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Dis Model Mech. 2018 Jan 29;11(1). pii: dmm031716. doi: 10.1242/dmm.031716.

Expanding primary cells from mucoepidermoid and other salivary gland neoplasms for genetic and chemosensitivity testing.

Author information

1
Oncology, Georgetown University, Washington, DC 20057, USA.
2
Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Khalid University, Abha, 61413, Saudi Arabia.
3
Pathology, Center for Cell Reprogramming, Georgetown University, Washington, DC 20057, USA.
4
Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC 20057, USA.
5
Biostatistics, Bioinformatics and Biomathematics, Georgetown University, Washington, DC 20057, USA.
6
Pathology, Georgetown University, Washington, DC 20057, USA.
7
Pathology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC 20057, USA.
8
Otolaryngology - Head and Neck Surgery, MedStar Georgetown University Hospital, Washington, DC 20007, USA.
9
Oncology and Medicine, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC 20057, USA paf3@georgetown.edu.

Abstract

Restricted availability of cell and animal models is a rate-limiting step for investigation of salivary gland neoplasm pathophysiology and therapeutic response. Conditionally reprogrammed cell (CRC) technology enables establishment of primary epithelial cell cultures from patient material. This study tested a translational workflow for acquisition, expansion and testing of CRC-derived primary cultures of salivary gland neoplasms from patients presenting to an academic surgical practice. Results showed that cultured cells were sufficient for epithelial cell-specific transcriptome characterization to detect candidate therapeutic pathways and fusion genes, and for screening for cancer risk-associated single nucleotide polymorphisms (SNPs) and driver gene mutations through exome sequencing. Focused study of primary cultures of a low-grade mucoepidermoid carcinoma demonstrated amphiregulin-mechanistic target of rapamycin-protein kinase B (AKT; AKT1) pathway activation, identified through bioinformatics and subsequently confirmed as present in primary tissue and preserved through different secondary 2D and 3D culture media and xenografts. Candidate therapeutic testing showed that the allosteric AKT inhibitor MK2206 reproducibly inhibited cell survival across different culture formats. By contrast, the cells appeared resistant to the adenosine triphosphate competitive AKT inhibitor GSK690693. Procedures employed here illustrate an approach for reproducibly obtaining material for pathophysiological studies of salivary gland neoplasms, and other less common epithelial cancer types, that can be executed without compromising pathological examination of patient specimens. The approach permits combined genetic and cell-based physiological and therapeutic investigations in addition to more traditional pathologic studies, and can be used to build sustainable bio-banks for future inquiries.This article has an associated First Person interview with the first author of the paper.

KEYWORDS:

AKT; Drug sensitivity; Mucoepidermoid carcinoma; Next-generation sequencing; Primary cell culture; Salivary gland neoplasms

PMID:
29419396
PMCID:
PMC5818080
DOI:
10.1242/dmm.031716
[Indexed for MEDLINE]
Free PMC Article

Conflict of interest statement

Competing interestsGeorgetown University has submitted a patent application for the cell reprogramming technology described, for which X.L. is an inventor. The intellectual property is under an exclusive license to Propagenix, in which Georgetown University has a founding equity interest. All other authors declare no competing interests.

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