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Principal Investigator: Ram I. Mahato
Organization: UNIVERSITY OF NEBRASKA MEDICAL CENTER
Fiscal Year: 2023
Award: $475,579
Funding agency: National Institute of Neurological Disorders and Stroke
PROJECT SUMMARY
Medulloblastoma (MB) is the most common childhood brain tumor arising from the cerebellum. Many factors
influence the proliferation, differentiation, and migration of cerebellar granular neuronal precursor (GNP). Among
them, MDM2 is a major nexus between tumor suppressor TP53 and hedgehog (Hh) signaling in GNPs and
promotes MB tumor growth and metastasis. In addition, PI3K and BRD4 signaling also play key roles in MB cell
growth, cancer stem cell (CSC) proliferation, and tumorigenesis. Further, MB treatment is challenging due to the
development of chemoresistance, inefficient drug transport across the blood brain barrier (BBB) and drug
induced neurotoxicity. Hh inhibitors are effective initially to treat SHH-MB, but their repeated use develops
chemoresistance due to mutations in smoothened (SMO) but can be overcome by modulating GLI, which is
downstream of SMO. In our preliminary studies, we synthesized a series of potent BRD4/PI3K dual inhibitors
by modifying structure of parent compound SF2523. One of the compounds 8-(2,3-dihydrobenzo[b][1,4]dioxin-
6-yl)-2-morpholino-4H-chromen-4one (abbreviated as MDP5) was found highly potent. We then determined X-
ray crystal structures of the recombinant BD1 and BD2 domains from BRD2 in complex with MDP5. While MDP5
showed higher potency in DOAY cells compared to SF2523 (12.6 µM), IC50 values for MDP5 and SF2523 were
similar potency on HD-MB03 MB (MYC amplified) cells. MDP5 decreased the target downstream proteins like
p-AKT, MYCN, Cyclin D1, and increased the degradation of MYCN protein indicated by p-MYCN (ser 54). We
also discovered a small molecule JW-475A which is a potent dual MDM2 and XIAP inhibitor. MDP5 and JW-
475A (a dual MDM2 and XIAP inhibitor) effectively inhibited the proliferation of MB cells in a dose dependent
manner, with significantly higher cell killing when these drugs were used in combination. Treatment of MB cells
with the combination of these two drugs significantly decreased the colony formation capacity compared to
individual drugs. We prepared PEG-DSPE based lipid nanoparticles (LNPs) with 4.9±0.1% and 4.8±0.1% loading
for MDP5 and JW-475A. BBB penetrating targeted LNPs were prepared by surface decorating with rabies virus
glycoprotein (RVG) peptide-peptide. Our hypothesis is that inhibition of BRD4/PI3K and MDM2/XIAP
simultaneously using MDP5 and JW-475A represents a promising strategy to inhibit MB tumor in vivo. Further,
we will use RVG-PEG-DSPE LNPs to encapsulate MDP5 and JW-475A, which have poor drug transport across
the BBB. Our specific aims are to i) Synthesize novel MDP5 derivatives as dual function BRD4/PI3K inhibitors
and characterize in vitro activity; ii) Evaluate anti-cancer efficacy of JW-475A in combination with MDP5 in vitro.;
iii) Formulate MDP5 and JW-475A into LNPs decorated with RVG peptide and determine their biodistribution,
therapeutic efficacy, and systemic/organ toxicity in in SHH and MYC driven cells and PDX-based orthotopic and
transgenic SmoA1 MB mouse models. Long-term significance. Successful completion of this project will
provide a platform technology for treating brain tumors using this innovative LNP-based combination therapy.
Terms: <1-Phosphatidylinositol 3-Kinase><AKT><Abbreviations><Akt protein><Antioncogene Protein p53><Apoptosis><Apoptosis Inhibitor><Apoptosis Inhibitor Gene><Apoptosis Pathway><Assay><Avian Myelocytomatosis Viral Oncogene Homolog><BBB penetration><BRD2><BRD2 gene><Binding><Bioassay><Biodistribution><Biologic Assays><Biological Assay><Blood - brain barrier anatomy><Blood-Brain Barrier><Brain><Brain Neoplasia><Brain Neoplasms><Brain Nervous System><Brain Tumors><CCND1 Protein><Cell Body><Cell Communication and Signaling><Cell Cycle><Cell Division Cycle><Cell Growth in Number><Cell Line><Cell Multiplication><Cell Proliferation><Cell Signaling><CellLine><Cells><Cellular Expansion><Cellular Growth><Cellular Proliferation><Cellular Tumor Antigen P53><Cerebellum><Chemoresistance><Childhood Brain Neoplasm><Childhood Brain Tumor><Cholesterol><Choline Chloride Dihydrogen Phosphate><Choline Phosphate><Choline Phosphate Chloride><Clinical><Combined Modality Therapy><Complex><Cyclin D1><D6S113E><Data><Development><Dioxin Compound><Dioxins><Dose><Drug Transport><Drugs><E coli><E. coli><Encapsulated><Encephalon><Erinaceidae><Escherichia coli><Exhibits><FSRG1><Female Sterile Homeotic-Related Gene 1><Film><G1/S-Specific Cyclin D1><Generalized Growth><Genetic Alteration><Genetic Change><Genetic defect><Goals><Growth><HDM2><Hedgehog (Hh) signal transduction pathway><Hedgehogs><Hemato-Encephalic