Not listed LEVERAGING A NEUROEPITHELIAL STEM CELL MODEL TO DETERMINE REGULATORS OF DIFFERENTIATION FAILURE IN NF2-MUTANT NEURAL PROGENITORS - PROJECT SUMMARY SPINAL EPENDYMOMA (SP-EPN) IS A RARE CENTRAL NERVOUS TUMOR THAT IS ASSOCIATED WITH NF2 MUTATIONS AND OCCURS IN NEARLY HALF OF PATIENTS WITH NF2-RELATED SCHWANNOMATOSIS (NF2). THE CURRENT STANDARD OF CARE IS SURGICAL RESECTION; HOWEVER, THIS CAN BE RISKY FOR PATIENTS, ESPECIALLY FOR PATIENTS WITH NF2 BECAUSE OF THEIR PREDISPOSITION TO DEVELOPING MULTIPLE SP-EPN AND DEVELOPING THEM AT AN EARLIER AGE. THERE ARE CURRENTLY NO EFFECTIVE MEDICAL OPTIONS, HIGHLIGHTING A CRITICAL NEED FOR THESE PATIENTS. VERY LITTLE IS KNOWN ABOUT SP-EPN DEVELOPMENT OTHER THAN THAT IT APPEARS TO ARISE FROM A NEURAL PROGENITOR-LIKE CELL IN THE RADIAL GLIA LINEAGE. OUR PRELIMINARY TRANSCRIPTOMIC DATA SHOWS THAT THERE MIGHT BE SMALL CLUSTERS OF NEURAL PROGENITORS WITHIN PATIENT SP-EPN. WHEN NF2 MUTATIONS ARE INDUCED IN OUR NEUROEPITHELIAL STEM (NES) CELL MODEL, THESE CELLS FAIL TO DIFFERENTIATE AND FORM CLUSTERS IN VITRO. WE HYPOTHESIZE THAT DIFFERENTIATION FAILURE IN NF2-MUTANT NES CELLS IS MEDIATED THROUGH DYSREGULATED DEVELOPMENTAL PATHWAYS, AND THAT TEAD INHIBITION WILL BE EFFECTIVE IN REVERSING THESE EFFECTS. THE CENTRAL GOAL OF THIS PROPOSAL IS TO DETERMINE REGULATORS OF THIS STALLED DIFFERENTIATION IN OUR NF2-MUTANT NES CELLS AND ATTEMPT TO RESCUE DIFFERENTIATION USING TEAD INHIBITORS, VT104 AND MGH-CP1, IN THESE CELLS. IN AIM 1, WE WILL PERFORM A CRISPR KINASE AND TRANSCRIPTION FACTOR KNOCKOUT SCREEN TO PINPOINT WHICH REGULATORY COMPONENTS ARE RESPONSIBLE FOR FAILED DIFFERENTIATION OF NF2-MUTANT NEURAL PROGENITORS. THIS WILL PROVIDE INSIGHT INTO POTENTIAL MECHANISMS DRIVING FORMATION OF NF2-MUTANT SP-EPN. IN AIM 2, WE WILL USE VT104 AND MGH-CP1 TO RESCUE DIFFERENTIATION IN OUR NF2-MUTANT CELLS. THESE INHIBITORS HAVE BEEN SOMEWHAT EFFECTIVE IN OTHER NF2-MUTANT TUMORS DUE TO THEIR EFFECTS ON TEAD TRANSCRIPTION FACTORS, WHICH ARE ABERRANTLY ACTIVATED IN THE CONTEXT OF NF2 LOSS AND PROMOTE THE EXPRESSION OF GENES CRITICAL FOR STEM CELL MAINTENANCE, PROLIFERATION, AND SURVIVAL. OVERALL, OUR LONG-TERM GOAL IS TO BETTER UNDERSTAND THE MOLECULAR PATHWAYS GOVERNING SP-EPN TUMORIGENESIS AND USING THIS INFORMATION TO IDENTIFY THERAPEUTICS THAT CAN TARGET THESE TUMORS. FUTURE STUDIES WILL INCLUDE TESTING THESE THERAPIES IN AN ORTHOTOPIC MOUSE MODEL USING THE NF2-MUTANT NES CELLS. THIS PROJECT IS HIGHLY TRANSLATABLE AND IS ESSENTIAL FOR FINDING BETTER MEDICAL OPTIONS AND IMPROVING OUTCOMES FOR PATIENTS WITH NF2-MUTANT SP-EPN. $56.6k 5/7/26 