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

> **NIH NIH F99** · UNIVERSITY OF COLORADO DENVER · 2022 · $36,896

## 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. Single-
cell 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:** 10528820
- **Project number:** 1F99CA274654-01
- **Recipient organization:** UNIVERSITY OF COLORADO DENVER
- **Principal Investigator:** John A DeSisto
- **Activity code:** F99 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $36,896
- **Award type:** 1
- **Project period:** 2022-08-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10528820

## Citation

> US National Institutes of Health, RePORTER application 10528820, Investigation of tumor stem cell maintenance and cellular hierarchy in pediatric high-grade glioma (1F99CA274654-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10528820. Licensed CC0.

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