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KR-20260068113-A - Chromosome detection analysis method

KR20260068113AKR 20260068113 AKR20260068113 AKR 20260068113AKR-20260068113-A

Abstract

Through the characterization of chromosomal interactions associated with aging, it was revealed that these interactions occur in clusters, and that each cluster reflects the underlying biological systems involved in the aging system. This study explains the multilayered and multidimensional nature of aging. Accordingly, the present invention provides a method for determining system-specific aging and further enables the adoption of intervention strategies to inhibit or reverse the aging process in a manner tailored to the individual's needs. Essentially, the present invention provides a method for determining aging by analyzing chromosomal interactions.

Inventors

  • 헌터, 이완
  • 아쿨리체프, 알렉상드르

Assignees

  • 옥스포드 바이오다이나믹스 피엘씨

Dates

Publication Date
20260513
Application Date
20240919
Priority Date
20230920

Claims (14)

  1. A method for determining the aging of an individual, comprising the step of detecting the presence or absence of one or more chromosomal interactions shown in any one of Tables 1 to 15, thereby determining the aging of said individual, wherein optionally, a multidimensional measure of aging is determined by detecting the presence or absence of one or more chromosomal interactions from more than one of Tables 1 to 15.
  2. A method according to claim 1, characterized in that the presence and/or absence of a chromosomal interaction is determined in any one of the following combinations of chromosomal interactions: - All of the first 10 chromosomal interactions listed in at least 5 of Tables 1 through 15, or - At least 5 of the first 10 chromosomal interactions listed in at least 5 of Tables 1 through 15, or - All of the first 10 chromosomal interactions listed in Tables 1 through 15.
  3. A method according to claim 1 or 2, characterized in that the presence and/or absence of a chromosomal interaction is determined in any one of the following combinations of chromosomal interactions: - Any one of the chromosomal interactions listed in Table 28 or 30, or - At least 5 of the chromosomal interactions listed in either Table 28 or 30, or - At least 5 of the chromosomal interactions listed in Table 28 and at least 5 of the chromosomal interactions listed in Table 30.
  4. A method according to any one of claims 1 to 3, characterized in that the chromosomal interaction is typed by the following: - From a sample from an object, and/or - By detecting the presence or absence of a DNA loop at the site of the chromosomal interaction above, and/or - By detecting the presence or absence of distal regions of chromosomes that come together in chromosomal form, and/or - By detecting the presence of ligated DNA, which includes two regions corresponding respectively to regions of a chromosome where the sequences are generated during the above typing and come together in chromosomal interactions, and/or - It is typed by a process of detecting the proximity of chromosomal regions gathered together in the above chromosomal interaction.
  5. In any one of claims 1 to 4, the detection of the presence or absence of the chromosomal interaction is: (i) a step of crosslinking the epigenetic chromosomal interactions of the above individual in vitro; (ii) a step of applying the cross-linked DNA to the cutting process; (iii) a step of forming linked DNA by connecting the cross-linked cleaved DNA ends; and (iv) a step of confirming the presence or absence of the linked DNA; comprising, A method characterized by determining the presence or absence of the above-mentioned chromosomal interaction.
  6. A method according to claim 4 or 5 in which the linked DNA is detected by PCR or by the use of a probe.
  7. In claim 6, a method characterized in that the detection of the linked DNA is by the following use: (i) As a probe, the probe has at least 70% identity with respect to any one of the probes shown in Table 16, and/or (ii) PCR, wherein the PCR uses a primer pair capable of amplifying the linked DNA, wherein one or two primers comprise a sequence that is part of at least 10 nucleic acid bases of one of the probes shown in Table 16, and/or (b) a sequence of 10 nucleic acid bases of at least 70% the same length as (a).
  8. In any one of paragraphs 1 through 7, (i) the above method is performed in the early stages of the aging process in which the individual does not have any symptoms caused by aging, preferably 40 years of age or younger, and/or, (ii) The above method is performed to select subjects to receive anti-aging intervention, preferably for therapeutic or cosmetic purposes, and/or (iii) The above method is performed on individuals pre-selected based on the presence of physical characteristics, risk factors, or symptoms associated with aging, and/or, (iv) The above method is performed to determine the prognosis for aging and/or, (v) A method characterized in that the above method is performed at multiple points in time, preferably at least two points in time with an interval of at least 30 days, on samples taken from an individual.
  9. In any one of paragraphs 1 to 8, the above entity, (i) suspected of having accelerated aging, and/or (ii) hospitalization, and/or (iii) A method characterized by being a human individual.
  10. In any one of claims 1 to 9, the step of detecting the presence or absence of the chromosomal interaction comprises specific detection of the linked product by quantitative PCR (qPCR) using a primer capable of amplifying the linked DNA and a probe that binds to the linkage site during the PCR reaction, wherein the probe comprises a sequence complementary to the sequence from each chromosomal region gathered together in the chromosomal interaction, and wherein, preferably, the probe is, - Oligonucleotides that specifically bind to the above-mentioned linked DNA, and/or - A fluorophore covalently attached to the 5' end of the above oligonucleotide, and/or - Includes a quencher attached by a covalent bond to the 3' end of the above oligonucleotide, Optionally - The above fluorescent dye is selected from HEX, Texas Red, and FAM; and/or - A method characterized in that the probe comprises a nucleic acid sequence of 10 to 40 nucleotide bases in length, preferably 20 to 30 nucleotide bases in length.
  11. A method according to any one of paragraphs 1 through 10, further comprising the following: (i) a step in which the above-mentioned individuals perform an intervention to reduce or reverse their aging, and/or (ii) A step in which advice regarding reducing or reversing their aging is provided to the above subjects, preferably in the form of a written report.
  12. In paragraph 11, a method characterized in that the above intervention and/or advice relates to the following: (i) as a step of changing their diet, preferably restricting their calorie intake, and/or (ii) As a step of changing their lifestyle, preferably increasing their exercise.
  13. As an anti-aging agent to be used in a method for treating accelerated aging, the above method, - A step of determining whether an individual has accelerated aging by the method of any one of claims 1 to 12, and - An anti-aging agent comprising the step of administering the above agent to any individual identified as having accelerated aging.
  14. As a cosmetic method to reduce or reverse the deterioration of appearance, - A step of determining whether an individual has accelerated aging by the method of any one of claims 1 to 12, and - A cosmetic method comprising the step of providing an anti-aging intervention to any individual identified as having accelerated aging, which preferably includes the step of administering an anti-aging agent.

