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EP-4737903-A2 - METHOD FOR DETERMINING A LIKELIHOOD OF A SUBJECT TO RESPOND TO LIPID LOWERING THERAPY

EP4737903A2EP 4737903 A2EP4737903 A2EP 4737903A2EP-4737903-A2

Abstract

A method for determining a likelihood of a subject to respond to lipid lowering therapy is disclosed. The method may comprise determining a quantitative value of the ability of cells obtained from a biological sample of the subject to take up low-density lipoprotein (LDL), a quantitative value of the expression of LDL receptor (LDLR) in the cells, a quantitative value of the lipid storage capability of the cells, and/or a quantitative value of the lipid mobilization capability of the cells; wherein the quantitative value of the ability of the cells to take up LDL, the quantitative value of the expression of LDLR in the cells, the quantitative value of the lipid storage capability of the cells, and/or the quantitative value of the lipid mobilization capability of the cells is/are indicative of the likelihood of the subject to respond to lipid lowering therapy.

Inventors

  • IKONEN, Elina
  • PFISTERER, SIMON

Assignees

  • MONCYTE Health Oy

Dates

Publication Date
20260506
Application Date
20211210

Claims (15)

  1. A method for determining whether a subject is at risk of developing or having atherosclerosis and/or a cardiovascular disease, wherein the method comprises determining a quantitative value of the ability of cells obtained from a biological sample of the subject to take up low-density lipoprotein (LDL), a quantitative value of the expression of LDL receptor (LDLR) in the cells, a quantitative value of the lipid storage capability of the cells, and/or a quantitative value of the lipid mobilization capability of the cells; wherein the quantitative value of the ability of the cells to take up LDL, the quantitative value of the expression of LDLR in the cells, the quantitative value of the lipid storage capability of the cells, and/or the quantitative value of the lipid mobilization capability of the cells is/are indicative of the likelihood of the subject having an increased risk of developing or having atherosclerosis and/or the cardiovascular disease.
  2. The method according to claim 1, wherein the cells comprise or are peripheral blood mononuclear cells (PBMCs).
  3. The method according to claim 1 or 2, wherein the cells comprise or are monocytes.
  4. The method according to any one of claims 1 - 3, wherein the subject has hyperlipidemia.
  5. The method according to any one of claims 1 - 4, wherein the cardiovascular disease comprises or is at least one of coronary artery disease, such as angina or myocardial infarction, stroke, heart attack, cerebrovascular disease, or peripheral artery disease.
  6. The method according to any one of claims 1 - 5, wherein the method is performed using high content imaging followed by automated image and data analysis.
  7. The method according to any one of claims 1 - 6, wherein the cells are immobilized on a surface, optionally as a monolayer.
  8. The method according to any one of claims 1 - 7, wherein the cells are grouped to two or more cell populations, and the quantitative value of the ability to take up LDL, the quantitative value of the expression of LDLR, the quantitative value of the lipid storage capability, and/or the quantitative value of the lipid mobilization capability of at least one of the cell populations is/are determined separately from the other cell populations.
  9. The method according to any one of claims 1 - 8, wherein the quantitative value of the ability of the cells of the subject to take up LDL is determined using labelled LDL particles, wherein the LDL particles are optionally fluorescently or radioactively labelled.
  10. The method according to any one of claims 1 - 9, wherein the quantitative value of the ability of the cells of the subject to take up LDL is determined using LDL particles preserved by snap freezing.
  11. The method according to any one of claims 1 - 10, wherein the quantitative value of the ability of the cells to take up LDL, the quantitative value of the expression of LDLR, the quantitative value of the lipid storage capability, and/or the quantitative value of the lipid mobilization capability is/are normalized using a standard comprising a mixture of cells obtainable from biological samples of a plurality of subjects.
  12. The method according to any one of claims 1 - 11, wherein the method further comprises determining whether the subject is at risk of developing or having the cardiovascular disease on the basis of at least one genetic marker associated with the cardiovascular disease or the risk thereof, and/or the method further comprises calculating a risk score based on the presence of at least one genetic marker associated with the cardiovascular disease or the risk thereof and on the quantitative value of the ability of the cells to take up LDL, the quantitative value of the expression of LDLR, the quantitative value of the lipid storage capability, and/or the quantitative value of the lipid mobilization capability.
  13. The method according to any one of claims 1 - 12, wherein the method is performed using high content imaging followed by automated image and data analysis; the cells are immobilized on a surface, optionally as a monolayer; the cells are grouped to two or more cell populations, and the quantitative value of the ability to take up LDL, the quantitative value of the expression of LDLR, the quantitative value of the lipid storage capability, and/or the quantitative value of the lipid mobilization capability of at least one of the cell populations is/are determined separately from the other cell populations; and the quantitative value of the ability of the cells to take up LDL, the quantitative value of the expression of LDLR, the quantitative value of the lipid storage capability, and/or the quantitative value of the lipid mobilization capability is/are normalized using a standard comprising a mixture of cells obtainable from biological samples of a plurality of subjects.
  14. The method according to claim 13, wherein the quantitative value of the ability of the cells of the subject to take up LDL is determined using LDL particles preserved by snap freezing.
  15. The method according to claim 13 or 14, wherein the cells comprise or are peripheral blood mononuclear cells (PBMCs), the PBMC cells are grouped to monocytes, lymphocytes and optionally to other cell types, and the quantitative value(s) are determined separately for the monocytes, lymphocytes, and any other optional cell populations.

