Grant 02309-T Update End Year 3: Targeting the Cancer Epigenome: The Effect of Specific Histone Lysine Methyltransferase Inhibition in Canine B-Cell Lymphoma

Update on targeting epigenome for treatment of B-cell lymphoma from Dr McCleary-Wheeler.

Lymphoma, particularly the large, B-cell subtype, is one of the most common malignancies in dogs. Canine lymphoma can be treated, but it is rarely cured. Novel therapeutic strategies are necessary to improve outcomes in dogs diagnosed with lymphoma. Recently, advances in the understanding of human lymphomas have focused on the area of epigenetics. One area of this research involves understanding how genes are turned on or off based on different modifications to histone proteins, a specific group of proteins that interact with DNA. Specific enzymes that modify these histone proteins have altered activity that can lead to lymphoma development in human lymphomas. One of these enzymes is EZH2. Increased activity of EZH2 has been shown to play an important role in the development of some human lymphomas. Data from a Phase I study of an EZH2 inhibitor, tazemetostat, in relapsed or refractory human B-cell, non-Hodgkin lymphoma has shown to be a safe, oral therapy with potential clinical benefit. The role of EZH2, however, has not been evaluated in canine B-cell lymphomas to date. Given the similarity between human and canine B-cell lymphoma, we seek to investigate whether EZH2 activity plays a role in canine B-cell lymphoma. To do this, we use canine lymphoma cells and specific EZH2 inhibitors, including the tazemetostat used in early human studies, to evaluate the effect of EZH2 inhibition on cell growth and survival. Our data suggest that this inhibitor is highly potent and effective for inhibiting EZH2 effects on histone modification in canine lymphoma. This is important as this inhibitor is an orally bioavailable drug with a good toxicity profile in humans, making this inhibitor a candidate for clinical trials in dogs with lymphoma. Initial data suggests that EZH2 inhibition may not impede lymphoma cell proliferation or survival. However, we have confirmed that some genes that regulate the ability of canine lymphoma cells to replicate are altered with EZH2 inhibition. Specifically, one gene, CDKN1a, is turned back on when EZH2 is inhibited. The activation of CDKN1a is repeatable and profound. We will be continuing this work with a sequencing approach to further understand what genes are regulated by EZH2 in canine B-cell lymphoma cells. Our findings are suggesting an importance for EZH2 in canine lymphoma and for continued investigations into cell cycle regulators that may be abnormal. By understanding the genes regulated by EZH2, we can characterize the role this histone modifying enzyme has in canine lymphoma. Moreover, we can assess how inhibition of EZH2 activity may be utilized clinical in dogs with lymphoma. We appreciate the continued generous support provided by this grant from the CHF and its supporting donors.

Research Update Dr. McCleary-Wheeler for targeting epigenome in treatment of B-cell lymphoma