Metabolic response of lung cancer cells to radiation in a paper-based 3D cell culture system.
This work demonstrates the application of a 3D culture system-Cells-in-Gels-in-Paper (CiGiP)-in evaluating the metabolic response of lung cancer cells to ionizing radiation. The 3D tissue-like construct-prepared by stacking multiple sheets of paper containing cell-embedded hydrogels-generates a gradient of oxygen and nutrients that decreases monotonically in the stack. Separating the layers of the stack after exposure enabled analysis of the cellular response to radiation as a function of oxygen and nutrient availability; this availability is dictated by the distance between the cells and the source of oxygenated medium. As the distance between the cells and source of oxygenated media increased, cells show increased levels of hypoxia-inducible factor 1-alpha, decreased proliferation, and reduced sensitivity to ionizing radiation. Each of these cellular responses are characteristic of cancer cells observed in solid tumors. With this setup we were able to differentiate three isogenic variants of A549 cells based on their metabolic radiosensitivity; these three variants have known differences in their metastatic behavior in vivo. This system can, therefore, capture some aspects of radiosensitivity of populations of cancer cells related to mass-transport phenomenon, carry out systematic studies of radiation response in vitro that decouple effects from migration and proliferation of cells, and regulate the exposure of oxygen to subpopulations of cells in a tissue-like construct either before or after irradiation.
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA.
Karen A
KA
Simon
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, 60 Oxford Street, Cambridge, MA 02138, USA; Dalio Institute of Cardiovascular Imaging, Department of Radiology, Weill Cornell Medicine, 413 E. 69th Street Suite BRB-108, New York, NY 10021, USA.
Bobak
B
Mosadegh
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA.
Kyaw Thu
KT
Minn
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA; Department of Chemistry, University of North Carolina at Chapel Hill, 125 South Road, Chapel Hill, NC 27599, USA.
Matthew R
MR
Lockett
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA.
Marym R
MR
Mohammady
Vertex Pharmaceuticals Incorporated, 50 Northern Blvd., Boston, MA 02210, USA.
Diane M
DM
Boucher
Vertex Pharmaceuticals Incorporated, 50 Northern Blvd., Boston, MA 02210, USA.
Amy B
AB
Hall
Vertex Pharmaceuticals Incorporated, 50 Northern Blvd., Boston, MA 02210, USA.
Shawn M
SM
Hillier
Vertex Pharmaceuticals Incorporated, 50 Northern Blvd., Boston, MA 02210, USA.
Taturo
T
Udagawa
Vertex Pharmaceuticals Incorporated, 50 Northern Blvd., Boston, MA 02210, USA. Electronic address: brenda_eustace@vrtx.com.
Brenda K
BK
Eustace
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, 60 Oxford Street, Cambridge, MA 02138, USA. Electronic address: gwhitesides@gmwgroup.harvard.edu.
George M
GM
Whitesides
D000072283:A549 Cells; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000528:radiation effects; D002470:Cell Survival / Q000528:radiation effects; D006801:Humans; D020100:Hydrogels; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D008175:Lung Neoplasms / Q000378:metabolism / Q000532:radiotherapy*; D010100:Oxygen / Q000378:metabolism; D010209:Paper; D011836:Radiation Tolerance; D000072258:Tumor Hypoxia / Q000528:radiation effects
D020100:Hydrogels; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D010100:Oxygen
3D cell culture; Oxygen gradients; Radiation response; Radioresistance; Tumor hypoxia