Targeted gene delivery to human airway epithelial cells with synthetic vectors incorporating novel targeting peptides selected by phage display

Michele J. Writer, Barry Marshall, Michael A. Pilkington-Miksa, Susie E. Barker, Marianne Jacobsen, Angelika Kritz, Paul C. Bell, Douglas H. Lester, Alethea B. Tabor, Helen C. Hailes, Nigel Klein, Stephen L. Hart

Research output: Contribution to journalArticle

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Abstract

Human airway epithelial cell targeting peptides were identified by biopanning on 1HAEo-cells, a well characterised epithelial cell line. Bound phage were recovered after three rounds of binding, high stringency washing and elution, leading to the production of an enriched phage peptide population. DNA sequencing of 56 clones revealed 14 unique sequences. Subsequent binding analysis revealed that 13 of these peptides bound 1HAEo-cells with high affinity. Three peptides, SERSMNF, YGLPHKF and PSGAARA were represented at high frequency. Three clearly defined families of peptide were identified on the basis of sequence motifs including R/KSM, LP/QHK and PSGA/TARA. Two peptides, LPHKSMP and LQHKSMP contained two motifs. Further detailed sequence analysis by comparison of peptide sequences with the SWISSPROT protein database revealed that some of the peptides closely resembled the cell binding proteins of viral and bacterial pathogens including Herpes Simplex Virus, rotavirus, Mycoplasma pneumoniae and rhinovirus, the latter two being respiratory pathogens, as well as peptide YGLPHKF having similarity to a protein of unknown function from the respiratory pathogen Legionella pneumophila. Peptides were incorporated into gene delivery formulations with the cationic lipid Lipofectin and plasmid DNA and shown to confer a high degree of transfection efficiency and specificity in 1HAEo-cells. Improved transfection efficiency and specificity was also observed in human endothelial cells, fibroblasts and keratinocytes. Therefore, on the basis of clone frequency after biopanning, cell binding affinity, peptide sequence conservation and pathogenic similarity, we have identified 3 novel peptide families and 5 specific peptides that have the potential for gene transfer to respiratory epithelium in vivo as well as providing useful in vitro transfection reagents for primary human cell types of scientific and commercial interest.
Original languageEnglish
Pages (from-to)185-193
Number of pages9
JournalJournal of Drug Targeting
Volume12
Issue number4
DOIs
StatePublished - May 2004

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Bacteriophages
Epithelial Cells
Peptides
Genes
Transfection
Protein Sequence Analysis
Clone Cells
Rhinovirus
Legionella pneumophila
Respiratory Mucosa
Mycoplasma pneumoniae
Protein Databases
Rotavirus
Simplexvirus
DNA Sequence Analysis
Keratinocytes
Carrier Proteins
Plasmids
Endothelial Cells
Fibroblasts

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Writer, M. J., Marshall, B., Pilkington-Miksa, M. A., Barker, S. E., Jacobsen, M., Kritz, A., ... Hart, S. L. (2004). Targeted gene delivery to human airway epithelial cells with synthetic vectors incorporating novel targeting peptides selected by phage display. Journal of Drug Targeting, 12(4), 185-193. DOI: 10.1080/10611860410001724459

Writer, Michele J.; Marshall, Barry; Pilkington-Miksa, Michael A.; Barker, Susie E.; Jacobsen, Marianne; Kritz, Angelika; Bell, Paul C.; Lester, Douglas H.; Tabor, Alethea B.; Hailes, Helen C.; Klein, Nigel; Hart, Stephen L. / Targeted gene delivery to human airway epithelial cells with synthetic vectors incorporating novel targeting peptides selected by phage display.

In: Journal of Drug Targeting, Vol. 12, No. 4, 05.2004, p. 185-193.

Research output: Contribution to journalArticle

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title = "Targeted gene delivery to human airway epithelial cells with synthetic vectors incorporating novel targeting peptides selected by phage display",
abstract = "Human airway epithelial cell targeting peptides were identified by biopanning on 1HAEo-cells, a well characterised epithelial cell line. Bound phage were recovered after three rounds of binding, high stringency washing and elution, leading to the production of an enriched phage peptide population. DNA sequencing of 56 clones revealed 14 unique sequences. Subsequent binding analysis revealed that 13 of these peptides bound 1HAEo-cells with high affinity. Three peptides, SERSMNF, YGLPHKF and PSGAARA were represented at high frequency. Three clearly defined families of peptide were identified on the basis of sequence motifs including R/KSM, LP/QHK and PSGA/TARA. Two peptides, LPHKSMP and LQHKSMP contained two motifs. Further detailed sequence analysis by comparison of peptide sequences with the SWISSPROT protein database revealed that some of the peptides closely resembled the cell binding proteins of viral and bacterial pathogens including Herpes Simplex Virus, rotavirus, Mycoplasma pneumoniae and rhinovirus, the latter two being respiratory pathogens, as well as peptide YGLPHKF having similarity to a protein of unknown function from the respiratory pathogen Legionella pneumophila. Peptides were incorporated into gene delivery formulations with the cationic lipid Lipofectin and plasmid DNA and shown to confer a high degree of transfection efficiency and specificity in 1HAEo-cells. Improved transfection efficiency and specificity was also observed in human endothelial cells, fibroblasts and keratinocytes. Therefore, on the basis of clone frequency after biopanning, cell binding affinity, peptide sequence conservation and pathogenic similarity, we have identified 3 novel peptide families and 5 specific peptides that have the potential for gene transfer to respiratory epithelium in vivo as well as providing useful in vitro transfection reagents for primary human cell types of scientific and commercial interest.",
author = "Writer, {Michele J.} and Barry Marshall and Pilkington-Miksa, {Michael A.} and Barker, {Susie E.} and Marianne Jacobsen and Angelika Kritz and Bell, {Paul C.} and Lester, {Douglas H.} and Tabor, {Alethea B.} and Hailes, {Helen C.} and Nigel Klein and Hart, {Stephen L.}",
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Writer, MJ, Marshall, B, Pilkington-Miksa, MA, Barker, SE, Jacobsen, M, Kritz, A, Bell, PC, Lester, DH, Tabor, AB, Hailes, HC, Klein, N & Hart, SL 2004, 'Targeted gene delivery to human airway epithelial cells with synthetic vectors incorporating novel targeting peptides selected by phage display' Journal of Drug Targeting, vol 12, no. 4, pp. 185-193. DOI: 10.1080/10611860410001724459

