Methods and computer program products for determining risk of developing type 2 diabetes and other insulin resistance related disorders

• US 6,518,069 B1
• Filed: 04/19/2000
• Issued: 02/11/2003
• Est. Priority Date: 04/22/1999
• Status: Expired due to Term

• Current Assignee: German University, LipoScience, Inc. (Laboratory Corporation Of America Holdings), North Carolina State University (University of North Carolina System)
• Original Assignee: LipoScience, Inc. (Laboratory Corporation Of America Holdings), North Carolina State University (University of North Carolina System)
• Inventors: Otvos, James D., Bennett, Dennis W.
• Primary Examiner(s): Soderquist, Arlen
• Application Number:
  US09/550,359
• Time in Patent Office: 1,028 Days
• Field of Search: 436/71, 436/95, 436/173
• US Class Current: 436/173
• CPC Class Codes: G01N 24/08 by using nuclear magnetic resonance G01N24/12 takes precedence
• CPC Class Codes: G01N 2405/00 Assays, e.g. immunoassays or enzyme assays, involving lipids lipopolysaccharides G01N2400/50
• CPC Class Codes: G01N 2800/042 Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
• CPC Class Codes: G01N 2800/324 Coronary artery diseases, e.g. angina pectoris, myocardial infarction
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
• CPC Class Codes: G01R 33/465 applied to biological material, e.g. in vitro testing
• CPC Class Codes: Y10T 436/144444 Glucose
• CPC Class Codes: Y10T 436/24 Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry

Abstract

Methods for assessing the risk of developing Type 2 diabetes and other related disorders include obtaining an NMR derived reference spectrum for a known glucose concentration sample and storing this information as a reference standard. A patient blood sample is collected and NMR derived patient spectrums for the blood sample are obtained. The two NMR data sets (the reference and the patient) are compared and a glucose concentration is determined for the patient sample. The glucose concentration can be evaluated with a blood sample undergoing lipoprotein cholesterol evaluation. The NMR based test can be used to concurrently provide a glucose concentration and lipoprotein constituent values based on a single testing event. The disclosure also includes a multi-purpose test, i.e., a test which concurrently provides lipoprotein screening and coronary heart disease risk evaluation along with a diabetes screening and risk assessment for developing Type 2 diabetes. A method for assessing diabetes includes identifying the presence of diabetic dyslipidemia based on the values of predetermined NMR measured lipoprotein constituents.

Claims

1. A method for assessing a patient'"'"'s risk of having or developing Type 2 diabetes and/or insulin resistance disorders based on NMR-derived lipoprotein-based information, comprising the steps of:

obtaining at least one proton NMR spectroscopic signal of a patient'"'"'s blood plasma or serum sample to perform a NMR spectral analysis thereof;

deriving a NMR-based lipoprotein measurement value for a plurality of selected lipoprotein subclass constituents of interest based on said obtaining step;

comparing the NMR-based measured lipoprotein subclass constituent values to predetermined test criteria, wherein the predetermined test criteria define at least one lipoprotein constituent value or range of values having an unfavorable or positive risk factor associated therewith, for each of the plurality of selected lipoprotein subclass constituents of interest; and

assessing a patient'"'"'s risk of having or developing at least one of Type 2 diabetes and/or insulin resistance disorders based on the number of and/or degree of risk associated with individual lipoprotein subclass constituent values having an unfavorable or positive risk factor identified in said comparing step.

2. A method according to claim 1, further comprising:

concurrently generating a NMR-based glucose measurement value for the patient'"'"'s blood plasma or serum sample by considering NMR data from a plurality of peaks in the 3.1-3.9 region of the proton-NMR spectrum;

comparing the NMR-based glucose measurement value to predetermined test criteria; and

determining the patient'"'"'s risk of developing or having insulin resistance or Type 2 diabetes based, at least in part, on the detection of an elevated glucose level.

3. A method according to claim 2, wherein said step of comparing comprises comparing the NMR-based measured values associated with the selected lipoprotein subclass constituents of at least a plurality of the subclass constituents of the group consisting of:

LDL particle size, large HDL concentration, LDL particle concentration, and large VLDL concentration to respective predetermined test criteria and wherein said assessing step is based on the presence of both the determination of the presence of an unfavorable or positive risk factor for a plurality of the selected lipoprotein subclass constituents and the detection of an elevated glucose level.

4. A method according to claim 3, wherein the unfavorable or positive risk factor for LDL particle size in the predetermined test criteria identifies the LDL size as Pattern B.

