Characterization of uniquely tumorigenic cancer stem cells in salivary gland adenoid cystic carcinoma

BackgroundCancer stem cells (CSC) are endowed with multipotency, self-renewal and unique tumorigenic potential. They have been identified as cells with high activity of aldehyde dehydrogenase (ALDH) and high expression of CD44 in head and neck squamous cell carcinoma (HNSCC). The objective of this w...

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Main Authors: Kristy A. Warner, Sosuke Sahara, Alexandra E. Herzog, Felipe Nör, Rogerio M. Castilho, Peter J. Polverini, Jacques E. Nör
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Oral Health
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Online Access:https://www.frontiersin.org/articles/10.3389/froh.2025.1570042/full
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author Kristy A. Warner
Sosuke Sahara
Alexandra E. Herzog
Felipe Nör
Rogerio M. Castilho
Peter J. Polverini
Jacques E. Nör
Jacques E. Nör
Jacques E. Nör
author_facet Kristy A. Warner
Sosuke Sahara
Alexandra E. Herzog
Felipe Nör
Rogerio M. Castilho
Peter J. Polverini
Jacques E. Nör
Jacques E. Nör
Jacques E. Nör
author_sort Kristy A. Warner
collection DOAJ
description BackgroundCancer stem cells (CSC) are endowed with multipotency, self-renewal and unique tumorigenic potential. They have been identified as cells with high activity of aldehyde dehydrogenase (ALDH) and high expression of CD44 in head and neck squamous cell carcinoma (HNSCC). The objective of this work is to understand whether salivary gland adenoid cystic carcinoma contain CSCs and whether they exhibit a unique tumorigenic activity in this cancer.MethodsWe used flow cytometry, salisphere and western blot assays with 3 human ACC cell lines (UM-HACC-2A, UM-HACC-14, UM-HACC-6) to characterize the impact of ALDH activity and CD44 expression. In vitro results were verified in vivo by orthotopic injection of cells retrieved from a patient-derived xenograft (PDX) model of ACC (UM-PDX-HACC-14) in the submandibular gland of SCID mice. Primary tumor and metastatic spread were evaluated by 2 pathologists blinded for experimental conditions.ResultsThe fraction of ALDHhighCD44high cells in UM-HACC-2A, UM-HACC-14 and UM-HACC-6 ranged from 3% to 8%. ALDHhighCD44high cells formed more salispheres and expressed higher levels of stem cell markers (e.g., Notch2, Bmi-1) compared to control ALDHlowCD44low cells. ALDHhighCD44high cells sorted from the ACC PDX tumors were more tumorigenic upon orthotopic transplantation into submandibular salivary glands and generated more lung metastases than control ACC cells. Strong ALDH1A1 staining was observed in the majority of salivary gland tumors and lung metastases generated by transplantation of ALDHhighCD44high cells.ConclusionsWe conclude that salivary gland adenoid cystic carcinoma contain a small population of uniquely tumorigenic cells characterized by high ALDH activity and expression.
