title: Essential role of aldehyde dehydrogenase 1A3 for the maintenance of non-small cell lung cancer stem cells is associated with the STAT3 pathway.
abstract: OBJECTIVE
  Lung cancer stem cells (CSC) with elevated aldehyde dehydrogenase (ALDH) activity are self-renewing, clonogenic, and tumorigenic. The purpose of our study is to elucidate the mechanisms by which lung CSCs are regulated.
  
  
  METHODS
  A genome-wide gene expression analysis was performed to identify genes differentially expressed in the ALDH(+) versus ALDH -: cells. RT-PCR, Western blot analysis, and Aldefluor assay were used to validate identified genes. To explore the function in CSCs, we manipulated their expression followed by colony and tumor formation assays.
  
  
  RESULTS
  We identified a subset of genes that were differentially expressed in common in ALDH(+) cells, among which ALDH1A3 was the most upregulated gene in ALDH(+) versus ALDH -: cells. shRNA-mediated knockdown of ALDH1A3 in non-small cell lung cancer (NSCLC) resulted in a dramatic reduction in ALDH activity, clonogenicity, and tumorigenicity, indicating that ALDH1A3 is required for tumorigenic properties. In contrast, overexpression of ALDH1A3 by itself it was not sufficient to increase tumorigenicity. The ALDH(+) cells also expressed more activated STAT3 than ALDH -: cells. Inhibition of STAT3 or its activator EZH2 genetically or pharmacologically diminished the level of ALDH(+) cells and clonogenicity. Unexpectedly, ALDH1A3 was highly expressed in female, never smokers, well-differentiated tumors, or adenocarcinoma. ALDH1A3 low expression was associated with poor overall survival.
  
  
  CONCLUSIONS
  Our data show that ALDH1A3 is the predominant ALDH isozyme responsible for ALDH activity and tumorigenicity in most NSCLCs, and that inhibiting either ALDH1A3 or the STAT3 pathway are potential therapeutic strategies to eliminate the ALDH(+) subpopulation in NSCLCs.
authors.affiliation: Hamon Center for Therapeutic Oncology Research, Simmons Comprehensive Cancer Center.
authors.forename: Chunli
authors.initials: C
authors.lastname: Shao
authors.affiliation: Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts;
authors.forename: James P
authors.initials: JP
authors.lastname: Sullivan
authors.affiliation: Hamon Center for Therapeutic Oncology Research, Simmons Comprehensive Cancer Center, Pharmacology, and.
authors.forename: Luc
authors.initials: L
authors.lastname: Girard
authors.affiliation: Hamon Center for Therapeutic Oncology Research, Simmons Comprehensive Cancer Center.
authors.forename: Alexander
authors.initials: A
authors.lastname: Augustyn
authors.affiliation: Hamon Center for Therapeutic Oncology Research, Simmons Comprehensive Cancer Center.
authors.forename: Paul
authors.initials: P
authors.lastname: Yenerall
authors.affiliation: Department of Translational Molecular Pathology, University of Texas M.D. Anderson Cancer Center, Houston; Departments of.
authors.forename: Jaime
authors.initials: J
authors.lastname: Rodriguez-Canales
authors.affiliation: Department of Translational Molecular Pathology, University of Texas M.D. Anderson Cancer Center, Houston; Departments of.
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.affiliation: Department of Translational Molecular Pathology, University of Texas M.D. Anderson Cancer Center, Houston; Departments of.
authors.forename: Carmen
authors.initials: C
authors.lastname: Behrens
authors.affiliation: Cell Biology.
authors.forename: Jerry W
authors.initials: JW
authors.lastname: Shay
authors.affiliation: Department of Translational Molecular Pathology, University of Texas M.D. Anderson Cancer Center, Houston; Departments of.
authors.forename: Ignacio I
authors.initials: II
authors.lastname: Wistuba
authors.affiliation: Hamon Center for Therapeutic Oncology Research, Simmons Comprehensive Cancer Center, Pharmacology, and Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas john.minna@utsouthwestern.edu.
authors.forename: John D
authors.initials: JD
authors.lastname: Minna
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D002289:Carcinoma, Non-Small-Cell Lung / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007124:Immunoenzyme Techniques; D008175:Lung Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014411:Neoplastic Stem Cells / Q000201:enzymology / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D046888:Tissue Array Analysis; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay; D023041:Xenograft Model Antitumor Assays
chemical_list: D014408:Biomarkers, Tumor; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D050796:STAT3 Transcription Factor; D000445:Aldehyde Oxidoreductases; C018699:aldehyde dehydrogenase (NAD(P)+)