A human ACTH-secreting corticotroph tumoroid model: Novel Human ACTH-Secreting Tumor Cell in vitro Model.

ACTH-Secreting Pituitary Adenoma / etiology Biomarkers Cell Line, Tumor Cell Proliferation Cells, Cultured Computational Biology / methods Gene Expression Profiling High-Throughput Nucleotide Sequencing Humans Pituitary ACTH Hypersecretion / etiology Single-Cell Analysis Spheroids, Cellular Tumor Cells, Cultured
3D culture Adrenocorticotropic hormone (ACTH) Cushing disease (CD) Proopiomelanocortin (POMC)

Journal

EBioMedicine
ISSN: 2352-3964
Titre abrégé: EBioMedicine
Pays: Netherlands
ID NLM: 101647039

Informations de publication

Date de publication:
Apr 2021
Historique:
received: 07 10 2020
revised: 06 03 2021
accepted: 08 03 2021
pubmed: 28 3 2021
medline: 30 11 2021
entrez: 27 3 2021
Statut: ppublish

Résumé

Cushing disease (CD), although rare, is a life-threatening disorder caused by an adrenocorticotropic hormone (ACTH)-secreting pituitary adenoma, which leads to excess adrenal-derived cortisol. Efficacious and safe medical therapies that control both hormonal hypersecretion and pituitary corticotroph tumor growth remain an unmet need in the management of CD. Translational research in pituitary tumors has been significantly hampered by limited quantities of surgically resected tissue for ex vivo studies, and unavailability of human pituitary tumor cell models. To characterize human corticotroph tumors at the cellular level, we employed single cell RNA-sequencing (scRNA-seq) to study 4 surgically resected tumors. We also used microarrays to compare individualized paired consecutive culture passages to understand transcriptional shifts as in vitro cultures lost ACTH secretion. Based on these findings, we then modified our in vitro culture methods to develop sustained ACTH-secreting human corticotroph tumoroid cultures. scRNA-seq identified 4 major cell populations, namely corticotroph tumor (73.6%), stromal (11.2%), progenitor (8.3%), and immune cells (6.8%). Microarray analysis revealed striking changes in extracellular matrix, cell adhesion and motility-related genes concordant with loss of ACTH secretion during conventional 2D culture. Based on these findings, we subsequently defined a series of crucial culture nutrients and scaffold modifications that provided a more favorable trophic and structural environment that could maintain ACTH secretion in in vitro human corticotroph tumor cultures for up to 4 months. Our human corticotroph tumoroid model is a significant advance in the field of pituitary tumors and will further enable translational research studies to identify critically needed therapies for CD. This work was partly funded by NCI P50-CA211015 and the Warley Trust Foundation.

Sections du résumé

BACKGROUND BACKGROUND
Cushing disease (CD), although rare, is a life-threatening disorder caused by an adrenocorticotropic hormone (ACTH)-secreting pituitary adenoma, which leads to excess adrenal-derived cortisol. Efficacious and safe medical therapies that control both hormonal hypersecretion and pituitary corticotroph tumor growth remain an unmet need in the management of CD. Translational research in pituitary tumors has been significantly hampered by limited quantities of surgically resected tissue for ex vivo studies, and unavailability of human pituitary tumor cell models.
METHODS METHODS
To characterize human corticotroph tumors at the cellular level, we employed single cell RNA-sequencing (scRNA-seq) to study 4 surgically resected tumors. We also used microarrays to compare individualized paired consecutive culture passages to understand transcriptional shifts as in vitro cultures lost ACTH secretion. Based on these findings, we then modified our in vitro culture methods to develop sustained ACTH-secreting human corticotroph tumoroid cultures.
FINDINGS RESULTS
scRNA-seq identified 4 major cell populations, namely corticotroph tumor (73.6%), stromal (11.2%), progenitor (8.3%), and immune cells (6.8%). Microarray analysis revealed striking changes in extracellular matrix, cell adhesion and motility-related genes concordant with loss of ACTH secretion during conventional 2D culture. Based on these findings, we subsequently defined a series of crucial culture nutrients and scaffold modifications that provided a more favorable trophic and structural environment that could maintain ACTH secretion in in vitro human corticotroph tumor cultures for up to 4 months.
INTERPRETATION CONCLUSIONS
Our human corticotroph tumoroid model is a significant advance in the field of pituitary tumors and will further enable translational research studies to identify critically needed therapies for CD.
FUNDING BACKGROUND
This work was partly funded by NCI P50-CA211015 and the Warley Trust Foundation.

