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Principal Investigator: ANTHONY P HEANEY
Organization: UNIVERSITY OF CALIFORNIA LOS ANGELES
Fiscal Year: 2024
Award: $339,008
Funding agency: National Cancer Institute
Abstract
Cushing Disease (CD) is a life-threatening “orphan disease” caused by an adrenocorticotropic hormone
(ACTH)-secreting pituitary adenoma driving excess adrenal cortisol production. There is a large unmet medical
need for CD treatment. However, translational research has been greatly hampered due to unavailability of any
human pituitary corticotroph tumor cell models. Using single cell RNA-sequencing (scRNAseq) and microarray
transcriptome analysis of surgically resected human corticotroph tumors, we observed that loss of pituitary
corticotroph tumor ACTH secretion coincided with reduced angiogenesis, survival signals and immune
responses in parallel with increased collagen catabolism, cell adhesion and extracellular matrix organization.
Guided by these findings, we developed a unique 3-dimensional (3D) pituitary tumor culture system and for the
first time, we have been able to generate 3D human corticotroph tumor cultures that secrete ACTH >4 months.
We have assembled an experienced multidisciplinary team to complete 3 focused specific aims using this first
of its kind resource. Firstly, we will use whole exome sequencing to characterize the genomic landscape of our
corticotroph 3D culture biobank and compare genomic and genetic fidelity between the original corticotroph
tumor, normal blood and matched 3D corticotroph tumoroid cultures from the same individual patient.
ScRNAseq analysis of serial passages of individual patient-derived corticotroph tumor cultures will monitor for
transcriptome changes in a temporal fashion over the course of culture. The histopathological structure of our
3D corticotroph cultures at the single cell level will quantify tissue architecture so we can map corticotroph
tumoroid cellular composition and distribution. A second aim will employ a miniaturized automated system to
conduct a high throughput drug screen in our 3D corticotroph tumor cultures. Compounds will be subjected to
rigorous evaluation to define primary “hits” and validated by re-screening in triplicate using 20 concentrations
from 100µM to 20pM (2-fold dilution) to reliably calculate an EC50 for each compound. Finally, three
complementary approaches, computational cheminformatic profiling, scRNAseq to delineate transcriptomic
changes at the single cell level following drug treatment and functional genomics will be employed to explore
the MOA of validated hit compounds. This integrated interrogation of our drug screen results and the genetic
features of our patient-derived 3D tumor cultures as well as that of the original tumor tissue, will allow us to
disentangle an individual drug's mode(s) of action, and directly document drug sensitivity of individualized
parental corticotroph tumors. In summary, we will use our unique biobank of comprehensively molecularly
characterized pituitary corticotroph tumor tissues and paired derived 3D corticotroph tumor cultures to test
libraries of clincially relevant compounds. This pituitary 3D tumor culture system is transformative in the field
due to the lack of any human pituitary corticotroph tumor cell models and will pave the path for much needed
improved therapy for patients with this dreadfully disabling and often fatal disorder, Cushing disease.
Terms: <3-D><3-Dimensional><3D><ACTH><Adrenal Glands><Adrenals><Adrenocorticotropic Hormone><Adrenocorticotropin><Aeroseb-HC><Architecture><Automobile Driving><Blood><Blood Reticuloendothelial System><Body Tissues><Catabolism><Cell Adhesion><Cell Body><Cell Communication and Signaling><Cell Signaling><Cell model><Cell-Extracellular Matrix><Cells><Cellular Adhesion><Cellular model><Cetacort><Clinical><Collagen><Cort-Dome><Cortef><Cortenema><Corticotropin><Cortisol><Cortispray><Cortril><Cushing Disease><Dermacort><Development><Disabling><Disease><Disorder><Drug Interactions><Drug Screening><Drug Therapy><Drugs><ECM><Eldecort><Engineering / Architecture><Evaluation><Extracellular Matrix><Generalized Growth><Genes><Genetic><Genomics><Growth><High Throughput Assay><Hormone secretion><Human><Hydrocortisone><Hydrocortone><Hypophysis><Hypophysis Cerebri><Hytone><Immune response><Immunological response><Individual><Intracellular Communication and Signaling><Libraries><Life><Maps><Medical><Medication><Methods><Modeling><Modern Man><Molecular><Monitor><Nutracort><Operative Procedures><Operative Surgical Procedures><Orphan Disease><Pathway interactions><Patients><Pattern Recognition><Pharmaceutical Preparations><Pharmacotherapy><Pituitary><Pituitary ACTH Hypersecretion><Pituitary Adenoma><Pituitary Gland><Pituitary Gland Adenoma><Pituitary Gland Neoplasm><Pituitary Gland Tumor><Pituitary Neoplasms><Pituitary Nervous System><Pituitary Tumors><Pituitary-dependent Cushing's disease><Pituitary-dependent Cushing's disorder><Pre-Clinical Model><Preclinical Models><Primary Neoplasm><Primary Tumor><Proctocort><Production><Proteins><Rare Diseases><Rare Disorder><Research Resources><Resected><Resources><Screening Result><Serial Passage><Signal Transduction><Signal Transduction Systems><Signaling><Structure><Surgical><Surgical Interventions><Surgical Procedure><System><Testing><Time><Tissue Growth><Tissues><Translational Research><Translational Science><Tumor Cell><Tumor Tissue><Validation><Visualization><angiogenesis><biobank><biological signal transduction><biorepository><chemical informatics><cheminformatics><developmental><driving><drug discovery><drug sensitivity><drug treatment><drug/agent><exome sequencing><exome-seq><experience><experiment><experimental research><experimental study><experiments><functional genomics><genomic platform><global gene expression><global transcription profile><high throughput screening><high-throughput drug screening><hormonal secretion><host response><immune system response><immunocytochemistry><immunoresponse><improved><individual patient><inhibitor><miniaturize><miniaturized><multidisciplinary><neoplastic cell><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><ontogeny><orphan disorder><pathway><personalization of treatment><personalized medicine><personalized therapy><personalized treatment><scRNA-seq><screening><screenings><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><small molecule><suprarenal gland><surgery><three dimensional><transcriptome><transcriptomics><translation research><translational investigation><tumor><validations>