Sphingo-lipotoxicity and Trans-differentiation of Adipose Tissue in Obesity (SFINGOTRANS) (SFINGOTRANS)

Relationship Between Sphingo-lipotoxicity and Trans-differentiation of Adipose Tissue: Cross-sectional Study in Subjects With Different Dysmetabolic Severity

After recruiting a population of subjects with different metabolic severity (subjects of normal weight and obese patients with and without metabolic syndrome), the objectives of the present research will be:

1. determine leukocyte mRNA levels of Cidea (gene associated with BAT functional status), Hoxc9 (gene associated with browning of WAT) and Cpt1a (gene associated with β-oxidation of fatty acids in both tissues, i.e. BAT and WAT) (secondary endpoint);
2. to determine energy expenditure with indirect calorimetric technique, body temperature and circulating catecholamine levels, which will be correlated to leukocyte levels of Cidea, Hoxc9 and Cpt1a mRNA (secondary endpoint);
3. determine the plasma levels of an extensive panel of sphingolipids, including in particular ceramides and sphingosine-1-phosphate which, by exerting a lipotoxic, lipoinflammatory and anti-adipogenic effect, will be correlated to the leukocyte levels of Cidea, Hoxc9 mRNA and Cpt1a (primary endpoint);
4. determine the erythrocyte, leukocyte and platelet levels of sphingolipids which, acting as peripheral biomarkers of cardiometabolic dysfunction (e.g., atherogenesis, thromboembolism, arterial hypertension, insulin resistance, low-grade chronic inflammation, etc.), could phenotypically identify patients with increased cardiovascular risk (e.g., obese patients with or without metabolic syndrome) (secondary endpoint).

Hypothesis: the existence of a relationship between sphingohypotoxicity and transdifferentiation of adipose tissue and a combination of sphingolipids (plasma/erythrocyte/platelet/leukocyte) and gene regulators (WAT/BAT-related) which, with sensitivity and specificity, is associated with diagnosis of metabolic syndrome.

Study Overview

• Status: Completed
• Conditions: Healthy; Obesity
• Intervention / Treatment: Diagnostic test: Blood sample collection

Detailed Description

Materials and methods Patients: 90 adults of both sexes will be recruited, of whom 30 of normal weight (age: 18-50 years; BMI < 25 kg/m2), 30 obese without metabolic syndrome (age: 18-35 years; BMI > 35 kg/m2) and 30 obese with metabolic syndrome (age: 18-35 years; BMI > 35 kg/m2), according to the 2009 IDF criteria. Subjects of normal weight will be recruited from medical/paramedical staff, while obese patients from hospitalized at the Division of Metabolic Diseases, Istituto Auxologico Italiano, Piancavallo (VB), Italy, for a 3-week multidisciplinary weight reduction program (BWRP), which includes low-calorie diet, physical exercise, psychological support and nutrition education.

Subjects with other pathologies other than obesity will be excluded from the study, including those treated with anticoagulant and antiplatelet drugs, since the evaluation of intraplatelet levels of sphingolipids will be foreseen.

In basal conditions, the main anthropometric data will be collected (weight, height, waist circumference, hip circumference, BMI), body composition will be evaluated with a bioimpedance technique, the main cardiovascular parameters will be recorded (blood pressure and heart rate), a calorimetric examination will be performed, collection of body temperature (morning and evening), request of the environmental temperature to which one is generally exposed during the day and determined, with automated clinical biochemistry techniques, the following biochemical parameters: glucose, total cholesterol, triglycerides, LDL, HDL, fatty acids non-esterified, insulin, glycated Hb, catecholamines and C-reactive protein.

Determination of the lipidomic profile in plasma and cell extracts A lipidomics will be performed in plasma and in cellular extracts from erythrocytes, leukocytes and platelets. The levels of the individual analytes will be determined with a technologically advanced analytical instrumentation, consisting of a triple quadruple hybrid mass spectrometer with linear ion trap (QTRAP 5500, AB Sciex), interfaced with an ultra-high performance liquid chromatograph (UHPLC).

