JP7442221B1 - Method for calculating biological age, method for evaluating aging status, method for evaluating CpG sites, program, and computer-readable storage medium - Google Patents

Abstract

An object of the present invention is to provide a novel method for calculating biological age. [Solution] The biological age calculation method according to the present invention is a method for calculating the biological age of an individual, in which numbers 1 to 235 listed in Table 1 are used in the genome of a cell population obtained from the individual. Among the CpG sites in the DNA at the chromosome location, (1) select 215 to 235 CpG sites, or (2) select 1 to 3 randomly selected CpG sites among the 235 CpG sites. A step of creating a pool of CpG sites by adding CpG sites, a step of examining methylation of the CpG sites in the pool, and calculating a methylation rate at each CpG site in the pool, using the following formula. the step of calculating the biological age of the individual; This is a calculation method that indicates the methylation rate of a certain CpG site, and Cn indicates a value between 50% and 150% of the coefficient for the n-th chromosome position listed in Table 1. [Selection diagram] None

Description

The present invention relates to a method for calculating biological age, a method for evaluating aging status, a method for evaluating CpG sites, a program, and a computer-readable storage medium.

As people age and age, they develop various diseases and exhibit declines in cognitive and physical functions. In modern society, where the average lifespan of humans is increasing, it is becoming increasingly important to delay the onset of aging-related diseases and the period during which functional decline occurs, that is, to extend healthy life expectancy. To this end, it is essential to evaluate the aging status of individuals, but since aging progresses at different rates depending on the individual, an index that can evaluate biological age rather than chronological age is needed.

For example, there has conventionally been the concept of an epigenetic clock that uses DNA methylation as a target for evaluating biological age. The DNA methylation state changes acquiredly due to environmental factors, and some CpGs show a certain change in DNA methylation rate with aging. Therefore, we identify CpGs associated with age and aging status from DNA methylation information derived from people of a wide range of chronological ages and aging status, and estimate the individual's biological age as a value calculated from DNA methylation information. A model has been developed that allows this. In this model, the value calculated from DNA methylation information is called epigenomic age. By comparing an individual's epigenome age calculated using this model with that individual's chronological age, it is also possible to evaluate the degree of aging of that individual.

An object of the present invention is to provide a novel method for calculating biological age, a novel method for evaluating aging status, a novel method for evaluating CpG sites, a novel program, and a novel computer-readable storage medium.

One embodiment of the present invention is a method for calculating the biological age of an individual, in which the positions of chromosomes 1 to 235 listed in Table 1 below (based on GRCh37) are determined in the genome of a cell population obtained from the individual. ) out of the CpG sites in the DNA of A step of creating a pool of CpG sites by adding calculating the biological age of the individual;

[Number 1]
(However, t = 79 to 85, DNAm _n indicates the methylation rate of the CpG site in the DNA at the position of the chromosome n listed in Table 1 below, and C _{n indicates the methylation rate of the CpG site in the DNA at the position of chromosome n} listed in Table 1 below. (indicates a value between 50% and 150% of the coefficient for the position of the nth chromosome). t may be 80 to 84, or t may be 81.946609. C _n may be a value between 80% and 120% of the coefficient for the position of the nth chromosome listed in Table 1 below; There may be. The 235 CpG sites listed in Table 1 may be selected to create a pool of CpG sites. The individual may be Asian. The cell population may be blood cells.

Another embodiment of the present invention is a method for evaluating the aging status of an individual, comprising the steps of calculating the biological age of the individual by any of the calculation methods described above; , and the chronological age of the individual.

A further embodiment of the present invention is a method for evaluating CpG sites for calculating the biological age of an individual, wherein CpG sites numbered 1 to 235 listed in Table 1 below are determined in the genome of a cell population obtained from a certain individual. Among the CpG sites in the DNA at the chromosomal location (based on GRCh37), (1) select between 200 and 214 CpG sites, or (2) select 4 to 6 CpG sites among the 235 CpG sites. A step of creating a pool of CpG sites by adding CpG sites, a step of examining methylation of the CpG sites in the pool, and calculating a methylation rate at each CpG site in the pool, using the following formula. calculating the biological age of the individual;

[Number 2]
(However, t = 79 to 85, DNAm _n indicates the methylation rate of the CpG site in the DNA at the position of the chromosome n listed in Table 1 below, and C _{n indicates the methylation rate of the CpG site in the DNA at the position of chromosome n} listed in Table 1 below. (indicates a value between 50% and 150% of the coefficient for the position of the nth chromosome). The individual may be Asian. The cell population may be blood cells.

