Expression of β-glucuronidase on the surface of bacteria enhances activation of glucuronide prodrugs

C. M. Cheng; F. M. Chen; Y. L. Lu; Shey-Cherng Tzou; J. Y. Wang; C. H. Kao; Kuang-Wen Liao; T. C. Cheng; C. H. Chuang; B. M. Chen; S. Roffler; T. L. Cheng

doi:10.1038/cgt.2013.17

Expression of β-glucuronidase on the surface of bacteria enhances activation of glucuronide prodrugs

C. M. Cheng, F. M. Chen, Y. L. Lu, Shey-Cherng Tzou, J. Y. Wang, C. H. Kao, Kuang-Wen Liao, T. C. Cheng, C. H. Chuang, B. M. Chen, S. Roffler^*, T. L. Cheng

^*Corresponding author for this work

Department of Biological Science and Technology

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

15 Scopus citations

Abstract

Extracellular activation of hydrophilic glucuronide prodrugs by β-glucuronidase (βG) was examined to increase the therapeutic efficacy of bacteria-directed enzyme prodrug therapy (BDEPT). βG was expressed on the surface of Escherichia coli by fusion to either the bacterial autotransporter protein Adhesin (membrane βG (mβG)/AIDA) or the lipoprotein (lpp) outermembrane protein A (mβG/lpp). Both mβG/AIDA and mβG/lpp were expressed on the bacterial surface, but only mβG/AIDA displayed enzymatic activity. The rate of substrate hydrolysis by mβG/AIDA-BL21cells was 2.6-fold greater than by pβG-BL21 cells, which express periplasmic βG. Human colon cancer HCT116 cells that were incubated with mβG/AIDA-BL21 bacteria were sensitive to a glucuronide prodrug (p-hydroxy aniline mustard β-D-glucuronide, HAMG) with an half maximal inhibitory concentration (IC₅₀) value of 226.53±45.4 μM, similar to the IC 50 value of the active drug (p-hydroxy aniline mustard, pHAM; 70.6±6.75 μM), indicating that mβG/AIDA on BL21 bacteria could rapidly and efficiently convert HAMG to an active anticancer agent. These results suggest that surface display of functional βG on bacteria can enhance the hydrolysis of glucuronide prodrugs and may increase the effectiveness of BDEPT.

Original language	English
Pages (from-to)	276-281
Number of pages	6
Journal	Cancer Gene Therapy
Volume	20
Issue number	5
DOIs	https://doi.org/10.1038/cgt.2013.17
State	Published - May 2013

Keywords

Autotransporter protein adhesin
Bacteria-directed enzyme prodrug therapy
Extracellular activation
Surface display
β-glucuronidase

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10.1038/cgt.2013.17

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@article{6240a13597f14b60a9acccfd701f29bd,

title = "Expression of β-glucuronidase on the surface of bacteria enhances activation of glucuronide prodrugs",

abstract = "Extracellular activation of hydrophilic glucuronide prodrugs by β-glucuronidase (βG) was examined to increase the therapeutic efficacy of bacteria-directed enzyme prodrug therapy (BDEPT). βG was expressed on the surface of Escherichia coli by fusion to either the bacterial autotransporter protein Adhesin (membrane βG (mβG)/AIDA) or the lipoprotein (lpp) outermembrane protein A (mβG/lpp). Both mβG/AIDA and mβG/lpp were expressed on the bacterial surface, but only mβG/AIDA displayed enzymatic activity. The rate of substrate hydrolysis by mβG/AIDA-BL21cells was 2.6-fold greater than by pβG-BL21 cells, which express periplasmic βG. Human colon cancer HCT116 cells that were incubated with mβG/AIDA-BL21 bacteria were sensitive to a glucuronide prodrug (p-hydroxy aniline mustard β-D-glucuronide, HAMG) with an half maximal inhibitory concentration (IC50) value of 226.53±45.4 μM, similar to the IC 50 value of the active drug (p-hydroxy aniline mustard, pHAM; 70.6±6.75 μM), indicating that mβG/AIDA on BL21 bacteria could rapidly and efficiently convert HAMG to an active anticancer agent. These results suggest that surface display of functional βG on bacteria can enhance the hydrolysis of glucuronide prodrugs and may increase the effectiveness of BDEPT.",

keywords = "Autotransporter protein adhesin, Bacteria-directed enzyme prodrug therapy, Extracellular activation, Surface display, β-glucuronidase",

author = "Cheng, {C. M.} and Chen, {F. M.} and Lu, {Y. L.} and Shey-Cherng Tzou and Wang, {J. Y.} and Kao, {C. H.} and Kuang-Wen Liao and Cheng, {T. C.} and Chuang, {C. H.} and Chen, {B. M.} and S. Roffler and Cheng, {T. L.}",

note = "Funding Information: This work was supported by grants from the National Research Program for Biopharmaceuticals, National Science Council, Taipei, Taiwan (NSC101-2325-B-037-001, NSC101-2321-B-037-001, NSC101-2313-B-022-001), the Department of Health, Executive Yuan, Taiwan (DOH100-TD-C-111-002) and the Grant of Biosignature in Colorectal Cancers, Academia Sinica, Taiwan.",

year = "2013",

month = may,

doi = "10.1038/cgt.2013.17",

language = "English",

volume = "20",

pages = "276--281",

journal = "Cancer Gene Therapy",

issn = "0929-1903",

publisher = "Nature Publishing Group",

number = "5",

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TY - JOUR

T1 - Expression of β-glucuronidase on the surface of bacteria enhances activation of glucuronide prodrugs

AU - Cheng, C. M.

