Bioengineered Scaffolds for 3D Analysis of Glioblastoma Proliferation and Invasion

Heffernan, John M.; Overstreet, Derek J.; Le, Duc Long; Vernon, Brent L.; Sirianni, Rachael W.

doi:10.1007/s10439-014-1223-1

Cited by 64 publications

(92 citation statements)

References 38 publications

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“…As a result, three-dimensional scaffolds have been routinely used to measure various interactions between GBM cell lines or GSCs and their local microenvironment by us [33,43] and others [24,29,30,44–52]. Although immortalized cell lines are proven tools in GBM research, they are poor biological models of the clinical condition and do not maintain functional GSCs [4,5,15,28].…”

Section: Discussionmentioning

confidence: 99%

PNIPAAm-co-Jeffamine® (PNJ) scaffolds as in vitro models for niche enrichment of glioblastoma stem-like cells

et al. 2017

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Glioblastoma (GBM) is the most common adult primary brain tumor, and the 5-year survival rate is less than 5%. GBM malignancy is driven in part by a population of GBM stem-like cells (GSCs) that exhibit indefinite self-renewal capacity, multipotent differentiation, expression of neural stem cell markers, and resistance to conventional treatments. GSCs are enriched in specialized niche microenvironments that regulate stem phenotypes and support GSC radioresistance. Therefore, identifying GSC-niche interactions that regulate stem phenotypes may present a unique target for disrupting the maintenance and persistence of this treatment resistant population. In this work, we engineered 3D scaffolds from temperature responsive poly(N-isopropylacrylamide-co-Jeffamine M-1000® acrylamide), or PNJ copolymers, as a platform for enriching stem-specific phenotypes in two molecularly distinct human patient-derived GSC cell lines. Notably, we observed that, compared to conventional neurosphere cultures, PNJ cultured GSCs maintained multipotency and exhibited enhanced self-renewal capacity. Concurrent increases in expression of proteins known to regulate self-renewal, invasion, and stem maintenance in GSCs (NESTIN, EGFR, CD44) suggest that PNJ scaffolds effectively enrich the GSC population. We further observed that PNJ cultured GSCs exhibited increased resistance to radiation treatment compared to GSCs cultured in standard neurosphere conditions. GSC radioresistance is supported in vivo by niche microenvironments, and this remains a significant barrier to effectively treating these highly tumorigenic cells. Taken in sum, these data indicate that the microenvironment created by synthetic PNJ scaffolds models niche enrichment of GSCs in patient-derived GBM cell lines, and presents tissue engineering opportunities for studying clinically important behaviors such as radioresistance in vitro.

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Section: Discussionmentioning

confidence: 99%

PNIPAAm-co-Jeffamine® (PNJ) scaffolds as in vitro models for niche enrichment of glioblastoma stem-like cells

et al. 2017

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“…9 In a HA and gelatin composite hydrogel system, U87 cells proliferated less than 3 fold after 14 days. 27 Our hydrogel platform may be more permissive for cell proliferation for a number of reasons, including differences in the hydrogel stiffness and mechanisms of MMP-mediated degradation of hydrogel crosslinks. First, in this study, the initial stiffness of hydrogels with varying MMP-degradability ranged from 1.2 to 2.0 kPa, which is within the range reported for normal human brain tissue.…”

Section: Discussionmentioning

confidence: 99%

“…24,25 Previous hydrogel systems had stiffnesses ranging from 2.0 kPa to over 50 kPa, which is significantly stiffer than brain tissue. 9,27 Moreover, in our hydrogel platform, a MMP-degradable peptide was used for hydrogel crosslinking and for cell-mediated matrix degradation. The peptide selected has been shown to be degraded by GBM-relevant MMPs, MMP1, MMP2, and MMP9.…”

Section: Discussionmentioning

confidence: 99%

Effect of matrix metalloproteinase‐mediated matrix degradation on glioblastoma cell behavior in 3D PEG‐based hydrogels

