Validating long-read single-cell PIPseq as a technology to study T Cell Variant Dynamics in Aging and Dementia in Women (RPA Pilot / Seed Project)

Neuro-immune reflex circuits between innate and adaptive immune cells and brain inflammation may enhance systemic immune system inflammation in dementia patients. A planned MPI R01 with a June 2024 deadline aims to understand the molecular mechanism governing dementia-associated immune response decline to improve dementia patients' health. This NRPA pilot project, a new collaboration with Mark Ebbert, uses single-cell nanopore long-read sequencing to characterize and quantify gene expression at the RNA isoform level rather than collapsing all RNAs for a gene into a single gene-expression measurement, which oversimplifies the biology. RNA transcriptomic variations in T cell subsets from older adults will be examined in our R01 application to uncover and test mechanisms linking the neuro-immune reflex arc to dementia-associated T cell alterations. Single-cell nanopore sequencing is a new technology with few studies and no standard techniques. Therefore, pilot data using human peripheral blood is needed to prove its superiority over scSeq. We hypothesize that single-cell nanopore sequencing of human T cells using PIPseq will reveal new mechanistic insights into the systemic impact of cognitive decline by allowing us to characterize and quantify T cell changes with much greater granularity than ever before.