A further embodiment of the present invention is a program for causing a computer to execute any of the calculation methods or evaluation methods described above, and a computer-readable storage medium storing the program.

The present invention makes it possible to provide a novel method for calculating biological age, a novel method for evaluating aging status, a novel method for evaluating CpG sites, a novel program, and a novel computer-readable storage medium. .

In the examples of the present invention, the chronological age (x-axis) and methylation rate (%) of the subject at the CpG sites located in the DNA of chromosomes 1 to 50 listed in Table 1 are shown in Table 1. y-axis). In the examples of the present invention, the chronological age (x-axis) of the test subject and the methylation rate (%) ( y-axis). In the examples of the present invention, the chronological age (x-axis) of the subject and the methylation rate (%) ( y-axis). In the examples of the present invention, the chronological age (x-axis) and methylation rate (%) of the subject at the CpG site located in the DNA at the 151st to 200th chromosome positions listed in Table 1. y-axis). In the examples of the present invention, the chronological age (x-axis) of the test subject and the methylation rate (%) ( y-axis). (a) It is a graph showing the correlation between epigenetic age (Epigenetic Age) and chronological age (Chronological Age) obtained using an epigenomic age calculation formula in an example of the present invention. (b) In the examples of the present invention, the value of the formula for calculating the aging evaluation degree for epigenetic age (Epigenetic Age) obtained using CpG sites showing blood cell type-specific DNA methylation rates (Epigenetic Age Acceleration) is a graph plotted against chronological age. In the example in which CpG sites were randomly excluded from the CpG sites numbered 1 to 235 listed in Table 1 and the epigenome age was calculated using the remaining CpG sites, the number of CpG sites excluded (x axis) was It is a graph plotting the absolute value (y axis) of the difference between epigenomic age and chronological age. The absolute value of the difference between epigenomic age and chronological age is shown as the median (point) and interquartile range (line segment). In the example in which CpG sites were randomly excluded from the CpG sites numbered 1 to 235 listed in Table 1 and the epigenomic age was calculated using the remaining CpG sites, the number of excluded CpG sites (x axis) was It is a graph plotting the correlation coefficient (y axis) between epigenomic age and chronological age. The correlation coefficient between epigenomic age and chronological age is shown as the median (point) and interquartile range (line segment). In an example in which randomly selected CpG sites not listed in Table 1 were added to the CpG sites numbered 1 to 235 listed in Table 1, and the epigenomic age was calculated using the obtained CpG sites, the number of added sites ( This is a graph in which the average absolute value of the difference between epigenomic age and chronological age (y-axis) is plotted against (x-axis). The absolute value of the difference between epigenomic age and chronological age is shown as the median (point) and interquartile range (line segment). In the example in which a randomly selected CpG site not listed in Table 1 was added to the CpG sites numbered 1 to 235 listed in Table 1, and the epigenomic age was calculated using the obtained CpG site, the added CpG site It is a graph plotting the correlation coefficient (y-axis) between epigenomic age and chronological age against the number of individuals (x-axis). The correlation coefficient between epigenomic age and chronological age is shown as the median (point) and interquartile range (line segment). In the formula for calculating the epigenomic age obtained in Example 1, the coefficients listed in Table 1 are multiplied by a random number between 1-x and 1+x (where x is 0, 0.50, or 1.00). This is a graph showing the correlation between epigenetic age and chronological age by calculating epigenomic age using the calculated coefficients. In the formula for calculating epigenome age obtained in Example 1, the coefficients listed in Table 1 are calculated from 1-x to 1+x (x is 0.00 to 1.00, 101 numbers in 0.01 increments). ), the epigenomic age is calculated using the coefficient obtained by multiplying by a random number between be. The absolute value of the difference between epigenomic age and chronological age is shown as the median (point) and interquartile range (line segment).