Identifiants

DOI: 10.1016/j.ebiom.2021.103294 PMID: 33773184 PMC: PMC8024915
pubmed: 33773184
pii: S2352-3964(21)00087-6
doi: 10.1016/j.ebiom.2021.103294
pmc: PMC8024915
pii:
doi:

Substances chimiques

Biomarkers 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

103294

Subventions

Organisme : NCI NIH HHS
ID : P30 CA016042
Pays : United States
Organisme : NCI NIH HHS
ID : P50 CA211015
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA251930
Pays : United States

Commentaires et corrections

Type : CommentIn

Informations de copyright

Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of Competing Interest The authors declare no competing interests.

Références

Endocrine. 2020 Oct;70(1):15-23
pubmed: 32613546
Nat Protoc. 2009;4(1):44-57
pubmed: 19131956
Proc Natl Acad Sci U S A. 1999 Dec 7;96(25):14378-82
pubmed: 10588713
Res Commun Chem Pathol Pharmacol. 1992 Jan;75(1):39-48
pubmed: 1352645
Clin Cancer Res. 2019 Apr 1;25(7):2194-2205
pubmed: 30385653
Mol Endocrinol. 2013 Mar;27(3):422-36
pubmed: 23371388
Stem Cells. 2009 May;27(5):1182-95
pubmed: 19418455
Cell. 2020 Nov 25;183(5):1420-1435.e21
pubmed: 33159857
Acta Anat (Basel). 1951;11(2-3):361-82
pubmed: 14829142
J Biol Chem. 2004 Feb 27;279(9):8063-9
pubmed: 14660570
J Immunol. 2006 Jun 1;176(11):6954-61
pubmed: 16709856
Cell Biol Int. 2000;24(1):1-7
pubmed: 10826768
Endocr Relat Cancer. 2015 Aug;22(4):481-504
pubmed: 25921430
Blood. 2017 Nov 9;130(19):2092-2100
pubmed: 28807980
Eur J Pharmacol. 1982 Sep 10;83(1-2):101-5
pubmed: 6290240
Genes Dev. 2006 Oct 15;20(20):2871-86
pubmed: 17043312
Acta Endocrinol (Copenh). 1982 Feb;99(2):174-8
pubmed: 6277127
Proc Natl Acad Sci U S A. 2012 Jan 10;109(2):E59-67
pubmed: 22203994
J Biol Chem. 2011 Mar 4;286(9):7070-81
pubmed: 21193407
J Endocrinol. 2019 Feb 1;240(2):287-308
pubmed: 30475227
Compr Physiol. 2018 Mar 13;8(2):631-709
pubmed: 29687901
Int J Gen Med. 2009 Dec 29;2:209-17
pubmed: 20360906
Nat Genet. 2015 Jan;47(1):31-8
pubmed: 25485838
Endocr Pathol. 1993 Jun;4(2):86-94
pubmed: 32138413
Brain Tumor Pathol. 2018 Apr;35(2):62-70
pubmed: 29500747
Endocr Rev. 1997 Apr;18(2):206-28
pubmed: 9101137
J Histochem Cytochem. 2001 Jun;49(6):783-90
pubmed: 11373325
Nat Genet. 2009 Jun;41(6):746-52
pubmed: 19465910
Agents Actions Suppl. 1993;42:103-10
pubmed: 8102834
J Neurosurg. 2014 Sep;121(3):527-35
pubmed: 24926650
Proc Natl Acad Sci U S A. 2013 May 21;110(21):8555-60
pubmed: 23653479
Front Endocrinol (Lausanne). 2020 May 22;11:313
pubmed: 32528411
Cell Res. 2015 Mar;25(3):306-17
pubmed: 25675982
Nucleic Acids Res. 2009 Jan;37(1):1-13
pubmed: 19033363
Oncotarget. 2016 Oct 25;7(43):69149-69158
pubmed: 27708250
Cell Res. 2015 Jun;25(6):649-50
pubmed: 25930709
Case Rep Neurol. 2017 Aug 31;9(2):216-221
pubmed: 28966590
Brain Res. 1999 Nov 27;848(1-2):45-62
pubmed: 10701998
Biomed Res Int. 2015;2015:824514
pubmed: 26339645
Neuroendocrinology. 2012;96(3):173-87
pubmed: 22538498
Cell. 2019 Jun 13;177(7):1888-1902.e21
pubmed: 31178118
J Biol Chem. 1999 Feb 26;274(9):5919-24
pubmed: 10026216
Front Endocrinol (Lausanne). 2019 Sep 18;10:623
pubmed: 31620083
Cancer Metastasis Rev. 2005 Sep;24(3):383-93
pubmed: 16258726
Methods Cell Biol. 1998;57:219-27
pubmed: 9648107
Mol Endocrinol. 2003 May;17(5):959-66
pubmed: 12576489
Endocr Pathol. 1995 Winter;6(4):289-299
pubmed: 12114811
In Vitro Cell Dev Biol Anim. 1997 Mar;33(3):158-60
pubmed: 9112122
J Biol Chem. 1998 Feb 6;273(6):3129-31
pubmed: 9452419
Eur J Endocrinol. 2016 Feb;174(2):213-26
pubmed: 26578638
Endocr Pathol. 1991 Dec;2(4):200-209
pubmed: 32138408
Mol Cell Endocrinol. 2004 Dec 30;228(1-2):1-21
pubmed: 15541569
Cell Stem Cell. 2013 Oct 3;13(4):433-45
pubmed: 24094324
Acta Neuropathol. 1986;71(1-2):111-8
pubmed: 3022531
Adv Protein Chem. 2005;70:437-61
pubmed: 15837523
eNeuro. 2020 Jan 17;7(1):
pubmed: 31915267
Physiol Rev. 2012 Jan;92(1):1-38
pubmed: 22298650

