# Investigation of tumor stem cell maintenance and cellular hierarchy in pediatric high-grade glioma

> **NIH NIH K00** · UNIVERSITY OF COLORADO DENVER · 2024 · $93,872

## Abstract

Project Summary
The project’s overall goal is to determine important biological characteristics and investigate therapeutic 
options for pediatric high-grade gliomas (PHGG), the most aggressive of childhood central nervous system 
tumors and most common cause of childhood cancer mortality. PHGG survival rates are less than 5% for the 
subtype diffuse midline glioma and 20% for hemispheric histone 3-wild type (H3-wt) PHGG. PHGG are highly 
invasive and often grow diffusely among normal cells, limiting surgery as a therapeutic option. Radiation 
therapy (RT) is transiently effective, but the tumors nearly always recur. Despite hundreds of clinical trials, no 
chemotherapy has shown a definitive survival benefit in PHGG. Effective PHGG therapies are critically 
needed.
PHGG likely originates from stemlike tumor initiating cells (PICs). PHGG tumors comprise several 
distinct cell types of glial origin, in varying proportions. This tumor heterogeneity complicates understanding 
PHGG tumor biology and designing therapies. Aim 1 will investigate how each distinct cell type in PHGG 
contributes to overall tumorigenesis in a mouse model. Single-cell RNA-Seq (scRNA-Seq) analysis of 
orthotopic patient derived PHGG xenografts (PDX) will be used to define the cell types present and identify 
differentially regulated oncogenic pathways that drive their growth. Pathway expression will be knocked down 
by targeting key effector genes with shRNA using stable lentiviral transduction. The effect on tumor growth will 
be evaluated using survival, histology and single-cell RNA-Seq. 
Aim 2 will perform lineage tracing to determine whether a single PIC cell type produces all of the 
proliferating cell types that comprise PHGG. Lineage tracing will be performed in a mouse PDX model. Singlecell genomic DNA sequencing will be performed on PDX tumors. Mutational signatures consisting of single and 
multiple nucleotide variations as well as copy number variation will be used to define each cell type. Conserved 
patterns of mutation among cell types will be used to determine the hierarchical relationships among cell types. 
Once the lineage relationships are worked out, resistance to RT will be studied in the PDX model. RT is the 
most consistently effective therapy against PHGG but works only temporarily before cells regrow. RT 
resistance by cell type will be determined based upon differential survival of cell types versus control following 
RT. Drug screening of resistant cell types to identify radiation sensitizers will be performed. The candidate 
drugs will be combined with RT to investigate their effectiveness at increasing the duration of the RT effect.

## Key facts

- **NIH application ID:** 10890190
- **Project number:** 5K00CA274654-03
- **Recipient organization:** UNIVERSITY OF COLORADO DENVER
- **Principal Investigator:** John A DeSisto
- **Activity code:** K00 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $93,872
- **Award type:** 5
- **Project period:** 2022-08-01 → 2027-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10890190

## Citation

> US National Institutes of Health, RePORTER application 10890190, Investigation of tumor stem cell maintenance and cellular hierarchy in pediatric high-grade glioma (5K00CA274654-03). Retrieved via AI Analytics 2026-06-11 from https://api.ai-analytics.org/grant/nih/10890190. Licensed CC0.

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