5. A method according to claim 3, wherein the large HDL concentration predetermined test criteria that defines an unfavorable or positive risk factor identifies a low level of large HDL concentration.

6. A method according to claim 3, wherein the large VLDL concentration predetermined test criteria identifies an elevated level of large VLDL concentration.

7. A method according to claim 3, wherein said comparing step used to identify the patient as having or being at risk for having Type-2 diabetes and/or insulin resistance disorders is based on a positive risk test match for at least three of the selected NMR based lipoprotein subclass constituent measured values.

8. A method according to claim 2, further comprising the step of determining whether the sample indicates the presence of diabetic dyslipidemia in the patient based on said comparing and assessing steps, and wherein the presence of diabetic dyslipidemia is determined based on an unfavorable value for at least two of the measured lipoprotein constituent subclass values as defined by the predetermined test criteria and when the NMR-based glucose measurement value indicates an elevated level of blood glucose.

9. A method according to claim 1, further comprising the step of determining whether the sample indicates the presence of Type 2 diabetes, insulin related disorders, and/or diabetic dyslipidemia in the patient based on said comparing and assessing steps, and wherein the NMR-based lipoprotein constituent values used includes values associated with positive risk factors for at least a plurality of the lipoprotein subclass constituents comprising LDL particle size, LDL particle concentration, large HDL concentration, and large VLDL concentration.

10. A method according to claim 1, wherein the selected lipoprotein subclass constituents of the deriving step comprise at least two of LDL particle size, LDL particle concentration, large HDL concentration, and large VLDL concentration, and wherein the positive or unfavorable predetermined test criteria of the comparing step identifies the LDL particle size test criteria as Pattern B, the large HDL concentration as a low level of large HDL concentration, and the large VLDL concentration as an elevated level of large VLDL concentration, and wherein the presence of an at-risk condition for Type 2 diabetes and/or insulin related disorders is determined in the assessing step based on the positive identification of at least two of the NMR lipoprotein based constituent values to the predetermined test criteria.

11. A method according to claim 1, wherein the assessing step is carried out so that an at-risk condition is identified when a plurality of lipoprotein subclass constituents have unfavorable or positive risk values.

12. A method according to claim 11, wherein the lipoprotein subclass constituents and associated risk values denoted with unfavorable or positive risk includes at least two of:

a low large HDL concentration, an elevated LDL particle concentration, a small LDL particle size, and an elevated large VLDL concentration.

13. A method for assessing a patient'"'"'s risk of developing or having insulin resistance syndrome, insulin resistance disorders, and/or Type-2 diabetes, based on NMR measured lipoprotein-based information, comprising the steps of:

obtaining at least one proton NMR spectroscopic signal of a patient'"'"'s blood plasma or serum sample to perform a NMR spectral analysis thereof;

deriving a NMR-based lipoprotein measurement value for a plurality of lipoprotein subclass constituents based on said obtaining step;

comparing the NMR-based measured lipoprotein subclass constituent values to predetermined test criteria, wherein the predetermined test criteria define unfavorable or positive risk factors for each of the lipoprotein subclass constituents of interest which are associated with insulin resistance syndrome, insulin resistance disorders and/or Type 2 diabetes; and

assessing a patient'"'"'s risk of developing or having insulin resistance syndrome, insulin resistance disorders, and/or Type-2 diabetes based on the number of and/or degree of risk associated with the plurality of individual lipoprotein subclass constituents that are identified as having an unfavorable or positive risk factor in said comparing step.

14. A method according to claim 13, further comprising:

concurrently generating at least one NMR-based glucose measurement value for the patient'"'"'s blood plasma or serum sample based on the NMR spectral analysis of said obtaining step, the glucose measurement comprises evaluating the amplitudes of a plurality of peaks in the portion of the NMR spectrum between about 3.1-3.9 ppm;

comparing the NMR-based glucose measurement value to predetermined test criteria that defines elevated levels of glucose; and

wherein said assessing step further considers the detection of elevated glucose from said comparing step.

15. A method according to claim 14, wherein the patient is identified as being at risk for having or developing insulin resistance disorders, insulin resistance syndrome and/or diabetes when both (a) the NMR-based glucose measurement value is an elevated level of blood glucose;

and (b) the concurrent positive identification that at least two of the lipoprotein based constituent subclass values are unfavorable according to the predetermined test criteria.

