Abstract 2159: Ikaros and Casein kinase II (CK2) regulate PI3K pathway in pediatric leukemia

Abstract

Abstract Ikaros (IKZF1) is a tumor suppressor whose function is impaired in high-risk pediatric B-cell acute lymphoblastic leukemia (B-ALL). IKZF1 encodes a DNA-binding, zinc finger protein that regulates expression of genes involved in important biological pathways. Using chromatin immunoprecipitation coupled with next-generation sequencing (ChIP-SEQ) we found that Ikaros binds to the upstream regulatory regions of multiple genes that regulate the phosphatidylinositol-3-Kinase (PI3K) pathway. Ikaros target genes include PIK3C2B and PI3KFYVE. We used gain-of-function and loss-of-function experiments to determine how Ikaros regulates transcription of its target genes. Overexpression of Ikaros by retroviral transduction in Nalm6 leukemia cells results in reduced transcription of PIK3C2B and PI3KFYVE as evidenced by qRT-PCR. Luciferase reporter assays with PIK3C2B and PI3KFYVE promoters showed that Ikaros can function as a transcriptional repressor of these genes. Transfection of Nalm6 cells with Ikaros shRNA resulted in increased expression of PIK3C2B and PI3KFYVE genes. These results suggest that Ikaros functions as a transcriptional repressor of PIK3C2B and PI3KFYVE genes in leukemia. Next, we studied signaling pathways that regulate the ability of Ikaros to transcriptionally repress the PIK3C2B and PI3KFYVE genes. We have previously shown that a pro-oncogenic Casein Kinase II (CK2) can directly phosphorylate Ikaros in vivo and that CK2-mediated phosphorylation impairs Ikaros function. We tested whether inhibition of CK2 activity affects Ikaros ability to regulate PIK3C2B and PI3KFYVE transcription in leukemia. Results show that molecular and pharmacological inhibition of CK2 have a very similar effect on transcription of Ikaros target genes and they result in transcriptional repression of both PIK3C2B and PI3KFYVE genes. Treatment of leukemia cell lines, as well as primary B-ALL cells, with different CK2 inhibitors resulted in enhanced Ikaros binding to its target genes, as evidenced by quantitative chromatin immunoprecipitation (qChIP). In summary, the presented data provide evidence that Ikaros and CK2 regulate the PI3K pathway via transcriptional regulation of the PIK3C2B and PI3KFYVE genes. Our results demonstrate that CK2 inhibition enhances Ikaros activity as a transcriptional repressor of genes that promote the PI3K pathway in primary B-ALL cells, and identify CK2 inhibitors as candidate drugs to therapeutically restore Ikaros function in B-ALL. Supported by the National Institutes of Health R01 HL095120, and the Four Diamonds Fund Endowment. Citation Format: Chandrika Gowda, Chunhua Song, Yali Ding, Sunil Muthusami, Xiaokang Pan, Dhimant Desai, Shantu G. Amin, Kimberly J. Payne, Sinisa Dovat. Ikaros and Casein kinase II (CK2) regulate PI3K pathway in pediatric leukemia. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2159. doi:10.1158/1538-7445.AM2015-2159