EPCO-37. IDENTIFICATION OF KEY EPIGENETIC REGULATORS FOR MEDULLOBLASTOMA INVASION AND METASTASIS

Abstract

Abstract Medulloblastoma (MB) is most common type of pediatric brain cancer. It starts from the hind brain or cerebellum. It is actually an embryonic malignant cancer of the cerebellum is mainly reported over 20% of all central nervous system (CNS) neoplasms in children. MB frequently affected the 3.8-6.9 per million children population in North America and Europe. MB embracing four subgroups: Sonic Hedgehog, Wingless, Group 3, and Group 4. However, the molecular and cellular factors and their mechanism in compelling the maintenance of malignancy are have not well understood. Invasion of tumor cells across the extracellular matrix to the other neighboring tissues is main key event critical for metastasis of the medulloblastoma metastasis. Several factors affect the medulloblastoma genesis, progression, invasion and metastasis. But epigenetic regulators are the crucial regulators MB progression, invasion and metastasis. In order to identify the key regulators for medulloblastoma progression, invasion and metastasis we are doing CRISPR epigenome library screening targeting more than 500 epigenetic genes and 4 guide RNAs targeting each for brain medulloblastoma cells (ONS76 and D425 cells). Initially we have transduced cells with epigenome CRISPR library with different titers to determine the effective viral titer to transfect 30% or lesser number of cells. So that each transduced cell has each guide RNA targeting each gene. Further the transduction was scaled up and after selection the cells were expanded. Further in vitro invasion assay the invaded and un-invaded cells were collected across the Matrigel matrix coated membrane. Along with further enrichment and expansion of these cells we isolated the genomic DNA and sequenced them. The present CRISPR epigenome sequencing analysis targeting invasion property of medulloblastoma cells may provide novel key epigenetic factors affecting the medulloblastoma invasion and eventually metastasis might be the novel and more effective therapeutic target for medulloblastoma treatment.