16. A method according to claim 13, wherein the NMR-based lipoprotein subclass constituent values of said deriving and comparing steps comprise the values associated with LDL particle size, LDL particle concentration, large HDL concentration, and large VLDL concentration.

17. A method according to claim 13, wherein said step of comparing a plurality of NMR-based lipoprotein subclass constituent values comprises comparing at least a plurality of the NMR measured values associated with LDL particle size, LDL particle concentration, large HDL concentration, and large VLDL concentration to respective predetermined test criteria.

18. A method according to claim 17, wherein the LDL particle size predetermined test criteria identifies the LDL particle size as Pattern B.

19. A method according to claim 17, wherein the large HDL concentration-predetermined test, criteria identifies a low level of large HDL concentration.

20. A method according to claim 17, wherein the large VLDL concentration predetermined test criteria identifies an elevated level of large VLDL concentration.

21. A method according to claim 17, wherein said comparing step is carried out to determine if there is a positive test match representative of an unfavorable risk factor for at least two of the identified NMR based lipoprotein subclass constituent values.

22. A method according to claim 17, wherein the step of comparing compares the measured values of LDL particle size, LDL particle concentration, large HDL concentration, and large VLDL concentration, and wherein the LDL particle size predetermined test criteria identifies the LDL particle size as pattern B, the large HDL concentration predetermined test criteria identifies a low level of the large HDL concentration, the LDL particle concentration predetermined test criteria identifies the LDL particle concentration as elevated, and the large VLDL concentration test criteria identifies an elevated level of large VLDL concentration, and wherein the assessing step is determined based on the identification that at least two of these measured lipoprotein subclass constituent values are unfavorable risk factors as defined by the predetermined test criteria.

23. A method of evaluating the efficacy of treatment of a subject undergoing treatment for insulin resistance disorders, insulin resistance syndrome, and/or Type 2 diabetes based on personalized NMR measured lipoprotein information, comprising:

obtaining a baseline profile including measured values for a plurality of lipoprotein subclass constituents in a patient'"'"'s blood plasma or serum sample, wherein the baseline profile is derived from a proton NMR spectral analysis of a NMR spectroscopic signal of the patient'"'"'s blood plasma or serum sample which measures lipoprotein constituent values of concentration or size, wherein the baseline profile includes a risk assessment analysis that considers the number of lipoprotein subclass constituent values that have an unfavorable risk factor associated therewith according to predetermined risk criteria to identify whether the patient is at risk for having or developing insulin resistance related disorders, insulin resistance syndrome, and/or Type-2 diabetes;

obtaining a second profile of proton NMR measured lipoprotein subclass constituent values for a patient'"'"'s blood plasma or serum sample after the subject has undergone treatment for insulin resistance related disorders, insulin resistance syndrome and/or Type 2 diabetes; and

automatically comparing the second profile with the baseline profile to identify differences in the lipoprotein subclass constituent measurements and/or risk analysis therebetween, wherein a difference between the second profile and the baseline profile of the subject provides an indication of the efficacy of treatment for insulin resistance related disorders, insulin resistance syndrome, and/or Type 2 diabetes of the subject.

24. The method according to claim 23, wherein the proton NMR spectral analysis of the baseline profile and the second profile are carried out to provide a glucose measurement value for the patient'"'"'s blood plasma or serum sample, wherein the glucose measurement is obtained concurrently with the proton NMR measured lipoprotein constituent values of the subject by evaluating the amplitudes of a plurality of peaks in the portion of the NMR spectrum between about 3.1-3.9 ppm.

25. An NMR-based method for measuring the concentration of glucose concentration of blood plasma or serum specimen, comprising the steps of:

acquiring a reference data spectrum in a region of interest corresponding to glucose in a reference blood plasma or serum specimen;

determining a reference glucose lineshape in the reference spectrum based on at least one signal in the reference spectrum, wherein the reference glucose lineshape spectrum in blood plasma comprises a plurality of peaks within a region extending between about 3.1-3.9 ppm of a proton NMR spectrum of the blood plasma or serum specimen;

assigning a reference coefficient to the reference glucose lineshape in the reference spectrum;

acquiring an NMR spectrum of a patient'"'"'s blood plasma or serum specimen corresponding to the region of interest in the reference specimen, the patient spectrum comprising a plurality of peaks within a region extending between about 3.1-3.9 ppm;

identifying a patient glucose lineshape with the plurality of peaks in about the 3.1-3.9 ppm region of the spectrum for the patient specimen in the patient NMR spectrum based on the same at least one signal used to determine the reference glucose lineshape in the reference spectrum;

comparing the reference glucose lineshape with the patient glucose lineshape;

fitting the amplitudes of the plurality of peaks in the reference glucose lineshape to the plurality of peaks in the patient glucose lineshape in the region extending between about 3.1-3.9 ppm;

calculating a scaling factor based on said fitting step; and

determining the concentration of glucose in the patient blood plasma or serum specimen as a function of the value of the scaling factor and reference coefficient.

