LSD1 Inhibition Induces Fetal Hemoglobin Expression and Provides a Novel Therapeutic Approach to Sickle Cell Disease

Abstract Sickle cell disease (SCD) is an autosomal recessive genetic disorder caused by a point mutation in the human β-globin gene. Patients harboring this mutation can exhibit long-chain polymers of hemoglobin and sickle-shaped red blood cells, and suffer from severe medical manifestations including hemolysis and vaso-occlusive crises. Multiple preclinical, clinical and epidemiologic studies have shown that the levels of unmutated fetal hemoglobin (HbF encoded by the γ-globin gene) correlate with less severe disease, validating HbF induction as a therapeutic approach in SCD. Treatment with hydroxyurea (HU), the only approved therapy for SCD, results in a variable induction of HbF and significant improvement in the frequency of pain crises. However, a significant percentage of patients treated with HU fail to exhibit durable benefit, necessitating the need for alternative therapeutic agents. The human γ-globin gene is repressed in the post-natal period by epigenetic mechanisms, and therefore may lend itself to pharmacological intervention aimed at derepressing gene expression. One of the most important of these epigenetic mechanisms is catalyzed by lysine-specific demethylase 1 (LSD1), a histone demethylase that removes mono-/dimethyl marks from the lysine 4 and 9 residues of histone H3 through an FAD-directed redox process. Here, we report the characterization of selective, potent, and orally bioavailable LSD1 inhibitors from two classes - FAD-directed inhibitors that achieve inhibitory activity through formation of covalent FAD-adducts and non-FAD-directed, reversible inhibitors - and demonstrate their ability to induce γ-globin gene expression in murine and primate preclinical models. In the Towne’s SCD mouse model, oral administration of LSD1 inhibitors significantly increased HbF+ cell (F cell) production. Concurrent with the increase in F cells, sickle cell numbers, reticulocyte counts, and bilirubin levels were all markedly reduced, indicating an amelioration of several pathophysiological features of SCD. FAD- and non-FAD-directed LSD1 inhibitors were more effective than HU in increasing F cells production, and the combination of HU and suboptimal doses of LSD1 inhibitors resulted in a greater induction of F cells and more pronounced reductions in reticulocyte counts and bilirubin levels. In addition to the humanized SCD model, HbF induction in response to LSD1 inhibitor treatment was evaluated in non-anemic cynomolgus monkeys. Oral administration of LSD1 inhibitors significantly induced F cells and HbF in a dose-dependent manner and over a sustained period (>50 days) following the discontinuation of treatment. The percentage of induced F cells in total RBCs was linearly correlated with the percentage of HbF protein induced by LSD1 inhibition. Taken together, these results support the potential utility of LSD1 inhibition as a novel therapeutic approach to increase HbF production. Disclosures Lee: Incyte Corporation: Employment, Other: Stock. Soloviev: Incyte Corporation: Employment, Other: Stock. Zhang: Incyte Corporation: Employment, Other: Stock. Roman: Incyte Corporation: Employment, Other: Stock. Yang: Incyte Corporation: Employment, Other: Stock. Bowman: Incyte Corporation: Employment, Other: Stock. Burke: Incyte Corporation: Employment, Other: Stock. Margulis: Incyte Corporation: Employment, Other: Stock. O'Connor: Incyte Corporation: Employment, Other: Stock. Yang: Incyte Corporation: Employment, Other: Stock. Wu: Incyte Corporation: Employment, Other: Stock. Wynn: Incyte Corporation: Employment, Other: Stock. Burn: Incyte Corporation: Employment, Other: Stock. Shuey: Incyte Corporation: Employment, Other: stock. Diamond: Incyte Corporation: Employment, Other: Stock. Yao: Incyte Corporation: Employment, Other: Stock. Hollis: Incyte Corporation: Employment, Other: Stock. Yeleswaram: Incyte Corporation: Employment, Other: Stocks. Roberts: Incyte Corporation: Employment, Other: Stock. Huber: Incyte Corporation: Employment, Other: Stock. Scherle: Incyte Corporation: Employment, Other: Stock. Ruggeri: Incyte Corporation: Employment, Other: Stock.