An interventional study of circadian gene expression evaluation in Cushing Syndrome, Adrenal Insufficiency and Cushing Disease, sponsored by University of Roma La Sapienza. Recruiting at 1 site in Italy. Open to participants aged 18 Years to 80 Years, including healthy volunteers. Per ClinicalTrials.gov, last updated 2026-10-20.
Sponsored by University of Roma La Sapienza · Not applicable, Interventional, and Other
This is a multicentric, prospective, intervention study on circadian genes expression in peripheral blood mononuclear cells as biomarkers of circadian rhythm derangement in patients affected by alterations of endogenous glucocorticoids secretion (Cushing's Syndrome during active phase, treatment and under remission and newly or on established glucocorticoid replacement therapy adrenal insufficiency)
This is an intervention, prospective, multicentric study.
Enrolled patients will undergo 4 visits:
Age-, sex- and BMI- matched healthy controls will be enrolled. Patients and controls will undergo the same procedures at baseline and after 1, 3 and 6 months.
The primary outcome measure will be the evaluation of circadian genes CLOCK and Aryl Hydrocarbon Receptor Nuclear Translocator Like (ARNTL) expression in peripheral blood mononuclear cells (PBMC) compared to healthy controls.
Secondary Outcomes measures will be:
Protocol Amendment (Version 2.1, in 2026)
The study protocol was amended to expand the investigation of circadian dysregulation to additional clinical populations and to introduce complementary physiological and molecular assessments, while maintaining the original primary research objective.
Additional study populations include individuals with obesity, with or without metabolic syndrome, and patients initiating systemic glucocorticoid therapy for non-endocrine conditions. Additional control populations may also be included according to the amended protocol.
The amendment introduces further assessments of circadian regulation, including clock-related microRNA expression, steroid profiling by LC-MS/MS, continuous glucose monitoring, wearable-derived sleep and circadian parameters, and dim light melatonin onset in selected participants.
Exploratory molecular assessments include DNA methylation-based epigenetic clocks, transcriptomic profiling, proteomics, and metabolomics, aimed at characterizing the relationships between glucocorticoid exposure, circadian disruption, immune function, and metabolic health.
Longitudinal follow-up may be extended to 12 months according to the relevant study cohort and assessment schedule.
130 studies on the registry are indexed under Cushing Syndrome; 40 are open to participants now.
This study's planned enrollment of 44 is below the median of 50 across 61 interventional studies indexed under Cushing Syndrome.
Browse Cushing Syndrome studies →University of Roma La Sapienza is the lead sponsor of 397 studies on the registry; 56 are open to participants now.
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ACTH-dependent or ACTH-independent Cushing's Syndrome diagnosis during active disease (new diagnosis or recidivating).
Exclusion Criteria:
- acute adrenal insufficiency;
Additional inclusion criteria introduced by Protocol Amendment Version 2.1 (approved 2026):
Adults with obesity (BMI ≥30 kg/m²), with or without metabolic syndrome. Adults initiating systemic glucocorticoid therapy for non-endocrine conditions, including respiratory or rheumatologic disorders.
Additional control participants, including euthyroid individuals with benign nodular thyroid disease, according to the amended protocol.
Patients with Adrenal Insufficiency established or newly diagnosed, under glucocorticoid replacement therapy.
Diagnostic Test: circadian gene expression evaluation
patients with adrenocorticotropic hormone (ACTH)-dependent or ACTH-independent Cushing's Syndrome diagnosis during active disease (new diagnosis or recidivating) at enrollment.
Diagnostic Test: circadian gene expression evaluation
Age-, sex- and BMI- matched patients referring to our center for diagnostic procedures not affected by Adrenal Insufficiency or Cushing's Syndrome.
Diagnostic Test: circadian gene expression evaluation
Adults with obesity (BMI ≥30 kg/m²), with or without metabolic syndrome, enrolled to investigate circadian, endocrine, immune, and metabolic alterations associated with obesity.
Diagnostic Test: circadian gene expression evaluation
Adults initiating systemic glucocorticoid therapy for non-endocrine conditions, including rheumatologic or respiratory disorders. Participants will undergo baseline assessment before treatment initiation, followed by assessments after 2-3 weeks and 2-3 months, with optional follow-up at 6 and 12 months.
Diagnostic Test: circadian gene expression evaluation
patients and controls will undergo circadian gene expression (CLOCK, ARNTL) evaluation at baseline and after 1, 3 and 6 months
Circadian genes CLOCK and ARNTL expression evaluation
Change in relative expression circadian genes of CLOCK and ARNLT from baseline compared to healthy controls. After PBMC isolation by Ficoll-Plaque gradient, complementary DNA (cDNA) pool will be extracted and used as the template for subsequent Polymerase Chain Reaction (PCR) amplification in Real time PCR; Gene expression will be quantified as relative expression compared to housekeeping genes.