Plasma and cellular levels of the following sphingolipids will be measured: ceramides and dihydroceramides from C16 to C24, including 2 unsaturated ones (C18:1 and C24:1), the sphingomyelins (those from C16 to C24 and the one C24:1), sphingosine, sphinganine , sphingosine-1-phosphate and sphinganine-1-phosphate.

Determination of leukocyte mRNA levels of gene regulators From an aliquot containing leukocytes, stored ad hoc, the total mRNA will be extracted and, with RT/PCR technique, the leukocyte mRNA levels of the following genes will be determined: Cidea, Hoxc9 and Cpt1a.

• Study Type: Observational
• Enrollment (Actual): 84

Contacts and Locations

Study Locations

• Italy
  • Verbania
    • Oggebbio, Verbania, Italy, 28824
      • Istituto Auxologico Italiano, Site Piancavallo

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 50 years (Adult)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

• Sampling Method: Non-Probability Sample

Study Population

Obese patients (BMI > 35 kg/m2) and age- and sex-matched healthy controls (normal weighted)

Description

Inclusion Criteria (obese subjects):

- obesity (BMI > 35 kg/m2)

Inclusion criteria (healthy controls):

- BMI < 25 kg/m2

Exclusion Criteria:

- patients/controls treated with anticoagulant and antiplatelet drugs

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Cases - obese with metabolic syndrome; Obese subjects with metabolic syndrome	Diagnostic test: Blood sample collection; Blood sample collection
Cases - obese without metabolic syndrome; Obese subjects without metabolic syndrome	Diagnostic test: Blood sample collection; Blood sample collection
Controls; Normal weight subjects	Diagnostic test: Blood sample collection; Blood sample collection

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Evaluation of sphingolipids	Determination of an extensive panel of sphingolipids, including ceramides and sphingosine-1-phosphate which, by exerting a lipotoxic, lipoinflammatory and anti-adipogenic effect, are correlated to the leukocyte levels of Cidea, Hoxc9 mRNA and Cpt1a	Baseline

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Determination of leukocyte mRNA levels of Cidea and Cpt1a	determination of leukocyte mRNA levels of Cidea (gene associated with BAT functional status), Hoxc9 (gene associated with browning of WAT) and Cpt1a (gene associated with β-oxidation of fatty acids in both tissues, i.e. BAT and WAT)	Baseline
Determination of the erythrocyte, leukocyte and platelet levels of sphingolipids	Determination of the erythrocyte, leukocyte and platelet levels of sphingolipids which, acting as peripheral biomarkers of cardiometabolic dysfunction (e.g., atherogenesis, thromboembolism, arterial hypertension, insulin resistance, low-grade chronic inflammation, etc.), could phenotypically identify patients with increased cardiovascular risk (e.g., obese patients with or without metabolic syndrome)	Baseline

Collaborators and Investigators

• Sponsor: Istituto Auxologico Italiano

Study record dates

Study Major Dates

• Study Start (Actual): June 15, 2021
• Primary Completion (Actual): March 23, 2022
• Study Completion (Actual): March 23, 2022

Study Registration Dates

• First Submitted: February 27, 2023
• First Submitted That Met QC Criteria: February 27, 2023
• First Posted (Actual): March 9, 2023

Study Record Updates

• Last Update Posted (Actual): March 13, 2023
• Last Update Submitted That Met QC Criteria: March 9, 2023
• Last Verified: March 1, 2023

More Information

Terms related to this study

• Keywords: Obesity; Metabolic syndrome; Adipose tissue; Normals; Sphyngolipids
• Additional Relevant MeSH Terms: Overnutrition; Nutrition Disorders; Overweight; Body Weight; Obesity
• Other Study ID Numbers: 01C126

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No