Organotypic microfluidic breast cancer model reveals starvation-induced spatial-temporal metabolic adaptations

Jose M Ayuso; Amani Gillette; Karina Lugo-Cintrón; Suehelay Acevedo-Acevedo; Ismael Gomez; Molly Morgan; Tiffany Heaster; Kari B Wisinski; Sean P Palecek; Melissa C Skala; David J Beebe

doi:10.1016/j.ebiom.2018.10.046

Organotypic microfluidic breast cancer model reveals starvation-induced spatial-temporal metabolic adaptations

EBioMedicine. 2018 Nov:37:144-157. doi: 10.1016/j.ebiom.2018.10.046. Epub 2018 Oct 26.

Authors

Affiliations

¹ Morgridge Institute for Research, 330 N Orchard street, Madison, WI, USA; Department of Biomedical Engineering, University of Wisconsin, Madison, WI, USA; The University of Wisconsin Carbone Cancer Center, University of Wisconsin, Madison, WI, USA. Electronic address: ayusodomingu@wisc.edu.
² Department of Biomedical Engineering, University of Wisconsin, Madison, WI, USA.
³ Department of Chemical and Biological Engineering, University of Wisconsin, Madison, USA.
⁴ Allergy research group, IdISSC. San Carlos Clinic Hospital, Madrid, Spain; Materials department, Carlos III University. Leganes, Spain.
⁵ The University of Wisconsin Carbone Cancer Center, University of Wisconsin, Madison, WI, USA.
⁶ The University of Wisconsin Carbone Cancer Center, University of Wisconsin, Madison, WI, USA; Department of Chemical and Biological Engineering, University of Wisconsin, Madison, USA.
⁷ Morgridge Institute for Research, 330 N Orchard street, Madison, WI, USA; Department of Biomedical Engineering, University of Wisconsin, Madison, WI, USA; The University of Wisconsin Carbone Cancer Center, University of Wisconsin, Madison, WI, USA.
⁸ Department of Biomedical Engineering, University of Wisconsin, Madison, WI, USA; The University of Wisconsin Carbone Cancer Center, University of Wisconsin, Madison, WI, USA; Department of Pathology & Laboratory Medicine, University of Wisconsin, MAdison, WI,USA. Electronic address: djbeebe@wisc.edu.

Abstract

Background: Ductal carcinoma in situ (DCIS) is the earliest stage of breast cancer. During DCIS, tumor cells remain inside the mammary duct, growing under a microenvironment characterized by hypoxia, nutrient starvation, and waste product accumulation; this harsh microenvironment promotes genomic instability and eventually cell invasion. However, there is a lack of biomarkers to predict what patients will transition to a more invasive tumor or how DCIS cells manage to survive in this harsh microenvironment.

Methods: In this work, we have developed a microfluidic model that recapitulates the DCIS microenvironment. In the microdevice, a DCIS model cell line was grown inside a luminal mammary duct model, embedded in a 3D hydrogel with mammary fibroblasts. Cell behavior was monitored by confocal microscopy and optical metabolic imaging. Additionally, metabolite profile was studied by NMR whereas gene expression was analyzed by RT-qPCR.

Findings: DCIS cell metabolism led to hypoxia and nutrient starvation; revealing an altered metabolism focused on glycolysis and other hypoxia-associated pathways. In response to this starvation and hypoxia, DCIS cells modified the expression of multiple genes, and a gradient of different metabolic phenotypes was observed across the mammary duct model. These genetic changes observed in the model were in good agreement with patient genomic profiles; identifying multiple compounds targeting the affected pathways. In this context, the hypoxia-activated prodrug tirapazamine selectively destroyed hypoxic DCIS cells.

Interpretation: The results showed the capacity of the microfluidic model to mimic the DCIS structure, identifying multiple cellular adaptations to endure the hypoxia and nutrient starvation generated within the mammary duct. These findings may suggest new potential therapeutic directions to treat DCIS. In summary, given the lack of in vitro models to study DCIS, this microfluidic device holds great potential to find new DCIS predictors and therapies and translate them to the clinic.

Keywords: 3D model; Breast cancer; DCIS; Microfluidics; Organotypic.

MeSH terms

Breast Neoplasms* / metabolism
Breast Neoplasms* / pathology
Cell Line, Tumor
Female
Gene Expression Regulation, Neoplastic*
Genomic Instability*
Humans
Hydrogels / chemistry
Microfluidic Analytical Techniques
Models, Biological*
Neoplasm Invasiveness
Tumor Microenvironment*

Substances

Hydrogels

Abstract

MeSH terms

Substances

Grants and funding