Description

TECHNICAL FIELD The present disclosure relates to a method for determining a likelihood of a subject to respond to lipid lowering therapy and to a method for determining whether a subject is at risk of developing or having atherosclerosis and/or a cardiovascular disease. BACKGROUND Hypercholesterolemia, i.e. an elevated serum cholesterol level in the blood, is currently a leading cause for cardiovascular disease (CVD) in many geographical areas. The regulation of serum cholesterol levels is controlled by dietary intake, endogenous production, and cellular metabolism. Statins are often used as the first line medication for subjects suffering from hypercholesterolemia. However, often subjects do not respond or achieve a target cholesterol level immediately or in the long term. Some patients do not adhere to statin therapy. Further, a significant portion of patients who have had a heart attack have a second attack within one year of the first attack. Thus there may be a need to lower the cholesterol level of such patients without waiting for a prolonged period to see if they respond to statin treatment. Further, most cholesterol lowering treatments and therapies affect cellular mechanisms, and therefore it may be highly beneficial to be able to quantify such cellular mechanisms. SUMMARY This Summary is provided to introduce a selection of concepts in a simplified form that are further described below in the Detailed Description. This Summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter. A method for determining a likelihood of a subject to respond to lipid lowering therapy is disclosed. The method may comprise determining a quantitative value of the ability of cells obtained from a biological sample of the subject to take up low-density lipoprotein (LDL), a quantitative value of the expression of LDL receptor (LDLR) in the cells, a quantitative value of the lipid storage capability of the cells, and/or a quantitative value of the lipid mobilization capability of the cells; wherein the quantitative value of the ability of the cells to take up LDL, the quantitative value of the expression of LDLR in the cells, the quantitative value of the lipid storage capability of the cells, and/or the quantitative value of the lipid mobilization capability of the cells is/are indicative of the likelihood of the subject to respond to lipid lowering therapy. A method for determining whether a subject is at risk of developing or having atherosclerosis and/or a cardiovascular disease is also disclosed. The method may comprise determining a quantitative value of the ability of cells obtained from a biological sample of the subject to take up low-density lipoprotein (LDL), a quantitative value of the expression of LDL receptor (LDLR) in the cells, a quantitative value of the lipid storage capability of the cells, and/or a quantitative value of the lipid mobilization capability of the cells; wherein the quantitative value of the ability of the cells to take up LDL, the quantitative value of the expression of LDLR in the cells, the quantitative value of the lipid storage capability of the cells, and/or the quantitative value of the lipid mobilization capability of the cells is/are indicative of the likelihood of the subject having an increased risk of developing or having atherosclerosis and/or the cardiovascular disease. BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are included to provide a further understanding of the invention and constitute a part of this specification, illustrate embodiments and together with the description help to explain the principles of the invention. In the drawings: Figure 1: Automated analysis pipeline for multiplex quantification of functional defects in PBMCs. A) Schematic presentation of the automated sample processing and image analysis pipeline. For each experiment cryopreserved PBMC samples were thawed, aliquoted into 96 wells and incubated overnight with 10% FBS (complete medium) or 5% LPDS (lipid starvation medium). Cells were labeled with fluorescent LDL (DiI-LDL) or directly transferred to 384 well imaging plates, automatically fixed, stained and subjected to automated high-content imaging. Images were quantified with CellProfiler and single cell data was processed with Python tools. B) Representative images showing DiI-LDL uptake in lymphocyte and monocyte populations after treatment with 5% LPDS. C) Automated quantification of mean DiI-LDL intensities in lymphocyte and monocyte populations from two standards (1, 2) each consisting of a mix of PBMCs from four individuals. n > 74000 lymphocytes and > 16100 monocytes for each treatment and standard from 10 experiments. D) DiI-LDL uptake in control EBV lymphoblasts (EBV) and monocytes after LPDS treatment. Representative images are shown. E) Quantification of mean DiI-LDL intensities in EBV lymphob