Three-dimensional organoid culture unveils resistance to clinical therapies in adult and pediatric glioblastoma

Sundar, Swetha J.; Shakya, Sajina; Barnett, Austin; Wallace, Lisa C.; Jeon, Hye-Min; Sloan, Andrew E.; Recinos, Violette Renard; Hubert, Christopher G.

doi:10.1016/j.tranon.2021.101251

Cited by 43 publications

(33 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…RNA sequencing of spatially isolated GBM cells from dissociated organoids reflects region-specific gene expression profiles from patient GBM tumors, and the SOX2-high organoid outer rim is functionally enriched for stem cell activity (Shakya et al, 2021). This SOX2+ GBM organoid rim is highly proliferative and the SOX2+ cells within this region are resistant to standard of care therapies including radiation therapy (Hubert et al, 2016), the chemotherapy temozolomide, and other clinically relevant therapeutics (Sundar et al, 2021). A wide range of patient-derived GBM organoid specimens demonstrated increased SOX2 expression within the highly proliferative rim region ( Fig.…”

Section: Resultsmentioning

confidence: 99%

WDR5 represents a therapeutically exploitable target for cancer stem cells in glioblastoma

Mitchell

Shakya

Silver

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Glioblastomas (GBMs) are heterogeneous, treatment-resistant tumors that are driven by populations of cancer stem cells (CSCs). Despite their importance for tumor growth, few molecular mechanisms critical for CSC population maintenance have been exploited for therapeutic development. We employed spatially resolved loss-of-function screening in GBM patient-derived organoids to identify essential epigenetic regulators and identified WDR5 as indispensable for CSCs. WDR5 is a component of the WRAD complex, which promotes SET1-family-mediated Lys4 methylation of histone H3, associated with positive regulation of transcription. Genetic and pharmacologic inhibition of WDR5 reduced growth and self-renewal of CSCs as well as CSC-mediated tumor growth. Further, WDR5 inhibitors partially blocked WRAD complex assembly, reduced H3K4 trimethylation, and inhibited tumor growth. These findings highlight the role of WDR5 and the WRAD complex in CSCs for maintaining the CSC state and provide a rationale for therapeutic development of WRAD complex inhibitors for GBM and other advanced cancers.

show abstract

Section: Resultsmentioning

confidence: 99%

WDR5 represents a therapeutically exploitable target for cancer stem cells in glioblastoma

Mitchell

Shakya

Silver

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…GBOs were developed by incorporating minced pieces of GBM specimen with Matrigel in serum-free media ( 107 ). GBOs have been shown to have similar histological and transcriptional profiles to their respective patient tumor and recapitulate patient-specific responses and resistance to standard of care therapy ( 108 , 109 ). A cryopreserved GBO repository has been developed and used to screen a library of 22 compounds that inhibit tumor invasion into surrounding tissue ( 110 ).…”

Section: Discussionmentioning

confidence: 99%

Receptor tyrosine kinase (RTK) targeting in pediatric high-grade glioma and diffuse midline glioma: Pre-clinical models and precision medicine

et al. 2022

View full text Add to dashboard Cite

Pediatric high-grade glioma (pHGG), including both diffuse midline glioma (DMG) and non-midline tumors, continues to be one of the deadliest oncologic diagnoses (both henceforth referred to as “pHGG”). Targeted therapy options aimed at key oncogenic receptor tyrosine kinase (RTK) drivers using small-molecule RTK inhibitors has been extensively studied, but the absence of proper in vivo modeling that recapitulate pHGG biology has historically been a research challenge. Thankfully, there have been many recent advances in animal modeling, including Cre-inducible transgenic models, as well as intra-uterine electroporation (IUE) models, which closely recapitulate the salient features of human pHGG tumors. Over 20% of pHGG have been found in sequencing studies to have alterations in platelet derived growth factor-alpha (PDGFRA), making growth factor modeling and inhibition via targeted tyrosine kinases a rich vein of interest. With commonly found alterations in other growth factors, including FGFR, EGFR, VEGFR as well as RET, MET, and ALK, it is necessary to model those receptors, as well. Here we review the recent advances in murine modeling and precision targeting of the most important RTKs in their clinical context. We additionally provide a review of current work in the field with several small molecule RTK inhibitors used in pre-clinical or clinical settings for treatment of pHGG.

show abstract

“…Neurobasal medium minus phenol red 500 mL B-27 supplement minus vitamin A (50x) them with a distinct purpose as to not waste lab resources (both materials and time) 4 . Organoid growth has been tested in multiple well sizes and formats, and shows that a 10 cm plate is the ideal setting for maintaining optimal cell viability, followed by a 6-well plate with three organoids per well 34 .…”

Section: Component Quantitymentioning

confidence: 99%

Maintaining Human Glioblastoma Cellular Diversity <em>Ex vivo</em> using Three-Dimensional Organoid Culture

Sundar

Shakya

Recinos

et al. 2022

JoVE

Self Cite

View full text Add to dashboard Cite

Glioblastoma (GBM) is the most commonly occurring primary malignant brain cancer with an extremely poor prognosis. Intra-tumoral cellular and molecular diversity, as well as complex interactions between tumor microenvironments, can make finding effective treatments a challenge. Traditional adherent or sphere culture methods can mask such complexities, whereas three-dimensional organoid culture can recapitulate regional microenvironmental gradients. Organoids are a method of three-dimensional GBM culture that better mimics patient tumor architecture, contains phenotypically diverse cell populations, and can be used for medium-throughput experiments.Although three-dimensional organoid culture is more laborious and time-consuming compared to traditional culture, it offers unique benefits and can serve to bridge the gap between current in vitro and in vivo systems. Organoids have established themselves as invaluable tools in the arsenal of cancer biologists to better understand tumor behavior and mechanisms of resistance, and their applications only continue to grow. Here, details are provided about methods for generating and maintaining GBM organoids. Instructions of how to perform organoid sample embedding and sectioning using both frozen and paraffin-embedding techniques, as well as recommendations for immunohistochemistry and immunofluorescence protocols on organoid sections, and measurement of total organoid cell viability, are all also described.

show abstract

Three-dimensional organoid culture unveils resistance to clinical therapies in adult and pediatric glioblastoma

Abstract: Highlights Glioblastoma organoid cultures preserve diversity of proliferative cell phenotypes. Heterogeneous 3D cultures recapitulate resistance to clinical GBM therapeutics. Patient specimens show different behavior depending on 2D vs 3D growth.

Cited by 43 publications

References 50 publications

WDR5 represents a therapeutically exploitable target for cancer stem cells in glioblastoma

WDR5 represents a therapeutically exploitable target for cancer stem cells in glioblastoma

Receptor tyrosine kinase (RTK) targeting in pediatric high-grade glioma and diffuse midline glioma: Pre-clinical models and precision medicine

Maintaining Human Glioblastoma Cellular Diversity <em>Ex vivo</em> using Three-Dimensional Organoid Culture

Contact Info

Product

Resources

About