MDB-41. TARGETING EPIGENETIC REGULATORS TO INHIBIT GROUP 3 MEDULLOBLASTOMA MIGRATION

Medulloblastoma highly malignant type of childhood brain cancer which develops at the cerebellum. Medulloblastoma can be grouped into four molecular subtypes; Sonic Hedgehog (Shh), Wnt, group 3, and group 4 based on their molecular development. Mostly group 3 medulloblastoma are highly malignant subtype. Medulloblastoma migration from the cerebellum to the leptomeningeal space and spinal cord is critical for worsening the symptoms. Targeting the medulloblastoma migration would potentially attenuate the symptoms. Identifying potential epigenetic regulators and targeting them might be an effective approach to cure the symptoms in medulloblastoma patients. CRISPR epigenome library screening targeting more than 500 epigenetic factors by using more than 3000 sgRNA can help to identify potential epigenetic regulators to medulloblastoma migration. Group 3 medulloblastoma tumor is highly malignant so, we have selected a group 3 medulloblastoma human cell line D425 for our study. CRISPR epigenome library or sg RNA targeting 500 epigenetic genes was introduced into D425 cells with the help of lentiviral particles. The cells were antibiotically selected and expanded after selection. In vitro, trans-well migration assays were performed with technical and biological replicates. The cells that migrated through the transmembrane were collected and the cells which did not show any migration were also collected followed by enrichment. The DNA isolation followed by amplification of barcoded sequences identifiers tagged to different sgRNAs were size selected and undergone for Illumina NGS sequencing. The NGS sequencing results will reveal the key epigenetic factors involved in the migration of Group 3 medulloblastoma. After confirmation of pathway analysis and in vitro and in vivo validation would identify potential factors. This study will ultimately help us to find a potential epigenetic regulator for targeted therapy to inhibit migration of group 3 medulloblastoma to reduce the symptoms and combine with other effective drugs to establish synergistic effects for treatment.