26. A method according to claim 25, wherein the value of the reference coefficient is based on an independent chemical measurement of glucose concentration of the reference specimen.

27. A method according to claim 25, wherein the NMR reference and patient spectrums are obtained at substantially the same magnetic field strength and same specimen temperature.

28. A method according to claim 25, wherein the glucose reference spectrum is within a region extending between about 3.64-3.90 ppm of the proton NMR spectrum of blood plasma.

29. A method according to claim 25, wherein the glucose reference spectrum is within a region extending between about 3.29-3.54 ppm of the proton NMR spectrum of blood plasma.

30. A method according to claim 25, wherein the glucose reference spectrum is within at least one of two regions of interest of the proton NMR spectrum of blood plasma or serum, a first region extending between about 3.64-3.90 ppm and a second region extending between about 3.29-3.54 ppm.

31. A computer program product for determining whether a subject is at risk for having or developing insulin resistance disorders and/or Type 2 diabetes, comprising a computer-readable storage medium having computer-readable program code means embodied in the medium, the computer-readable program code means comprising:

computer code means for generating a proton NMR spectrum and measuring the values associated with concentration or size of a plurality of lipoprotein subclass constituents for a patient'"'"'s blood plasma or serum sample;

computer code means for comparing a plurality of NMR-measured lipoprotein subclass constituent values to predetermined test criteria defined as an unfavorable value or range of values for selected lipoprotein subclass constituents; and

computer code means for assessing a patient'"'"'s risk of developing or having Type 2 diabetes and/or insulin resistance disorders based on the number of individual lipoprotein subclass constituents identified as having an unfavorable or positive risk factor associated with the measured values.

32. The computer program product of claim 31, further comprising:

computer code means for concurrently generating a proton NMR spectral analysis of a proton NMR spectrum to define the glucose level for the patient'"'"'s blood plasma or serum sample, wherein the computer code means comprises generating a patient glucose lineshape having a plurality of peaks that rise and fall in relation to the concentration of glucose, the peaks residing in a portion of the spectrum between about 3.1 to 3.9 ppm, comparing the glucose spectrum to a glucose reference lineshape having an associated amplitude, fitting the amplitude of the reference glucose lineshape to the patient glucose lineshape and determining the concentration of glucose in the patient blood plasma or serum specimen;

computer code means for comparing the measured glucose concentration to predetermined test criteria to assess whether the glucose measurement is elevated; and

computer code means for assessing the patient'"'"'s risk of having or developing insulin resistance disorders and/or Type 2 diabetes which considers the presence of an elevated glucose level.

33. A screening test method for concurrently screening a patient for the risk of coronary heart disease and insulin resistance or Type-2 diabetes, comprising the steps of:

performing a proton NMR spectral analysis of a patient'"'"'s blood plasma or serum sample, the spectral analysis producing a spectrum having a plurality of peaks in the region between about 3.1-3.9 ppm;

measuring a plurality of lipoprotein subclass constituent values based on said performing step;

assessing the patient'"'"'s risk of coronary heart disease based on said measuring step;

concurrently measuring a glucose level in the sample based on said performing step by evaluating a plurality of the peaks in the 3.1-3.9 ppm region of the spectrum;

determining the patient'"'"'s risk of developing or having Type 2 diabetes or insulin resistance disorders based on said measuring steps; and

generating an individualized patient report presenting the risk of having or developing coronary heart disease and insulin resistance and/or Type-2 diabetes.

