Multi-omics profiling reveals microbiota, metabolite, lipid, and immunological heterogeneity underlying distinct pathophysiological mechanisms of age-related endotypes in type 1 diabetes

Type 1 diabetes (T1D) is an autoimmune disease characterized by marked heterogeneity in age at diagnosis, clinical progression, and immune pathology. Increasing evidence suggests that age-related T1D endotypes may reflect distinct underlying molecular mechanisms; however, these mechanisms remain incompletely characterized at the cellular and transcriptional levels. To investigate age-associated immune heterogeneity in T1D, peripheral blood mononuclear cells (PBMCs) were collected from a selected cohort of newly diagnosed pediatric individuals with T1D and healthy controls. Single-cell RNA sequencing was performed on PBMCs from 27 individuals with T1D and 27 age- and sex-matched healthy controls to profile peripheral immune cell populations and characterize transcriptional differences associated with age-related T1D endotypes. Overall design: PBMCs were isolated from peripheral blood samples collected from 27 individuals with type 1 diabetes and 27 age- and sex-matched healthy controls. Single-cell suspensions were processed using the 10x Genomics Chromium Single Cell 3' Reagent Kits v3.1, and sequencing libraries were generated according to the manufacturer's instructions. Libraries were sequenced on the Illumina NovaSeq 6000 platform to generate single-cell RNA-seq data.