Production of Human Acid-Alpha Glucosidase With a Paucimannose Structure by Glycoengineered Arabidopsis Cell Culture

Sariyatun, Ratna; Florence,; Kajiura, Hiroyuki; Ohashi, Takao; Misaki, Ryo; Fujiyama, Kazuhito

doi:10.3389/fpls.2021.703020

Cited by 5 publications

(8 citation statements)

References 56 publications

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“…However, the use of this platform is hindered by the generation of plant-specific N -glycans, the inability to produce essential N -glycans for cellular delivery of biopharmaceutics and low productivity. Sariyatun et al ( 117 ) producted a hGAA for ERT with a paucimannose structure-Man3GlcNAc2 (M3), without the plant-specific N -glycans by glycoengineered in Arabidopsis alg3 cell culture.…”

Section: Discussionmentioning

confidence: 99%

Preclinical studies with ground germinated barley (GGB) for oral enzyme replacement therapy (Oral-ERT) in Pompe disease knockout mice

Martiniuk,

Mack,

Martiniuk

et al. 2023

Preprint

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Genetic deficiency of lysosomal acid maltase or acid α-glucosidase (GAA) results in the orphan disease known as glycogen storage disease type II or acid maltase deficiency (AMD) or Pompe disease (PD), encompassing at least four clinical subtypes of varying severity. PD results from mutations in theGAAgene and deficient GAA activity, resulting in the accumulation of glycogen in tissues (primarily muscle) and characterized by progressive skeletal muscle weakness and respiratory insufficiency. The current approved enzyme replacement therapy (ERT) for PD is via intravenous infusion of a recombinant human GAA (rhGAA) secreted by CHO cells (Myozyme, Sanofi-Genzyme) given once every 2 weeks and has shown varying efficacy in patients. Although the current ERT has proven to be very efficient in rescuing cardiac abnormalities and extending the life span of infants, the response in skeletal muscle is variable. In late-onset patients, only mild improvements in motor and respiratory functions have been achieved and the current ERT is unsatisfactory in the reversal of skeletal muscle pathology. Additional challenges for ERT include insufficient targeting/uptake of enzyme into disease-relevant tissues, poor tolerability due to severe ERT-mediated anaphylactic and immunologic reactions and the prohibitively high cost of lifelong ERT ($250-500K/year adult patient). A consensus at a Nov.-2019 US Acid Maltase Deficiency Association conference suggested that a multi-pronged approach including gene therapy, diet, exercise, etc. must be evaluated for a successful treatment. Our objective is to develop an innovative and affordable approach via barley GAA (bGAA) from ground germinated barley (GGB) or liquid GGB (L-GGB) for Oral-ERT for PD or as a daily supplement to Myozyme. To this end, we hypothesize that a bGAA produced in germinated barley can be ingested daily that allows the maintenance of a therapeutic level of enzyme. We have shown in extensive preliminary data thatGGBorL-GGBwas (1) enzymatically active, (2) was taken up by GAA KO mice and human WBCs to reverse the enzyme defect that was blocked by mannose-6-phosphate, (3) hydrolyzed glycogen, (4) increased significant changes in the clinical phenotype towards the WT levels in GAA KO mice dose-dependently, (5) taken up by PD myoblasts, lymphoid/fibroblasts cells to reverse the defect, (6) bGAA was ∼70kD, (7) Km, Vmax, pH optima, inhibitors and kinetics was similar to human placental GAA and an rhGAA and (8) was strain specific.

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Section: Discussionmentioning

confidence: 99%

Preclinical studies with ground germinated barley (GGB) for oral enzyme replacement therapy (Oral-ERT) in Pompe disease knockout mice

Martiniuk,

Mack,

Martiniuk

et al. 2023

Preprint

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“…Interestingly, some mammal native SPs can also be recognized and accurately processed in plant cells. For instance, the native SP of human growth hormone (hGH) led to the correct processing and apoplast targeting of active hGH in N. benthamiana [88], and the native human acid-alpha glucosidase (GAA) SP was shown to be fully capable of facilitating the entrance of GAA into the ER lumen, leading to the eventual secretion into the Arabidopsis cell culture medium [89]. The native murine mAb24 heavy chain-derived signal peptide (LPH) was also frequently used for the expression of viral glycoproteins [90].…”

