Cushing SyndromeCHROnOSNCT04374721
Clinical Study on Circadian Genes Dysregulation in Patients With Glucocorticoid Disorders
A study testing circadian gene expression evaluation in people with Cushing Syndrome and Adrenal Insufficiency, run by University of Roma La Sapienza at 1 site in Italy.
Trial summary
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… Read more about this study
- Who is it for?
- Ages 18 to 80 yearsHealthy volunteers can take part
- Conditions the study is for
- Cushing Syndrome, Adrenal Insufficiency, Cushing Disease
- What is being tested?
- circadian gene expression evaluation
- Could I receive a placebo?
- No, everyone receives an active treatment
- How many people
- 44 participants
- Where
- 1 siteItaly
- Study dates
- 4 Jul 2018to 1 Dec 2029
- Phase
- Interventional
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 80 years
- All sexes
- Healthy volunteers accepted
- Adult, Older adult
Who may be able to join
- Primary or secondary chronic adrenal insufficiency, previously or newly diagnosed.
- ACTH-dependent or ACTH-independent Cushing's Syndrome diagnosis during active disease (new diagnosis or recidivating).
- Signed informed consent to participate in the study.
Who cannot join
- acute adrenal insufficiency;
- clinical or laboratory signs of significant respiratory, hepatobiliary, or pancreatic disease;
- pregnancy;
- severe infections, surgery, trauma requiring hospitalization within 3 months before study entry;
- any active blood or rheumatic disorders, and active liver disease in the previous 5 years;
- clinically significant chronic kidney disease;
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- severe psychiatric diseases;
- history of neoplasms in the last 5 years (except for adrenal or pituitary adenoma in Cushing Syndrome, pituitary adenoma or related neolpasms in secondary adrenal insufficiency);
- heart disease with a class III or class IV functional capacity;
- BMI greater than 40 kg/m²;
- use of medication that interferes with cortisol metabolism within 1 month before study entry;
- treatment with systemic Glucocorticoid (GC) therapy other than hydrocortisone (HC), or cortisone acetate (CA); (except for the exogenous glucocorticoid cohort)
- alcoholism and/or drug addictions;
- night-shift workers;
- use of melatonin, antipsychotic medications, estroprogestinic preparations
- 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.
Trial locations and contacts
1 site in 1 country. Contact the study team or a site near you to ask about taking part.
Italy1 site, 1 recruiting
- Recruiting
Department of Experimental Medicine, "Sapienza" University of Rome
Roma, Roma, 00161
Andrea M Isidori, MD,PhD, 0039649970711, andrea.isidori@uniroma1.it
About this study
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)
Read the detailed description
This is an intervention, prospective, multicentric study.
Enrolled patients will undergo 4 visits:
- Adrenal insufficiency (AI) patients: patients affected by primary or secondary adrenal insufficiency, whether newly diagnosed or on established glucocorticoid therapy, will be evaluated at baseline and after one, three and six months.
- Cushing's Syndrome (CS) patients: patients affected by Cushing's Syndrome will be evaluated at baseline during active phase of the disease and one, three and six months after treatment or remission. Patients affected by Cushing's Syndrome who will require glucocorticoid replacement therapy after remission will be evaluated three and six months after remission and then three and six months after the eventual glucocorticoid replacement therapy withdrawal. CS treatment will be surgery or medical therapy according to guidelines. Timing of medical therapy administration will change during protocol according to circadian rhythms.
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:
- Circadian genes expression assessment compared to healthy controls at 7:00-8:00 Ante Meridiem (AM) (before breakfast), 12:00 AM (before lunch), 3:00-4:00 Post Meridiem (PM) (after lunch), 7:00 PM (before dinner), 12:00 PM;
- Immune profiling compared to healthy controls by the quantification of peripheral blood mononuclear cells (PBMC) subpopulations assessed by flow cytometry 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;
- Evaluation of inflammatory cytokines and adipokines production 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;
- 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;
- Sleep disturbances evaluation by The Pittsburgh Sleep Quality Index (PSQI) self reported questionnaire
- Evaluation of infectious diseases frequencies and severity compared to healthy controls. Infectious diseases will be evaluated by an adaptation of Infectious Diseases Questionnaire (GNC)
- Circadian blood pressure using ambulatory blood pressure monitoring blood pressure waves for a noninvasive estimation
- Quality of life using SF-36-Item Health Survey questionnaire
- Psychometric Evaluation using Beck Depression Inventory questionnaire
- Evaluation of sexual dysfunction using FSFI questionnaire in woman and IIEF questionnaire in man
- Evaluation of insuline resistance calculated with HOMA index
- Evaluation of body weight (kg)
- Evaluation of blood lipid profile
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.