Time frame: baseline, +1 month, +3 months, +6 months
Circadian gene expression profile
Change from baseline in relative expression and variations throughout the day of circadian genes (CLOCK and ARNTL) compared to healthy controls at 7:00-8:00 AM (before breakfast), 12:00 AM (before lunch), 3:00-4:00 PM (after lunch), 7:00 PM (before dinner), 12:00 PM. After PBMC isolation by Ficoll-Plaque gradient, cDNA pool will be extracted and used as the template for subsequent Polymerase Chain Reaction (PCR) amplification in Real time PCR; Gene expression will be quantified as relative expression compared to housekeeping genes.
Time frame: baseline, +1 month, +3 months, +6 months
Peripheral Blood Mononuclear Cells circadian profiling
Number of cells (number per mm3) of peripheral blood mononuclear cell subpopulations at 7:00-8:00 AM (before breakfast), 12:00 AM (before lunch), 3:00-4:00 PM (after lunch), 7:00 PM (before dinner), 12:00 PM
Time frame: baseline, +1 month, +3 months, +6 months
Circadian cortisol rhythm
Circadian cortisol rhythm by serum and salivary dosage at 7:00-8:00 AM (before breakfast), 12:00 AM (before lunch), 3:00-4:00 PM (after lunch), 7:00 PM (before dinner), 12:00 PM.
Time frame: baseline, +1 month, +3 months, +6 months
Sleep Disturbances
Sleep disturbances will be evaluated by The Pittsburgh Sleep Quality Index (PSQI). This questionnaire contains 19 self-related questions which are combined to create 7 component scores with a range of 0-3 points (0:no difficulty, 3: severe difficulty). Global score is the result to the addition of all component scores with a range of 0-21 points (0:no difficulty, 21: severe difficulty).
Time frame: baseline, +1 month, +3 months, +6 months
Infectious Diseases Frequency and Severity
Frequencies and severity of infectious diseases will be evaluated by modified Infectious Diseases Questionnaire (GNC). This questionnaire includes questions on infectious diseases of upper and lower respiratory tract, gastrointestinal tract, skin and urogenital tract contracted during the previous 12 months. Questions investigate on the number and duration of infections, necessity of antibiotic or antifungal therapy, hospital stay and days of absence from work. Final score represents the frequency of infections. Moreover, some questions investigate possible susceptible or protective factors for infectious diseases: vaccinations, use of corticosteroids, concomitant diseases, previous appendectomy, tonsillectomy, adenoidectomy, splenectomy or thymectomy.
Time frame: baseline, +1 month, +3 months, +6 months
Circadian blood pressure
Circadian blood pressure will be evaluated using ambulatory blood pressure monitoring blood pressure waves for a noninvasive estimation
Time frame: baseline, +1 month, +3 months, +6 months
Quality of life evaluation
Quality of life will be evaluated by the Physical Component score and the Mental Component score of the self-administered questionnaire SF-36-Item Health Survey questionnaire. This questionnaire measures eight scales: physical functioning, role physical, bodily pain, general health (physical component) and vitality, social functioning, role emotional, mental health (mental component). Interpretation of the score will be the following: at each item of the questionnaire corresponds a percentage value (from 0% to 100%). The average of the single items constitutes the scale total percentage (from 0% to 100%); missing data are not considered during calculation. High score defines a more favorable health state.
Time frame: baseline, +1 month, +3 months, +6 months
Psychometric Evaluation
Psychometric Evaluation will be evaluated using Beck Depression Inventory questionnaire. : a 21-item measure of depressive symptoms. Each answer is scored on a scale value (from 0 to 3 points). The global score is obtained adding all single scores, with a range of 0-63. Higher score constitutes worse burden of symptoms (0-13: minimal depression, 14-19: mild depression, 20-28: moderate depression, 29-63: severe depression).
Time frame: baseline, +1 month, +3 months, +6 months
Sexual dysfunction in women
Evaluation of sexual dysfunction will be evaluated in women using FSFI questionnaire. This is a 19-item questionnaire that covers six domains: desire, arousal, lubrication, and orgasm, satisfaction, and pain. The ranges of the domain scores are as follows: for desire 1.2 - 6.0, for arousal, lubrication, orgasm and pain 0 - 6.0 and for satisfaction 0.8 - 6.0. The higher scores of the six domains indicate better sexual functioning about that domain. The overall FSFI score is obtained by summing the six domain scores. The overall FSFI scores range from 2.0 to 36.0, with higher scores indicating better sexual functioning.
