Multi-omics Mechanisms of Lifestyle Intervention in Regulating Blood Glucose

A composite outcome summarizing glucose metabolism indicators, including:

HbA1c (%), Fasting plasma glucose (mmol/L), 60- and 120-minute plasma glucose during OGTT (mmol/L), Fasting and post-load insulin (μU/mL), Fasting and post-load C-peptide (ng/mL), Homeostatic Model Assessment of Insulin Resistance (HOMA-IR, index), etc.

A composite outcome summarizing kidney function indicators, including:

Serum creatinine (mg/dL), Blood urea nitrogen (mg/dL), Estimated glomerular filtration rate (eGFR, mL/min/1.73m²), etc.

A composite outcome summarizing liver function biomarkers, including: Alanine aminotransferase (ALT, U/L), Aspartate aminotransferase (AST, U/L), Gamma-glutamyl transferase (GGT, U/L), Alkaline phosphatase (ALP, U/L), Total bilirubin (μmol/L), Albumin (g/L), etc.

Total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, etc.

High-sensitivity C-reactive protein, etc.

Retinopathy grading assessed by standardized digital retinal fundus images.

A composite outcome summarizing dietary intake indicators, including:

Total daily energy intake (kcal), Macronutrient distribution and intake (percentage and intake (gram) of total energy from carbohydrate, protein, fat, etc.), Selected micronutrient intakes (mg/day), etc.

This composite outcome includes multiple physiological parameters measured via wearable devices to assess rest and recovery status. The parameters include: Sleep duration (hours), Sleep stages (percent time in REM, deep, light), Respiratory rate (breaths per minute), Heart rate (n per minute), Other relevant physiologic signals (as available from device).

A structured lifestyle management questionnaire will be administered to assess multiple aspects of participants' personal health-related behaviours and preferences. This includes: food preference, exercise preference, past medical histories, health-related personal behaviours, etc.

Body weight measured in kilograms using a calibrated digital scale.

Standing height measured in meters using a stadiometer.

Calculated as weight in kilograms divided by height in meters squared (kg/m²).

Measured in centimeters

Systolic and diastolic blood pressure will be measured in millimeters of mercury (mmHg) using an automated sphygmomanometer. Participants will be seated quietly for at least 5 minutes before measurement, and two readings will be taken 1-2 minutes apart; the average will be recorded.

Resting heart rate will be measured in beats per minute (bpm) using the automated sphygmomanometer's pulse reading or a validated heart rate monitor. Measurements will be taken in the seated position after at least 5 minutes of rest.

Whole-genome sequencing for variant analysis.

Untargeted metabolomics profiling using LC-MS/MS.

Quantitative proteomics using mass spectrometry.

Shotgun metagenomic sequencing of fecal samples to characterize gut microbiome taxonomic composition and functional gene profiles.

Untargeted metabolomics profiling of fecal samples using liquid chromatography-mass spectrometry (LC-MS/MS) to identify and quantify metabolites.

Targeted lipidomics analysis of serum lipids using mass spectrometry to quantify lipid species and subclasses.

Peripheral blood mononuclear cells (PBMCs) will be analyzed using multiple omics platforms to comprehensively profile molecular changes. Analyses will include:

Transcriptomics: Bulk RNA sequencing to quantify gene expression. Epigenomics: Genome-wide DNA methylation profiling using bisulfite sequencing. Phosphoproteomics: Quantitative analysis of phosphorylated proteins using mass spectrometry-based phosphoproteomics to assess signaling pathway activation.

Single-cell Transcriptomics: Single-cell RNA sequencing to characterize cell type-specific transcriptional changes.

Results from these platforms will be integrated to generate a composite PBMC multi-omics dataset.

Glycemic excursions will be evaluated using a composite measure derived from continuous glucose monitoring (CGM) data. The composite score will integrate multiple CGM-derived metrics, including:

Mean glucose (mg/dL) Standard deviation of glucose (mg/dL) Coefficient of variation (%) Mean amplitude of glycemic excursions (MAGE, mg/dL)

Average heart rate during each exercise session.

Number of completed exercise sessions.

Energy expenditure estimated by wearable device.

Duration of each exercise session.

Total sets and repetitions performed during exercise.

Intensity or load during exercise session (self-reported using Borg RPE scale or device-recorded).

Post-exercise oxygen consumption recorded by wearable device.

Total fat mass measured by bioelectrical impedance analysis.

Fat-free mass measured by bioelectrical impedance analysis.

Percentage of total body weight as fat.

Estimated visceral fat area.

Skeletal muscle mass measured by bioelectrical impedance analysis.