Defining cellular responses to HDAC-selective inhibitors reveals that efficient targeting of HDAC3 is required to elicit cytotoxicity and overcome na?ve resistance to pan-HDACi in diffuse large B cell lymphoma

Approved histone deacetylase (HDAC) inhibitors have low efficacy against the most commonly-diagnosed nonHodgkin lymphoma, diffuse large B cell lymphoma (DLBCL), but the mechanisms underlying clinical resistance are poorly understood. Using a DLBCL cell-based model, we previously demonstrated that resistance to panHDAC inhibitors (HDACi) is characterized by reversible growth arrest and sensitivity by mitotic arrest and apoptosis. The goal of the current study is to better define mechanisms of sensitivity and resistance to the cytotoxic effects of HDACi by using HDAC-selective inhibitors to determine which HDACs need to be targeted to achieve the sensitive and resistant phenotypes. We find that an inhibitor selective for HDACs 1 and 2 induces G1 arrest across DLBCL cell lines used, which is consistent with the resistant phenotype. In contrast an HDAC3selective inhibitor induces DNA damage and cytotoxicity in a cell line that is sensitive to pan-HDACi but has no effect on resistant cell lines. RNAi-mediated depletion of HDAC3 indicate the presence of a long-lived population of HDAC3 in DLBCL cell lines. Finally, doses of pan-HDACi 3-5 times higher than the IC50 established for reversible growth inhibition induce the sensitive phenotype in resistant cell lines, suggesting that resistance may be associated with failure to efficiently inhibit HDAC3. Our findings indicate that selective inhibition of HDACs 1 and 2 is associated with G1 arrest and resistance to pan-HDACi while efficient targeting of HDAC3 could be key to achieving a cytotoxic response. Thus, our work reveals a potential novel mechanism of resistance to panHDACi.