Auteurs

Dongyun Zhang (D)

Departments of Medicine, David Geffen School of Medicine, University of California, Los Angeles, United States.

Willy Hugo (W)

Departments of Medicine, David Geffen School of Medicine, University of California, Los Angeles, United States.

Peter Redublo (P)

Departments of Medicine, David Geffen School of Medicine, University of California, Los Angeles, United States.

Hui Miao (H)

Departments of Medicine, David Geffen School of Medicine, University of California, Los Angeles, United States.

Marvin Bergsneider (M)

Departments of Neurosurgery, David Geffen School of Medicine, University of California, Los Angeles, United States.

Marilene B Wang (MB)

Departments of Head and Neck Surgery, David Geffen School of Medicine, University of California, Los Angeles, United States.

Won Kim (W)

Departments of Neurosurgery, David Geffen School of Medicine, University of California, Los Angeles, United States.

William H Yong (WH)

Departments of Pathology and Lab Medicine, David Geffen School of Medicine, University of California, Los Angeles, United States.

Anthony P Heaney (AP)

Departments of Medicine, David Geffen School of Medicine, University of California, Los Angeles, United States; Departments of Neurosurgery, David Geffen School of Medicine, University of California, Los Angeles, United States. Electronic address: aheaney@mednet.ucla.edu.

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Classifications MeSH

questionsmedicales.fr › Maladies endocriniennes › Maladies de l'hypophyse › Tumeurs de l'hypophyse › Adénome à ACTH › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Facteurs biologiques › Marqueurs biologiques › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Cellules › Cellules cultivées › Cellules cancéreuses en culture › Lignée cellulaire tumorale › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Phénomènes physiologiques › Croissance et développement › Croissance › Processus de croissance cellulaire › Prolifération cellulaire › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Cellules › Cellules cultivées › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Sciences de l'information › Informatique › Biologie informatique › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Analyse de profil d'expression de gènes › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Analyse de séquence › Séquençage nucléotidique à haut débit › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Maladies endocriniennes › Maladies de l'hypophyse › Hyperpituitarisme › Hypersécrétion hypophysaire d'ACTH › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Techniques d'investigation › Techniques cytologiques › Analyse sur cellule unique › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Cellules › Cellules cultivées › Sphéroïdes de cellules › A human ACTH-secreting corticotroph tumoroid...
questionsmedicales.fr › Cellules › Cellules cultivées › Cellules cancéreuses en culture › A human ACTH-secreting corticotroph tumoroid...