Methods of identifying and monitoring mitochondrial dysfunction using monocyte screening

• US 10,627,392 B2
• Filed: 06/17/2015
• Issued: 04/21/2020
• Est. Priority Date: 06/17/2014
• Status: Active Grant

• Current Assignee: Stealth BioTherapeutics, Inc. (Morningside Group Ltd.)
• Original Assignee: Stealth BioTherapeutics Corp. (Morningside Group Ltd.)
• Inventors: Wilson, D. Travis
• Primary Examiner(s): Steele, Amber D
• Application Number:
  US15/318,876
• Publication Number:
  US 20170146521A1
• Time in Patent Office: 1,770 Days
• CPC Class Codes: G01N 2800/2835 Movement disorders, e.g. Parkinson, Huntington, Tourette
• CPC Class Codes: G01N 2800/2871 Cerebrovascular disorders, e.g. stroke, cerebral infarct, cerebral haemorrhage, transient ischemic event
• CPC Class Codes: G01N 2800/325 Heart failure or cardiac arrest, e.g. cardiomyopathy, congestive heart failure
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80

Abstract

The present technology provides methods for detecting and diagnosing diseases and conditions characterized by mitochondrial dysfunction using monocytes as an indicator of the dysfunction.

Claims

1. A method for evaluating the therapeutic efficacy of an aromatic-cationic peptide on a disease or condition characterized by mitochondrial dysfunction in a subject, comprising:

assaying the level of a population of activated monocytes present in a biological sample obtained from the subject; and

comparing the level of the population of activated monocytes observed in step (a) with the level of a corresponding population of activated monocytes observed in a biological sample obtained from the subject following administration of a dose of an aromatic-cationic peptide,

wherein the aromatic-cationic peptide is identified as having a therapeutic effect on the disease or condition characterized by mitochondrial dysfunction if the level of the population of activated monocytes in the biological sample following the administration of the aromatic-cationic peptide is reduced compared to the level of the population of activated monocytes observed in step (a).

2. The method of claim 1, wherein the aromatic-cationic peptide is Phe-D-Arg-Phe-Lys-NH₂ or D-Arg-2′

6′

Dmt-Lys-Phe-NH₂ or a pharmaceutically acceptable salt thereof.

3. The method of claim 1, wherein the level of classical monocytes (CD14^high CD16⁻

) in the biological sample following the administration of the aromatic-cationic peptide is reduced compared to the level of classical monocytes observed in step (a).

4. The method of claim 1, wherein the level of intermediate monocytes (CD14^highCD16⁺) in the biological sample following the administration of the aromatic-cationic peptide is reduced compared to the level of intermediate monocytes observed in step (a).

5. The method of claim 1, wherein the level of non-classical monocytes (CD14^lowCD16^high) in the biological sample following the administration of the aromatic-cationic peptide is increased compared to the level of non-classical monocytes observed in step (a).

6. The method of claim 1, wherein the disease or condition characterized by mitochondrial dysfunction is ischemia, stroke, renal injury, neurodegenerative disease, atherosclerosis, metabolic syndrome, acute myocardial infarction, heart failure, ischemia reperfusion, ureteral obstruction, diabetic nephropathy, diabetes, Leber'"'"'s Heredity Optic Neuropathy (LHON), Dominant Optic Atrophy (DOA), Barth'"'"'s syndrome, POLG disease, or Leigh'"'"'s disease.

7. The method of claim 6, wherein the neurodegenerative disease is Alzheimer'"'"'s disease, Amyotrophic Lateral Sclerosis (ALS), Huntington'"'"'s disease, Friedreich'"'"'s ataxia or Multiple Sclerosis.

8. A method for evaluating the therapeutic efficacy of an aromatic-cationic peptide on a disease or condition characterized by mitochondrial dysfunction in a subject, comprising:

a. assaying the ratio of different monocyte types present in a biological sample obtained from the subject; and

b. comparing the ratio of different monocyte types observed in step (a) with the ratio of corresponding monocyte types observed in a biological sample obtained from the subject following administration of a dose of an aromatic-cationic peptide,

wherein the aromatic-cationic peptide is identified as having a therapeutic effect on the disease or condition characterized by mitochondrial dysfunction if the ratio of different monocyte types in the biological sample following the administration of the aromatic-cationic peptide is altered compared to the ratio of different monocyte types observed in step (a).