Wang

Tong

Jiang

et al. 2016

J Biomedical Materials Res

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Glioblastoma (GBM) is the most common and aggressive form of primary brain tumor with median survival of 12 months. To improve clinical outcomes, it is critical to develop in vitro models that support GBM proliferation and invasion for deciphering tumor progression and screening drug candidates. A key hallmark of GBM cells is their extreme invasiveness, a process mediated by matrix metalloproteinase (MMP)-mediated degradation of the extracellular matrix. We recently reported the development of a MMP-degradable, poly(ethylene-glycol)-based hydrogel platform for culturing GBM cells. In the present study, we modulated the percentage of MMP-degradable crosslinks in 3D hydrogels to analyze the effects of MMP-degradability on GBM fates. Using an immortalized GBM cell line (U87) as a model cell type, our results showed that MMP-degradability was not required for supporting GBM proliferation. All hydrogel formulations supported robust GBM proliferation, up to 10 fold after 14 days. However, MMP-degradability was essential for facilitating tumor spreading, and 50% MMP-degradable hydrogels were sufficient to enable both robust tumor cell proliferation and spreading in 3D. The findings of this study highlight the importance of modulating MMP-degradability in engineering 3D in vitro brain cancer models and may be applied for engineering in vitro models for other cancer types.

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“…To date, 3D in vitro models for GBM primarily incorporate only tumor cells, despite the demonstrated contributions of additional cell types to in vivo GBM behavior [6,[24][25][26][27][28][29][30]. A few studies have examined the interactions between GBM cells, perivascular niche cells, and macrophages within 3D in vitro platforms, but these models feature spatial segregation between the different cell types that is not reminiscent of the in vivo microenvironment [20,[31][32][33][34]. Mixed cultures of tumor cells, endothelial cells, and stromal cells have been achieved within biomaterials for breast, lung, and prostate cancers, and these models have been used to investigate how non-tumor cells impact tumor proliferation, phenotype, and therapeutic response [35][36][37][38][39].…”

Section: Introductionmentioning

confidence: 99%

Perivascular signals alter global gene expression profile of glioblastoma and response to temozolomide in a gelatin hydrogel

Harley

Ngo

2018

Preprint

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Glioblastoma (GBM) is the most common primary malignant brain tumor, with patients exhibiting poor survival (median survival time: 15 months). Difficulties in treating GBM include not only the inability to resect the diffusively-invading tumor cells but also therapeutic resistance. The perivascular niche within the GBM tumor microenvironment contributes significantly to tumor cell invasion, cancer stem cell maintenance, and has been shown to protect tumor cells from radiation and chemotherapy. In this study, we describe the development of an in vitro artificial perivascular niche (PVN), culturing endothelial and stromal cells along with GBM cells within a methacrylamide-functionalized gelatin hydrogel, in order to examine howthe presence of the PVN alters global genomic expression profiles. Using RNA-seq, we demonstrate that the inclusion of perivascular niche cells with GBM cells upregulates genes related to angiogenesis and remodeling, while downregulated genes are related to cell cycle and DNA damage repair.Signaling pathways and genes commonly implicated in GBM malignancy, such as MGMT, EGFR, PI3K-Akt signaling, and Ras/MAPK signaling are also upregulated in the presence of perivascular niche cells. We describe the kinetics of gene expression within the PVN hydrogels over a course of 14 days, observing the patterns associated with previously-observed GBMmediated endothelial network co-option and regression that are altered in the presence of covalently-bound hyaluronic acid. We finally examine the effect of PVN culture on GBM cell responsiveness to temozolomide, a frontline chemotherapy used clinically against GBM.Notably, the presence of a PVN leads to significantly increased TMZ resistance compared to hydrogels containing only GBM cells. Overall, these results demonstrate that inclusion of cellular and matrix-associated elements of the PVN within in vitro models of GBM provide critical signals to regulate the phenotype and therapeutic responsiveness of GBM cells.

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Bioengineered Scaffolds for 3D Analysis of Glioblastoma Proliferation and Invasion

Cited by 64 publications

References 38 publications

PNIPAAm-co-Jeffamine® (PNJ) scaffolds as in vitro models for niche enrichment of glioblastoma stem-like cells

PNIPAAm-co-Jeffamine® (PNJ) scaffolds as in vitro models for niche enrichment of glioblastoma stem-like cells

Effect of matrix metalloproteinase‐mediated matrix degradation on glioblastoma cell behavior in 3D PEG‐based hydrogels

Perivascular signals alter global gene expression profile of glioblastoma and response to temozolomide in a gelatin hydrogel

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