The objects, features, advantages, and ideas of the present invention will be apparent to those skilled in the art from the description of this specification, and from the description of this specification, those skilled in the art will be able to easily reproduce the present invention. The embodiments and specific examples of the invention described below are indicative of preferred embodiments of the invention, and are provided for illustration or explanation, and the invention is not limited thereto. It is not limited. It will be apparent to those skilled in the art that various changes and modifications can be made based on the description herein within the spirit and scope of the present invention disclosed herein. Note that in this specification, the "Nth number" with respect to a table refers to the Nth number counted from the top of the table.

==How to calculate biological age==
The method for calculating the biological age of an individual disclosed herein includes the steps of obtaining a cell population from the individual, extracting a genome from the cell population, and 1 to 1 listed in Table 1 in the genome of the cell population. Among the CpG sites in the DNA at chromosome number 235,

by (1) selecting a first number of CpG sites or (2) adding a second randomly selected number of CpG sites to the 235 CpG sites.

A step of creating a pool of CpG sites, a step of examining the methylation of CpG sites in the created pool and calculating the methylation rate at each CpG site in the pool, and a step of calculating the biological age of an individual using the following formula. and calculating.

[Number 3]
(In the formula, t is the first coefficient, DNAm _n indicates the methylation rate of the CpG site in the DNA at the nth chromosome position listed in Table 1, and C _n is the (Indicates the second coefficient for the position of the nth chromosome.)

In addition, in this specification, "biological age" means the epigenome age which has the average aging degree of people whose age is a chronological age. Furthermore, in this specification, the location on the chromosome is specified by the location on GRCh37.

The race, nationality, and group to which the individual belongs are not particularly limited, but the individual is preferably Asian, more preferably East Asian, and even more preferably Japanese. Here, Asian, East Asian, and Japanese are defined as clusters consisting of the majority of Asians, East Asians, and Japanese, respectively, based on racial analysis based on genomic information or mitochondrial DNA information. refers to the individual that enters the The analysis method is not particularly limited, and any method that is academically recognized may be used, such as a method using microsatellite DNA polymorphism.

The cell type of the cell population is not particularly limited, and may be composed of a single cell type or a plurality of cell types. Blood cells are preferred because they are easy to obtain.

The first coefficient t is preferably 77 to 87, more preferably 79 to 85, even more preferably 80 to 84, and even more preferably 81.946609.

The second coefficient C _n is preferably a value in the range of 50 to 150%, more preferably 80 to 120%, and 95% of the value listed in Table 1. More preferably, the value is in the range from 105% to 105%, and even more preferably 100% (ie, the values listed in Table 1).

Among the CpG sites located in the DNA at chromosome positions 1 to 235 listed in Table 1, the first number of CpG sites selected is preferably 185 to 235, and 215 to 235. It is more preferably the following, more preferably 225 or more and 235 or less, even more preferably 230 or more and 235 or less, even more preferably all 235.

Furthermore, the number of randomly selected CpG sites to be added to the 235 CpG sites among the CpG sites in the DNA at chromosome positions 1 to 235 listed in Table 1 is 1 to 9. The number is preferably from 1 to 6, even more preferably from 1 to 3.

The method for analyzing methylation of CpG sites is not particularly limited, and any known method may be used. For example, in the bisulfite method, when DNA is treated with bisulfite, unmethylated cytosine is converted to uracil, and methylated cytosine remains as cytosine and is not converted. Therefore, after the treatment, the base sequence is determined and the DNA before treatment is Comparing the sequence and the DNA sequence after treatment, we identified the base that was cytosine before treatment and uracil after treatment as unmethylated cytosine, and the base that was cytosine before treatment and remained cytosine after treatment was identified as methylated cytosine. It can be determined that

The methylation rate at each CpG site can be calculated as the ratio of methylated CpG sites to the total number of CpG sites in a cell population from which the genome in which methylation at CpG sites was investigated is derived. For example, in the bisulfite method described above, the methylation rate can be calculated at each CpG site by comparing the strength of cytosine and uracil signals during sequencing. Specifically, the methylation rate is the ratio of the cytosine signal strength to the sum of the cytosine signal strength and uracil signal strength.

