Glioblastoma (GBM) displays diffusive invasion throughout the brain microenvironment, which is partially responsible for its short median survival rate (<15 months). Stem-like subpopulations (GBM stem-like cells, GSCs) are believed to play a central role in therapeutic resistance and poor patient prognosis. Given the extensive tissue remodeling and processes such as vessel co-option and regression that occur in the tumor microenvironment, it is essential to understand the role of metabolic constraint such as hypoxia on GBM cell populations. This work describes the use of a multidimensional gelatin hydrogel to culture patient-derived GBM cells, to evaluate the influence of hypoxia and the inclusion brain-mimetic hyaluronic acid on the relative activity of GSCs versus overall GBM cells. Notably, CD133+ GBM cell fraction is crucial for robust formation of tumor spheroids in multidimensional cultures. In addition, while the relative size of the CD133+ GBM subpopulation increased in response to both hypoxia and matrix-bound hyaluronan, we did not observe cell subtype-specific changes in invasion signaling pathway activation. Taken together, this study highlights the potential of biomimetic culture systems for resolving changes in the population dynamics and behavior of subsets of GBM specimens for the future development of precision medicine applications.
Impact Statement
This study describes a gelatin hydrogel platform to investigate the role of extracellular hyaluronic acid and hypoxia on the behavior of a CD133+ subset of cells within patient-derived glioblastoma (GBM) specimens. We report that the relative expansion of the CD133+ GBM stem cell-like population is strongly responsive to extracellular cues, highlighting the significance of biomimetic hydrogel models of the tumor microenvironment to investigate invasion and therapeutic response.
Get full access to this article
View all access options for this article.
References
1.
AnneC., CatherineG., RogatienF., et al.Glioblastoma-associated stromal cells (GASCs) from histologically normal surgical margins have a myofibroblast phenotype and angiogenic properties. J Pathol, 233, 74, 2014.
2.
PaolilloM., BoselliC., and SchinelliS.Glioblastoma under siege: an overview of current therapeutic strategies. Brain Sci, 8, 15, 2018.
3.
JohnsonD.R., and O'NeillB.P.Glioblastoma survival in the United States before and during the temozolomide era. J Neurooncol, 107, 359, 2012.
4.
JacksonC., RuzevickJ., PhallenJ., BelcaidZ., and LimM.Challenges in immunotherapy presented by the glioblastoma multiforme microenvironment. Clin Dev Immunol, 2011, 732413, 2011.
5.
StuppR., MasonW.P., van den BentM.J., et al.Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med, 352, 987, 2005.
6.
MehtaA.I., LinningerA., LesniakM.S., and EngelhardH.H.Current status of intratumoral therapy for glioblastoma. J Neurooncol, 125, 1, 2015.
7.
WiranowskaM.R., and RojianiM.V.Extracellular Matrix Microenvironment in Glioma Progression. Rijeka, Croatia: InTech, 2011.
8.
SykováE. Plasticity of extracellular space. In: WalzW., ed. The Neuronal Environment: Brain Homeostasis in Health and Disease. Totowa, NJ: Humana Press, 2002, p. 57.
9.
Quirico-SantosT., FonsecaC.O., and Lagrota-CandidoJ.Brain sweet brain: importance of sugars for the cerebral microenvironment and tumor development. Arq Neuro Psiquiatr, 68, 799, 2010.
10.
ChenJ.-W.E., PedronS., and HarleyB.A.C.The combined influence of hydrogel stiffness and matrix-bound hyaluronic acid content on glioblastoma invasion. Macromol Biosci, 17, 1700018, 2017.
11.
ChenJ.-W.E., PedronS., ShyuP., HuY., SarkariaJ.N., and HarleyB.A.C.Influence of hyaluronic acid transitions in tumor microenvironment on glioblastoma malignancy and invasive behavior. Front Mater, 5, 39, 2018.
12.
MunsonJ.M., BellamkondaR.V., and SwartzM.A.Interstitial flow in a 3D microenvironment increases glioma invasion by a CXCR4-dependent mechanism. Cancer Res, 73, 1536, 2013.
