JPWO2020240511A5

JPWO2020240511A5 -

Info

Publication number: JPWO2020240511A5
Application number: JP2021570246A
Authority: JP
Publication date: 2023-04-18

Claims

Steps below:
(a) extracting DNA from the subject's matrix;
(b) obtaining a DNA methylation profile by measuring DNA methylation of DNA extracted from the substrate;
(c) obtaining a polygenic score by analysis of said DNA methylation profile; and (d) determining the subject's biological age from said polygenic score;
A method for calculating the biological age of a subject comprising
A method wherein extracting the DNA comprises extracting genomic DNA from saliva or blood obtained from the subject.

Said DNA methylation measurements are targeted amplicon next-generation bisals on a platform selected from the group of DNA pyrosequencing, mass spectrometry-based (Epityper ^™ ), PCR-based methylation assays, HiSeq, MiniSeq, MiSeq and NextSeq sequencers. performed using methods including phytosequencing, ion torrent sequencing, methylated DNA immunoprecipitation (MeDIP) sequencing, or hybridization with oligonucleotide arrays , or
The DNA methylation measurement was performed by SEQ
performed for polygenic DNA methylation biomarkers comprising measuring the methylation status of CG sites within any one of the human CG sites located in the ElovL2 AS1 region in ID NO:1 and combinations thereof; or,
Said DNA methylation measurement was performed using a biotinylated forward primer denoted by SEQ ID NO:2, a reverse primer denoted by SEQ ID NO:3, and a
is performed using DNA pyrosequencing comprising the pyrosequencing primer designated NO: 4, or
Said DNA methylation measurements were performed on a platform selected from the group of HiSeq, MiniSeq, MiSeq, and NextSeq sequencers using the forward primer designated SEQ ID NO:5 and the reverse primer designated SEQ ID NO:6.
carried out using target amplicon next-generation bisulfite sequencing containing mers, or
2. The method of claim 1, wherein said DNA methylation measurement is performed using a PCR-based methylation assay selected from the group of methylation-specific PCR and digital PCR.

2. The method of claim 1, wherein analyzing DNA methylation profiles to obtain the polygenic score comprises using multiple linear regression equations or neural network analysis.

Steps below:
(a) extracting DNA from multiple substrates of multiple subjects;
(b) obtaining a DNA methylation profile by measuring DNA methylation of DNA extracted from said plurality of substrates;
(c) analyzing said DNA methylation profile to obtain a polygenic score; and (d) determining biological age across a plurality of subjects from said polygenic score;
A method for calculating biological age across multiple subjects comprising
A method, wherein extracting the DNA comprises extracting genomic DNA from saliva or blood obtained from the subject.

Measuring DNA methylation of DNA extracted from said plurality of substrates comprises the following steps:
(a) amplifying genomic DNA extracted from multiple substrates with target-specific primers to obtain PCR product 1;
(b) amplifying PCR product 1 of step (a) by barcoding primers to obtain PCR product 2;
(c) performing multiplex sequencing in a single next-generation Miseq sequencing reaction using PCR product 2 from step (b);
(d) extracting data from the multiplexed sequencing of step (c) and (e) quantifying DNA methylation from the extracted data of step (d) to obtain a DNA methylation profile for each substrate. matter,
including
extracting the DNA comprises extracting genomic DNA from saliva or blood obtained from the subject;
The target-specific primers for obtaining said PCR product 1 include the forward primer indicated by SEQ ID NO: 5 and the reverse primer indicated by SEQ ID NO: 6, barcode for obtaining said PCR product 2 5. The method of claim 4, wherein the primers are barcode index primers and include a forward primer designated SEQ ID NO:7 and a reverse primer designated SEQ ID NO:8.

A combination of DNA methylation biomarkers for calculating biological age, comprising:
Said DNA methylation biomarker combinations comprise a combination of their human CG sites, which are the ElovL2 AS1 regions mapped to putative antisense regions to the ElovL2 gene and designated SEQ ID NO:1.

A kit for determining the biological age of a subject, comprising:
means and reagents for collecting and stabilizing a subject's substrate;
a scanner for reading the barcode of the kit;
instructions for substrate collection and stabilization;
including
the substrate is saliva or blood of a subject;
Stabilization of the substrate includes mailing the substrate collected to extract the DNA for the purpose of measuring the DNA methylation of the DNA extracted from the substrate to obtain the subject's DNA methylation profile, and the subject's biological A kit for determining age.

