Defining the pathway for Tat-mediated delivery of β-glucuronidase in cultured cells and MPS VII mice

Koji O. Orii; Jeffrey H. Grubb; Carole Vogler; Beth Levy; Yun Tan; Kamelia Markova; Beverly L. Davidson; Q. Mao; Tadao Orii; Naomi Kondo; William S. Sly

doi:10.1016/j.ymthe.2005.02.031

Defining the pathway for Tat-mediated delivery of β-glucuronidase in cultured cells and MPS VII mice

Koji O. Orii, Jeffrey H. Grubb, Carole Vogler, Beth Levy, Yun Tan, Kamelia Markova, Beverly L. Davidson, Q. Mao, Tadao Orii, Naomi Kondo, William S. Sly^*

^*Corresponding author for this work

Pathology

Research output: Contribution to journal › Article › peer-review

37 Scopus citations

Abstract

We used recombinant forms of human β-glucuronidase (GUS) purified from secretions from stably transfected CHO cells to compare the native enzyme to a GUS-Tat C-terminal fusion protein containing the 11-amino-acid HIV Tat protein transduction domain for: (1) susceptibility to endocytosis by cultured cells, (2) rate of clearance following intravenous infusion, and (3) tissue distribution and effectiveness in clearing lysosomal storage following infusion in the MPS VII mouse. We found: (1) Native GUS was more efficiently taken up by cultured human fibroblasts and its endocytosis was exclusively mediated by the M6P receptor. The GUS-Tat fusion protein showed only 30-50% as much M6P-receptor-mediated uptake, but also was taken up by adsorptive endocytosis through binding of the positively charged Tat peptide to cell surface proteoglycans. (2) GUS-Tat was less rapidly cleared from the circulation in the rat (t_1/2 = 13 min vs 7 min). (3) Delivery to most tissues of the MPS VII mouse was similar, but GUS-Tat was more efficiently delivered to kidney. Histology showed that GUS-Tat more efficiently reduced storage in renal tubules, retina, and bone. These studies demonstrate that Tat modification can extend the range of tissues corrected by infused enzyme.

Original language	English (US)
Pages (from-to)	345-352
Number of pages	8
Journal	Molecular Therapy
Volume	12
Issue number	2
DOIs	https://doi.org/10.1016/j.ymthe.2005.02.031
State	Published - Aug 2005

Funding

This work was supported by a fellowship grant to K.O.O. from the National Organization for Rare Disorders, Inc., and by National Institutes of Health Grant GM34182 to W.S.S. We gratefully acknowledge Abdul Waheed, Ph.D. (Saint Louis University School of Medicine), for discussion and editing of the manuscript and Dr. George Vogler for expert assistance with the study of enzyme clearance from plasma in the rat.

Keywords

Adsorptive endocytosis
Enzyme replacement therapy
Lysosomal storage disease
MPS VII mouse
Mannose 6-phosphate
Receptor-mediated endocytosis
Sly syndrome
Tat peptide
β-glucuronidase

ASJC Scopus subject areas

Drug Discovery
Genetics
Molecular Medicine
Molecular Biology
Pharmacology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.ymthe.2005.02.031

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@article{3cc9c547432044f699d452979535c2ef,

title = "Defining the pathway for Tat-mediated delivery of β-glucuronidase in cultured cells and MPS VII mice",

abstract = "We used recombinant forms of human β-glucuronidase (GUS) purified from secretions from stably transfected CHO cells to compare the native enzyme to a GUS-Tat C-terminal fusion protein containing the 11-amino-acid HIV Tat protein transduction domain for: (1) susceptibility to endocytosis by cultured cells, (2) rate of clearance following intravenous infusion, and (3) tissue distribution and effectiveness in clearing lysosomal storage following infusion in the MPS VII mouse. We found: (1) Native GUS was more efficiently taken up by cultured human fibroblasts and its endocytosis was exclusively mediated by the M6P receptor. The GUS-Tat fusion protein showed only 30-50% as much M6P-receptor-mediated uptake, but also was taken up by adsorptive endocytosis through binding of the positively charged Tat peptide to cell surface proteoglycans. (2) GUS-Tat was less rapidly cleared from the circulation in the rat (t1/2 = 13 min vs 7 min). (3) Delivery to most tissues of the MPS VII mouse was similar, but GUS-Tat was more efficiently delivered to kidney. Histology showed that GUS-Tat more efficiently reduced storage in renal tubules, retina, and bone. These studies demonstrate that Tat modification can extend the range of tissues corrected by infused enzyme.",

keywords = "Adsorptive endocytosis, Enzyme replacement therapy, Lysosomal storage disease, MPS VII mouse, Mannose 6-phosphate, Receptor-mediated endocytosis, Sly syndrome, Tat peptide, β-glucuronidase",