13.
AtayN., YuanJ., CornelisonC., and MunsonJ.Tami-15. The effect of interstitial fluid flow and astrocytes/microglia on invasion, proliferation and stemness of patient-derived glioma stem cells. Neurooncology, 22, ii216, 2020.
14.
HoI.A.W., and ShimW.S.N.Contribution of the microenvironmental niche to glioblastoma heterogeneity. BioMed Res Int, 2017, 13, 2017.
15.
MichielsC.Physiological and pathological responses to hypoxia. Am J Pathol, 164, 1875, 2004.
16.
VartanianA., SinghS.K., AgnihotriS., et al.GBM's multifaceted landscape: highlighting regional and microenvironmental heterogeneity. Neuro Oncol, 16, 1167, 2014.
BindaE., VisioliA., ReynoldsB., and VescoviA.L.Heterogeneity of cancer-initiating cells within glioblastoma. Front Biosci (Schol Ed), 4, 1235, 2012.
19.
ReynoldsB.A., and VescoviA.L.Brain cancer stem cells: think twice before going flat. Cell Stem Cell, 5, 466, 2009.
20.
LathiaJ.D., MackS.C., Mulkearns-HubertE.E., ValentimC.L., and RichJ.N.Cancer stem cells in glioblastoma. Genes Dev, 29, 1203, 2015.
21.
UemaeY., IshikawaE., OsukaS., et al.CXCL12 secreted from glioma stem cells regulates their proliferation. J Neurooncol, 117, 43, 2014.
22.
SunL., HuiA.M., SuQ., et al.Neuronal and glioma-derived stem cell factor induces angiogenesis within the brain. Cancer Cell, 9, 287, 2006.
23.
BaysanM., WoolardK., BozdagS., et al.Micro-environment causes reversible changes in DNA methylation and mRNA expression profiles in patient-derived glioma stem cells. PLoS One, 9, e94045, 2014.
24.
BindaE., ReynoldsB.A., and VescoviA.L.Glioma stem cells: turpis omen in nomen? (The evil in the name?). J Intern Med, 276, 25, 2014.
25.
LemkeD., WeilerM., BlaesJ., et al.Primary glioblastoma cultures: can profiling of stem cell markers predict radiotherapy sensitivity? J Neurochem, 131, 251, 2014.
26.
HambardzumyanD., ChengY.-K., HaenoH., HollandE.C., and MichorF.The probable cell of origin of NF1- and PDGF-driven glioblastomas. PLoS One, 6, e24454, 2011.
27.
PranatharthiA., RossC., and SrivastavaS.Cancer stem cells and radioresistance: Rho/ROCK pathway plea attention. Stem Cells Int, 2016, 5785786, 2016.
28.
KalkanR.Hypoxia is the driving force behind GBM and could be a new tool in GBM treatment. Crit Rev Eukaryot Gene Expr, 25, 363, 2015.
PersanoL., RampazzoE., BassoG., and ViolaG.Glioblastoma cancer stem cells: role of the microenvironment and therapeutic targeting. Biochem Pharmacol, 85, 612, 2013.
31.
SakariassenP.Ø., ImmervollH., and ChekenyaM.Cancer stem cells as mediators of treatment resistance in brain tumors: status and controversies. Neoplasia, 9, 882, 2007.
ClarkeM.F., DickJ.E., DirksP.B., et al.Cancer stem cells—perspectives on current status and future directions: AACR workshop on cancer stem cells. Cancer Res, 66, 9339, 2006.
34.
MeiX., ChenY.-S., ChenF.-R., XiS.-Y., and ChenZ.-P.Glioblastoma stem cell differentiation into endothelial cells evidenced through live-cell imaging. Neurooncology, 19, 1109, 2017.
35.
FilatovaA., AckerT., and GarvalovB.K.The cancer stem cell niche(s): the crosstalk between glioma stem cells and their microenvironment. Biochim Biophys Acta, 1830, 2496, 2013.
36.