Time frame: baseline, +1 month, +3 months, +6 months
Sexual dysfunction in men
Evaluation of sexual dysfunction will be evaluated in men using IIEF questionnaire. The IIEF is an internationally validated test. It is used to assign a score of erectile dysfunction: severe score 1 to 10; moderate score 11 to 16; low to moderate score 17 to 21; Low score 22 to 25; no erectile dysfunction score 26 to 30
Time frame: baseline, +1 month, +3 months, +6 months
insuline resistance
Measurement of HOMA index calculated according to the formula: fasting insulin (microU/L) x fasting glucose (nmol/L)/22.5.
Time frame: baseline, +1 month, +3 months, +6 months
blood lipid profile
Measurement of Total Cholesterol, LDL cholesterol, HDL cholesterol, Triglyceredes
Time frame: baseline, +1 month, +3 months, +6 months
Inflammatory cytokines levels
Evaluation of Tumor Necrosis Factor alpha (TNFα), Transforming Growth Factor beta (TGF-β), Interferon gamma (IFN-γ), Leptin, Resistin, Adiponectin, Adipsin, Monocyte Chemoattractant Protein-1 (MCP-1), Serpin, C Reactive Protein (CRP), Interleukin 6 (IL-6), Interleukin 10 (IL-10) serum concentrations at 7:00-8:00 AM (before breakfast), 12:00 AM (before lunch), 3:00-4:00 PM (after lunch), 7:00 PM (before dinner), 12:00 PM
Time frame: baseline, +1 month, +3 months, +6 months
Steroid profiling LC-MS/MS
Assessment of steroid metabolism using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in blood, saliva, and 24-hour urine samples. Steroid concentrations and selected metabolite ratios will be evaluated longitudinally.
Time frame: Baseline and months 3, 6, and 12.
DNA Methylation-Based Epigenetic Age
Exploratory assessment of DNA methylation patterns and epigenetic age using validated DNA methylation-based epigenetic clock algorithms. Epigenetic age estimates and derived measures of epigenetic age acceleration will be calculated to investigate associations with glucocorticoid exposure, metabolic status, and circadian dysregulation.
Time frame: Baseline and after remission or therapy change
Continuous Glucose Monitoring-Derived Glycemic Parameters
Assessment of interstitial glucose fluctuations using a commercially available continuous glucose monitoring device worn for approximately 7 days. Glucose profiles and their temporal variation will be analyzed to investigate circadian glucose regulation.
Time frame: Approximately 7 days during selected study visits, at baseline and follow-up assessments.
Wearable-Derived Sleep and Circadian Rhythm Parameters
Assessment of sleep duration, sleep timing, midsleep point, heart rate, heart rate variability, and other available physiological parameters using a commercially available wearable device worn for 7 days. Derived parameters will be used to characterize circadian rhythmicity.
Time frame: Seven-day monitoring periods at selected study visits.
Dim Light Melatonin Onset
Assessment of dim light melatonin onset (DLMO) using repeated salivary samples collected under dim-light conditions. Salivary melatonin concentrations will be measured by LC-MS/MS to estimate the timing of central circadian phase.
Time frame: Evening salivary sampling during selected study visits.
Transcriptomic Signatures of Circadian Dysregulation
Exploratory assessment of circadian gene expression and clock-controlled gene signatures using targeted gene panels or high-throughput transcriptomic approaches in peripheral blood mononuclear cells and other available biological samples.
Time frame: Baseline and available follow-up visits.
Circulating Proteomic Profiles
Exploratory profiling of circulating proteins involved in endocrine, immune, metabolic, and circadian pathways using multiplex immunoassays or other validated proteomic platforms.
Time frame: Baseline and available follow-up visits.
Metabolomic Profiles Associated With Circadian Regulation
Exploratory characterization of circulating and/or urinary metabolites associated with circadian metabolic regulation using targeted or untargeted metabolomic approaches.
Time frame: Baseline and available follow-up visits
Plan to share: Undecided
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Started Jul 2018 (actual). Primary completion is estimated for Dec 2027, 112 months after the start.
Since we started tracking on Sep 25, 2026, the dates moved once, on Oct 20, 2026: primary completion Jun 2023 → Dec 2027 and completion Dec 2023 → Dec 2029.
Earlier changes are on the record history on ClinicalTrials.gov ↗
How these figures are worked out, and which studies are left out of them.
From the registry record's own update history. This site started tracking changes on Sep 25, 2026; for anything earlier, see the record history on ClinicalTrials.gov ↗
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