Oncotarget

Research Papers:

A 3D biofabricated cutaneous squamous cell carcinoma tissue model with multi-channel confocal microscopy imaging biomarkers to quantify antitumor effects of chemotherapeutics in tissue

James R. Browning, Paige Derr, Kristy Derr, Nicole Doudican, Sam Michael, Samantha R. Lish, Nicholas A. Taylor, James G. Krueger, Marc Ferrer, John A. Carucci and Daniel S. Gareau _

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Oncotarget. 2020; 11:2587-2596. https://doi.org/10.18632/oncotarget.27570

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Abstract

James R. Browning1, Paige Derr2, Kristy Derr2, Nicole Doudican3, Sam Michael2, Samantha R. Lish1, Nicholas A. Taylor3, James G. Krueger1, Marc Ferrer2, John A. Carucci3 and Daniel S. Gareau1

1 Laboratory for Investigative Dermatology, The Rockefeller University, New York, New York, USA

2 National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland, USA

3 The Ronald O. Perelman Department of Dermatology, New York University School of Medicine, New York, New York, USA

Correspondence to:

Daniel S. Gareau,email: dgareau@rockefeller.edu

Keywords: squamous cell carcinoma; screening; 3D printing; in vitro model; confocal microscopy

Received: January 05, 2020     Accepted: April 03, 2020     Published: July 07, 2020

ABSTRACT

Cutaneous squamous cell carcinoma (cSCC) causes approximately 10,000 deaths annually in the U. S. Current therapies are largely ineffective against metastatic and locally advanced cSCC. There is a need to identify novel, effective, and less toxic small molecule cSCC therapeutics. We developed a 3-dimensional bioprinted skin (3DBPS) model of cSCC tumors together with a microscopy assay to test chemotherapeutic effects in tissue. The full thickness SCC tissue model was validated using hematoxylin and eosin (H&E) and immunohistochemical histological staining, confocal microscopy, and cDNA microarray analysis. A nondestructive, 3D fluorescence confocal imaging assay with tdTomato-labeled A431 SCC and ZsGreen-labeled keratinocytes was developed to test efficacy and general toxicity of chemotherapeutics. Fluorescence-derived imaging biomarkers indicated that 50% of cancer cells were killed in the tissue after 1?M 5-Fluorouracil 48-hour treatment, compared to a baseline of 12% for untreated controls. The imaging biomarkers also showed that normal keratinocytes were less affected by treatment (11% killed) than the untreated tissue, which had no significant killing effect. Data showed that 5-Fluorouracil selectively killed cSCC cells more than keratinocytes. Our 3DBPS assay platform provides cellular-level measurement of cell viability and can be adapted to achieve nondestructive high-throughput screening (HTS) in bio-fabricated tissues.


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