GuelfiS., DuffauH., BauchetL., RothhutB., and HugnotJ.P.Vascular transdifferentiation in the CNS: a focus on neural and glioblastoma stem-like cells. Stem Cells Int, 2016, 2759403, 2016.
37.
DirksP.B.Cancer: stem cells and brain tumours. Nature, 444, 687, 2006.
38.
GilbertsonR.J., and RichJ.N.Making a tumour's bed: glioblastoma stem cells and the vascular niche. Nat Rev Cancer, 7, 733, 2007.
GoffartN., KroonenJ., and RogisterB.Glioblastoma-initiating cells: relationship with neural stem cells and the micro-environment. Cancers (Basel), 5, 1049, 2013.
41.
BayinN.S., ModrekA.S., and PlacantonakisD.G.Glioblastoma stem cells: molecular characteristics and therapeutic implications. World J Stem Cells, 6, 230, 2014.
42.
JooK.M., KimS.Y., JinX., et al.Clinical and biological implications of CD133-positive and CD133-negative cells in glioblastomas. Lab Invest, 88, 808, 2008.
43.
RappaG., FodstadO., and LoricoA.The stem cell-associated antigen CD133 (Prominin-1) is a molecular therapeutic target for metastatic melanoma. Stem Cells, 26, 3008, 2008.
44.
YaoJ., ZhangT., RenJ., YuM., and WuG.Effect of CD133/prominin-1 antisense oligodeoxynucleotide on in vitro growth characteristics of Huh-7 human hepatocarcinoma cells and U251 human glioma cells. Oncol Rep, 22, 781, 2009.
45.
GuvencH., PavlyukovM.S., JoshiK., et al.Impairment of glioma stem cell survival and growth by a novel inhibitor for survivin–ran protein complex. Clin Cancer Res, 19, 631, 2013.
46.
ChenJ.-W.E., LumibaoJ., BlazekA., GaskinsH.R., and HarleyB.Hypoxia activates enhanced invasive potential and endogenous hyaluronic acid production by glioblastoma cells. Biomater Sci, 6, 854, 2018.
47.
MinetE., ArnouldT., MichelG., et al.ERK activation upon hypoxia: involvement in HIF-1 activation. FEBS Lett, 468, 53, 2000.
48.
GuoG., YaoW., ZhangQ., and BoY.Oleanolic acid suppresses migration and invasion of malignant glioma cells by inactivating MAPK/ERK signaling pathway. PLoS One, 8, e72079, 2013.
49.
KimJ.-Y., KimY.-J., LeeS., and ParkJ.-H.The critical role of ERK in death resistance and invasiveness of hypoxia-selected glioblastoma cells. BMC Cancer, 9, 27, 2009.
50.
DuR., PetritschC., LuK., et al.Matrix metalloproteinase-2 regulates vascular patterning and growth affecting tumor cell survival and invasion in GBM. Neurooncology, 10, 254, 2008.
51.
PedronS., WolterG.L., ChenJ.-W.E., LakenS., SarkariaJ.N., and HarleyB.A.C.Hyaluronic acid-functionalized gelatin hydrogels reveal extracellular matrix signals temper the efficacy of erlotinib against patient-derived glioblastoma specimens. Biomaterials, 219, 119371, 2019.
52.
PedronS., BeckaE., and HarleyB.A.C.Regulation of glioma cell phenotype in 3D matrices by hyaluronic acid. Biomaterials, 34, 7408, 2013.
53.
ChenJ.-W.E., LumibaoJ., LearyS., et al.Crosstalk between microglia and patient-derived glioblastoma cells inhibit invasion in a three-dimensional gelatin hydrogel model. J Neuroinflamm, 17, 346, 2020.
54.
CarlsonB.L., PokornyJ.L., SchroederM.A., and SarkariaJ.N.Establishment, maintenance and in vitro and in vivo applications of primary human glioblastoma multiforme (GBM) xenograft models for translational biology studies and drug discovery. Curr Protoc Pharmacol Chapter 14, Unit 16, 2011.
55.