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spelling doaj-art-b1842a4a7bcd4fa499cdfc2af32339fb2025-08-20T03:53:39ZengFrontiers Media S.A.Frontiers in Oral Health2673-48422025-04-01610.3389/froh.2025.15700421570042Characterization of uniquely tumorigenic cancer stem cells in salivary gland adenoid cystic carcinomaKristy A. Warner0Sosuke Sahara1Alexandra E. Herzog2Felipe Nör3Rogerio M. Castilho4Peter J. Polverini5Jacques E. Nör6Jacques E. Nör7Jacques E. Nör8Department of Cariology, Restorative Sciences, Endodontics, University of Michigan School of Dentistry, Ann Arbor, MI, United StatesDepartment of Cariology, Restorative Sciences, Endodontics, University of Michigan School of Dentistry, Ann Arbor, MI, United StatesDepartment of Cariology, Restorative Sciences, Endodontics, University of Michigan School of Dentistry, Ann Arbor, MI, United StatesDepartment of Periodontics and Oral Medicine, School of Dentistry, Ann Arbor, MI, United StatesDepartment of Periodontics and Oral Medicine, School of Dentistry, Ann Arbor, MI, United StatesDepartment of Periodontics and Oral Medicine, School of Dentistry, Ann Arbor, MI, United StatesDepartment of Cariology, Restorative Sciences, Endodontics, University of Michigan School of Dentistry, Ann Arbor, MI, United StatesDepartment of Otolaryngology, University of Michigan School of Medicine, Ann Arbor, MI, United StatesRogel Cancer Center, University of Michigan, Ann Arbor, MI, United StatesBackgroundCancer stem cells (CSC) are endowed with multipotency, self-renewal and unique tumorigenic potential. They have been identified as cells with high activity of aldehyde dehydrogenase (ALDH) and high expression of CD44 in head and neck squamous cell carcinoma (HNSCC). The objective of this work is to understand whether salivary gland adenoid cystic carcinoma contain CSCs and whether they exhibit a unique tumorigenic activity in this cancer.MethodsWe used flow cytometry, salisphere and western blot assays with 3 human ACC cell lines (UM-HACC-2A, UM-HACC-14, UM-HACC-6) to characterize the impact of ALDH activity and CD44 expression. In vitro results were verified in vivo by orthotopic injection of cells retrieved from a patient-derived xenograft (PDX) model of ACC (UM-PDX-HACC-14) in the submandibular gland of SCID mice. Primary tumor and metastatic spread were evaluated by 2 pathologists blinded for experimental conditions.ResultsThe fraction of ALDHhighCD44high cells in UM-HACC-2A, UM-HACC-14 and UM-HACC-6 ranged from 3% to 8%. ALDHhighCD44high cells formed more salispheres and expressed higher levels of stem cell markers (e.g., Notch2, Bmi-1) compared to control ALDHlowCD44low cells. ALDHhighCD44high cells sorted from the ACC PDX tumors were more tumorigenic upon orthotopic transplantation into submandibular salivary glands and generated more lung metastases than control ACC cells. Strong ALDH1A1 staining was observed in the majority of salivary gland tumors and lung metastases generated by transplantation of ALDHhighCD44high cells.ConclusionsWe conclude that salivary gland adenoid cystic carcinoma contain a small population of uniquely tumorigenic cells characterized by high ALDH activity and expression.https://www.frontiersin.org/articles/10.3389/froh.2025.1570042/fullsalivary gland cancercancer stemnesstumor-initiating cellsaldehyde dehydrogenasetumor cell heterogeneity
spellingShingle Kristy A. Warner
Sosuke Sahara
Alexandra E. Herzog
Felipe Nör
Rogerio M. Castilho
Peter J. Polverini
Jacques E. Nör
Jacques E. Nör
Jacques E. Nör
Characterization of uniquely tumorigenic cancer stem cells in salivary gland adenoid cystic carcinoma
Frontiers in Oral Health
salivary gland cancer
cancer stemness
tumor-initiating cells
aldehyde dehydrogenase
tumor cell heterogeneity
title Characterization of uniquely tumorigenic cancer stem cells in salivary gland adenoid cystic carcinoma
title_full Characterization of uniquely tumorigenic cancer stem cells in salivary gland adenoid cystic carcinoma
title_fullStr Characterization of uniquely tumorigenic cancer stem cells in salivary gland adenoid cystic carcinoma
title_full_unstemmed Characterization of uniquely tumorigenic cancer stem cells in salivary gland adenoid cystic carcinoma
title_short Characterization of uniquely tumorigenic cancer stem cells in salivary gland adenoid cystic carcinoma
title_sort characterization of uniquely tumorigenic cancer stem cells in salivary gland adenoid cystic carcinoma
topic salivary gland cancer
cancer stemness
tumor-initiating cells
aldehyde dehydrogenase
tumor cell heterogeneity
url https://www.frontiersin.org/articles/10.3389/froh.2025.1570042/full
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