Laura López de Frutos1, Jorge J Cebolla2, Pilar Irún3, Ralf Köhler4, Pilar Giraldo5. 1. Instituto de Investigación Sanitaria Aragón (IIS Aragón) GIIS-012, Unidad de Investigación Traslacional, Hospital Universitario Miguel Servet, Zaragoza 50009, Spain; Fundación para el Estudio y la Terapéutica de la Enfermedad de Gaucher y Otras Lisosomales (FEETEG), Zaragoza 50009, Spain. Electronic address: l_lopezdefrutos@sehh.es. 2. Instituto de Investigación Sanitaria Aragón (IIS Aragón) GIIS-012, Unidad de Investigación Traslacional, Hospital Universitario Miguel Servet, Zaragoza 50009, Spain; Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER) U-752, Zaragoza 50009, Spain; Universidad de Zaragoza, Departamento de Bioquímica, Biología Molecular y Celular, Zaragoza 50009, Spain. 3. Instituto de Investigación Sanitaria Aragón (IIS Aragón) GIIS-012, Unidad de Investigación Traslacional, Hospital Universitario Miguel Servet, Zaragoza 50009, Spain; Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER) U-752, Zaragoza 50009, Spain. 4. Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER) U-752, Zaragoza 50009, Spain; Fundación Agencia Aragonesa para la Investigación y el Desarrollo (ARAID), Zaragoza 50009, Spain. 5. Instituto de Investigación Sanitaria Aragón (IIS Aragón) GIIS-012, Unidad de Investigación Traslacional, Hospital Universitario Miguel Servet, Zaragoza 50009, Spain; Fundación para el Estudio y la Terapéutica de la Enfermedad de Gaucher y Otras Lisosomales (FEETEG), Zaragoza 50009, Spain; Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER) U-752, Zaragoza 50009, Spain.
Abstract
BACKGROUND: Erythrocyte volume regulation and membrane elasticity are essential for adaptation to osmotic and mechanical stress, and life span. Here, we evaluated whether defective cholesterol trafficking caused by the rare lysosomal storages diseases (LSDs), Niemann-Pick type C (NPC) and Lysosomal acid lipase (LAL) deficiency (LALD) impairs these properties. Moreover, we tested whether measurements of cholesterol membrane content and osmotic resistance serve as a screening test for these LSDs. METHODS: Patients were genotyped for mutations in NPC1, NPC2, or LIPA genes. We measured LSD plasma biomarkers and LAL activity. Red blood cells (RBC) membrane cholesterol content was evaluated in 73 subjects. Osmotic resistance tests (ORT) were conducted in 121 blood samples from LSD suspected patients and controls. RESULTS: We did not find statistically significant differences between RBC cholesterol content between subjects and controls. However, the ORT, particularly at 0.49% (w/v) hypotonic sodium chloride solution, revealed a significant higher osmotic resistance in LSDs patients than in controls. We established a cut-off value of ≤51% of haemolysis with sensibility and specificity values of 80% and 70%, respectively. CONCLUSIONS: NPC and LALD do not alter cholesterol content in the RBC membrane but increase osmotic resistance. Therefore, ORT serves as screening test for the studied LSDs.
BACKGROUND: Erythrocyte volume regulation and membrane elasticity are essential for adaptation to osmotic and mechanical stress, and life span. Here, we evaluated whether defective cholesterol trafficking caused by the rare lysosomal storages diseases (LSDs), Niemann-Pick type C (NPC) and Lysosomal acid lipase (LAL) deficiency (LALD) impairs these properties. Moreover, we tested whether measurements of cholesterol membrane content and osmotic resistance serve as a screening test for these LSDs. METHODS:Patients were genotyped for mutations in NPC1, NPC2, or LIPA genes. We measured LSD plasma biomarkers and LAL activity. Red blood cells (RBC) membrane cholesterol content was evaluated in 73 subjects. Osmotic resistance tests (ORT) were conducted in 121 blood samples from LSD suspected patients and controls. RESULTS: We did not find statistically significant differences between RBC cholesterol content between subjects and controls. However, the ORT, particularly at 0.49% (w/v) hypotonicsodium chloride solution, revealed a significant higher osmotic resistance in LSDs patients than in controls. We established a cut-off value of ≤51% of haemolysis with sensibility and specificity values of 80% and 70%, respectively. CONCLUSIONS:NPC and LALD do not alter cholesterol content in the RBC membrane but increase osmotic resistance. Therefore, ORT serves as screening test for the studied LSDs.
Authors: Maria Aparecida Knychala; Mario da Silva Garrote-Filho; Breno Batista da Silva; Samantha Neves de Oliveira; Sarah Yasminy Luz; Manuela Ortega Marques Rodrigues; Nilson Penha-Silva Journal: J Cell Mol Med Date: 2021-02-16 Impact factor: 5.310