9. The method of claim 8, wherein the ratio of classical monocytes to non-classical monocytes in the biological sample following the administration of the aromatic-cationic peptide is reduced compared to the ratio of classical monocytes to non-classical monocytes observed in step (a).

10. The method of claim 8, wherein the ratio of intermediate monocytes to non-classical monocytes in the biological sample following the administration of the aromatic-cationic peptide is reduced compared to the ratio of intermediate monocytes to non-classical monocytes observed in step (a).

11. The method of claim 8, wherein the ratio of classical monocytes to intermediate monocytes in the biological sample following the administration of the aromatic-cationic peptide is reduced compared to the ratio of classical monocytes to intermediate monocytes observed in step (a).

12. The method of claim 8, wherein the disease or condition characterized by mitochondrial dysfunction is ischemia, stroke, renal injury, neurodegenerative disease, atherosclerosis, metabolic syndrome, acute myocardial infarction, heart failure, ischemia reperfusion, ureteral obstruction, diabetic nephropathy, diabetes, Leber'"'"'s Heredity Optic Neuropathy (LHON), Dominant Optic Atrophy (DOA), Barth'"'"'s syndrome, POLG disease, or Leigh'"'"'s disease.

13. The method of claim 12, wherein the neurodegenerative disease is Alzheimer'"'"'s disease, Amyotrophic Lateral Sclerosis (ALS), Huntington'"'"'s disease, Friedreich'"'"'s ataxia or Multiple Sclerosis.

14. A method for evaluating the therapeutic efficacy of an aromatic-cationic peptide on a disease or condition characterized by mitochondrial dysfunction in a subject, comprising:

a. assaying at least one biomarker of mitochondrial physiology of a population of monocytes present in a biological sample obtained from the subject; and

b. comparing the biomarker of mitochondrial physiology of the population of monocytes observed in step (a) with the biomarker of mitochondrial physiology of a corresponding population of monocytes observed in a biological sample obtained from the subject following administration of a dose of an aromatic-cationic peptide,

wherein the aromatic-cationic peptide is identified as having a therapeutic effect on the disease or condition characterized by mitochondrial dysfunction if the biomarker of mitochondrial physiology of the population of monocytes in the biological sample following the administration of the aromatic-cationic peptide is similar to the biomarker of mitochondrial physiology of a corresponding population of monocytes in a reference sample obtained from a healthy subject.

15. The method of claim 14, wherein the aromatic-cationic peptide is Phe-D-Arg-Phe-Lys-NH₂ or D-Arg-2′

6′

Dmt-Lys-Phe-NH₂ or a pharmaceutically acceptable salt thereof.

16. The method of claim 14, wherein the alterations in biomarkers of mitochondrial physiology is determined using assays that measure disruption in oxidative phosphorylation.

17. The method of claim 16, wherein the disruption in oxidative phosphorylation is determined using assays that measure CoQ10 levels, uncoupling ratio, net routine flux control ratio, leak flux control ratio or phosphorylation respiratory control ratio.

18. The method of claim 14, wherein the alterations in biomarkers of mitochondrial physiology of monocytes is determined by measuring alterations in the level of one or more biomarkers of mitochondrial physiology in a sample of monocytes.

19. The method of claim 18, wherein the biomarkers of mitochondrial physiology are selected from the group consisting of consisting of lactic acid (lactate) levels, pyruvic acid (pyruvate) levels, lactate/pyruvate ratios, phosphocreatine levels, NADH (NADH+H³⁰) or NADPH (NADPH+H³⁰) levels;

NAD or NADP levels;

ATP levels;

reduced coenzyme Q (CoQred) levels;

oxidized coenzyme Q (CoQox) levels;

total coenzyme Q (CoQtot) levels;

oxidized cytochrome C levels;

reduced cytochrome C levels;

oxidized cytochrome C/reduced cytochrome C ratio;

acetoacetate levels;

beta-hydroxy butyrate levels;

acetoacetate/beta-hydroxy butyrate ratio;

8-hydroxy-2′

-deoxyguanosine (8-OHdG) levels;

levels of reactive oxygen species;

oxygen consumption (VO2), carbon dioxide output (VCO2), and respiratory quotient (VCO2/VO2).

20. The method of claim 14, wherein alterations in the biomarkers of mitochondrial physiology are determined using high throughput bioenergetics platforms.