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Numerous pathologies (sickle cell disease, thalassemia, spherocytosis, etc.) lead to changes in the rheological properties of the blood, in particular via alterations in the deformability of red blood cells. These alterations lead to circulatory complications of which an emblematic example is the sickle cell crisis which manifests itself by microcirculatory occlusions. Several authors suggest that the deformability of erythrocytes is a key parameter for the diagnosis and monitoring of patients. Numerous studies, especially in vitro, show that the mechanical properties of the red blood cell significantly influence its dynamics in flow (blood viscosity, distribution in capillary networks). Moreover, concerning the specific problem of vaso-occlusion, the proportion of the most rigid red blood cells is a determining factor of the probability of occlusion more than the average value of this rigidity which can hide great disparities.
There is no clinically usable test to assess the alteration of the fine rheology of the red blood cell in a patient. Functional tests such as ektacytometry require heavy equipment and teams of specialized biologists; this technique is therefore only available in 3 biological reference centers in France. "Mechanical phenotyping" seems to be a potentially simpler and more accessible technique, and has already shown promising prospects in other nosological settings than red blood cell pathologies.
Today, there is no specific marker of sickle cell vaso-occlusive crisis, nor marker of severity, that would be useful for pathophysiological understanding but also for clinical management.
This study aims to characterize the microfluidic flow and intra-erythrocyte viscosity of sickle cell red blood cells, and to identify specific biological phenotype or clinical severity profiles. The techniques used are microfluidic circuits for the study of flow and molecular rotors for the measurement of intra-erythrocyte viscosity, using deoxygenation cycles in order to model physio-pathological situations.
The first part will allow the calibration of the microfluidic techniques used (microfluidic circuit and molecular rotors), testing blood from healthy subjects (without constitutional or acquired red blood cell pathology) and blood from SCD patients. The aim is to define the reproducibility and sensitivity of the techniques.
A second part is aimed at establishing a rheological profile of the blood of patients with SCD in comparison with blood from control subjects, i.e. with other constitutional or acquired red blood cell pathology.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Healthy subjects | Subjects with no documented hematological pathology (neither constitutional nor acquired). From the recruitment of living kidney donors, transplantation unit of Grenoble Alpes University Hospital |
| |
| SCD patients | Patients with SCD |
| |
| Control patients | With a constitutional non-sickle cell disease of the red blood cell, or an acquired red blood cell disease. |
|
| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Blood sample collection | Other | Blood sample collection |
|
| Measure | Description | Time Frame |
|---|---|---|
| Study of the intra-erythrocyte viscosity dispersion and rheological profile of red blood cells | Measure of the intra-erythrocyte viscosity dispersion using molecular rotors technique, study of rheological profile of red blood cells in microfluidic circuit : measure of the speed of flowing, and DI deformability Index [DI = (L-W)/(L+W)] of each red blood cell, DI dispersion in each sample, in basal state and after exposure to deoxygenation cycles of blood SCD patients versus control subjects. | 30 months |
| Measure | Description | Time Frame |
|---|---|---|
| Study of the intra-erythrocyte viscosity dispersion and rheological profile of red blood cells | Measure of the intra-erythrocyte viscosity dispersion using molecular rotors technique, study of rheological profile of red blood cells in microfluidic circuit : measure of the speed of flowing, and DI deformability Index [DI = (L-W)/(L+W)] of each red blood cell, DI dispersion in each sample, in basal state and after exposure to deoxygenation cycles in different conditions : congenital red blood cell disorders, acquired red blood cell disorders and clinical events (vasoocclusive crisis, pregnancy, infection). |
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Inclusion Criteria for healthy volunteers :
Inclusion Criteria for SDC patient :
Inclusion Criteria for patient with a constitutional non-sickle cell disease of the red blood cell, or an acquired red blood cell disease :
Patient age ≥ 18 years
With social care protection
With any of the following conditions :
Exclusion Criteria for all patients:
Patient age < 18 years
Subject under guardianship, or subject deprived of freedom
Linguistic or literacy status not allowing for informed consent despite patient information in "Easy to Read and Understand" format
Known history of HIV, HTLV, syphilis, or positive serology and active viral hepatitis B or C.
Additional Exclusion Criteria for healthy volunteers :
Abnormal blood count, or possible martial deficiency with ferritin levels below 50µg/l, or current treatment with hydroxycarbamide, or transfusion within 4 months prior to inclusion.
Additional Exclusion Criteria for SCD patient :
5) Treatment with hydroxycarbamide started less than 6 months ago 6) Anemia with hemoglobin level <60g/l in the absence of cardiorespiratory pathology, <70g/l in pregnancy, or in the presence of cardiorespiratory pathology that may alter the tolerance of anemia.
Additional Exclusion Criteria for patient with a constitutional non-sickle cell disease of the red blood cell, or an acquired red blood cell disease :
5) Anemia with hemoglobin level <60g/l, <70g/l in pregnancy, or in the presence of cardio-respiratory pathology that may alter the tolerance of anemia.
6) Diagnosis not finalized (in progress), or uncertain nosological framework, or diagnostic wandering.
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Study population will be patient with sickle cell disease (SCD) in comparaison of two populations : healthy volunteers (living donor of kidney) and patient with a constitutional non-sickle cell disease of the red blood cell, or an acquired red blood cell disease
| Name | Role | Phone | Extension | |
|---|---|---|---|---|
| Caroline MAKOWSKI, Md | Contact | +33476767640 | cmakowski@chu-grenoble.fr | |
| Bruna DUCOTTERD, CRA | Contact | +3376767838 | bducotterd@chu-grenoble.fr |
| Name | Affiliation | Role |
|---|---|---|
| Caroline MAKOWSKI, mD | CHU Grenoble Alpes | Principal Investigator |
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| PubMed Identifier | Type | Citation | Retractions |
|---|---|---|---|
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| 24 months |
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| 31053255 | Background | Woodcock EM, Girvan P, Eckert J, Lopez-Duarte I, Kubankova M, van Loon JJWA, Brooks NJ, Kuimova MK. Measuring Intracellular Viscosity in Conditions of Hypergravity. Biophys J. 2019 May 21;116(10):1984-1993. doi: 10.1016/j.bpj.2019.03.038. Epub 2019 Apr 8. |
| 24404073 | Background | Yaginuma T, Oliveira MS, Lima R, Ishikawa T, Yamaguchi T. Human red blood cell behavior under homogeneous extensional flow in a hyperbolic-shaped microchannel. Biomicrofluidics. 2013 Sep 24;7(5):54110. doi: 10.1063/1.4820414. eCollection 2013. |
| ID | Term |
|---|---|
| D000755 | Anemia, Sickle Cell |
| ID | Term |
|---|---|
| D000745 | Anemia, Hemolytic, Congenital |
| D000743 | Anemia, Hemolytic |
| D000740 | Anemia |
| D006402 | Hematologic Diseases |
| D006425 | Hemic and Lymphatic Diseases |
| D006453 | Hemoglobinopathies |
| D030342 | Genetic Diseases, Inborn |
| D009358 | Congenital, Hereditary, and Neonatal Diseases and Abnormalities |
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