Clinical Trial Details
— Status: Recruiting
Administrative data
| NCT number |
NCT05761210 |
| Other study ID # |
01C217 |
| Secondary ID |
|
| Status |
Recruiting |
| Phase |
|
| First received |
|
| Last updated |
|
| Start date |
April 19, 2022 |
| Est. completion date |
December 31, 2023 |
Study information
| Verified date |
March 2023 |
| Source |
Istituto Auxologico Italiano |
| Contact |
Alessandro Sartorio |
| Phone |
+390261911 |
| Email |
sartorio[@]auxologico.it |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Observational
|
Clinical Trial Summary
With the project proposal we aim to identify serum markers for the characterization of
steatosis in subjects affected by essential obesity at a young age. In this case the markers
could be useful not only for the development of new diagnostic scores, or for combining them
with diagnostic imaging technologies, but also for understanding the metabolic alterations
according to the patient's gender, extremely important data in this disease to which only
recently has biomedical research started.
Description:
Experimental design The study includes a discovery phase in which a serum proteomic analysis
of the subjects included in the study will be performed. This phase will make it possible to
identify a small number of possible candidate markers, which will be validated in a
subsequent phase with a greater number of subjects.
Patients:
In the discovery phase, 60 pubescent subjects (30 males and 30 females) will be taken into
consideration. The subjects will be divided into two groups:
- control group including subjects with absence of steatosis, verified by ultrasound
method according to standardized criteria [10,11], and with APRI score < 0.3;
- study group with subjects with the proven presence of steatosis using ultrasound
techniques, and with APRI score > 0.50.
For each patient, the following variables will be considered in baseline conditions, i.e. at
the time of sample collection.
- anthropometric parameters (height, weight, BMI, waist circumference);
- body composition (lean mass, fat mass), using the impedance measurement technique (BIA);
- levels of steatosis measured with ultrasound according to standard criteria
- blood chemistry parameters
- basic metabolism, using indirect calorimetry;
- biochemical parameters determining the metabolic syndrome (HDL cholesterol,
triglycerides, blood sugar);
- biochemical parameters for the prediction of hepatic steatosis using surrogate markers
(AST, ALT, GGT, platelets, alkaline phosphatase, albumin);
- blood pressure;
- concomitant hormonal therapies (growth hormone therapy, sex steroids, and/or
l-thyroxine).
Serum collection Fasting blood samples will be collected by standard venipuncture into BD
Vacutainer® serum separation tubes (BD - Plymouth PL6 7BP, UK) and centrifuged at 1900g at
4°C for 10 min and at 16,000g at 4°C for a further 10 min. The supernatants will be
transferred to new tubes and subsequently frozen at -80°C for long-term storage.
Proteomic analysis Before the proteomic analysis, the presence of hemoglobin will be
evaluated by spectrophotometer Beckman Coulter® DU®730 (Beckman Coulter, Brea, CA, USA) using
the direct spectrophotometric method of Harboe with Allen correction: Hb ( g / L) = (167, 2 ×
A415 - 83.6 × A380 - 83.6 × A450) x 1/1000 × 1 / dilution in dH2O. The cut-off considered for
serum will be 0.020 g/L.
For the proteomic characterization of serum, a new high-throughput omics technology based on
the SomaLogic SOMAscan platform will be used. The system is based on the detection of
SOMAmers (Slow Off-rate Modified Aptamers), which are aptameric forms of DNA with high
affinity and specificity for the protein target. Furthermore, these aptamers are resistant to
nucleotidase activity and demonstrate higher affinity than normal antibodies. The panel used
in this study will allow us to evaluate 1500 protein targets that are involved in the
development of metabolic diseases, are part of the general inflammatory response, and are of
interest in the study of cardiovascular diseases and also in oncology.
In general, the sensitivity and performance of the test are comparable to that of a normal
ELISA kit, with limits for quantification around 100 fM and detection of 40 fM, and the need
for only 65 µL of the sample.
Validation The validation of 2 markers selected based on the significance and differential
expression between the groups will be validated by the ELISA method. In this phase, each
marker will be validated by determining its concentration in the serum of 60 subjects
(included in the discovery phase) and in other 32 complementary subjects to reach the sample
size useful for obtaining an appropriate statistical significance.
Furthermore, the experiments will be set up to include positive and negative controls and
controls to check for inter-plate variability, as required by ELISA assay set-up procedures.
Standard curves will be calculated to determine the absolute quantification of the markers of
interest in serum.