34. A screening test method of claim 33, further comprising assessing the risk of developing gestational diabetes.

35. A screening test method of claim 33, further comprising assessing the risk of developing juvenile diabetes.

36. A method for concurrently obtaining measurement data to assess both the risk of CHD and insulin resistance or related disorders and/or Type-2 diabetes for a patient, comprising:

obtaining data associated with at least one NMR spectroscopic signal of an in vitro sample of a patient'"'"'s blood or serum to perform a NMR spectral analysis thereof;

deriving a plurality of NMR-based measurements for a plurality of different lipoprotein subclass constituents comprising at least two of LDL particle size, LDL particle concentration, large HDL concentration, and large VLDL concentration;

evaluating the patient'"'"'s risk of having or developing coronary heart disease (CHD) based on selected measurements obtained from said obtaining and deriving steps based on predetermined CHD test criteria; and

concurrently determining the patient'"'"'s risk of having or developing insulin resistance related disorders and/or Type-2 diabetes based on selected measurements obtained from said deriving step and based on predetermined test criteria for insulin resistance related disorders and/or Type-2 diabetes, wherein the test criteria defines values and their associated degree of risk, including increased and decreased values, for the selected lipoprotein subclass constituent measurements and the risk is determined to be present when a plurality of the selected lipoprotein subclass constituents present with increased risk values.

37. A method according to claim 36, wherein the evaluating and determining steps consider at least one common lipoprotein particle subclass constituent value.

38. A method according to claim 36, further comprising concurrently generating a NMR-based glucose measurement value for the patient'"'"'s blood plasma or serum sample by evaluating NMR data from a plurality of peaks in the 3.1-3.9 region of the proton NMR spectrum;

and

comparing the NMR-based glucose measurement value to predetermined test criteria;

wherein said determining step considers the glucose value in establishing the patient'"'"'s risk of having or developing insulin resistance related disorders and/or Type-2 diabetes.

39. A method according to claim 36, wherein said evaluating and determining steps are performed based on NMR data gathered simultaneously on the same patient sample.

40. A screening method for concurrently assessing a patient'"'"'s risk of having or developing coronary heart disease and/or insulin resistance related disorders and/or Type-2 diabetes, comprising:

measuring a plurality of lipoprotein subclass constituents in a patient'"'"'s blood or plasma sample using NMR spectroscopy-derived data, the lipoprotein subclass constituent measurements being carried out to measure at least two of;

LDL particle size, LDL particle concentration, large HDL concentration, and/or large VLDL concentration;

evaluating the patient'"'"'s risk of having or developing coronary heart disease (CHD) based on predetermined CHD test criteria using selected measurements obtained from said measuring step;

determining the patient'"'"'s risk of having or developing insulin resistance related disorders and/or Type-2 diabetes based on predetermined insulin resistance related disorders and/or Type-2 diabetes test criteria using selected measurements obtained from said measuring step; and

repeating said measuring, evaluating, and determining steps for a plurality of different patients to provide a mass screening test that is able to identify at-risk patients.

41. A method according to claim 40, further comprising measuring glucose in the patient'"'"'s blood or plasma sample using NMR spectroscopy derived data that includes data that corresponds to a plurality of peaks in about the 3.1-3.9 portion of a NMR proton spectrum.

42. A method according to claim 40, wherein the lipoprotein subclass constituents and associated risk values denoted with unfavorable or positive risk includes at least two of:

a low large HDL concentration, an elevated LDL particle concentration, a small LDL particle size, and an elevated large VLDL concentration.

43. A mass screening method for assessing a patient'"'"'s risk of having or developing Type 2 diabetes and/or insulin resistance disorders, comprising:

deriving a respective No spectroscopy based lipoprotein measurement value for each of a plurality of selected lipoprotein subclass constituents of interest in a blood plasma or serum sample of a patient;

comparing the measured lipoprotein subclass constituent values to predetermined test criteria, wherein the predetermined test criteria define a respective lipoprotein subclass constituent value or range of values having an unfavorable or positive risk factor associated therewith for each of the plurality of selected lipoprotein subclass constituents of interest;

assessing the patient'"'"'s risk of having or developing at least one of Type 2 diabetes and/or insulin resistance disorders based on the number of individual lipoprotein subclass constituents having an unfavorable or positive risk factor identified in said comparing step; and

automatically repeating the deriving, comparing, and assessing steps for a plurality of different patient samples to thereby perform a mass screening test that is able to identify at-risk patients.

44. A method according to claim 43, further comprising measuring glucose in the patient'"'"'s blood or plasma sample using NMR spectroscopy derived data that includes data that corresponds to a plurality of peaks in about the 3.1-3.9 portion of a NMR proton spectrum.

45. A method according to claim 43, wherein the lipoprotein subclass constituents and associated risk values denoted with unfavorable or positive risk includes at least two of:

a low large HDL concentration, an elevated LDL particle concentration, a small LDL particle size, and an elevated large VLDL concentration.