IP Library Granted Patent US 12,474,341
Granted Patent B2
US 12,474,341 · App. 17/675,194 · Granted Nov 18, 2025

Method for determining onset risk of cardiovascular disease in subject, and method for determining vascular calcification in subject

Inventors: Maria Kiriyama (Kobe, JP); Eiya Tamada (Kobe, JP); Elnaz Nakhaei (Kobe, IR); Mami Onishi (Kobe, JP)
Assignee: SYSMEX CORPORATION
G01N33/573G01N2333/916G01N2800/32G01N2800/50
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,474,341
App. No.
17/675,194
Granted
Nov 18, 2025
Kind
B2
Abstract

Disclosed is a method of determining onset risk of cardiovascular disease in a subject, the method comprising: capturing, on a solid phase, an extracellular vesicle derived from a blood sample collected from the subject; and measuring alkaline phosphatase activity of the extracellular vesicle, a measured result of the alkaline phosphatase activity being directed for use as an index of the onset risk of cardiovascular disease in the subject.

Claims (23)

1 . A method of determining onset risk of cardiovascular disease in a subject, the method comprising:

identifying a subject having or at risk of developing cardiovascular disease, wherein said subject is not a chronic renal failure patient or a dialysis patient, by performing the following steps (i) and (ii):

(i) capturing, on a solid phase, an extracellular vesicle derived from a blood sample collected from the subject; and

(ii) measuring alkaline phosphatase activity of the extracellular vesicle,

wherein a measured result of the alkaline phosphatase activity is an index of the onset risk of cardiovascular disease in the subject, wherein when the measured result of the alkaline phosphatase activity is equal to or higher than a predetermined threshold value, the subject is identified as having or at risk of developing cardiovascular disease; and

treating the identified subject by a treatment comprising antihypertensive therapy and/or a medicine suitable for treating cardiovascular disease.

2 . The method according to claim 1 , wherein the blood sample is whole blood, plasma, or serum.

3 . The method according to claim 1 , wherein the capturing comprises contacting the blood sample of the subject, the solid phase, and a capture antibody capable of capturing the extracellular vesicle to form a complex of the extracellular vesicle and the capture antibody on the solid phase.

4 . The method according to claim 3 , wherein the capture antibody is at least one selected from the group consisting of anti-CD9 antibody, anti-CD81 antibody, anti-CD63 antibody, anti-Annexin VI antibody, and anti-Pit1 antibody.

5 . The method according to claim 1 , wherein the measuring comprises contacting a luminescent substrate with alkaline phosphatase of the extracellular vesicle captured on the solid phase, and detecting a resultant chemiluminescent signal.

6 . The method according to claim 1 , wherein the cardiovascular disease is at least one selected from the group consisting of ischemic heart disease, ischemic brain disease, and ischemic intestinal disorder.

7 . A method for determining vascular calcification in a subject, the method comprising:

identifying a subject having vascular calcification, wherein said subject is not a chronic renal failure patient or a dialysis patient, by performing the steps (i) and (ii):

(i) capturing, on a solid phase, an extracellular vesicle derived from a blood sample collected from the subject; and

(ii) measuring alkaline phosphatase activity of the extracellular vesicle,

wherein a measured result of the alkaline phosphatase activity is an index of vascular calcification in the subject, wherein when the measured result is equal to or higher than a predetermined threshold value, the subject is identified as having vascular calcification; and

treating the identified subject having vascular calcification by a treatment comprising antihypertensive therapy and/or medicine suitable for treating vascular calcification.

8 . The method according to claim 7 , wherein the blood sample is whole blood, plasma, or serum.

9 . The method according to claim 7 , wherein the capturing comprises contacting the blood sample of the subject, the solid phase, and a capture antibody capable of capturing the extracellular vesicle to form a complex of the extracellular vesicle and the capture antibody on the solid phase.