As shown in the examples, by using this method, the biological age of an individual can be calculated.

Further, a program for causing a computer to execute the calculation method described herein, and a computer-readable storage medium storing the program are also embodiments of the present invention.

==Method for evaluating individual aging status==
The method for evaluating the aging status of an individual disclosed herein includes the steps of calculating the biological age of the individual using the biological age calculation method described above, calculating the biological age of the individual, and determining the biological age of the individual. and a step of comparing the chronological age. Here, the "aging state" of a certain individual does not refer to a specific aspect of aging, but is defined by age and refers to the standard physical condition of a person of that age. A specific evaluation method is to compare an individual's biological age with the individual's chronological age, and if the biological age is greater than the chronological age, the person is aging faster than normal and the biological age is If the age is equal to the chronological age, the person is considered to be aging normally; if the biological age is smaller than the chronological age, the person is considered to be aging less than the normal age. Here, "normal" means the average aging state of a person of that age.

For example, the aging evaluation level may be defined as (biological age - chronological age), and it may be determined that the larger the value, the higher the aging level, and the smaller the value, the lower the aging level.

Alternatively, the aging evaluation degree may be calculated using the following formula, and it may be determined that the larger the value, the higher the aging degree, and the smaller the value, the lower the aging degree.

[Number 4]
Aging evaluation level=p1+chronological age×p2−epigenome age In the formula, the first constant p1 is preferably 7.5 to 7.8, more preferably 7.677139. The second constant p2 is preferably 0.8 to 0.9, more preferably 0.8507557.

Furthermore, if the cell population is blood cells, the composition of the cell type is determined, the DNA methylation rate is calculated for each blood cell type, and the aging evaluation degree is determined by the following formula. The higher the value, the higher the aging degree. However, it may be determined that the smaller the value, the smaller the degree of aging. Analysis of the composition of cell types may be performed by separating blood into individual cells, or may be performed using blood cell type-specific DNA methylation rates.

[Number 5]
Aging evaluation level = q1 + chronological age × q2 + CD8 × q3 + CD4 × q4 + NK × q5 + B × q6 + M × q7 + N × q8 − epigenome age In the formula, CD8, CD4, NK, B, M, and N are CD8 ⁺ T cells and CD4 ⁺ T cells, respectively. DNA methylation rates in cells, NK cells, B cells, monocytes, and neutrophils. Further, the first constant q1 is preferably 64 to 65, and more preferably 64.30288. The second constant q2 is preferably 0.8 to 0.9, more preferably 0.8674881. The third constant q3 is preferably -40 to -50, more preferably -44.36649. The fourth constant q4 is preferably -70 to -80, more preferably -76.08973. The fifth constant q5 is preferably -45 to -55, more preferably -50.9861. The sixth constant q6 is preferably -47.5 to -57.5, more preferably -52.4978. The seventh constant q7 is preferably -77.5 to -87.5, more preferably -82.13944. The eighth constant q8 is preferably -50 to -60, more preferably -55.28485.

Furthermore, by introducing such an aging evaluation level, it becomes possible to determine whether aging is progressing faster than average or slower than average over time. Specifically, if the aging evaluation level at a certain point decreases after a predetermined period of time, it can be determined that aging is progressing more slowly than average, and if it increases after a predetermined period of time, it can be determined that aging is progressing faster than average. It can be determined that

== CpG site evaluation method for calculating biological age ==
The CpG site evaluation method for calculating biological age disclosed herein includes the steps of obtaining a cell population from a plurality of individuals, extracting a genome from the cell population, and in the genome of the cell population. Among the CpG sites located in the DNA at chromosome positions 1 to 235 listed in Table 1 above,

by (1) selecting a first number of CpG sites or (2) adding a second randomly selected number of CpG sites to the 235 CpG sites.

A process of creating a pool of CpG sites, a process of examining the methylation of the CpG sites in the created pool, and calculating the methylation rate at each CpG site in the pool, and an individual's biological calculating the age.

[Number 6]
(In the formula, t is the first coefficient, DNAm _n indicates the methylation rate of the CpG site in the DNA at the nth chromosome position listed in Table 1, and C _n is the (Indicates the second coefficient for the position of the nth chromosome.)

