The heat shock protein 70 (Hsp70) family of chaperones is essential for rhinovirus replication

<jats:p>Rhinoviruses (RVs) are the predominant cause of the common cold and a major trigger of acute asthma exacerbations. Yet, unlike related enteroviruses such as poliovirus (PV) and enterovirus A71 (EV-A71), no approved vaccines or antivirals exist. Because enteroviruses depend heavily on host factors for replication, cellular proteins that support the replication of multiple enteroviruses have emerged as attractive broad-spectrum antiviral targets that may offer a higher barrier to resistance than virus-targeted therapies. The 70-kDa heat shock protein (Hsp70) family, a highly conserved class of molecular chaperones, is required for the replication of several enteroviruses, including EV-A71 and coxsackievirus A16, but whether RVs share this dependency was unknown. Here, we show that two mechanistically distinct small-molecule inhibitors of the Hsp70 family abolish replication of RV-A16, a clinically relevant RV type widely used in asthma research. Using siRNA knockdown, we demonstrate a role for HSPA8, the major constitutively expressed Hsp70 isoform, in RV replication. We further demonstrate that Hsp70 activity is indispensable for viral translation, identifying this as a key Hsp70-dependent step in the RV replication cycle. Together, these findings establish Hsp70 chaperones as essential host factors for RV replication and strengthen the rationale for targeting them as a broad-spectrum antiviral strategy.</jats:p>