1 International Res. Center for Med. Sci., Kumamoto Univ.2 Kozo Keikaku Eng. Inc.A machine learning pipeline uncovers LAG-3-mediated immunoregulation in inflammatory HSCs49 Inflammatory signals trigger emergency hematopoiesis for host immune defence but impair HSC self-renewal when sustained chronically. Understanding how HSCs resolve inflammatory stress is critical for maintaining hematopoietic homeostasis. Single-cell multi-omics analysis is a powerful tool to examine molecular dynamics in individual cells, yet current techniques cannot extract multiple biological features from the same cell simultaneously. To overcome this, we developed a machine learning-based pipeline to generate in silico bridge data that predicts inflammation-associated regulatory factors across modalities. Inflammation often alters HSC immunophenotypes. So, we employed Hlf-tdTomato reporter mice, which label functional HSCs in the RFPhigh fraction even under inflammatory stress. RFPhigh HSCs were isolated from mice treated with PBS or LPS, a bacterial molecule inducing systemic inflammation, and subjected to scRNA-seq, scATAC-seq, and CyTOF analysis. Using a transfer learning model, we aligned data in a shared latent space to generate the integrated in silico bridge data. This approach predicted key regulators of LPS-induced immune responses in HSCs, with 80% reproducibility in qPCR. Notably, expression of LAG-3 and IL2RG was increased on LPS-treated HSC surface, confirmed by FACS. Functionally, the LAG-3 on LPS-treated HSCs could suppress OT-II CD4+ T cell activation in co-culture with CD11c+ dendritic cells, which was reversed by anti-LAG-3 neutralizing antibody.In summary, our machine learning approach provides an analytic pipeline that outperforms the conventional gene filtering method for identifying key inflammation regulators. The discovery of LAG-3 upregulation on inflamed HSCs suggests a novel immunosuppressive mechanism to facilitate resolution of inflammation.○Nicole Pui-Yu Ho1 Minato Aihara2 Taichi Shimura2Daisuke Kurotaki1 Hitoshi Takizawa1P 019
元のページ ../index.html#52