Targeted gene delivery to human airway epithelial cells with synthetic vectors incorporating novel targeting peptides selected by phage display. / Writer, Michele J.; Marshall, Barry; Pilkington-Miksa, Michael A.; Barker, Susie E.; Jacobsen, Marianne; Kritz, Angelika; Bell, Paul C.; Lester, Douglas H.; Tabor, Alethea B.; Hailes, Helen C.; Klein, Nigel; Hart, Stephen L.

In: Journal of Drug Targeting, Vol. 12, No. 4, 05.2004, p. 185-193.

Research output: Contribution to journalArticle

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AU - Writer,Michele J.

AU - Marshall,Barry

AU - Pilkington-Miksa,Michael A.

AU - Barker,Susie E.

AU - Jacobsen,Marianne

AU - Kritz,Angelika

AU - Bell,Paul C.

AU - Lester,Douglas H.

AU - Tabor,Alethea B.

AU - Hailes,Helen C.

AU - Klein,Nigel

AU - Hart,Stephen L.

PY - 2004/5

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N2 - Human airway epithelial cell targeting peptides were identified by biopanning on 1HAEo-cells, a well characterised epithelial cell line. Bound phage were recovered after three rounds of binding, high stringency washing and elution, leading to the production of an enriched phage peptide population. DNA sequencing of 56 clones revealed 14 unique sequences. Subsequent binding analysis revealed that 13 of these peptides bound 1HAEo-cells with high affinity. Three peptides, SERSMNF, YGLPHKF and PSGAARA were represented at high frequency. Three clearly defined families of peptide were identified on the basis of sequence motifs including R/KSM, LP/QHK and PSGA/TARA. Two peptides, LPHKSMP and LQHKSMP contained two motifs. Further detailed sequence analysis by comparison of peptide sequences with the SWISSPROT protein database revealed that some of the peptides closely resembled the cell binding proteins of viral and bacterial pathogens including Herpes Simplex Virus, rotavirus, Mycoplasma pneumoniae and rhinovirus, the latter two being respiratory pathogens, as well as peptide YGLPHKF having similarity to a protein of unknown function from the respiratory pathogen Legionella pneumophila. Peptides were incorporated into gene delivery formulations with the cationic lipid Lipofectin and plasmid DNA and shown to confer a high degree of transfection efficiency and specificity in 1HAEo-cells. Improved transfection efficiency and specificity was also observed in human endothelial cells, fibroblasts and keratinocytes. Therefore, on the basis of clone frequency after biopanning, cell binding affinity, peptide sequence conservation and pathogenic similarity, we have identified 3 novel peptide families and 5 specific peptides that have the potential for gene transfer to respiratory epithelium in vivo as well as providing useful in vitro transfection reagents for primary human cell types of scientific and commercial interest.

AB - Human airway epithelial cell targeting peptides were identified by biopanning on 1HAEo-cells, a well characterised epithelial cell line. Bound phage were recovered after three rounds of binding, high stringency washing and elution, leading to the production of an enriched phage peptide population. DNA sequencing of 56 clones revealed 14 unique sequences. Subsequent binding analysis revealed that 13 of these peptides bound 1HAEo-cells with high affinity. Three peptides, SERSMNF, YGLPHKF and PSGAARA were represented at high frequency. Three clearly defined families of peptide were identified on the basis of sequence motifs including R/KSM, LP/QHK and PSGA/TARA. Two peptides, LPHKSMP and LQHKSMP contained two motifs. Further detailed sequence analysis by comparison of peptide sequences with the SWISSPROT protein database revealed that some of the peptides closely resembled the cell binding proteins of viral and bacterial pathogens including Herpes Simplex Virus, rotavirus, Mycoplasma pneumoniae and rhinovirus, the latter two being respiratory pathogens, as well as peptide YGLPHKF having similarity to a protein of unknown function from the respiratory pathogen Legionella pneumophila. Peptides were incorporated into gene delivery formulations with the cationic lipid Lipofectin and plasmid DNA and shown to confer a high degree of transfection efficiency and specificity in 1HAEo-cells. Improved transfection efficiency and specificity was also observed in human endothelial cells, fibroblasts and keratinocytes. Therefore, on the basis of clone frequency after biopanning, cell binding affinity, peptide sequence conservation and pathogenic similarity, we have identified 3 novel peptide families and 5 specific peptides that have the potential for gene transfer to respiratory epithelium in vivo as well as providing useful in vitro transfection reagents for primary human cell types of scientific and commercial interest.

U2 - 10.1080/10611860410001724459

DO - 10.1080/10611860410001724459

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Writer MJ, Marshall B, Pilkington-Miksa MA, Barker SE, Jacobsen M, Kritz A et al. Targeted gene delivery to human airway epithelial cells with synthetic vectors incorporating novel targeting peptides selected by phage display. Journal of Drug Targeting. 2004 May;12(4):185-193. Available from, DOI: 10.1080/10611860410001724459