Proteomic Analysis in Morquio A Cells Treated with Immobilized Enzymatic Replacement Therapy on Nanostructured Lipid Systems

Abstract

Morquio A syndrome, or mucopolysaccharidosis type IVA (MPS IVA), is a lysosomal storage disease due to mutations in the N-acetylgalactosamine-6-sulfatase (GALNS) gene. Systemic skeletal dysplasia and the related clinical features of MPS IVA are due to disruption of cartilage and its extracellular matrix, leading to an imbalance of growth. Enzyme replacement therapy (ERT) with recombinant human GALNS, alpha elosulfase, provides a systemic treatment. However, this therapy has a limited impact on skeletal dysplasia because the infused enzyme cannot penetrate cartilage and bone. Therefore, an alternative therapeutic approach to reach the cartilage is an unmet challenge. We have developed a new drug delivery system based on a nanostructure lipid carrier with the capacity to immobilize enzymes used for ERT and to target the lysosomes. This study aimed to assess the effect of the encapsulated enzyme in this new delivery system, using in vitro proteomic technology. We found a greater internalization of the enzyme carried by nanoparticles inside the cells and an improvement of cellular protein routes previously impaired by the disease, compared with conventional ERT. This is the first qualitative and quantitative proteomic assay that demonstrates the advantages of a new delivery system to improve the MPS IVA ERT.

Characterization of New Proteomic Biomarker Candidates in Mucopolysaccharidosis Type IVA

“…Interestingly, the inflammation pathway was activated only in samples from untreated MPS IV patients. In a previous study, we demonstrated qualitative and quantitative changes in proteins expressed by fibroblasts from MPS IVA patients following ERT and encapsulated ERT in nanoparticles [ 61 ].…”

“…We postulate that these proteins expressed in leukocytes may also be expressed in bone cells, given that macrophages and osteoclasts originate from the same cell line and may share similar proteins [ 65 ]. In a previous proteomic analysis of fibroblasts, we observed upregulation of TRFL in untreated MPS IVA patients and upregulation of GANAB in ERT-treated versus untreated MPS IVA patients [ 61 ].…”

“…This specific pyrimidine nuclease shows a slight preference for cytotoxin and helminthotoxin, is located in the lysosome, and is described as a chemotactic factor for dendritic cells. Several other proteins showed partial or complete normalization of expression after ERT, including proteins implicated in the pyruvate metabolic pathway (LDHA, LDHB) and in cytoskeletal organization (MOES) [ 61 ]. Inflammation-associated proteins that were dysregulated in untreated patients included OLFM4, TGFB1, THAS, PGH1, CAP7, andCHIT1 [ 70 , 75 ].…”

Mucopolysaccharidosis IVA: Diagnosis, Treatment, and Management

“…This cholesterol reserve blocks a complex necessity to direct the lysosome [155]. Regarding of MPS IVA, there has been little study on the effect of nanomedicine [142,156]. In 2019, Alvarez et al developed a NLC containing elosulfase alfa and studied its efficacy using in vitro cellular models and its in vivo biodistribution in mice.…”

A Case Report of a Japanese Boy with Morquio A Syndrome: Effects of Enzyme Replacement Therapy Initiated at the Age of 24 Months

“…Monitoring of the elosulfase alfa dose by using the urine KS will facilitate more precise management, although it was performed only once in this patient, where the urine KS decreased by nearly 40% after the ERT. Research is underway regarding new therapies (e.g., pentosane polysulfate) and improvements to existing drugs (e.g., immobilized ERT on nanostructured lipid systems) [26,27]. It is expected that a combination of these drugs can achieve better prognosis for patients with MPS IVA.…”