Steps below:
(a) assessing entries in a computer readable medium obtained through the sharing of user data from subjects;
(b) matching the entries of step (a) to the kit of claim 7 obtained from said subject to determine the biological age of said subject;
(c) calculating the subject's biological age using the method of claim 1 or 4 to obtain a calculated biological age;
(d) integrating the calculated biological age of step (c) into a machine learning model of said subject by performing a statistical analysis using the assessment of step (a) to obtain an integrated data report; ,
(e) analyzing the consolidated data report of step (d) along with progress in responding to questionnaires obtained by sharing user data from said subjects over time and comparing them to the National Association's recommendations; and (f) sharing the dynamic report of step (e) with the subject on a computer readable medium to provide lifestyle change recommendations;
including,
A computer-implemented method for providing lifestyle change recommendations.

The computer-readable medium is
an open source development tool containing information on a test for calculating biological age based on the method of claim 1 or claim 4 ;
a virtual shopping cart for ordering said test;
a scanning facility for scanning the barcode of the kit of claim 7 and receiving test results from the lab;
A computer-implemented method comprising:
Said open source development tools include basic physiological measurements, weight, height, blood pressure, heart rate, self-assessment of mood, McGill pain, and weight, height, blood pressure, heart rate, self-assessment of mood, contained in a computer-readable medium for investigating lifestyle features that impact healthy aging. 9. The computer-implemented method of claim 8 , comprising questionnaires including questionnaires, diet and nutrition questionnaires, exercise questionnaires and lifestyle questionnaires including alcohol, drugs, smoking, and combinations thereof.

creating a health ecosystem focused on normalizing or slowing biological aging in subjects using Android, Apple, or We Chat mini-programs for personalized lifestyle recommendations; or , using standard data pipelines across multiple subjects and management systems such as Cloud data prep, storing data in object storage enterprises on servers or cloud servers such as Amazon, Ali cloud, Microsoft Azure, etc.
10. The computer-implemented method of claim 9 , comprising:

Random Forests (RF), Support Vector Machines (SVM), Linear Discriminant Analysis (LDA), Generalized Linear Models ( GLM), and the use of a set of artificial intelligence algorithms such as deep learning (DL), which are dynamically updated to provide personalized lifestyle recommendations for lifestyle changes. The described computer-implemented method.

Steps below:
(a) storing data obtained from multiple subjects;
(b) analyzing the stored data of step (a); and (c) building a model;
A method for developing a computer readable medium comprising
storing the data obtained from the plurality of users is a cloud-based SQL database;
analyzing the stored data comprises a group selected from deep machine learning, reinforcement learning, and machine learning, or a combination thereof;
The step of building the model correlates input questionnaire measurements with DNA methylation age and chronological age differences as outputs, and other physiological and psychological outputs such as pain, blood pressure, BMI, and mood. including, method.

The machine learning is a data mining algorithm including random forest analysis or a data mining algorithm including K-Means cluster analysis, or Amazon
Machine Learning (AML) or on platforms including Apache Hadoop Distributed File System, Amazon EC2 Google Compute Engine and Microsoft Azure. 13. The method for developing a computer readable medium of claim 12 , comprising a group selected from software including ai products and combinations thereof.

Steps below:
(a) calculating the subject's biological age using the method of claim 1 or claim 4 to obtain an initial biological age prior to biological intervention;
(b) performing a biological intervention on said subject;
(c) repeating step (a) on a subsequent substrate obtained from said subject after step (b) has been performed to obtain the post-biological intervention biological age;
(d) integrating post-biological intervention biological age into a machine learning model of said subject to assess the effect of biological intervention on said subject's biological age;
5. The method of claim 1 or claim 4 for use in a method of assessing the effect of a biological intervention on the biological age of a subject comprising
The biological intervention of step (b) includes nutritional supplements, vitamins, therapy, test substance administration, dietary manipulation, metabolic manipulation, surgical manipulation, social manipulation, behavioral manipulation, environmental manipulation, sensory manipulation, hormonal manipulation. and epigenetic manipulation, or a combination thereof;
extracting the DNA comprises extracting genomic DNA from saliva or blood obtained from the subject;
Integrating post-biological intervention biological age in the subject's machine learning model combines the biological age assessed in step (c) and physiological parameters obtained through sharing of user data from the subject. including, method.

Steps below:
(a) calculating the age of the substrate obtained from the subject using the method of claim 1 or claim 4 to obtain the initial biological age prior to administration of the test agent;
(b) administering a test agent to the subject;
(c) repeating step (a) on a subsequent substrate obtained from said subject after step (b) has been performed to obtain the biological age after administration of the test agent;
(d) integrating biological age after administration of a test agent into a machine learning model of said subject; assessing whether a reduction in age was calculated by integrating into the machine learning model; determining as an anti-aging agent;
including,
5. A method according to claim 1 or claim 4 for use in a method of screening drugs that are anti-aging agents,
extracting the DNA comprises extracting genomic DNA from saliva or blood obtained from the subject;
Integrating the post-administration biological age of the test agent into the subject's machine learning model was obtained through the sharing of the biological age assessed in step (c) and the user data from the subject. A method comprising a physiological parameter.