The individual's race, nationality, and group affiliation are not particularly limited, but it is preferable that the individual be Asian, and more preferably Japanese.

The cell type of the cell population is not particularly limited, and may be composed of a single cell type or a plurality of cell types. Blood cells are preferred because they are easy to obtain.

The first coefficient t is preferably 77 to 87, more preferably 79 to 85, even more preferably 80 to 84, and even more preferably 81.946609.

The second coefficient C _n is preferably a value in the range of 50 to 150%, more preferably 80 to 120%, and 95% of the value listed in Table 1. More preferably, the value is in the range from 105% to 105%, and even more preferably 100% (ie, the values listed in Table 1).

Among the CpG sites located in the DNA at chromosome positions 1 to 235 listed in Table 1, the first number of CpG sites selected is preferably 180 to 214, and preferably 200 to 214. It is more preferable that it is below.

Furthermore, the number of randomly selected CpG sites to be added to the 235 CpG sites among the CpG sites in the DNA at chromosome positions 1 to 235 listed in Table 1 is 4 to 12. The number is preferably 4 to 9, even more preferably 4 to 6.

By using this method, it can be investigated whether biological age can be calculated appropriately even if specific CpG sites are added or subtracted from the CpG sites listed in Table 1. . Moreover, it is possible to find suitable conditions more frequently than by using random sites.

[Example 1]
DNA was extracted from the blood of 85 healthy Japanese men and women in their 20s to 70s. After bisulfite conversion, the methylation rate of cytosine was measured at CpG sites located in DNA at chromosome positions 1 to 235 listed in Table 1. Figure 1 shows the relationship between each person's chronological age and methylation rate at each position.

The obtained results were substituted into Equation 3 below to calculate the epigenome age of each person.

[Number 7]
(In the formula, t' is 81.946609, DNAm _n is the methylation rate of the CpG site in the DNA at the position of the chromosome n listed in Table 1, and C' _n is n (Indicates the coefficient for the position of the chromosome number.)

Figure 2a shows the correlation between epigenomic age and chronological age. From the obtained results, we calculated the correlation coefficients for men and women and the overall correlation coefficient, and found that the overall correlation coefficient was 0.964, the correlation coefficient based on male data was 0.961, and the correlation coefficient based on female data was: 0.970, a very high correlation was observed, and it can be seen that the above epigenomic age can be regarded as a biological age.

Next, we conducted a simple regression analysis using each person's epigenome age obtained in this way as the objective variable and chronological age as an explanatory variable. The regression equation thus obtained is shown below.

[Number 8]
Regression formula: 7.677139 + chronological age x 0.8507557 - epigenomic age By using this regression formula as a standard formula for calculating epigenetic age acceleration (EAA), the degree of aging of each person can be determined. .

Furthermore, as a tool for estimating the composition of blood cell types from DNA methylation information, we used the estimateCellCounts function in the Minfi package (Bioconductor) to estimate the composition of blood cells from CpG sites that show blood cell type-specific DNA methylation rates. Species composition was estimated. We then performed multiple regression analysis using chronological age and estimated cell type composition as explanatory variables and epigenomic age as the objective variable. The regression equation thus obtained is shown below.

[Number 9]
Regression equation: 64.30288 + chronological age x 0.8674881 + CD8 x (-44.36649) + CD4 x (-76.08973) + NK x (-50.9861) + B x (-52.4978) + M x (-82.13944 ) + N × (-55.28485) - epigenomic age where CD8, CD4, NK, B, M, and N are CD8 ⁺ T cells, CD4 ⁺ T cells, NK cells, B cells, monocytes, and This is the DNA methylation rate in neutrophils.

A graph in which the value of this regression equation is plotted on the y-axis and the chronological age is plotted on the x-axis is shown in FIG. 2b. As a result, EAA is approximately 0 regardless of chronological age, so by using this regression equation as a reference equation for calculating epigenomic age acceleration, the degree of aging of each person can be determined.