Description

Chromosome detection analysis method The present invention relates to the aging process. Many current theories of aging focus on the 'damage concept,' which posits that the accumulation of damage (e.g., DNA oxidation) can lead to the failure of biological systems. Aging results in a progressive decline in physical and mental abilities, an increased risk of disease, and ultimately, death. These changes are not linear or consistent and are only loosely associated with a person's age. Figure 1 shows 1,096 CCS found to meet the statistical criteria of this study. The figure shows 150 CCS whose abundance increased over time, while the remaining 946 decreased over time. The X-axis represents time, and the Y-axis represents FC (increase in CCS over time). Figure 2 shows 150 CCS that increase over time, further subdivided using Quality Threshold (QT) clustering to create 5 clusters (QT radius set to 1). The Slope Coef is related to the slope of change of each set, and Cluster 3 shows the largest change over time. The X-axis represents time, and the Y-axis represents FC. A represents Cluster 1. The Slope Coef is 0.173, and there are 64 CCS. B represents Cluster 2. The Slope Coef is 0.17, and there are 51 CCS. C represents Cluster 3. The Slope Coef is 0.176, and there are 15 CCS. D represents Cluster 4. The Slope Coef is 0.168, and there are 14 CCS. E represents Cluster 5. The Slope Coef is 0.161, and there are 6 CCS. Figure 3 shows 946 CCS whose abundance decreased with age. Most of the 1,096 significant CCS decrease over time. The X-axis represents time, and the Y-axis represents FC. Figure 4 shows 946 CCS that decrease with age, further subdivided using Quality Threshold (QT) clustering to generate 10 clusters (QT radius set to 1.1). The Slope Coef is related to the slope of change for each set, and Cluster 5 represents the largest change with age in this set. The X-axis represents time, and the Y-axis represents FC. A represents Cluster 1. The Slope Coef is -0.177, and there are 231 CCS. B represents Cluster 2. The Slope Coef is -0.179, and there are 226 CCS. C represents Cluster 3. The Slope Coef is -0.19, and there are 157 CCS. D represents Cluster 4. The Slope Coef is -0.187, and there are 122 CCS. E represents Cluster 5. The Slope Coef is -0.21, and there are 61 CCS. Figure 5 shows the remaining 5 clusters among the 10 clusters (QT radius set to 1.1) formed using 946 CCS whose abundance decreases with age. Cluster 7 represents the greatest change with age in this set. The X-axis represents time, and the Y-axis represents FC. A represents Cluster 6. The Slope Coef is -0.178, and there are 53 CCS. B represents Cluster 7. The Slope Coef is -0.204, and there are 50 CCS. C represents Cluster 8. The Slope Coef is -0.183, and there are 33 CCS. D represents Cluster 9. The Slope Coef is -0.183, and the number of CCS is 7. E represents Cluster 10. The Slope Coef is -0.182, and there are 6 CCS. Figure 6 illustrates a preferred method for typing chromosome interactions. Explanation of the table Tables 1 to 5 show chromosomal interactions that increase with age. Tables 6 to 15 show chromosomal interactions that decrease with age. Table 16 shows probes corresponding to the chromosome interactions of Tables 1 to 15. Table 17 shows the groups of individuals investigated in this study. Table 18 shows the mapping of 150 age-increasing CCS to the nearest coding gene locations (upstream, downstream, and overlap). Biological pathway and disease enrichment analyses were performed based on identified coding gene locations (genes). Table 19 shows disease ontologies associated with gene locations enriched in 150 CCS that increase over time. Table 20 shows the path enrichment for 64 CCS belonging to cluster 1 of CCS that increases over time. Table 21 shows the pathway enrichment for 51 CCS belonging to Cluster 2 of CCS that increases over time. [NRF2 is involved in redox regulation in aging and disease.] Table 22 shows the path enrichment for the 35 combined CCS of clusters 3, 4, and 5. Table 23 shows pathway enrichment for coding gene locations mapped to 946 CCS that decrease with age. Table 24 shows the path enrichment for CCS (cluster 5) that decreases the most with age. Table 25 shows the path enrichment for CCS (cluster 7), which decreases significantly with age. Table 26 shows the cluster 2 'age down' (decreasing with age) CCS used in LDA. Tables 27 and 28 show 135 markers of the same group that decrease with age. Tables 29 and 30 show 42 markers of the same group that increase with age. Terms Used Herein The method of the present invention may be referred to herein as the 'process' of the invention. Chromosomal interactions being typed may be referred to herein as 'markers', 'CCS', 'chromosome conformation signatures', 'epigenetic interactions', or 'EpiSwitch markers'. These are discussed in more detail below and represent the aggregation of different regions of chromosomes in a stable manner detectable, for example, using the