Aptamer targeting HMGB1 attenuates inflammatory disease via domain-specific antagonism

Damage-associated molecular patterns (DAMPs) are key mediators of inflammatory disease, among which HMGB1 is a prototypical extracellular alarmin and an attractive therapeutic target. Here, we report ZH-1a, a high-affinity DNA aptamer (Kd = 2.1 nM) identified through SELEX and sequence optimization, that preferentially recognizes the proinflammatory B-box region of HMGB1. ZH-1a functions as an extracellular HMGB1-neutralizing aptamer and suppresses HMGB1-induced inflammatory signaling, including cytokine secretion and NF-κB activation in macrophages. In vivo, ZH-1a reduced late-phase systemic inflammation and multiorgan injury in LPS-induced endotoxemia, improved survival in polymicrobial sepsis, and attenuated inflammatory responses and organ damage in an HMGB1-challenge model. In addition, ZH-1a alleviated joint inflammation and structural damage in collagen-induced arthritis, and further enhanced the therapeutic efficacy of methotrexate. Together, these findings establish ZH-1a as a promising anti-inflammatory aptamer targeting HMGB1 and support aptamer-based neutralization of pathogenic extracellular HMGB1 as a therapeutic strategy for inflammatory disease.