Anthrax Protective Antigen Retargeted with Single-Chain Variable Fragments Delivers Enzymes to Pancreatic Cancer Cells

Alexander R. Loftis; Michael S. Santos; Nicholas L. Truex; Marco Biancucci; Karla J.F. Satchell; Bradley L. Pentelute

doi:10.1002/cbic.202000201

Anthrax Protective Antigen Retargeted with Single-Chain Variable Fragments Delivers Enzymes to Pancreatic Cancer Cells

Alexander R. Loftis, Michael S. Santos, Nicholas L. Truex, Marco Biancucci, Karla J.F. Satchell, Bradley L. Pentelute^*

^*Corresponding author for this work

Microbiology-Immunology

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

13 Scopus citations

Abstract

The nontoxic, anthrax protective antigen/lethal factor N-terminal domain (PA/LF_N) complex is an effective platform for translocating proteins into the cytosol of cells. Mutant PA (mPA) was recently fused to epidermal growth factor (EGF) to retarget delivery of LF_N to cells bearing EGF receptors (EGFR), but the requirement for a known cognate ligand limits the applicability of this approach. Here, we render practical protective antigen retargeting to a variety of receptors with mPA single-chain variable fragment (scFv) fusion constructs. Our design enables the targeting of two pancreatic cancer-relevant receptors, EGFR and carcinoembryonic antigen. We demonstrate that fusion to scFvs does not disturb the basic functions of mPA. Moreover, mPA−scFv fusions enable cell-specific delivery of diphtheria toxin catalytic domain and Ras/Rap1-specific endopeptidase to pancreatic cancer cells. Importantly, mPA−scFv fusion-based treatments display potent cell-specific toxicity in vitro, opening fundamentally new routes toward engineered immunotoxins and providing a potential solution to the challenge of targeted protein delivery to the cytosol of cancer cells.

Original language	English (US)
Pages (from-to)	2772-2776
Number of pages	5
Journal	ChemBioChem
Volume	21
Issue number	19
DOIs	https://doi.org/10.1002/cbic.202000201
State	Published - Oct 1 2020

Funding

Financial support for this work was provided by an NSF CAREER grant (CHE\u20101351807 to B.L.P.), the MIT/NIGMS Biotechnology Training Program (T32 GM008334\u201028 to A.R.L.), an NIH postdoctoral fellowship (F32 CA239362 to N.L.T.), a PanCan/FNLCR KRAS postdoctoral fellowship (to M.B.), NIH grant R01 AI092825 and the Northwestern Medicine Catalyst Fund (to K.J.F.S.). Financial support for this work was provided by an NSF CAREER grant (CHE-1351807 to B.L.P.), the MIT/NIGMS Biotechnology Training Program (T32 GM008334-28 to A.R.L.), an NIH postdoctoral fellowship (F32 CA239362 to N.L.T.), a PanCan/FNLCR KRAS postdoctoral fellowship (to M.B.), NIH grant R01 AI092825 and the Northwestern Medicine Catalyst Fund (to K.J.F.S.).

Keywords

anthrax toxin
intracellular delivery
pancreatic cancer
protein therapeutics
scFv

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Organic Chemistry

UN SDGs

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

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10.1002/cbic.202000201

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title = "Anthrax Protective Antigen Retargeted with Single-Chain Variable Fragments Delivers Enzymes to Pancreatic Cancer Cells",

abstract = "The nontoxic, anthrax protective antigen/lethal factor N-terminal domain (PA/LFN) complex is an effective platform for translocating proteins into the cytosol of cells. Mutant PA (mPA) was recently fused to epidermal growth factor (EGF) to retarget delivery of LFN to cells bearing EGF receptors (EGFR), but the requirement for a known cognate ligand limits the applicability of this approach. Here, we render practical protective antigen retargeting to a variety of receptors with mPA single-chain variable fragment (scFv) fusion constructs. Our design enables the targeting of two pancreatic cancer-relevant receptors, EGFR and carcinoembryonic antigen. We demonstrate that fusion to scFvs does not disturb the basic functions of mPA. Moreover, mPA−scFv fusions enable cell-specific delivery of diphtheria toxin catalytic domain and Ras/Rap1-specific endopeptidase to pancreatic cancer cells. Importantly, mPA−scFv fusion-based treatments display potent cell-specific toxicity in vitro, opening fundamentally new routes toward engineered immunotoxins and providing a potential solution to the challenge of targeted protein delivery to the cytosol of cancer cells.",

keywords = "anthrax toxin, intracellular delivery, pancreatic cancer, protein therapeutics, scFv",

author = "Loftis, {Alexander R.} and Santos, {Michael S.} and Truex, {Nicholas L.} and Marco Biancucci and Satchell, {Karla J.F.} and Pentelute, {Bradley L.}",

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AU - Biancucci, Marco

AU - Satchell, Karla J.F.

AU - Pentelute, Bradley L.

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AB - The nontoxic, anthrax protective antigen/lethal factor N-terminal domain (PA/LFN) complex is an effective platform for translocating proteins into the cytosol of cells. Mutant PA (mPA) was recently fused to epidermal growth factor (EGF) to retarget delivery of LFN to cells bearing EGF receptors (EGFR), but the requirement for a known cognate ligand limits the applicability of this approach. Here, we render practical protective antigen retargeting to a variety of receptors with mPA single-chain variable fragment (scFv) fusion constructs. Our design enables the targeting of two pancreatic cancer-relevant receptors, EGFR and carcinoembryonic antigen. We demonstrate that fusion to scFvs does not disturb the basic functions of mPA. Moreover, mPA−scFv fusions enable cell-specific delivery of diphtheria toxin catalytic domain and Ras/Rap1-specific endopeptidase to pancreatic cancer cells. Importantly, mPA−scFv fusion-based treatments display potent cell-specific toxicity in vitro, opening fundamentally new routes toward engineered immunotoxins and providing a potential solution to the challenge of targeted protein delivery to the cytosol of cancer cells.

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Anthrax Protective Antigen Retargeted with Single-Chain Variable Fragments Delivers Enzymes to Pancreatic Cancer Cells

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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