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PMID: 19549879 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

An engineered substance P variant for receptor-mediated delivery of synthetic antibodies into tumor cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 106 ·No. 27 ·2009-07-07 ·Pages 11011-5

Rizk SS, Luchniak A, Uysal S, Brawley CM, Rock RS, Kossiakoff AA

Abstract

We have developed and tested a robust delivery method for the transport of proteins to the cytoplasm of mammalian cells without compromising the integrity of the cell membrane. This receptor-mediated delivery (RMD) technology utilizes a variant of substance P (SP), a neuropeptide that is rapidly internalized upon interaction with the neurokinin-1 receptor (NK1R). Cargos in the form of synthetic antibody fragments (sABs) were conjugated to the engineered SP variant (SPv) and efficiently internalized by NK1R-expressing cells. The sABs used here were generated to bind specific conformational forms of actin. The internalized proteins appear to escape the endosome and retain their binding activity within the cells as demonstrated by co-localization with the actin cytoskeleton. Further, since the NK1R is over-expressed in many cancers, SPv-mediated delivery provides a highly specific method for therapeutic utilization of affinity reagents targeting intracellular processes in diseased tissue.

MeSH Terms
Actins/ultrastructure Amino Acid Sequence Cell Line, Tumor Cell Survival Drug Delivery Systems Endocytosis Humans Immunoglobulin Fragments/chemistry,metabolism,ultrastructure Molecular Sequence Data Mutant Proteins/chemistry,metabolism Neoplasms/metabolism,pathology Protein Binding Protein Engineering Receptors, Neurokinin-1/metabolism Substance P/chemistry,metabolism
Chemicals
Actins Immunoglobulin Fragments Mutant Proteins Receptors, Neurokinin-1 Substance P
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Rizk Shahir S
Department of Biochemistry and Molecular Biology, University of Chicago, 929 E. 57th Street, Chicago, IL 60637, USA.
Luchniak Anna
Uysal Serdar
Brawley Crista M
Rock Ronald S
Kossiakoff Anthony A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
DOI
10.1073/pnas.0904907106
Published
2009-07-07
Epub
2009-00-22
Pages
11011-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2708752
Subset
IM
Grants
NIDDK NIH HHS · F32DK080619-02 · United States
NCI NIH HHS · T32 CA009594 · United States
NIGMS NIH HHS · R01 GM078450 · United States
NIDDK NIH HHS · F32 DK080619 · United States
NCI NIH HHS · T32CA009594 · United States
NIGMS NIH HHS · GM078450 · United States
External Links
PubMed source DOI: 10.1073/pnas.0904907106 PMC Full Text View journal details
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