Machine Learning Integration of Bulk and Single-Cell RNA-Seq Data Reveals Cathepsin B as a Central PANoptosis...

Area of Science:

• Immunology
• Molecular Biology
• Virology

Background:

• Influenza A virus (IAV) infection causes severe lung injury due to excessive PANoptosis, a programmed cell death pathway.
• The molecular mechanisms driving PANoptosis during IAV infection are not fully understood.

Purpose of the Study:

• To investigate the cellular heterogeneity and transcriptional dynamics of PANoptosis in influenza-infected lungs.
• To identify key molecular regulators of PANoptosis during IAV infection.

Main Methods:

• Integrated bulk and single-cell RNA sequencing (scRNA-seq) data.
• Utilized computational algorithms (AUCell, ssGSEA, AddModuleScore) to quantify PANoptosis activity.
• Employed machine learning (SVM, RF, LASSO) to identify regulatory genes.
• Performed in vivo validation in a mouse model.

Main Results:

• PANoptosis activity was mainly observed in macrophages and neutrophils.
• Cathepsin B (CTSB) was identified as a central regulator of PANoptosis.
• In vivo studies confirmed elevated PANoptosis markers and CTSB expression in IAV-infected mice.
• CTSB may promote NLRP3 inflammasome activation and lysosomal dysfunction-induced cell death.

Conclusions:

• CTSB is a critical mediator linking lysosomal integrity to innate immune-driven lung injury during IAV infection.
• Targeting CTSB may offer a therapeutic strategy to mitigate influenza-associated immunopathology.