[Example 2]
In this example, CpG sites were randomly excluded from the CpG sites numbered 1 to 235 listed in Table 1, and the remaining CpG sites were applied to the following formula 4 to calculate the epigenomic age and compared with the chronological age. Specifically, 1 to 100 CpG sites were randomly selected 100 times, and the epigenomic age was calculated using the following formula 4 a total of 10,000 times using the combination of CpG sites from which each was removed. At the same time, the epigenome age was calculated 100 times using all 235 genes.

[Number 10]
(In the formula, t' is 81.946609, DNAm _k indicates the methylation rate of the CpG site in the DNA at the position of the chromosome k listed in Table 1, and C' _k is k listed in Table 1. It shows the coefficient for the position of the chromosome No. 1. k is a numerical value representing the number of the remaining CpG site after 1 to 100 CpG sites are randomly removed from the CpG sites No. 1 to 235.)

For each epigenome age, calculate the absolute value of the difference from the chronological age, and plot the number of excluded items on the x-axis and the median (point) and interquartile range (line segment) of the absolute value of the difference on the y-axis. Plotted and shown in Figure 3A. When 20 CpG sites were excluded, the difference between epigenomic age and chronological age became approximately 5 years. For each number of excluded CpG sites, Pearson's correlation coefficient was calculated for the correlation between epigenomic age and chronological age, and the x-axis shows the number of excluded CpG sites, and the y-axis shows the median of Pearson's correlation coefficients (points). ) and the interquartile range (line segment) are plotted in a graph and shown in FIG. 3B. As a result, the correlation coefficient was approximately 0.96 when 50 CpG sites were excluded.

Therefore, even if at least 185 CpG sites, preferably at least 215 CpG sites are randomly selected from 235 CpG sites and applied to Equation 4, biological age that has an extremely high correlation with chronological age can be determined.

[Example 3]
In this example, among the CpG sites numbered 1 to 235 listed in Table 1, a new CpG site located in DNA at a chromosomal position not listed in Table 1 is randomly added, and the obtained CpG site is converted to the following formula. 5 to calculate the epigenomic age and compare it with the chronological age. Specifically, 1 to 100 CpG sites other than those in Table 1 were randomly selected 100 times, and each added combination of CpG sites was used to calculate the epigenome age a total of 10,000 times using Formula 5 below. did. Note that as the coefficient for the position of the newly added chromosome, a random number within the range of the minimum value to maximum value of the coefficient (ie, -0.100 to 0.067) listed in Table 1 was randomly applied. At the same time, the epigenome age was calculated 100 times using all 235 genes.

[Number 11]
(In the formula, t' is 81.946609, DNAm _k indicates the methylation rate of the CpG site in the DNA at the chromosomal position listed in Table 1 and the newly added chromosomal position, and C' _k is the methylation rate shown in Table 1. The coefficients for the position of the k-th chromosome described in 1 and the coefficients for the position of the newly added chromosome are shown. k is a value obtained by adding the number of newly added CpG sites to 235.)

For each epigenomic age, the absolute value of the difference from the chronological age and the correlation between the epigenomic age and the chronological age were analyzed in the same manner as in Example 2. The results are shown in FIGS. 4A and 4B.

As shown in Figure 4A, the difference between epigenomic age and chronological age became approximately 5 years when three CpG sites were added. As shown in Figure 4B, the correlation coefficient reached approximately 0.96 when nine CpG sites were added.

Therefore, even if you randomly select and add 9, preferably 3, CpG sites to 235 CpG sites and apply it to Equation 5, you will find a biological age that has an extremely high correlation with chronological age. be able to.

[Example 4]
In this example, the epigenomic age is calculated by multiplying the coefficients listed in Table 1 by random numbers within a predetermined range and the obtained coefficients are applied to Equation 6 below. The absolute value of the difference and the correlation between epigenomic age and chronological age were analyzed in the same manner as in Example 2.

Specifically, for the coefficients listed in Table 1, random numbers between 1-x and 1+x (x is 101 numbers in increments of 0.01, from 0.00 to 1.00) are shown in Table 1. The epigenomic age was calculated using the following formula 6 using the coefficient obtained by multiplying by the described coefficient.

