Chemotherapeutic resistance in 3D biomaterials
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1
University of Massachusetts - Amherst, Chemical Engineering, United States
Introduction: In early stages of drug-development pipelines, high-throughput technologies are being used to test potential pharmaceutical candidates. Drug screening tests are performed in tissue culture polystyrene (TCPS) surfaces that do not account the cell’s native microenvironment. It has been reported that cells exhibit distinctive morphology, motility, and commitment to live or die based on the chemical stimuli received from the place they are seeded[1]. Recently, our lab created a high-throughput 2D biomaterial testing platform that shows drug resistance is regulated by integrin binding and substrate stiffness[2]. Due to TCPS constraints and 2D platforms findings, here we designed a biomaterial platform in a 3D high-throughput manner simulating matrix cues (stiffness and cell-binding peptides) relevant to what cancer cells experiences in vivo. Moreover, a drug-response curve was obtained to measure how breast cancer cells encapsulated in single-cells and spheroids responded within different stiffness after doxorubicin treatment, a powerful chemotherapeutic.
Materials and Methods: Breast cancer MDA-MB-231 cells were cultured in DMEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. The formation of MDA-MB-231 cell spheroids was achieved by encapsulating cells in poly(N-isopropylacrylamide) hydrogels (P-NiPAAM) for 14 days (Fig. 1A). MDA-MB-231 single-cells and spheroids suspensions were mixed in serum free medium with 20K poly(ethylene glycol)-4-arm-Maleimide (PEG-Mal) previously conjugated with RGD, peptide sequence used to control adhesivity of cells (Fig. 1B). The polymerization of PEG-Mal was induced following the addition of 1K PEG-dithiol dissolved in triethanolamine. Since, PEG-Mal cannot be degraded by enzymes secreted from cells, its weight percentage (wt.%) was varied among platforms to control the stiffness of the materials (Fig. 1C). Drug-response of single-cells and spheroids was measured 48h after doxorubicin treatment using CellTiter-Glo ATP assay (Fig. 1D-E).
Results and Discussion: Doxorubicin is a potent chemotherapeutic agent that interchalates among DNA of dividing cells. The effect of this chemotherapeutic was evaluated upon MDA-MB-231 single-cells and spheroids within 3D hydrogels at different stiffness. Results suggested that either single-cells or spheroids does not exhibit a material-acquired resistance after drug treatment. It was expected at least that single-cells because of their higher cell-superficial contact with the materials would resist the treatment in stiff environments in comparisson to spheroids, but this hypothesis was rejected. This 3D platform suggested that MDA-MB-231 cells may resist doxorubic treatment by other means, such as integrin binding and other intercellular effectors. On the other hand, PEG-Mal 3D hydrogels proved to be adaptable to a high-throughput format with a facile control of the biomechanical and biochemical properties over the range of in vivo cancer tissue.
Conclusion: Even though MDA-MB-231 cells are an useful model for the characterization of aggressive breast cancer subtypes, we will increase the panel of cell lines to discern whether the extent of these findings are generalizable.
References:
[1] Levental, I., et al. Soft Matter 2007;3:299-306.
[2] Nguyen, T. V., et al. Biomaterials 2014;35:5749- 5759.
Keywords:
biomaterial,
RGD peptide,
Cell response,
Drug testing
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
Poster
Topic:
Biomaterials for cancer therapy
Citation:
Negron-Pineiro
LJ,
Brooks
EA,
Nguyen
TV,
Jansen
LE,
Barney
LE and
Peyton
SR
(2016). Chemotherapeutic resistance in 3D biomaterials.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.01131
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.
*
Correspondence:
Dr. Lenny J Negron-Pineiro, University of Massachusetts - Amherst, Chemical Engineering, Amherst, MA, United States, lenny.negron@upr.edu
Dr. Elizabeth A Brooks, University of Massachusetts - Amherst, Chemical Engineering, Amherst, MA, United States, eabrooks@engin.umass.edu
Dr. Thuy V Nguyen, University of Massachusetts - Amherst, Chemical Engineering, Amherst, MA, United States, thuyn@umass.edu
Dr. Lauren E Jansen, University of Massachusetts - Amherst, Chemical Engineering, Amherst, MA, United States, ljansen@ecs.umass.edu
Dr. Lauren E Barney, University of Massachusetts - Amherst, Chemical Engineering, Amherst, MA, United States, lbarney@umass.edu
Dr. Shelly R Peyton, University of Massachusetts - Amherst, Chemical Engineering, Amherst, MA, United States, speyton@engin.umass.edu