author = "Orii, {Koji O.} and Grubb, {Jeffrey H.} and Carole Vogler and Beth Levy and Yun Tan and Kamelia Markova and Davidson, {Beverly L.} and Q. Mao and Tadao Orii and Naomi Kondo and Sly, {William S.}",

note = "Funding Information: This work was supported by a fellowship grant to K.O.O. from the National Organization for Rare Disorders, Inc., and by National Institutes of Health Grant GM34182 to W.S.S. We gratefully acknowledge Abdul Waheed, Ph.D. (Saint Louis University School of Medicine), for discussion and editing of the manuscript and Dr. George Vogler for expert assistance with the study of enzyme clearance from plasma in the rat.",

year = "2005",

month = aug,

doi = "10.1016/j.ymthe.2005.02.031",

language = "English (US)",

volume = "12",

pages = "345--352",

journal = "Molecular Therapy",

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T1 - Defining the pathway for Tat-mediated delivery of β-glucuronidase in cultured cells and MPS VII mice

AU - Orii, Koji O.

AU - Grubb, Jeffrey H.

AU - Vogler, Carole

AU - Levy, Beth

AU - Tan, Yun

AU - Markova, Kamelia

AU - Davidson, Beverly L.

AU - Mao, Q.

AU - Orii, Tadao

AU - Kondo, Naomi

AU - Sly, William S.

N1 - Funding Information: This work was supported by a fellowship grant to K.O.O. from the National Organization for Rare Disorders, Inc., and by National Institutes of Health Grant GM34182 to W.S.S. We gratefully acknowledge Abdul Waheed, Ph.D. (Saint Louis University School of Medicine), for discussion and editing of the manuscript and Dr. George Vogler for expert assistance with the study of enzyme clearance from plasma in the rat.

PY - 2005/8

Y1 - 2005/8

N2 - We used recombinant forms of human β-glucuronidase (GUS) purified from secretions from stably transfected CHO cells to compare the native enzyme to a GUS-Tat C-terminal fusion protein containing the 11-amino-acid HIV Tat protein transduction domain for: (1) susceptibility to endocytosis by cultured cells, (2) rate of clearance following intravenous infusion, and (3) tissue distribution and effectiveness in clearing lysosomal storage following infusion in the MPS VII mouse. We found: (1) Native GUS was more efficiently taken up by cultured human fibroblasts and its endocytosis was exclusively mediated by the M6P receptor. The GUS-Tat fusion protein showed only 30-50% as much M6P-receptor-mediated uptake, but also was taken up by adsorptive endocytosis through binding of the positively charged Tat peptide to cell surface proteoglycans. (2) GUS-Tat was less rapidly cleared from the circulation in the rat (t1/2 = 13 min vs 7 min). (3) Delivery to most tissues of the MPS VII mouse was similar, but GUS-Tat was more efficiently delivered to kidney. Histology showed that GUS-Tat more efficiently reduced storage in renal tubules, retina, and bone. These studies demonstrate that Tat modification can extend the range of tissues corrected by infused enzyme.

AB - We used recombinant forms of human β-glucuronidase (GUS) purified from secretions from stably transfected CHO cells to compare the native enzyme to a GUS-Tat C-terminal fusion protein containing the 11-amino-acid HIV Tat protein transduction domain for: (1) susceptibility to endocytosis by cultured cells, (2) rate of clearance following intravenous infusion, and (3) tissue distribution and effectiveness in clearing lysosomal storage following infusion in the MPS VII mouse. We found: (1) Native GUS was more efficiently taken up by cultured human fibroblasts and its endocytosis was exclusively mediated by the M6P receptor. The GUS-Tat fusion protein showed only 30-50% as much M6P-receptor-mediated uptake, but also was taken up by adsorptive endocytosis through binding of the positively charged Tat peptide to cell surface proteoglycans. (2) GUS-Tat was less rapidly cleared from the circulation in the rat (t1/2 = 13 min vs 7 min). (3) Delivery to most tissues of the MPS VII mouse was similar, but GUS-Tat was more efficiently delivered to kidney. Histology showed that GUS-Tat more efficiently reduced storage in renal tubules, retina, and bone. These studies demonstrate that Tat modification can extend the range of tissues corrected by infused enzyme.

KW - Adsorptive endocytosis

KW - Enzyme replacement therapy

KW - Lysosomal storage disease

KW - MPS VII mouse

KW - Mannose 6-phosphate

KW - Receptor-mediated endocytosis

KW - Sly syndrome

KW - Tat peptide

KW - β-glucuronidase

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U2 - 10.1016/j.ymthe.2005.02.031

DO - 10.1016/j.ymthe.2005.02.031

M3 - Article

C2 - 16043103

AN - SCOPUS:22644447758

SN - 1525-0016

VL - 12

SP - 345

EP - 352

JO - Molecular Therapy

JF - Molecular Therapy

IS - 2

ER -

Defining the pathway for Tat-mediated delivery of β-glucuronidase in cultured cells and MPS VII mice

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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