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


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Dipartimento di Medicina Sperimentale, Sezione di Patologia Generale e Clinica, Università degli Studi di Parma, Parma, Italy.
Amelia
A
Barilli
Bianca Maria
BM
Rotoli
Rossana
R
Visigalli
Ovidio
O
Bussolati
Gian C
GC
Gazzola
Zamir
Z
Kadija
Giuseppe
G
Rodi
Francesca
F
Mariani
Maria Lorena
ML
Ruzza
Maurizio
M
Luisetti
Valeria
V
Dall'Asta
D000328:Adult; D000592:Amino Acid Metabolism, Inborn Errors / Q000150:complications*; D027241:Amino Acid Transport System y+L; D002454:Cell Differentiation; D002478:Cells, Cultured; D027301:Fusion Regulatory Protein 1, Light Chains / Q000235:genetics / Q000378:metabolism*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology* / Q000378:metabolism / Q000627:therapeutic use; D006801:Humans; D008239:Lysine / Q000378:metabolism; D016676:Macrophages, Alveolar / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D008297:Male; D009000:Monocytes / Q000276:immunology / Q000378:metabolism*; D011649:Pulmonary Alveolar Proteinosis / Q000276:immunology / Q000503:physiopathology* / Q000628:therapy; D055815:Young Adult
D027241:Amino Acid Transport System y+L; D027301:Fusion Regulatory Protein 1, Light Chains; C421261:SLC7A7 protein, human; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D008239:Lysine