Not listed LEVERAGING A NEUROEPITHELIAL STEM CELL MODEL TO DETERMINE REGULATORS OF DIFFERENTIATION FAILURE IN NF2-MUTANT NEURAL PROGENITORS - PROJECT SUMMARY SPINAL EPENDYMOMA (SP-EPN) IS A RARE CENTRAL NERVOUS TUMOR THAT IS ASSOCIATED WITH NF2 MUTATIONS AND OCCURS IN NEARLY HALF OF PATIENTS WITH NF2-RELATED SCHWANNOMATOSIS (NF2). THE CURRENT STANDARD OF CARE IS SURGICAL RESECTION; HOWEVER, THIS CAN BE RISKY FOR PATIENTS, ESPECIALLY FOR PATIENTS WITH NF2 BECAUSE OF THEIR PREDISPOSITION TO DEVELOPING MULTIPLE SP-EPN AND DEVELOPING THEM AT AN EARLIER AGE. THERE ARE CURRENTLY NO EFFECTIVE MEDICAL OPTIONS, HIGHLIGHTING A CRITICAL NEED FOR THESE PATIENTS. VERY LITTLE IS KNOWN ABOUT SP-EPN DEVELOPMENT OTHER THAN THAT IT APPEARS TO ARISE FROM A NEURAL PROGENITOR-LIKE CELL IN THE RADIAL GLIA LINEAGE. OUR PRELIMINARY TRANSCRIPTOMIC DATA SHOWS THAT THERE MIGHT BE SMALL CLUSTERS OF NEURAL PROGENITORS WITHIN PATIENT SP-EPN. WHEN NF2 MUTATIONS ARE INDUCED IN OUR NEUROEPITHELIAL STEM (NES) CELL MODEL, THESE CELLS FAIL TO DIFFERENTIATE AND FORM CLUSTERS IN VITRO. WE HYPOTHESIZE THAT DIFFERENTIATION FAILURE IN NF2-MUTANT NES CELLS IS MEDIATED THROUGH DYSREGULATED DEVELOPMENTAL PATHWAYS, AND THAT TEAD INHIBITION WILL BE EFFECTIVE IN REVERSING THESE EFFECTS. THE CENTRAL GOAL OF THIS PROPOSAL IS TO DETERMINE REGULATORS OF THIS STALLED DIFFERENTIATION IN OUR NF2-MUTANT NES CELLS AND ATTEMPT TO RESCUE DIFFERENTIATION USING TEAD INHIBITORS, VT104 AND MGH-CP1, IN THESE CELLS. IN AIM 1, WE WILL PERFORM A CRISPR KINASE AND TRANSCRIPTION FACTOR KNOCKOUT SCREEN TO PINPOINT WHICH REGULATORY COMPONENTS ARE RESPONSIBLE FOR FAILED DIFFERENTIATION OF NF2-MUTANT NEURAL PROGENITORS. THIS WILL PROVIDE INSIGHT INTO POTENTIAL MECHANISMS DRIVING FORMATION OF NF2-MUTANT SP-EPN. IN AIM 2, WE WILL USE VT104 AND MGH-CP1 TO RESCUE DIFFERENTIATION IN OUR NF2-MUTANT CELLS. THESE INHIBITORS HAVE BEEN SOMEWHAT EFFECTIVE IN OTHER NF2-MUTANT TUMORS DUE TO THEIR EFFECTS ON TEAD TRANSCRIPTION FACTORS, WHICH ARE ABERRANTLY ACTIVATED IN THE CONTEXT OF NF2 LOSS AND PROMOTE THE EXPRESSION OF GENES CRITICAL FOR STEM CELL MAINTENANCE, PROLIFERATION, AND SURVIVAL. OVERALL, OUR LONG-TERM GOAL IS TO BETTER UNDERSTAND THE MOLECULAR PATHWAYS GOVERNING SP-EPN TUMORIGENESIS AND USING THIS INFORMATION TO IDENTIFY THERAPEUTICS THAT CAN TARGET THESE TUMORS. FUTURE STUDIES WILL INCLUDE TESTING THESE THERAPIES IN AN ORTHOTOPIC MOUSE MODEL USING THE NF2-MUTANT NES CELLS. THIS PROJECT IS HIGHLY TRANSLATABLE AND IS ESSENTIAL FOR FINDING BETTER MEDICAL OPTIONS AND IMPROVING OUTCOMES FOR PATIENTS WITH NF2-MUTANT SP-EPN. $56.6k 5/7/26 