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Updated: Jul 19, 2025
Evaluation of a Reliable Biomarker in a Cecal Ligation and Puncture-Induced Mouse Model of Sepsis
Published on: December 9, 2022
A Synthetic Poly(A) Tail Targeting Extracellular CIRP Inhibits Sepsis
Atsushi Murao1, Alok Jha1, Gaifeng Ma1
1Center for Immunology and Inflammation, The Feinstein Institutes for Medical Research, Manhasset, NY.
Abstract:
Sepsis is an infectious inflammatory disease that often results in acute lung injury (ALI). Cold-inducible RNA-binding protein (CIRP) is an intracellular RNA chaperon that binds to mRNA's poly(A) tail. However, CIRP can be released in sepsis, and extracellular CIRP (eCIRP) is a damage-associated molecular pattern, exaggerating inflammation, ALI, and mortality. In this study, we developed an engineered poly(A) mRNA mimic, AAAAAAAAAAAA, named A12, with 2'-O-methyl ribose modification and terminal phosphorothioate linkages to protect it from RNase degradation, exhibiting an increased half-life. A12 selectively and strongly interacted with the RNA-binding motif of eCIRP, thereby preventing eCIRP's binding to its receptor, TLR4. In vitro treatment with A12 significantly decreased eCIRP-induced macrophage MAPK and NF-κB activation and inflammatory transcription factor upregulation. A12 also attenuated proinflammatory cytokine production induced by eCIRP in vitro and in vivo in macrophages and mice, respectively. We revealed that treating cecal ligation and puncture-induced sepsis with A12 significantly reduced serum organ injury markers and cytokine levels and ALI, and it decreased bacterial loads in the blood and peritoneal fluid, ultimately improving their survival. Thus, A12's ability to attenuate the clinical models of sepsis sheds lights on inflammatory disease pathophysiology and prevention of the disease progress.
Insights
A novel engineered mRNA mimic, A12, effectively neutralizes extracellular cold-inducible RNA-binding protein (eCIRP), significantly reducing sepsis-induced inflammation, acute lung injury, and improving survival rates in preclinical models.
Area of Science:
- Molecular Biology
- Immunology
- Pathophysiology
Background:
- Sepsis is a life-threatening inflammatory condition often leading to acute lung injury (ALI).
- Extracellular cold-inducible RNA-binding protein (eCIRP) acts as a damage-associated molecular pattern, exacerbating sepsis-related inflammation and ALI.
- Current therapeutic strategies for sepsis-induced ALI are limited.
Purpose of the Study:
- To develop a therapeutic agent targeting eCIRP to mitigate sepsis-induced inflammation and ALI.
- To investigate the mechanism of action of the engineered molecule A12 in neutralizing eCIRP.
- To evaluate the efficacy of A12 in preclinical models of sepsis and ALI.
Main Methods:
- Development of A12, an engineered poly(A) mRNA mimic with enhanced stability.
- Assessment of A12's binding affinity to eCIRP and its ability to block eCIRP-TLR4 interaction.
- In vitro and in vivo experiments to evaluate A12's effects on inflammatory pathways, cytokine production, and sepsis outcomes.
Main Results:
- A12 selectively bound to eCIRP, preventing its interaction with TLR4.
- A12 suppressed eCIRP-induced activation of MAPK and NF-κB signaling pathways in macrophages.
- A12 treatment reduced pro-inflammatory cytokine levels, ALI, bacterial load, and improved survival in a mouse model of sepsis.
Conclusions:
- A12 is a potent inhibitor of eCIRP activity, demonstrating significant therapeutic potential for sepsis and associated ALI.
- The study elucidates a novel therapeutic strategy targeting DAMPs in sepsis.
- A12 offers a promising avenue for the prevention and treatment of inflammatory diseases like sepsis.
