Pituitary adenomas evade apoptosis via noxa deregulation in Cushing's disease

David T Asuzu; Reinier Alvarez; Patrick A Fletcher; Debjani Mandal; Kory Johnson; Weiwei Wu; Abdel Elkahloun; Paul Clavijo; Clint Allen; Dragan Maric; Abhik Ray-Chaudhury; Sharika Rajan; Zied Abdullaev; Diana Nwokoye; Kenneth Aldape; Lynnette K Nieman; Constantine Stratakis; Stanko S Stojilkovic; Prashant Chittiboina

doi:10.1016/j.celrep.2022.111223

Pituitary adenomas evade apoptosis via noxa deregulation in Cushing's disease

Cell Rep. 2022 Aug 23;40(8):111223. doi: 10.1016/j.celrep.2022.111223.

Authors

David T Asuzu¹, Reinier Alvarez², Patrick A Fletcher³, Debjani Mandal⁴, Kory Johnson⁵, Weiwei Wu⁶, Abdel Elkahloun⁶, Paul Clavijo⁷, Clint Allen⁷, Dragan Maric⁸, Abhik Ray-Chaudhury⁹, Sharika Rajan¹⁰, Zied Abdullaev¹⁰, Diana Nwokoye⁴, Kenneth Aldape¹⁰, Lynnette K Nieman¹¹, Constantine Stratakis¹², Stanko S Stojilkovic¹³, Prashant Chittiboina¹⁴

Affiliations

¹ Neurosurgery Unit for Pituitary and Inheritable Diseases, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 10 Center Drive, Room 3D20, Bethesda, MD 20892, USA; Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, Bethesda, MD, USA; Department of Neurosurgery, University of Virginia, Charlottesville, VA, USA.
² Neurosurgery Unit for Pituitary and Inheritable Diseases, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 10 Center Drive, Room 3D20, Bethesda, MD 20892, USA; Florida International University Herbert Wertheim College of Medicine, Miami, FL, USA.
³ Laboratory of Biological Modeling, National Institute of Diabetes, Digestive and Kidney Diseases, Bethesda, MD, USA.
⁴ Neurosurgery Unit for Pituitary and Inheritable Diseases, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 10 Center Drive, Room 3D20, Bethesda, MD 20892, USA.
⁵ DIR Bioinformatics Section, National Institute of Neurological Disorders and Stroke, Bethesda, MD, USA.
⁶ Cancer Genetics and Comparative Genomics Branch, National Human Genome Research Institute, Bethesda, MD, USA.
⁷ Translational Tumor Immunology Program, National Institute on Deafness and Other Communication Disorders, Bethesda, MD, USA.
⁸ Flow and Imaging Cytometry Core Facility, National Institute of Neurological Disorders and Stroke, Bethesda, MD, USA.
⁹ Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, Bethesda, MD, USA; Laboratory of Pathology, National Cancer Institute, Bethesda, MD, USA.
¹⁰ Laboratory of Pathology, National Cancer Institute, Bethesda, MD, USA.
¹¹ Section on Translational Endocrinology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD, USA.
¹² Section on Endocrinology and Genetics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD, USA.
¹³ Section on Cellular Signaling, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD, USA.
¹⁴ Neurosurgery Unit for Pituitary and Inheritable Diseases, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 10 Center Drive, Room 3D20, Bethesda, MD 20892, USA; Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, Bethesda, MD, USA. Electronic address: prashant.chittiboina@nih.gov.

Abstract

Sporadic pituitary adenomas occur in over 10% of the population. Hormone-secreting adenomas, including those causing Cushing's disease (CD), cause severe morbidity and early mortality. Mechanistic studies of CD are hindered by a lack of in vitro models and control normal human pituitary glands. Here, we surgically annotate adenomas and adjacent normal glands in 25 of 34 patients. Using single-cell RNA sequencing (RNA-seq) analysis of 27594 cells, we identify CD adenoma transcriptomic signatures compared with adjacent normal cells, with validation by bulk RNA-seq, DNA methylation, qRT-PCR, and immunohistochemistry. CD adenoma cells include a subpopulation of proliferating, terminally differentiated corticotrophs. In CD adenomas, we find recurrent promoter hypomethylation and transcriptional upregulation of PMAIP1 (encoding pro-apoptotic BH3-only bcl-2 protein noxa) but paradoxical noxa downregulation. Using primary CD adenoma cell cultures and a corticotroph-enriched mouse cell line, we find that selective proteasomal inhibition with bortezomib stabilizes noxa and induces apoptosis, indicating its utility as an anti-tumor agent.

Keywords: CP: Cancer; CP: Molecular biology; Cushing’s disease; DNA methylation; bortezomib; preclinical testing; proteasome; single-cell RNA-seq; sporadic pituitary adenoma.

Publication types

Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural

MeSH terms

Adenoma* / metabolism
Adrenocorticotropic Hormone / metabolism
Animals
Apoptosis
Humans
Mice
Pituitary ACTH Hypersecretion* / genetics
Pituitary ACTH Hypersecretion* / pathology
Pituitary Neoplasms* / genetics
Pituitary Neoplasms* / metabolism
Pituitary Neoplasms* / pathology

Substances

Adrenocorticotropic Hormone

Grants and funding

ZIA NS003150/ImNIH/Intramural NIH HHS/United States