Not listed LEVERAGING A NEUROEPITHELIAL STEM CELL MODEL TO DETERMINE REGULATORS OF DIFFERENTIATION FAILURE IN NF2-MUTANT NEURAL PROGENITORS - PROJECT SUMMARY SPINAL EPENDYMOMA (SP-EPN) IS A RARE CENTRAL NERVOUS TUMOR THAT IS ASSOCIATED WITH NF2 MUTATIONS AND OCCURS IN NEARLY HALF OF PATIENTS WITH NF2-RELATED SCHWANNOMATOSIS (NF2). THE CURRENT STANDARD OF CARE IS SURGICAL RESECTION; HOWEVER, THIS CAN BE RISKY FOR PATIENTS, ESPECIALLY FOR PATIENTS WITH NF2 BECAUSE OF THEIR PREDISPOSITION TO DEVELOPING MULTIPLE SP-EPN AND DEVELOPING THEM AT AN EARLIER AGE. THERE ARE CURRENTLY NO EFFECTIVE MEDICAL OPTIONS, HIGHLIGHTING A CRITICAL NEED FOR THESE PATIENTS. VERY LITTLE IS KNOWN ABOUT SP-EPN DEVELOPMENT OTHER THAN THAT IT APPEARS TO ARISE FROM A NEURAL PROGENITOR-LIKE CELL IN THE RADIAL GLIA LINEAGE. OUR PRELIMINARY TRANSCRIPTOMIC DATA SHOWS THAT THERE MIGHT BE SMALL CLUSTERS OF NEURAL PROGENITORS WITHIN PATIENT SP-EPN. WHEN NF2 MUTATIONS ARE INDUCED IN OUR NEUROEPITHELIAL STEM (NES) CELL MODEL, THESE CELLS FAIL TO DIFFERENTIATE AND FORM CLUSTERS IN VITRO. WE HYPOTHESIZE THAT DIFFERENTIATION FAILURE IN NF2-MUTANT NES CELLS IS MEDIATED THROUGH DYSREGULATED DEVELOPMENTAL PATHWAYS, AND THAT TEAD INHIBITION WILL BE EFFECTIVE IN REVERSING THESE EFFECTS. THE CENTRAL GOAL OF THIS PROPOSAL IS TO DETERMINE REGULATORS OF THIS STALLED DIFFERENTIATION IN OUR NF2-MUTANT NES CELLS AND ATTEMPT TO RESCUE DIFFERENTIATION USING TEAD INHIBITORS, VT104 AND MGH-CP1, IN THESE CELLS. IN AIM 1, WE WILL PERFORM A CRISPR KINASE AND TRANSCRIPTION FACTOR KNOCKOUT SCREEN TO PINPOINT WHICH REGULATORY COMPONENTS ARE RESPONSIBLE FOR FAILED DIFFERENTIATION OF NF2-MUTANT NEURAL PROGENITORS. THIS WILL PROVIDE INSIGHT INTO POTENTIAL MECHANISMS DRIVING FORMATION OF NF2-MUTANT SP-EPN. IN AIM 2, WE WILL USE VT104 AND MGH-CP1 TO RESCUE DIFFERENTIATION IN OUR NF2-MUTANT CELLS. THESE INHIBITORS HAVE BEEN SOMEWHAT EFFECTIVE IN OTHER NF2-MUTANT TUMORS DUE TO THEIR EFFECTS ON TEAD TRANSCRIPTION FACTORS, WHICH ARE ABERRANTLY ACTIVATED IN THE CONTEXT OF NF2 LOSS AND PROMOTE THE EXPRESSION OF GENES CRITICAL FOR STEM CELL MAINTENANCE, PROLIFERATION, AND SURVIVAL. OVERALL, OUR LONG-TERM GOAL IS TO BETTER UNDERSTAND THE MOLECULAR PATHWAYS GOVERNING SP-EPN TUMORIGENESIS AND USING THIS INFORMATION TO IDENTIFY THERAPEUTICS THAT CAN TARGET THESE TUMORS. FUTURE STUDIES WILL INCLUDE TESTING THESE THERAPIES IN AN ORTHOTOPIC MOUSE MODEL USING THE NF2-MUTANT NES CELLS. THIS PROJECT IS HIGHLY TRANSLATABLE AND IS ESSENTIAL FOR FINDING BETTER MEDICAL OPTIONS AND IMPROVING OUTCOMES FOR PATIENTS WITH NF2-MUTANT SP-EPN. $0 5/7/26