Inhibition of High Mobility Group Box-1 (HMGB1) Eradicates Human Myelodysplastic Syndrome

Myelodysplastic syndrome (MDS) is a heterogeneous and pre-leukemic clonal stem cell disorder characterized by aberrant hematopoiesis and bone marrow failure. Since MDS is associated with a dysregulated innate immune system, we performed a genetic screen of the human toll-like receptor (TLR) pathway and found that high mobility group box-1 (HMGB1) was abundantly expressed in MDS. HMGB1 is a ubiquitously expressed, non-histone chromatin-binding protein. In solid cancers, high expression of HMGB1 was associated with both shortened progression-free and overall survival, but its role in hematologic malignancies is not well defined. We hypothesized that targeting HMGB1 could be therapeutic in MDS. To study HMGB1 in human MDS, we utilized a cell line, MDS-L, derived from a patient with myelodysplastic syndrome with ring sideroblasts. Morphologically, the cells display dysplastic features, including cytoplasmic vacuolation and prominent, irregular nucleoli. We found that HMGB1 was detected at high levels in both MDS-L and in primary CD34+ MDS compared to CD34+ cord blood and healthy human marrow (*p Next, we sought to pharmacologically inhibit HMGB1 signaling with sivelestat, which is a small molecule inhibitor for neutrophil elastase and HMGB1. Since sivelestat has not been studied in either normal or malignant hematopoiesis, whether it or other HMGB1 inhibitors could impact MDS is not yet defined. Following 72 h culture, HMGB1 protein levels were decreased in both MDS-L cells and in primary CD34+ MDS cells treated with 300 mg/ml sivelestat compared to cultures with vehicle alone (*p Since treatment with sivelestat could decrease both MDS cell expansion and impair the ability to generate colonies in methylcellulose assays, we sought to determine whether blocking HMGB1 could decrease MDS engraftment in NOD-scid ILR2gnull (NSG) mice. At 17 weeks, irradiated NSG mice that were transplanted with MDS-L cells treated with sivelestat demonstrated a 3.2-fold decrease in human CD45 marrow engraftment with MDS-L compared to control cultures (*p Mechanistically, sivelestat increased cellular apoptosis in cultures of MDS cells compared to control cultures (*p=0.001). This was done in part by increasing levels of p53 upregulated modulator of apoptosis (PUMA, *p=0.0003), activated caspase 3 (*p Disclosures No relevant conflicts of interest to declare.