Designer cell-penetrating peptides for the treatment of cervical cancer
Disciplines
Cell Biology
Abstract (300 words maximum)
Human papillomaviruses are the causative agents of cervical cancer. Cancer arises after integration of viral oncoproteins into the host genome with subsequent loss of regulatory viral protein E2. Reintroduction of E2 into cervical cancer cells can reduce cell proliferation and promote apoptosis in vitro. However, E2’s therapeutic potential is dampened by the need for impractical delivery mechanisms such as gene transfection or viral vector delivery. An alternative approach is to deliver the protein directly to cells. Cell penetrating peptides (CPPs) are a promising means for the development and delivery of protein-based therapeutics. We previously developed a novel CPP-adaptor system for the rapid and effective delivery of bioactive proteins into living cells. While highly effective at delivering E2 for the induction of senescence or apoptosis in cervical cancer cell lines, our CPP can also be taken up by normal cells. The goal of this work was to devise a new CPP-adaptor for the targeted delivery of viral protein E2 into cervical cancer cells. We created chimeric cancer-specific CPP constructs that can readily bind E2 in presence of calcium but dissociate rapidly in its absence, representative of the extracellular and intracellular environments, respectively. Constructs were expressed in E.coli and purified via immobilized metal affinity chromatography. Binding kinetics of our cancer-specific CPP with various cargos was determined utilizing optical biosensing. Our CPP bound protein constructs with nanomolar affinity in the presence of calcium and not at all in the absence of calcium. Currently, we are utilizing mixed normal/cancer cell cultures to verify our adaptor will only target cancer cells. This work aims to validate that our methodology can be used for direct and targeted delivery of functional protein for the induction of senescence/apoptosis in cervical cancer.
Academic department under which the project should be listed
CSM - Molecular and Cellular Biology
Primary Investigator (PI) Name
Jonathan L. McMurry
Additional Faculty
Julia C. LeCher, Department of Molecular and Cellular Biology, jwand@kennesaw.edu
Designer cell-penetrating peptides for the treatment of cervical cancer
Human papillomaviruses are the causative agents of cervical cancer. Cancer arises after integration of viral oncoproteins into the host genome with subsequent loss of regulatory viral protein E2. Reintroduction of E2 into cervical cancer cells can reduce cell proliferation and promote apoptosis in vitro. However, E2’s therapeutic potential is dampened by the need for impractical delivery mechanisms such as gene transfection or viral vector delivery. An alternative approach is to deliver the protein directly to cells. Cell penetrating peptides (CPPs) are a promising means for the development and delivery of protein-based therapeutics. We previously developed a novel CPP-adaptor system for the rapid and effective delivery of bioactive proteins into living cells. While highly effective at delivering E2 for the induction of senescence or apoptosis in cervical cancer cell lines, our CPP can also be taken up by normal cells. The goal of this work was to devise a new CPP-adaptor for the targeted delivery of viral protein E2 into cervical cancer cells. We created chimeric cancer-specific CPP constructs that can readily bind E2 in presence of calcium but dissociate rapidly in its absence, representative of the extracellular and intracellular environments, respectively. Constructs were expressed in E.coli and purified via immobilized metal affinity chromatography. Binding kinetics of our cancer-specific CPP with various cargos was determined utilizing optical biosensing. Our CPP bound protein constructs with nanomolar affinity in the presence of calcium and not at all in the absence of calcium. Currently, we are utilizing mixed normal/cancer cell cultures to verify our adaptor will only target cancer cells. This work aims to validate that our methodology can be used for direct and targeted delivery of functional protein for the induction of senescence/apoptosis in cervical cancer.