There are currently no reliable in vitro models for brain tumors that predict drug response in humans. Clinical and research evidence indicates that a small population of cancer stem cells (CSCs) in tumors are primarily responsible for tumor initiation, progression, recurrence, and resistance to therapeutics. These cells have self- renewal capacity and unlimited proliferative potential. Although the targeting of CSCs represents a potential treatment approach, it is very challenging to isolate CSCs from human cell lines or primary cancer specimens since they represent such a small proportion of the entire tumor cell population. Existing methods for isolation and expansion of CSCs are ineffective, cumbersome, expensive, and unreliable. Here we aim to develop clinically relevant and predicative in vitro human tumor models for rapid and low-cost drug assessment for anti- CSC therapy. The proposed research uses an advanced 3D system made of complex scaffold of chitosan and alginate (CA), two naturally occurring polymers that bear proxy structure of glycosaminoglycans, a major component of native extracellular matrix (ECM). These CA scaffolds will serve as a niche to selectively renew and rapidly enrich CSCs. Once the preliminary system is established with optimal structural and mechanical properties that demonstrate CSC renewal, we plan to further improve and fine-tune our tumor model by introducing environmental factors such as biochemical coatings, hypoxia, and human stromal signaling factors into CA scaffolds for human glioblastoma culture. The established optimal CA scaffold model is expected to support the formation of CSC-enriched tumor spheroids from cell lines, primary GBM cells, and freshly resected GBM tissue. A small number of cells from the spheroids implanted in nude mice are expected to form orthotopic tumors and recapitulate GBM phenotypes. GBM is selected as the target tumor for the proposed study due to its fatality and dismal prognosis and our extensive clinical and research experience of GBM treatments.
Specific aims of the proposed research are to: 1) determine the role of microstructure and mechanical properties of CA scaffolds on enrichment of CSCs and establish CSC-enriched tumor spheroid models;2) investigate the role of microenvironmental cues of CA scaffolds in CSC enrichment, and tumorigenic capacities of CSC-enriched tumor spheroids;and 3) use the tumor models in drug tests for personalized cancer treatment and design of therapeutic strategies that specifically target CSCs for GBM therapy. This research will provide a new platform for effectively evaluating potential therapeutic drugs by providing a more accurate and stable tumor microenvironment, thus considerably shortening the time and reducing the cost of drug development. The developed tumor models will also allow researchers and medical practitioners to study molecular mechanisms that regulate self-renewal and differentiation of CSCs, and provide insight into the origin of tumor formation and resistance to treatments.

Public Health Relevance

There are currently no reliable in vitro models that predict drug response of human tumors. We propose to develop an in vitro model that can be used both to expand and enrich cancer stem cells for downstream analysis and to better predict how drug candidates will act in patients. The success of the research will open a new avenue to rapidly evaluate potential therapeutic drugs at low cost and provide a technological platform to study cancer stem cell biology and resistance to treatments.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA172455-01A1
Application #
8577052
Study Section
Drug Discovery and Molecular Pharmacology Study Section (DMP)
Program Officer
Knowlton, John R
Project Start
2013-09-23
Project End
2017-07-31
Budget Start
2013-09-23
Budget End
2014-07-31
Support Year
1
Fiscal Year
2013
Total Cost
$266,843
Indirect Cost
$84,253
Name
University of Washington
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Kievit, Forrest M; Wang, Kui; Erickson, Ariane E et al. (2016) Modeling the tumor microenvironment using chitosan-alginate scaffolds to control the stem-like state of glioblastoma cells. Biomater Sci 4:610-3
Wang, Kui; Kievit, Forrest M; Erickson, Ariane E et al. (2016) Culture on 3D Chitosan-Hyaluronic Acid Scaffolds Enhances Stem Cell Marker Expression and Drug Resistance in Human Glioblastoma Cancer Stem Cells. Adv Healthc Mater :
Chang, Fei-Chien; Tsao, Ching-Ting; Lin, Anqi et al. (2016) PEG-chitosan hydrogel with tunable stiffness for study of drug response of breast cancer cells. Polymers (Basel) 8:
Jana, Soumen; Levengood, Sheeny K Lan; Zhang, Miqin (2016) Anisotropic Materials for Skeletal-Muscle-Tissue Engineering. Adv Mater :
Wang, Kui; Kievit, Forrest M; Florczyk, Stephen J et al. (2015) 3D Porous Chitosan-Alginate Scaffolds as an In Vitro Model for Evaluating Nanoparticle-Mediated Tumor Targeting and Gene Delivery to Prostate Cancer. Biomacromolecules 16:3362-72
Erickson, Ariane E; Edmondson, Dennis; Chang, Fei-Chien et al. (2015) High-throughput and high-yield fabrication of uniaxially-aligned chitosan-based nanofibers by centrifugal electrospinning. Carbohydr Polym 134:467-74
Tsao, Ching Ting; Hsiao, Meng Hsuan; Zhang, Mengying Y et al. (2015) Chitosan-PEG hydrogel with sol-gel transition triggerable by multiple external stimuli. Macromol Rapid Commun 36:332-8
Wang, Kui; Kievit, Forrest M; Jeon, Mike et al. (2015) Nanoparticle-Mediated Target Delivery of TRAIL as Gene Therapy for Glioblastoma. Adv Healthc Mater 4:2719-26
Tsao, Ching-Ting; Leung, Matthew; Chang, Julia Yu-Fong et al. (2014) A simple material model to generate epidermal and dermal layers in vitro for skin regeneration. J Mater Chem B Mater Biol Med 2:5256-5264
Tsao, Ching-Ting; Kievit, Forrest M; Ravanpay, Ali et al. (2014) Thermoreversible poly(ethylene glycol)-g-chitosan hydrogel as a therapeutic T lymphocyte depot for localized glioblastoma immunotherapy. Biomacromolecules 15:2656-62

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