In Lysinuric Protein Intolerance system y+L activity is defective in monocytes and in GM-CSF-differentiated macrophages

• Published in: Orphanet journal of rare diseases Vol. 5; no. 1; p. 32
• Main Authors: Barilli, Amelia, Rotoli, Bianca Maria, Visigalli, Rossana, Bussolati, Ovidio, Gazzola, Gian C, Kadija, Zamir, Rodi, Giuseppe, Mariani, Francesca, Ruzza, Maria Lorena, Luisetti, Maurizio, Dall'Asta, Valeria
• Format: Journal Article
• Language: English
• Published: England BioMed Central 26.11.2010; BioMed Central Ltd; BMC
• Subjects: Adult; Amino Acid Metabolism, Inborn Errors - complications; Blood; Cell Differentiation; Cells, Cultured; Disease; Fusion Regulatory Protein 1, Light Chains - genetics; Fusion Regulatory Protein 1, Light Chains - metabolism; Genes; Granulocyte-Macrophage Colony-Stimulating Factor - immunology; Granulocyte-Macrophage Colony-Stimulating Factor - metabolism; Granulocyte-Macrophage Colony-Stimulating Factor - therapeutic use; Humans; Lungs; Lysine - metabolism; Macrophages, Alveolar - cytology; Macrophages, Alveolar - immunology; Macrophages, Alveolar - metabolism; Male; Medical research; Metabolic disorders; Monocytes - immunology; Monocytes - metabolism; Mutation; Pathogenesis; Pulmonary Alveolar Proteinosis - immunology; Pulmonary Alveolar Proteinosis - physiopathology; Pulmonary Alveolar Proteinosis - therapy; Rare diseases; Young Adult
• ISSN: 1750-1172; 1750-1172
• DOI: 10.1186/1750-1172-5-32

In the recessive aminoaciduria Lysinuric Protein Intolerance (LPI), mutations of SLC7A7/y+LAT1 impair system y+L transport activity for cationic amino acids. A severe complication of LPI is a form of Pulmonary Alveolar Proteinosis (PAP), in which alveolar spaces are filled with lipoproteinaceous material because of the impaired surfactant clearance by resident macrophages. The pathogenesis of LPI-associated PAP remains still obscure. The present study investigates for the first time the expression and function of y+LAT1 in monocytes and macrophages isolated from a patient affected by LPI-associated PAP. A comparison with mesenchymal cells from the same subject has been also performed. Monocytes from peripheral blood were isolated from a 21-year-old patient with LPI. Alveolar macrophages and fibroblastic-like mesenchymal cells were obtained from a whole lung lavage (WLL) performed on the same patient. System y+L activity was determined measuring the 1-min uptake of [3H]-arginine under discriminating conditions. Gene expression was evaluated through qRT-PCR. We have found that: 1) system y+L activity is markedly lowered in monocytes and alveolar macrophages from the LPI patient, because of the prevailing expression of SLC7A7/y+LAT1 in these cells; 2) on the contrary, fibroblasts isolated from the same patient do not display the transport defect due to compensation by the SLC7A6/y+LAT2 isoform; 3) in both normal and LPI monocytes, GM-CSF induces the expression of SLC7A7, suggesting that the gene is a target of the cytokine; 4) GM-CSF-induced differentiation of LPI monocytes is comparable to that of normal cells, demonstrating that GM-CSF signalling is unaltered; 5) general and respiratory conditions of the patient, along with PAP-associated parameters, markedly improved after GM-CSF therapy through aerosolization. Monocytes and macrophages, but not fibroblasts, derived from a LPI patient clearly display the defect in system y+L-mediated arginine transport. The different transport phenotypes are referable to the relative levels of expression of SLC7A7 and SLC7A6. Moreover, the expression of SLC7A7 is regulated by GM-CSF in monocytes, pointing to a role of y+LAT1 in the pathogenesis of LPI associated PAP.