Targeting the MAP4K4 pathway to inhibit pediatric high-grade glioma invasion

Hemispheric high-grade gliomas (HGGs) in children are fast-growing and become rapidly widespread; the median survival rates for these diagnoses are measured in months, not years. It is therefore critical to discover more about the pathophysiology of pediatric HGGs (pHGGs), particularly their highly invasive nature. Investigating this characteristic to identify drug targets that would slow the speed of tumor invasion would significantly benefit our patients. MAP4K4 is a kinase with diverse biochemical and physiological roles in normal cell physiology. Previous data have suggested the MAP4K4 pathway may be a cornerstone in tumor invasiveness in multiple central nervous system and peripheral cancer types. We hypothesize that changes in function and expression of MAP4K4 play a significant role in the invasive ability of pHGGs. To determine whether MAP4K4 is altered in specific subtypes of hemispheric pHGG specimens from Stanford/Lucile Packard Children’s Hospital, our team will perform high- throughput methylation in combination with genomic sequencing to test for variants of MAP4K4 (and associated genes) (Aim 1). To determine whether MAP4K4 is necessary for pHGGs invasion, we will use pharmacological inhibition of (PF-06260933) and genetic (CRISPR/Cas9) silencing of MAP4K4 and downstream targets to regulate function and assess invasion in spheroid invasion assays (Aim 2). To evaluate whether inhibiting MAP4K4 reduces tumor cell invasion in vivo and prolongs survival, we will use MAP4K4 drug inhibitors in a pHGGs-luciferase orthotopic mouse model, and image the tumors at defined time points. We will also identify the cellular specificity of the MAP4K4 inhibitor in vivo with clearing-assisted tissue click chemistry (Aim 3). In conjunction with the Prolo laboratory, our team includes experts in CRISPR/Cas9 technology to probe molecular pathways genetically (Michael Bassik, PhD), leaders in neuro-oncology and xenograft mouse modeling of high-grade gliomas (Michelle Monje, MD, PhD), genomic and methylomic profiling of tumors (Matija Snuderl, MD) and click-chemistry (Li Ye, PhD). Analyzing these results will help to inform us of the molecular mechanisms underlying pHGG invasion. By creating a database detailing the profiles of our pHGG cohort, which will be published and made widely available for the broader scientific community, we hope to accelerate translational research to design treatments for this devastating cancer. Project Number: 1K08NS140556-01A1 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of Neurological Disorders and Stroke (NINDS) | Principal Investigator: Laura Prolo | Institution: STANFORD UNIVERSITY, STANFORD, CA | Award Amount: $209,177 | Activity Code: K08 | Study Section: Special Emphasis Panel[ZRG1 CTH-K (83)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11300011