Elucidation of mechanisms underlying the therapeutic effects of cordycepin on pulmonary hypertension, with a focus on cell senescence and gut microbiota.

RNA sequencing analysis revealed that cordycepin activates p53 signaling in pulmonary artery smooth muscle cells (PASMCs) from patients with pulmonary hypertension.

• Transcriptome analysis was performed on PASMCs derived from pulmonary hypertension patients.
• p53 signaling pathway was identified as a key mechanism through which cordycepin exerts antiproliferative effects.
• Cordycepin is a natural adenosine analogue derived from Cordyceps militaris.
• The analysis highlighted the p53-CDK1/pTERT axis as a central mechanistic pathway.

Cordycepin suppressed CDK1 expression and TERT phosphorylation at threonine 249 in PASMCs.

• CDK1 (cyclin-dependent kinase 1) suppression was identified as a downstream effect of p53 activation by cordycepin.
• TERT phosphorylation at the specific residue threonine 249 was reduced by cordycepin treatment.
• These molecular changes are consistent with inhibition of aberrant PASMC proliferation characteristic of pulmonary hypertension.
• The p53-CDK1/pTERT axis was described as the mechanistic pathway underlying cordycepin's antiproliferative effects.

Cordycepin ameliorated vascular and cardiac remodeling in both rat and mouse models of pulmonary hypertension.

• Therapeutic effects were demonstrated in rodent PH models including both rat and mouse species.
• Amelioration of vascular remodeling reflects reduction of excessive pulmonary vascular remodeling characteristic of PH.
• Cardiac remodeling was also improved, addressing the cardiopulmonary nature of the disorder.
• Both in vitro (PASMCs) and in vivo (rodent models) systems were used to evaluate cordycepin's effects.

Cordycepin induced M1-like macrophage senescence in p16Ink4a reporter mouse lungs and rat bone marrow-derived macrophages (BMDMs).

• Cellular senescence was analyzed using p16Ink4a-CreERT2 reporter mice, a specialized transgenic model for tracking senescent cells.
• Senescence was also confirmed in rat bone marrow-derived macrophages (BMDMs) in vitro.
• The macrophages affected were specifically M1-like (proinflammatory) phenotype.
• Induction of proinflammatory macrophage senescence represents a distinct immune-modulatory mechanism of cordycepin in PH.

Cordycepin significantly reshaped gut microbiota composition in rodent PH models, increasing beneficial genera and reducing proinflammatory taxa.

• Metagenomic analysis was used to assess gut microbiota changes in rodent PH models.
• Beneficial genera increased included Alistipes and Acetatifactor.
• Proinflammatory taxa reduced included Ruminococcus.
• Cordycepin modulated key metabolic pathways including short-chain fatty acid, tryptophan, and vitamin K2 metabolism.
• These findings implicate the gut-lung axis as a therapeutic target of cordycepin in PH.

Gut microbiota dysbiosis is implicated as a contributing factor to pulmonary hypertension development.

• Emerging evidence cited in the paper suggests gut microbiota dysbiosis contributes to PH development.
• The gut-lung axis was identified as one of three therapeutic axes (vascular, immune, and gut-lung) targeted by cordycepin.
• Metagenomic analysis was specifically employed to investigate microbiota changes in the context of PH treatment.
• This background framing motivated the investigation of cordycepin's microbiota-modulating properties as part of its mechanism of action.