AU - Chen, F. M.

AU - Lu, Y. L.

AU - Tzou, Shey-Cherng

AU - Wang, J. Y.

AU - Kao, C. H.

AU - Liao, Kuang-Wen

AU - Cheng, T. C.

AU - Chuang, C. H.

AU - Chen, B. M.

AU - Roffler, S.

AU - Cheng, T. L.

N1 - Funding Information: This work was supported by grants from the National Research Program for Biopharmaceuticals, National Science Council, Taipei, Taiwan (NSC101-2325-B-037-001, NSC101-2321-B-037-001, NSC101-2313-B-022-001), the Department of Health, Executive Yuan, Taiwan (DOH100-TD-C-111-002) and the Grant of Biosignature in Colorectal Cancers, Academia Sinica, Taiwan.

PY - 2013/5

Y1 - 2013/5

N2 - Extracellular activation of hydrophilic glucuronide prodrugs by β-glucuronidase (βG) was examined to increase the therapeutic efficacy of bacteria-directed enzyme prodrug therapy (BDEPT). βG was expressed on the surface of Escherichia coli by fusion to either the bacterial autotransporter protein Adhesin (membrane βG (mβG)/AIDA) or the lipoprotein (lpp) outermembrane protein A (mβG/lpp). Both mβG/AIDA and mβG/lpp were expressed on the bacterial surface, but only mβG/AIDA displayed enzymatic activity. The rate of substrate hydrolysis by mβG/AIDA-BL21cells was 2.6-fold greater than by pβG-BL21 cells, which express periplasmic βG. Human colon cancer HCT116 cells that were incubated with mβG/AIDA-BL21 bacteria were sensitive to a glucuronide prodrug (p-hydroxy aniline mustard β-D-glucuronide, HAMG) with an half maximal inhibitory concentration (IC50) value of 226.53±45.4 μM, similar to the IC 50 value of the active drug (p-hydroxy aniline mustard, pHAM; 70.6±6.75 μM), indicating that mβG/AIDA on BL21 bacteria could rapidly and efficiently convert HAMG to an active anticancer agent. These results suggest that surface display of functional βG on bacteria can enhance the hydrolysis of glucuronide prodrugs and may increase the effectiveness of BDEPT.

AB - Extracellular activation of hydrophilic glucuronide prodrugs by β-glucuronidase (βG) was examined to increase the therapeutic efficacy of bacteria-directed enzyme prodrug therapy (BDEPT). βG was expressed on the surface of Escherichia coli by fusion to either the bacterial autotransporter protein Adhesin (membrane βG (mβG)/AIDA) or the lipoprotein (lpp) outermembrane protein A (mβG/lpp). Both mβG/AIDA and mβG/lpp were expressed on the bacterial surface, but only mβG/AIDA displayed enzymatic activity. The rate of substrate hydrolysis by mβG/AIDA-BL21cells was 2.6-fold greater than by pβG-BL21 cells, which express periplasmic βG. Human colon cancer HCT116 cells that were incubated with mβG/AIDA-BL21 bacteria were sensitive to a glucuronide prodrug (p-hydroxy aniline mustard β-D-glucuronide, HAMG) with an half maximal inhibitory concentration (IC50) value of 226.53±45.4 μM, similar to the IC 50 value of the active drug (p-hydroxy aniline mustard, pHAM; 70.6±6.75 μM), indicating that mβG/AIDA on BL21 bacteria could rapidly and efficiently convert HAMG to an active anticancer agent. These results suggest that surface display of functional βG on bacteria can enhance the hydrolysis of glucuronide prodrugs and may increase the effectiveness of BDEPT.

KW - Autotransporter protein adhesin

KW - Bacteria-directed enzyme prodrug therapy

KW - Extracellular activation

KW - Surface display

KW - β-glucuronidase

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DO - 10.1038/cgt.2013.17

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C2 - 23598434

AN - SCOPUS:84878013881

SN - 0929-1903

VL - 20

SP - 276

EP - 281

JO - Cancer Gene Therapy

JF - Cancer Gene Therapy

IS - 5

ER -

Expression of β-glucuronidase on the surface of bacteria enhances activation of glucuronide prodrugs

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Keywords

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