[Number 12]
(In the formula, t' is 81.946609, DNAm _n indicates the methylation rate of the CpG site in the DNA at the nth chromosome position listed in Table 1, and C' _n is the coefficient listed in Table 1. , the coefficient obtained by multiplying a random number between 1-x and 1+x (x is a number between 0.00 and 1.00 in 101 increments of 0.01) by the coefficient listed in Table 1. Figure 5 shows a graph in which the obtained epigenomic age (y-axis) is plotted against the chronological age (x-axis).

In Figure 5, as typical cases, (A) x = 0, (B) x = 0.5, and (C) x = 1.0 are shown, but the coefficients are changed up to x = 0.5. Even though the difference between chronological age and epigenomic age was small, at x=1.0, a considerable discrepancy was observed between chronological age and epigenomic age.

Therefore, FIG. 6 shows a graph in which the absolute value of the difference between chronological age and epigenomic age (y-axis) is plotted for each x (x-axis). The absolute value of the difference is expressed as median (point) ± interquartile range (line segment).

As shown in FIG. 6, when x is about 0.2, the difference between chronological age and epigenomic age is about 4 years, and when x is about 0.5, the difference is about 5 years.

In this way, even if the epigenomic age is calculated by giving a certain range to the coefficients listed in Table 1, there is sufficient accuracy for estimating the chronological age.

Claims (14)

A method of calculating an individual's biological age, the method comprising:
In the genome of a cell population obtained from an individual, among the CpG sites located in the DNA of chromosomes 1 to 235 listed in the table below,
By (1) selecting 215 to 235 CpG sites, or (2) adding 1 to 3 randomly selected CpG sites to the 235 CpG sites,
A process of creating a pool of CpG sites,
examining the methylation of CpG sites in the pool and calculating the methylation rate at each CpG site in the pool;
calculating the biological age of the individual using the following formula;

(However, t = 79 to 85, DNAm _n indicates the methylation rate of the CpG site in the DNA at the position of the chromosome n listed in the table below, and C _n indicates the methylation rate of the CpG site in the DNA at the location n of the chromosome listed in the table below (Indicates a value between 50% and 150% of the coefficient for the location of the chromosome.)
including calculation methods.
[table]






The calculation method according to claim 1, wherein t=80 to 84. The calculation method according to claim 1, wherein t=81.946609. The calculation method according to claim 1, wherein C _n indicates a value between 80% and 120% of the coefficient for the nth chromosome position listed in the table. The calculation method according to claim 1, wherein C _n is a coefficient for the position of the nth chromosome listed in the table. The calculation method according to any one of claims 1 to 5, wherein 235 CpG sites listed in the table are selected to create a pool of CpG sites. The calculation method according to any one of claims 1 to 5 , wherein the individual is Asian. The calculation method according to any one of claims 1 to 5 , wherein the cell population is blood cells. A method for evaluating an individual's aging condition, the method comprising:
Calculating the biological age of the individual by the calculation method according to any one of claims 1 to 5 ;
comparing the biological age of the individual and the chronological age of the individual;
evaluation methods, including A method for evaluating CpG sites for calculating an individual's biological age, the method comprising:
In the genome of a cell population obtained from a certain individual, among the CpG sites located in the DNA of chromosomes 1 to 235 listed in the table below,
By (1) selecting between 200 and 214 CpG sites, or (2) adding 4 to 6 CpG sites to the 235 CpG sites,
A process of creating a pool of CpG sites,
examining the methylation of CpG sites in the pool and calculating the methylation rate at each CpG site in the pool;
calculating the biological age of the individual using the following formula;

(However, t = 79 to 85, DNAm _n indicates the methylation rate of the CpG site in the DNA at the position of the chromosome n listed in the table below, and C _n indicates the methylation rate of the CpG site in the DNA at the location n of the chromosome listed in the table below (Indicates a value between 50% and 150% of the coefficient for the location of the chromosome.)
evaluation methods, including
[table]






11. The evaluation method according to claim 10, wherein the individual is Asian. The evaluation method according to claim 10 or 11, wherein the cell population is blood cells. A program for causing a computer to execute the calculation method according to any one of claims 1 to 5 or the evaluation method according to claim 10 or 11 . A computer readable storage medium storing the program according to claim 13.

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