An interfacial and comparative in vitro study of gastrointestinal lipases and Yarrowia lipolytica LIP2 lipase, a candidate for enzyme replacement therapy

Background: The current effective delivered dose is a quality indicator for continuous renal replacement therapy. Its periodic assessment might enable physicians to deliver personalised treatments. Yet, its quantification as by extracorporeal urea clearance (Cl) is cumbersome and thus often neglected in routine practice. The aim of this in vitro study is to demonstrate the non-inferior effectiveness of assessing the current effective delivered dose using a simpler, cheaper and faster approach based on measurement of fluoride rather than urea extracorporeal Cl. Methods: We compared urea and fluoride removal in 3 post-dilution continuous veno-venous haemofiltration (CVVH) and 3 continuous veno-venous haemodialysis (CVVHD) in vitro experimental models. Experiments ran for 180 min, using 3 L of human blood, heparin anticoagulation and a machine dose of 30 mL/kg/h. Urea and fluoride were measured in the inflow, outflow and effluent lines to compare sieving coefficients (SC), saturation coefficients (SA) and transmembrane Cls. Results: In CVVH, the median SC values were 1.06 (1.02–1.07) and 1.02 (1.01–1.04) for fluoride and urea, respectively (discrepancy of 4.3%), while transmembrane convective Cls were 31.28 (30.01–31.31) mL/kg/h and 30.30 (29–31.85) mL/kg/h (discrepancy of 3.13%), respectively. In CVVHD, the median SA values were 1.01 (0.96–1.02) and 1 (0.95–1.01) for fluoride and urea, respectively (discrepancy of 1.6%), while transmembrane dialytic Cls were 30.26 (29.52–31.32) mL/kg/h and 31.16 (30–31.75) mL/kg/h (discrepancy of –2.97%), respectively. Conclusion: Fluoride transmembrane removal was close to that observed with urea, in terms of SC, SA and transmembrane Cl. Fluoride seems as much accurate as urea in assessing the current effective delivered dose during both CVVH and CVVHD and might therefore be adopted for dose measurement. Besides accuracy, fluoride bedside assessment could present many advantages over urea, particularly in terms of availability, costs, time requirement and rapidity of assessment.

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α-1,4-Glucosidase–albumin polymers : in vitro properties and advantages for enzyme replacement therapy

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y80-056 ◽

1980 ◽

Vol 58 (3) ◽

pp. 322-325 ◽

Cited By ~ 14

Author(s):

Mark J. Poznansky ◽

Damyanti Bhardwaj

Keyword(s):

Enzyme Replacement Therapy ◽

Replacement Therapy ◽

Half Life ◽

Metabolic Diseases ◽

Average Molecular Weight ◽

Human Albumin ◽

Free Enzyme ◽

Heat Denaturation ◽

Enzyme Replacement

Soluble polymers of rat (or human) albumin and α-1,4-glucosidase are prepared using the cross-linking agent glutaraldehyde. The resulting polymer has an average molecular weight of 800 000 indicating an average composition of 12 albumin molecules for each enzyme molecule. Compared with an equivalent amount of free enzyme, the enzyme–albumin polymer has an increased resistance to heat denaturation (half-life of 15 h compared with 1 h for free enzyme at 37 °C) and to proteolysis by trypsin (half-life of 180 min compared with 10 min). The high degree of resistance to bioinactivation of the enzyme–albumin polymer is discussed in relation to requirements for enzyme replacement therapy in a range of metabolic diseases including type II glycogenosis (Pompe's disease) where α-1,4-glucosidase is the defective enzyme.

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A first approach for an evidence-based in vitro digestion method to adjust pancreatic enzyme replacement therapy in cystic fibrosis

PLoS ONE ◽

10.1371/journal.pone.0212459 ◽

2019 ◽

Vol 14 (2) ◽

pp. e0212459 ◽

Cited By ~ 6

Author(s):

Joaquim Calvo-Lerma ◽

Victoria Fornés-Ferrer ◽

Irene Peinado ◽

Ana Heredia ◽

Carmen Ribes-Koninckx ◽

...

