CHROnOS: Clinical Study on Circadian Genes Dysregulation in Patients With Glucocorticoid Disorders

Sponsor

University of Roma La Sapienza (Other)

Overall Status

Unknown status

CT.gov ID

NCT04374721

Collaborator

(none)

Enrollment

Location

Arms

22.9

Anticipated Duration (Months)

1.9

Patients Per Site Per Month

Study Details

Study Description

Brief Summary

This is a monocentric, 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 and under remission and newly or on established glucocorticoid replacement therapy adrenal insufficiency)

Condition or Disease	Intervention/Treatment	Phase
Cushing Syndrome Adrenal Insufficiency Cushing Disease Addison Disease	Diagnostic Test: circadian gene expression evaluation	N/A

Detailed Description

This is an intervention, prospective, monocentric study.

Enrolled patients will undergo 3 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 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 three and six months after 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.

Age-, sex- and BMI- matched healthy controls will be enrolled. Patients and controls will undergo the same procedures at baseline and after 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).

Study Design

Study Type:

Interventional

Anticipated Enrollment :

44 participants

Allocation:

Non-Randomized

Intervention Model:

Parallel Assignment

Intervention Model Description:

patients will undergo circadian genes evaluation in peripheral blood mononuclear cellspatients will undergo circadian genes evaluation in peripheral blood mononuclear cells

Masking:

None (Open Label)

Primary Purpose:

Other

Official Title:

Clinical Study on Circadian Genes Dysregulation in Patients With Glucocorticoid Disorders

Actual Study Start Date :

Jul 4, 2018

Anticipated Primary Completion Date :

Jun 1, 2020

Anticipated Study Completion Date :

Jun 1, 2020

Arms and Interventions

Arm	Intervention/Treatment
Experimental: Patients with Adrenal Insufficiency Patients with Adrenal Insufficiency established or newly diagnosed, under glucocorticoid replacement therapy.	Diagnostic Test: circadian gene expression evaluation patients and controls will undergo circadian gene expression (CLOCK, ARNTL) evaluation at baseline and after 3 and 6 months
Experimental: Patients with Cushing's Syndrome 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 patients and controls will undergo circadian gene expression (CLOCK, ARNTL) evaluation at baseline and after 3 and 6 months
Experimental: Healthy Controls 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 patients and controls will undergo circadian gene expression (CLOCK, ARNTL) evaluation at baseline and after 3 and 6 months

Arm

Intervention/Treatment

Experimental: Patients with Adrenal Insufficiency

Patients with Adrenal Insufficiency established or newly diagnosed, under glucocorticoid replacement therapy.

Diagnostic Test: circadian gene expression evaluation

patients and controls will undergo circadian gene expression (CLOCK, ARNTL) evaluation at baseline and after 3 and 6 months

Experimental: Patients with Cushing's Syndrome

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

patients and controls will undergo circadian gene expression (CLOCK, ARNTL) evaluation at baseline and after 3 and 6 months

Experimental: Healthy Controls

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

patients and controls will undergo circadian gene expression (CLOCK, ARNTL) evaluation at baseline and after 3 and 6 months

Outcome Measures

Primary Outcome Measures

Circadian genes CLOCK and ARNTL expression evaluation [baseline, +3 months, +6 months]
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.

Secondary Outcome Measures

Circadian gene expression profile [baseline, +3 months, +6 months]
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.
Cytokines levels Inflammatory cytokines levels [baseline, +3 months, +6 months]
Composite outcomes consisting of simultaneous chemiluminescence measurement 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 (ng/mL) 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
Peripheral Blood Mononuclear Cells circadian profiling [baseline, +3 months, +6 months]
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
Circadian cortisol rhythm [baseline, +3 months, +6 months]
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 [baseline, +3 months, +6 months]
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).
Infectious Diseases Frequency and Severity [baseline, +3 months, +6 months]
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.

Eligibility Criteria

Criteria

Ages Eligible for Study:

18 Years to 80 Years

Sexes Eligible for Study:

All

Accepts Healthy Volunteers:

Yes

Inclusion Criteria:

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.

Exclusion Criteria:

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;
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);
alcoholism and/or drug addictions;
night-shift workers;
use of melatonin, antipsychotic medications, estroprogestinic preparations