Histone Mimetic Peptide Mediated Nuclear Delivery Of Therapeutic Protein To Inflammatory Breast Cancer Cells
Date
2021-05
Authors
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Journal ISSN
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Publisher
University of Delaware
Abstract
Therapeutic proteins are a novel class of biopharmaceuticals, ideal for disease
treatment due to their versatility and sophisticated functionality. A major challenge in
utilization of therapeutic proteins is delivery of the protein, especially when the
protein needs to reach a specific sub-cellular compartment such as the nucleus. This
study seeks to improve nuclear delivery of proteins into inflammatory breast cancer
(IBC) cells. A histone mimetic peptide (histone H3 tail) was used to target caveolar
uptake pathways to gain entry to the nucleus. Click chemistry was used to attach
synthetic alkyne-functionalized histone mimetic peptides to azide functionalized
fluorescent protein. The protein construct was functionalized either through unnatural
amino acids (UAA) incorporation or ester-amine chemistry. The protein also
contained a ligation system for easy attachment of therapeutic protein. Additional
fluorescent proteins were constructed to add GALA peptide functionalization to
promote endosomal escape. Delivery of the H3-conjugated protein construct to IBC
cells demonstrated the peptide’s ability to enhance protein uptake. Without GALA
functionalization, most proteins were trapped in lysosome. Constructs with GALA
functionalization exhibited endosomal escape activity based on reporter assays but
may require multiple peptides in order to function well. Additional experiments are
necessary to determine the relative uptake in IBC cells as compared to normal cells.
Description
Keywords
Breast cancer, Therapeutic proteins, Biopharmaceuticals