- Conditions
- Cushing Syndrome, Adrenal Insufficiency, Cushing Disease, Addison Disease, Obesity (BMI>30)
- Being tested
- circadian gene expression evaluation
- Keywords
- circadian rhythm, circadian genes, circadian clock, glucocorticoids, cortisol, obesity, Epigenetic Aging, DNA Methylation, Circadian Misalignment, Wearable Devices, Steroid Metabolomics
- Medical subject headings
- Cushing Syndrome, Adrenal Insufficiency, Pituitary ACTH Hypersecretion, Addison Disease, Obesity
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
circadian gene expression evaluationDiagnostic test
patients and controls will undergo circadian gene expression (CLOCK, ARNTL) evaluation at baseline and after 1, 3 and 6 months
| Study group | Role | What they receive |
|---|---|---|
| Patients with Adrenal Insufficiency | Experimental | Patients with Adrenal Insufficiency established or newly diagnosed, under glucocorticoid replacement therapy. Receives: Diagnostic Test: circadian gene expression evaluation |
| Patients with Cushing's Syndrome | Experimental | patients with adrenocorticotropic hormone (ACTH)-dependent or ACTH-independent Cushing's Syndrome diagnosis during active disease (new diagnosis or recidivating) at enrollment. Receives: Diagnostic Test: circadian gene expression evaluation |
| Healthy Controls | Experimental | Age-, sex- and BMI- matched patients referring to our center for diagnostic procedures not affected by Adrenal Insufficiency or Cushing's Syndrome. Receives: Diagnostic Test: circadian gene expression evaluation |
| Arm 4 - Participants with Obesity | Experimental | Adults with obesity (BMI ≥30 kg/m²), with or without metabolic syndrome, enrolled to investigate circadian, endocrine, immune, and metabolic alterations associated with obesity. Receives: Diagnostic Test: circadian gene expression evaluation |
| Arm 5 - Patients Receiving Exogenous Glucocorticoids | Experimental | 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. Receives: Diagnostic Test: circadian gene expression evaluation |
What is being measured
The primary outcome is the main question the study is designed to answer.
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.
Measured: baseline, +1 month, +3 months, +6 months
Secondary outcomes (14)
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.
Measured: 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
Measured: 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.
Measured: 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).
Measured: 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.
Measured: 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
Measured: 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.
Measured: 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).
Measured: 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.
Measured: 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
Measured: 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.
Measured: baseline, +1 month, +3 months, +6 months
blood lipid profile
Measurement of Total Cholesterol, LDL cholesterol, HDL cholesterol, Triglyceredes
Measured: 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
Measured: 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.
Measured: Baseline and months 3, 6, and 12.
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted5 May 2020
- Study start4 Jul 2018
- Primary completion1 Dec 2027 (estimated)
- Study completion1 Dec 2029 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for Cushing SyndromeStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Interventional
- Phase
- Phase N/A
- Primary purpose
- Other
- Allocation
- Non-randomised
- Intervention model
- Parallel groups
- Model description
- patients will undergo circadian genes evaluation in peripheral blood mononuclear cells
- Masking
- None (open label)
- Enrolment
- 44 participants (estimated)
Sponsor and investigators
- Lead sponsor
- University of Roma La Sapienza
- Sponsor type
- University, hospital or other
- Responsible party
- Principal investigator, Andrea M. Isidori, University of Roma La Sapienza
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT04374721
- Sponsor's protocol ID
- CHROnOS
- Acronym
- CHROnOS
- Status last verified
- Oct 2026
- First submitted
- 23 Apr 2020
- Last updated
- 20 Oct 2026
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- Undecided