Gut microbiome analysis of metagenome under the treatment of mannose or not from HM1 tumors in mice Raw sequence reads

The gut microbiome plays a critical role in enhancingtheeffectiveness of immune checkpoint blockade (ICB) therapy. Here, we demonstrate that oral supplementation with the prebiotics mannoseslows tumor growth in immunocompetent miceina manner dependent on the gut microbiota.This effect is associated with an increase in the abundance of Faecalibaculum. Mannose generates an immune-stimulatory tumor microenvironment (TME), characterized by a higher percentage of effector and memory CD8+ T cells, alongwith a notable immunoreactivegeneexpressionsignature. Crucially, mannose modulates the stemness program of CD8+ T cells and promotes the generation of progenitor exhausted CD8+T cells (Tpex). Mechanistically, mannose enhances theproduction of the short-chain fatty acids (SCFA) propionate and butyrate by the gut microbiota. Administration of propionate and butyrate suppresses tumor progression and stimulates the expansion and differentiation of Tpex both in vivo and in vitro. Furthermore,mannose workssynergistically with PD-1 blockade, resulting intumor regression and enhanced differentiation of intratumoral Tpex into intermediate exhausted CD8+ T cells (Tex-int) and terminally exhausted CD8+T cells (Tex-term). We also identified a gene signature related to mannosetherapy that correlateswith favorable responses to ICB across various cancers. Overall, our findings support the combination of mannose dietary supplementation with anti-PD-1 immunotherapy in treating ovarian cancer and suggest its potential forclinical application.