PenceJ.C., ClancyK.B.H., and HarleyB.A.C.The induction of pro-angiogenic processes within a collagen scaffold via exogenous estradiol and endometrial epithelial cells. Biotechnol Bioeng, 112, 2185, 2015.
56.
CaliariS.R., and HarleyB.A.C.The effect of anisotropic collagen-GAG scaffolds and growth factor supplementation on tendon cell recruitment, alignment, and metabolic activity. Biomaterials, 32, 5330, 2011.
57.
PedronS., BeckaE., and HarleyB.A.Spatially gradated hydrogel platform as a 3D engineered tumor microenvironment. Adv Mater, 27, 1567, 2015.
58.
CaliariS.R., WeisgerberD.W., GrierW.K., MahmassaniZ., BoppartM.D., and HarleyB.A.C.Collagen scaffolds incorporating coincident gradations of instructive structural and biochemical cues for osteotendinous junction engineering. Adv Healthcare Mater, 4, 831, 2015.
59.
ZambutoS.G., ClancyK.B.H., and HarleyB.A.C.Tuning trophoblast motility in a gelatin hydrogel via soluble cues from the maternal-fetal interface. Tissue Eng Part A, 27, 1064, 2020.
60.
ZambutoS., ClancyK.B.H., and HarleyB.A.C.A gelatin hydrogel to study endometrial angiogenesis and trophoblast invasion. Interface Focus, 9, 20190016, 2019.
61.
KimY., ParkY.W., LeeY.S., and JeoungD.Hyaluronic acid induces transglutaminase II to enhance cell motility; role of Rac1 and FAK in the induction of transglutaminase II. Biotechnol Lett, 30, 31, 2008.
62.
LeeS.H., LeeY.J., SongC.H., AhnY.K., and HanH.J.Role of FAK phosphorylation in hypoxia-induced hMSCS migration: involvement of VEGF as well as MAPKS and eNOS pathways. Am J Physiol Cell Physiol, 298, C847, 2010.
63.
MonaghanR.M., and WhitmarshA.J.Mitochondrial proteins moonlighting in the nucleus. Trends Biochem Sci, 40, 728, 2015.
64.
ArasS., BaiM., LeeI., SpringettR., HuttemannM., and GrossmanL.I.MNRR1 (formerly CHCHD2) is a bi-organellar regulator of mitochondrial metabolism. Mitochondrion, 20, 43, 2015.
65.
ArasS., PakO., SommerN., et al.Oxygen-dependent expression of cytochrome c oxidase subunit 4–2 gene expression is mediated by transcription factors RBPJ, CXXC5 and CHCHD2. Nucleic Acids Res, 41, 2255, 2013.
66.
GrossmanL.I., PurandareN., ArshadR., et al.MNRR1, a biorganellar regulator of mitochondria. Oxid Med Cell Longev, 2017, 6739236, 2017.
67.
CenL., CarlsonB.L., SchroederM.A., et al.p16-Cdk4-Rb axis controls sensitivity to a cyclin-dependent kinase inhibitor PD0332991 in glioblastoma xenograft cells. Neuro Oncol, 14, 870, 2012.
68.
LoebelC., KwonM.Y., WangC., HanL., MauckR.L., and BurdickJ.A.Metabolic labeling to probe the spatiotemporal accumulation of matrix at the chondrocyte–hydrogel interface. Adv Funct Mater, 30, 1909802, 2020.
69.
IzziV., DavisM.N., and NabaA.Pan-cancer analysis of the genomic alterations and mutations of the matrisome. Cancers (Basel), 12, 2046, 2020.
HynesR.O., and NabaA.Overview of the matrisome—an inventory of extracellular matrix constituents and functions. Cold Spring Harb Perspect Biol, 4, a004903, 2012.
72.
BaiL., YuZ., QianG., et al.SOCS3 was induced by hypoxia and suppressed STAT3 phosphorylation in pulmonary arterial smooth muscle cells. Respir Physiol Neurobiol, 152, 83, 2006.
73.
SeoM., LeeW.-H., and SukK.Identification of novel cell migration-promoting genes by a functional genetic screen. FASEB J, 24, 464, 2010.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
