Abstract LB-82: MLL2 maintains neoplastic cell growth and global histone H3 lysine 4 monomethylation
Authors: Guo Changcun , Chen Lee H. , Huang Yafen , Chang Chun-chi , Wang Ping , Pirozzi Christopher J. , Qin Xiaoxia , Bao Xuhui , Greer Paula K. , McLendon Roger E. , Yan Hai , Keir Stephen T. , Bigner Darell D. , He Yiping 
Published: May 2015

One striking theme emerging from recent findings in cancer genetics is that chromatin remodeling and histone methylation modifiers are frequently altered in human cancers. Among these newly identified cancer genes, one of the best examples is the gene encoding MLL2 (KMT2D, a.k.a. ALR/MLL4), a histone lysine methyltransferase that plays an important role in regulating gene transcription. Genetic alterations suggestive of a functional deficiency in MLL2 and other genes in the same pathways are common. The identification of alterations in MLL2 suggests potential new opportunities for therapeutics, and highlights an urgent need to understand the underlying tumorigenic mechanism. Filling such a knowledge gap has been challenging, due to the lack of appropriate assays for the gigantic (∼600 kDa), understudied MLL2 protein.

To overcome the difficulty, we have used innovative somatic gene editing-based assays to determine the effect of an MLL2 deficiency on neoplastic cells. In particular, we have used homologous recombination- and nuclease-mediated gene editing approaches to generate a panel of isogenic human cancer cell lines that differ with respect to their endogenous MLL2 status.

Our studies found that an MLL2 deficiency results in attenuated cancer cell proliferation and defective cell migration. We identified direct transcriptional target genes and revealed the connection of MLL2 to multiple cellular signaling pathways. Analysis of histone H3 modifications revealed that MLL2 is essential for maintaining the level of global histone H3 lysine 4 (H3K4) monomethylation and that its enzymatic SET domain is directly responsible for this function. Furthermore, we found that a majority of MLL2 binding sites are located in regions of potential enhancer elements. The finding concerning enhancer elements is significant, as enhancer elements have increasingly been recognized as critically involved in tumorigenesis.

Together, these findings revealed the role of MLL2 in mediating diverse signaling pathways and regulating enhancer elements in human cells, and shed light on the tumorigenic role of its deficiency. Our study supports that MLL2 has distinct roles in neoplastic cells, as opposed to pre-neoplastic cells, and that inhibiting MLL2 may be a viable strategy for cancer therapeutics.

Citation Format: Changcun Guo, Lee H. Chen, Yafen Huang, Chun-chi Chang, Ping Wang, Christopher J. Pirozzi, Xiaoxia Qin, Xuhui Bao, Paula K. Greer, Roger E. McLendon, Hai Yan, Stephen T. Keir, Darell D. Bigner, Yiping He. MLL2 maintains neoplastic cell growth and global histone H3 lysine 4 monomethylation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-82. doi:10.1158/1538-7445.AM2014-LB-82