10 . The method according to claim 9 , wherein the capture antibody is at least one selected from the group consisting of anti-CD9 antibody, anti-CD81 antibody, anti-CD63 antibody, anti-Annexin VI antibody, and anti-Pit1 antibody.

11 . The method according to claim 7 , wherein the measuring comprises contacting a luminescent substrate with alkaline phosphatase of the extracellular vesicle captured on the solid phase, and detecting a resultant chemiluminescent signal.

12 . The method according to claim 1 , wherein the treatment comprises administering to said subject one or more of a phosphorus adsorbent, a cholesterol treating agent, a thrombolytic agent, an antithrombotic agent, and antiplatelet agent, and a vasodilator.

13 . The method according to claim 7 , wherein the treatment comprises administering to said subject one or more of a phosphorus adsorbent, a cholesterol treating agent, a thrombolytic agent, an antithrombotic agent, and antiplatelet agent, and a vasodilator.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 26, 2023
From: KIRIYAMA, MARIA; TAMADA, EIYA; NAKHAEI, ELNAZ; ONISHI, MAMI
To: SYSMEX CORPORATION
Reel/Frame 062493/0594 →
Priority Claims (1)
JP 2021-050256 · Mar 24, 2021 · national
Continuity (1)
Related Publication 20220308057A1 · Sep 29, 2022
References Cited (16)
US 8993247B2 · Shanahan et al. · 2015 [cited by applicant]
US 10139402B2 · Fujii et al. · 2018 [cited by applicant]
US 20090098539A1 · Wilson · 2009 [cited by examiner]
WO 2020033467A1 · 2020 [cited by applicant]
Bengtsson et al., Clinical Oral Investigations (2019) 23:1171-1179, Carotid calcifications in panoramic radiographs are associated with future stroke or ischemic heart diseases: a long-term follow-up study. (Year: 2019). [cited by examiner]
Mobeirek et al, International Journal of Clinical Medicine, 2014, 5, 249-259. (Year: 2014). [cited by examiner]
Shanahan et, Circulation, vol. 100, No. 21, Nov. 23, 1999, p. 2168-2176. (Year: 1999). [cited by examiner]
Bilal Mir et al: “Extracellular Vesicles as Delivery Vehicles of Specific Cellular Cargo”, Cells, Jul. 2, 2020, vol. 9, Issue No. 7, Article No. 1601, 19 pages. [cited by applicant]
Wenling Yang et al: “Extracellular vesicles in vascular calcification”, Clinica Chimica Acta, 2019, vol. 499, pp. 118-122, Elsevier B.V. [cited by applicant]
The extended European search report issued on Aug. 18, 2022 in a counterpart European patent application No. 22163963.6. [cited by applicant]
Ryuichi Watanabe et al., “Mechanism of calcification in vivo”, The Japanese Journal of Nephrology, 2014, pp. 1196-1200, vol. 56, No. 8. [cited by applicant]
Jona B. Krohn et al., “Extracellular vesicles in cardiovascular calcification: expanding current paradigms”, The Journal of Physiology, 2016, pp. 2895-2903, vol. 594, No. 11, The Physiological Society. [cited by applicant]
Elena Aikawa, “Extracellular vesicles in cardiovascular disease: focus on vascular calcification”, The Journal of Physiology, 2016, pp. 2877-2880, vol. 594, No. 11, The Physiological Society. [cited by applicant]
Amirala Bakhshian Nik et al., “Extracellular Vesicles As Mediators of Cardiovascular Calcification”, Frontiers in Cardiovascular Medicine, Dec. 11, 2017, vol. 4, Article 78, 7 pages in total. [cited by applicant]
Claudia Goettsch et al., “Sortilin mediates vascular calcification via its recruitment into extracellular vesicles”, The Journal of Clinical Investigation, Apr. 2016, pp. 1323-1336, vol. 126, No. 4. [cited by applicant]
Japanese Office Action issued on Sep. 10. 2024, in a counterpart Japanese patent application No. 2021-050256. [cited by applicant]