Molecular endotypes in sepsis: integration of multicohort transcriptomics based on RNA sequencing.

Background: The heterogeneity of host responses in sepsis has hindered efforts to develop targeted therapies for this large patient population. Although growing evidence has identified sepsis endotypes based on the microarray data, studies using RNA-seq data-which offers higher sensitivity and a broader dynamic range-remain limited. We hypothesized that integrating RNA-seq data from patients with sepsis would reveal molecular endotypes with distinct biological and clinical signatures.

Methods: In this meta-analysis, we systematically searched for publicly available RNA-seq datasets of sepsis. Using identified datasets, we applied a consensus clustering algorithm to identify distinct endotypes. To characterize the biological differences between these endotypes, we performed gene-set enrichment analysis and immune cell deconvolution. Next, we investigated the association between these endotypes and mortality risks. We finally developed gene classifiers for endotype stratification and validated our endotype classification by applying these classifiers to an external cohort.

Results: A total of 280 adults with sepsis from four datasets were included in this analysis. Using an unsupervised approach, we identified three distinct endotypes: coagulopathic (n = 83, 30%), inflammatory (n = 118, 42%), and adaptive endotype (n = 79, 28%). The coagulopathic endotype exhibited upregulated coagulation signaling, along with an increased monocyte and neutrophil composition, although the adaptive endotype demonstrated enhanced adaptive immune cell responses, marked by elevated T and B cell compositions. The inflammatory endotype was characterized by upregulated TNF-α/NF-κB signaling and IL-6/JAK/STAT3 pathways with an increased neutrophil composition. Patients with the coagulopathic endotype had a significantly higher risk of mortality than those with the adaptive endotype (30% vs. 16%, odds ratio 2.19, 95% confidence interval 1.04-4.78, p = 0.04). To enable the practical application of these findings, we developed endotype classification models and identified 14 gene classifiers. In a validation cohort of 123 patients, we consistently identified these three endotypes. Furthermore, the mortality risk pattern was reproduced, with the coagulopathic endotype showing greater mortality risk than the adaptive endotype (34% vs 18%, p = 0.10).

Conclusions: This multicohort RNA-seq meta-analysis identified three biologically and clinically distinct sepsis endotypes characterized by coagulopathic, adaptive, and inflammatory responses. This endotype-based approach to patient stratification may facilitate the development of more precise therapeutic strategies for sepsis.