Keyword(s):

Cystic Fibrosis ◽

Enzyme Replacement Therapy ◽

Replacement Therapy ◽

Pancreatic Enzyme ◽

In Vitro Digestion ◽

Evidence Based ◽

Digestion Method ◽

Enzyme Replacement ◽

Pancreatic Enzyme Replacement Therapy

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The effects of long-term migalastat treatment in Fabry disease patients previously treated with enzyme replacement therapy who have migalastat-amenable variants with low alpha-galactosidase A response in the in vitro migalastat amenability assay

Molecular Genetics and Metabolism ◽

10.1016/j.ymgme.2018.12.272 ◽

2019 ◽

Vol 126 (2) ◽

pp. S108 ◽

Cited By ~ 1

Author(s):

Kathleen Nicholls ◽

Iacopo Olivotto ◽

Toya Ohashi ◽

Hadis Williams ◽

Vipul Jain ◽

...

Keyword(s):

Enzyme Replacement Therapy ◽

Fabry Disease ◽

Replacement Therapy ◽

Enzyme Replacement ◽

Previously Treated ◽

Alpha Galactosidase

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Intracerebroventricular enzyme replacement therapy with β-galactosidase reverses brain pathologies due to GM1 gangliosidosis in mice

Journal of Biological Chemistry ◽

10.1074/jbc.ra119.009811 ◽

2019 ◽

Vol 295 (39) ◽

pp. 13532-13555 ◽

Cited By ~ 5

Author(s):

Joseph C. Chen ◽

Amanda R. Luu ◽

Nathan Wise ◽

Rolando De Angelis ◽

Vishal Agrawal ◽

...

Keyword(s):

Enzyme Replacement Therapy ◽

Replacement Therapy ◽

Chinese Hamster Ovary Cells ◽

Chinese Hamster ◽

Lysosomal Storage ◽

Gm1 Gangliosidosis ◽

Enzyme Replacement ◽

Mannose 6 Phosphate ◽

The Unfolded Protein Response

Autosomal recessive mutations in the galactosidase β1 (GLB1) gene cause lysosomal β-gal deficiency, resulting in accumulation of galactose-containing substrates and onset of the progressive and fatal neurodegenerative lysosomal storage disease, GM1 gangliosidosis. Here, an enzyme replacement therapy (ERT) approach in fibroblasts from GM1 gangliosidosis patients with recombinant human β-gal (rhβ-gal) produced in Chinese hamster ovary cells enabled direct and precise rhβ-gal delivery to acidified lysosomes. A single, low dose (3 nm) of rhβ-gal was sufficient for normalizing β-gal activity and mediating substrate clearance for several weeks. We found that rhβ-gal uptake by the fibroblasts is dose-dependent and saturable and can be competitively inhibited by mannose 6-phosphate, suggesting cation-independent, mannose 6-phosphate receptor–mediated endocytosis from the cell surface. A single intracerebroventricularly (ICV) administered dose of rhβ-gal (100 μg) resulted in broad bilateral biodistribution of rhβ-gal to critical regions of pathology in a mouse model of GM1 gangliosidosis. Weekly ICV dosing of rhβ-gal for 8 weeks substantially reduced brain levels of ganglioside and oligosaccharide substrates and reversed well-established secondary neuropathology. Of note, unlike with the ERT approach, chronic lentivirus-mediated GLB1 overexpression in the GM1 gangliosidosis patient fibroblasts caused accumulation of a prelysosomal pool of β-gal, resulting in activation of the unfolded protein response and endoplasmic reticulum stress. This outcome was unsurprising in light of our in vitro biophysical findings for rhβ-gal, which include pH-dependent and concentration-dependent stability and dynamic self-association. Collectively, our results highlight that ICV-ERT is an effective therapeutic intervention for managing GM1 gangliosidosis potentially more safely than with gene therapy approaches.

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