--------------------------------------------- # Development of human pituitary adenoma organoids to facilitate effective targeted treatments of Cushing's disease Preferred citation (DataCite format): Chakrabarti, Jayati; Pandey, Ritu; Churko, Jared; Eschbacher, Jennifer; Mallick, Saptarshi; Chen, Yuliang; et al. (2022): Development of human pituitary adenoma organoids to facilitate effective targeted treatments of Cushing's disease. University of Arizona Research Data Repository. Dataset. https://doi.org/10.25422/azu.data.19755244 Corresponding Author: Yana Zavros, Cellular and Molecular Medicine, yzavros@arizona.edu License: CC BY 4.0 DOI: https://doi.org/10.25422/azu.data.19755244 --------------------------------------------- ## Summary Cushing's disease (CD) is a serious endocrine disorder caused by an unregulated adrenocorticotropic hormone (ACTH)-secreting pituitary tumor that stimulates the adrenal glands to overproduce cortisol. Chronic exposure to excess cortisol has detrimental effects on health, including increased stroke rates, diabetes, obesity, cognitive impairment, anxiety, depression, and death. The first-line treatment for CD is pituitary surgery, which is followed by disease remission in only 47% of patients. The lack of specificity of current standard of care treatments with low efficacy and tolerability makes CD a medical therapeutic challenge. One major limitation that hinders the development of specific medical therapies is the lack of human relevant model systems that recapitulate the cellular composition of pituitary adenomas. Human pituitary adenoma tissue was harvested during transsphenoidal surgery from CD patients to generate organoids (hPITOs). hPITOs generated from corticotroph, lactotroph, gonadotroph and somatotroph adenomas exhibited morphological diversity among the organoid lines between individual patients and amongst subtypes. The similarity in cell lineages between the organoid line and the patient’s tumor was validated by comparing the neuropathology report, to the expression pattern of pituitary adenoma specific markers using spectral flow cytometry and exome sequencing. A high-throughput drug screen demonstrated patient-specific drug responses of hPITOs amongst each adenoma subtype. Generation of induced pluripotent stem cells (iPSCs) from patients carrying germline mutations relevant to CD exhibited dysregulated cell lineage commitment. The development of the human pituitary adenoma organoids represent a paradigm shift in how we model complex pathologies in CD patients that may alter patient care. --------------------------------------------- ## Files and Folders Figures (PowerPoint/PDF) and raw data files (Excel, CSV) FCS files: - Fig2_HPITO25, Fig2_HPITO3, Fig2_HPITO24 is related to Fig 2a - Fig2_HPITO37N, Fig2_HPITO37 is related to Fig 2c - Fig2_HPITO38N, Fig2_HPITO38 is related to Fig 2d - Rest of all the .fcs files were used to generate the sub figures in Fig 2 We are using Cytek Aurora Spectrum Viewer software to generate the .fcs files. These files can be analyzed using Standard flow cytometric analysis package, like FlowJo, CytoBank, OMIQ etc. --------------------------------------------- ## Materials and Methods Microsoft Word, Excel, PowerPoint R 4.2 Thermal cycler, Incubator, Bio Hood, Spectral flow cytometer (Aurora), Nikon Spinning Disc Confocal Microscope, Zeiss 880 Confocal microscope, Agarose Gel electrophoresis apparatus, GelDoc imager, Rhapsody Single Cell Analyzer --------------------------------------------- ## Contributor Roles The roles are defined by the CRediT taxonomy http://credit.niso.org/ - Jayati Chakrabarti, University of Arizona: Conceptualization, Data Curation, Formal Analysis, Methodology, Validation, Writing – Drafting, review & editing and the manuscript - Ritu Pandey, University of Arizona: Formal Analysis, review & editing the manuscript - Jared M. Churko, University of Arizona: Formal Analysis, review & editing the manuscript - Jennifer Eschbacher, Barrow Neurological Institute, Phoenix, Arizona: Material sourcing review & editing of the manuscript - Saptarshi Mallick, University of Arizona: Conceptualization, Formal Analysis, Methodology, Writing - drafting, review & editing of the manuscript - Yuliang Chen - University of Arizona: Formal Analysis, review & editing of the manuscript - Beth Hermes, Barrow Neurological Institute, Phoenix, Arizona: Material sourcing, review & editing of the manuscript - Palash Mallick, University of Arizona: Formal Analysis, review & editing of the manuscript - Ben Stansfield, University of Arizona: Formal Analysis, review & editing of the manuscript - Kelvin W. Pond, University of Arizona: Methodology, Writing - review & editing of the manuscript - Curtis Thorne, University of Arizona: Methodology, review & editing of the manuscript - Kevin Yuen, Barrow Neurological Institute, Phoenix, Arizona: Conceptualization, Material sourcing, Writing – review & editing of the manuscript - Andrew S. Little, Barrow Neurological Institute, Phoenix, Arizona: Conceptualization, Material sourcing, Writing – review & editing of the manuscript - Yana Zavros, University of Arizona: Conceptualization, Data Curation, Funding acquisition, Investigation, Formal Analysis, Methodology, Project administration, Supervision, Validation, Visualization, Writing – original draft, review & editing the manuscript