treatment with a HFpEF modifying strategy, eg treatment with SGLT2 inhibitors. The therapy of the patient is adapted accordingly.

[0211] A 81 year old male patient with hypertension presents with a history of atrial fibrillation and shortness of breath. NTproBNP, IGFBP-7 and CRP are determined in a serum sample obtained from the patient (with the kits described above). The observed ejection fraction is in the grey zone (50%). Since NTproBNP values may as well be elevated because of the presence of atrial fibrillation differential diagnosis of heart failure is performed. The NTproBNP value is 3289 pg/mL, the IGFBP-7 value 250.7 ng/ml and the CRP value is 10.8 ng/mL. The calculated index (CRP+IGFBP-7): NTproBNP is 0.79 which is elevated versus a calculated reference ratio (0.09). The reference ratio was calculated as Median ratio in a reference cohort. The obtained ratio (0.79) is indicative of heart failure with preserved ejection fraction and the patient has been identified to benefit more from a treatment with a HFpEF modifying strategy, eg treatment with SGLT2 inhibitors. The therapy of the patient is adapted accordingly.

[0212] A 68 year old obese female patient with Diabetes Mellitus and hypertension presents with symptoms of exercise intolerance. Since symptoms of obesity and symptoms of heart failure with preserved ejection fraction are similar, eg exercise intolerance differential diagnosis of heart failure is performed. NTproBNP, IGFBP-7 and CRP are determined in a serum sample obtained from the patient (with the kits described above). The NTproBNP value is 4643 pg/mL, the IGFBP-7 value 245.5 ng/ml and the CRP value is 5.6 ng/ml. The calculated index (CRP+IGFBP-7):NTproBNP is 0.05 which is reduced versus the reference ratio (0.09). This result is indicative of heart failure with reduced ejection fraction and the patient has been identified not to benefit more from a treatment with a HFpEF modifying strategy, eg treatment with SGLT2 inhibitors. Thus the therapy of the patient is not adapted accordingly.