Section: Apoplastmentioning

confidence: 99%

“…Previous results have shown that the Arabidopsis HEXO mutant and N. benthamiana HEXO RNAi knockout line effectively prevent the formation of the paucimannosidic MMXF N-glycan [281,282]. However, the paucimannosidic structure MM (Man 3 GlcNAc 2 or M3) has been reported to be indispensable and superior in mannose receptor (MR)-mediated cellular delivery of biopharmaceutics because of its high MR-binding efficiency and relatively low clearance rate [89]. Carrot suspension cell-produced Elelyso (i.e., recombinant human β-glucocerebrosidase) is an example of a recombinant protein with the paucimannosidic type N-glycan [131].…”

Section: Elimination Of Plant Specific N-glycan Structuresmentioning

confidence: 99%

“…The human acid-alpha glucosidase (GAA) is also a therapeutic protein used for enzyme replacement therapy of Pompe disease. Using the glycoengineered Arabidopsis cell culture derived from a mutant plant lacking functional α1,3-mannosyltransferase (ALG3), the enzyme involved in the early assembly of the oligosaccharide precursor, the recombinant human GAA was produced with MM comprising the majority of the N-glycan structures [89]. Therefore, for therapeutic proteins designed for cellular delivery through MR, such as those used for enzyme replacement therapies, the paucimannosidic N-glycan modification is considered to be beneficial.…”

Section: Elimination Of Plant Specific N-glycan Structuresmentioning

confidence: 99%

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Improving Protein Quantity and Quality—The Next Level of Plant Molecular Farming

Liu

Timko

2022

IJMS

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Plants offer several unique advantages in the production of recombinant pharmaceuticals for humans and animals. Although numerous recombinant proteins have been expressed in plants, only a small fraction have been successfully put into use. The hugely distinct expression systems between plant and animal cells frequently cause insufficient yield of the recombinant proteins with poor or undesired activity. To overcome the issues that greatly constrain the development of plant-produced pharmaceuticals, great efforts have been made to improve expression systems and develop alternative strategies to increase both the quantity and quality of the recombinant proteins. Recent technological revolutions, such as targeted genome editing, deconstructed vectors, virus-like particles, and humanized glycosylation, have led to great advances in plant molecular farming to meet the industrial manufacturing and clinical application standards. In this review, we discuss the technological advances made in various plant expression platforms, with special focus on the upstream designs and milestone achievements in improving the yield and glycosylation of the plant-produced pharmaceutical proteins.

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“…On the other hand, the secretion mechanisms, which are highly conserved among eukaryotes, allow SPs of a heterologous origin—for example, animal SPs in a plant expression system—to be employed for targeting recombinant proteins to the apoplast. In particular, the authentic SP of human growth hormone ensures the correct processing and translocation of human growth hormone to the apoplast of N. benthamiana [ 16 , 119 ], as does the authentic SP of human acid α-glucosidase in A. thaliana cell culture [ 120 ]. Various SPs yield different secretion efficiency levels; hence, these peptides should be individually selected for each expression system.…”

Section: Intracellular Transport Of Proteins To the Er And Its Deriva...mentioning

confidence: 99%

Increasing the Efficiency of the Accumulation of Recombinant Proteins in Plant Cells: The Role of Transport Signal Peptides

Розов

Дейнеко

2022

Plants

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The problem with increasing the yield of recombinant proteins is resolvable using different approaches, including the transport of a target protein to cell compartments with a low protease activity. In the cell, protein targeting involves short-signal peptide sequences recognized by intracellular protein transport systems. The main systems of the protein transport across membranes of the endoplasmic reticulum and endosymbiotic organelles are reviewed here, as are the major types and structure of the signal sequences targeting proteins to the endoplasmic reticulum and its derivatives, to plastids, and to mitochondria. The role of protein targeting to certain cell organelles depending on specific features of recombinant proteins and the effect of this targeting on the protein yield are discussed, in addition to the main directions of the search for signal sequences based on their primary structure. This knowledge makes it possible not only to predict a protein localization in the cell but also to reveal the most efficient sequences with potential biotechnological utility.

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Production of Human Acid-Alpha Glucosidase With a Paucimannose Structure by Glycoengineered Arabidopsis Cell Culture

Cited by 5 publications

References 56 publications

Preclinical studies with ground germinated barley (GGB) for oral enzyme replacement therapy (Oral-ERT) in Pompe disease knockout mice

Preclinical studies with ground germinated barley (GGB) for oral enzyme replacement therapy (Oral-ERT) in Pompe disease knockout mice

Improving Protein Quantity and Quality—The Next Level of Plant Molecular Farming

Increasing the Efficiency of the Accumulation of Recombinant Proteins in Plant Cells: The Role of Transport Signal Peptides

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