Barrier><Human><Hydration><Hydration status><Hydrophobicity><IAP Family Gene><IAP Family Protein><IQ Deficit><Impairment><In Vitro><Individual><Intracellular Communication and Signaling><KIAA9001><Lipids><MB03><MDM2><MDM2 gene><MDMX protein><MYC gene><MYCN><MYCN Protein><MYCN gene><Macrogols><Mdm-2 protein><Medication><Metastasis><Metastasize><Metastatic Lesion><Metastatic Mass><Metastatic Neoplasm><Metastatic Tumor><Modern Man><Molecular Interaction><Multimodal Therapy><Multimodal Treatment><Mutate><Mutation><N-Myc Protein><NMYC><NMYC Gene><NMYC Gene Product><Names><Neoplasm Metastasis><Nerve Cells><Nerve Unit><Neural Cell><Neural Stem Cell><Neuroblastoma MYC><Neurocognitive Deficit><Neurocyte><Neurons><Oncogenesis><Oncoprotein MDM2><Oncoprotein p53><Organ><Outcome><P53><PI-3 Kinase><PI3-Kinase><PI3CG><PI3KGamma><PI3k><PIK3><PIK3CG><PIK3CG gene><PRAD1 Protein><Parents><Particle Size><Pathway interactions><Patients><Peptides><Pharmaceutic Preparations><Pharmaceutical Preparations><Phosphatidylinositol 3-Kinase><Phosphatidylinositol-3-OH Kinase><Phosphocholine><Phosphoinositide 3-Hydroxykinase><Phosphoprotein P53><Phosphoprotein pp53><Phosphorylcholine><Phosphorylcholine Chloride><Plasmids><Play><Polyethylene Glycols><Polyethylene Oxide><Polyethyleneoxide><Polyoxyethylenes><Progenitor Cells><Programmed Cell Death><Proliferating><Protein Kinase B><Protein TP53><Proteins><Proto-Oncogene Proteins c-akt><Proto-Oncogene Proteins c-bcl-1><PtdIns 3-Kinase><RAC-PK protein><RING3><RNF3><Recombinant Proteins><Recombinants><Resistance><Roentgen Rays><Role><SHH><SHH gene><Secondary Neoplasm><Secondary Tumor><Series><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Single Crystal Diffraction><Sonic Hedgehog><Strains Cell Lines><Structure><Surface><TP53><TP53 gene><TRP53><Tissue Growth><Toxic effect><Toxicities><Transgenic Organisms><Treatment Efficacy><Tumor Cell><Tumor Protein p53><Tumor Protein p53 Gene><Tumor Suppressor Proteins><Type I Phosphatidylinositol Kinase><Type III Phosphoinositide 3-Kinase><Wound Repair><X Ray Crystallographies><X-Radiation><X-Ray Crystallography><X-Ray Diffraction Crystallography><X-Ray Radiation><X-Ray/Neutron Crystallography><X-ray><Xray><Xray Crystallography><analog><anti-cancer><anticancer><bcl-1 Proto-Oncogene Products><bcl-1 Proto-Oncogene Proteins><bcl1 Proto-Oncogene Proteins><biological signal transduction><blood-brain barrier penetration><bloodbrain barrier><bloodbrain barrier penetration><c-akt protein><c-bcl-1 Proteins><cancer metastasis><cancer progenitor cells><cancer stem cell><cell growth><cell killing><chemoresistant><chemotherapy><chemotherapy resistance><chemotherapy resistant><combination therapy><combined modality treatment><combined treatment><cultured cell line><cyclin D><design><designing><determine efficacy><developmental><drug/agent><efficacy analysis><efficacy assessment><efficacy determination><efficacy evaluation><efficacy examination><ethanolamine phosphate><evaluate efficacy><examine efficacy><genome mutation><hedgehog signaling><hedgehog signaling pathway><hh signaling pathway><in vitro activity><in vivo><inhibitor><inhibitor-of-apoptosis protein><innovate><innovation><innovative><intelligence quotient deficit><intervention efficacy><lipid based nanoparticle><lipid nanoparticle><malignant stem cell><mdm-2 oncogene protein><mdm2 protein><medulloblastoma><medulloblastoma cell line><meter><migration><mouse model><multi-modal therapy><multi-modal treatment><multidrug use><multiple drug use><murine model><mutant><myc Oncogenes><name><named><naming><nano medicinal><nano medicine><nanomedicinal><nanomedicine><neoplastic cell><nerve stem cell><neural precursor><neural precursor cell><neural progenitor><neural progenitor cells><neurocognitive decline><neurocognitive impairment><neuron progenitors><neuron toxicity><neuronal><neuronal progenitor><neuronal progenitor cells><neuronal stem cells><neuronal toxicity><neuroprogenitor><neurotoxicity><novel><ontogeny><p53 Antigen><p53 Genes><p53 Tumor Suppressor><p53-Binding Protein MDM2><parent><pathway><pediatric brain neoplasm><pediatric brain tumor><phosphoethanolamine><phosphorylethanolamine><poly drug use><polydrug use><progenitor cell proliferation><protein p53><protein purification><proto-oncogene protein RAC><proto-oncogene protein akt><rabies virus G protein><rabies virus glycoprotein><rabies virus glycoprotein G><rac protein kinase><related to A and C-protein><resistant><small molecule><smoothened signaling pathway><social role><stem cell proliferation><stem cells><synergism><systemic toxicity><technology platform><technology system><therapeutic efficacy><therapy efficacy><transgenic><treatment strategy><tumor><tumor cell metastasis><tumor growth><tumor suppressor><tumorigenesis><tumorigenic><tumors in the brain><wound healing><wound resolution><x-linked inhibitor of apoptosis protein>