Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteins
Active tumor targeting of nanomedicines has recently shown significant improvements in the therapeutic activity of currently existing drug delivery systems, such as liposomal doxorubicin (Doxil/Caelyx/Lipodox). Previously, we have shown that isolated pVIII major coat proteins of the fd tet filamento...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2015-06-01
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| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fmicb.2015.00628/full |
| _version_ | 1828286891304157184 |
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| author | James W. Gillespie Amanda L. Gross Anatoliy T. Puzyrev Deepa eBedi Valery A. Petrenko |
| author_facet | James W. Gillespie Amanda L. Gross Anatoliy T. Puzyrev Deepa eBedi Valery A. Petrenko |
| author_sort | James W. Gillespie |
| collection | DOAJ |
| description | Active tumor targeting of nanomedicines has recently shown significant improvements in the therapeutic activity of currently existing drug delivery systems, such as liposomal doxorubicin (Doxil/Caelyx/Lipodox). Previously, we have shown that isolated pVIII major coat proteins of the fd tet filamentous phage vector, containing cancer cell-specific peptide fusions at their N terminus, can be used as active targeting ligands in a liposomal doxorubicin delivery system in vitro and in vivo. Here, we show a novel major coat protein isolation procedure in 2-propanol that allows spontaneous incorporation of the hydrophobic protein core into preformed liposomal doxorubicin with minimal damage or drug loss while still retaining the targeting ligand exposed for cell-specific targeting. Using a panel of 12 structurally unique ligands with specificity towards breast, lung, and/or pancreatic cancer, we showed the feasibility of pVIII major coat proteins to significantly increase the throughput of targeting ligand screening in a common nanomedicine core. Phage protein-modified Lipodox samples showed an average doxorubicin recovery of 82.8% across all samples with 100% of protein incorporation in the correct orientation (N-terminus exposed). Following cytotoxicity screening in a doxorubicin-sensitive breast cancer line (MCF-7), three major groups of ligands were identified. Ligands showing the most improved cytotoxicity included: DMPGTVLP, ANGRPSMT, VNGRAEAP, and ANDVYLD showing a 25-fold improvement (p < 0.05) in toxicity. Similarly DGQYLGSQ, ETYNQPYL, and GSSEQLYL ligands with specificity towards a doxorubicin-insensitive pancreatic cancer line (PANC-1) showed significant increases in toxicity (2-fold; p < 0.05). Thus, we demonstrated proof-of-concept that pVIII major coat proteins can be screened in significantly higher throughput to identify novel ligands displaying improved therapeutic activity in a desired cancer phenotype. |
| first_indexed | 2024-04-13T09:39:40Z |
| format | Article |
| id | doaj.art-37d38e1312b442c5a080a1e62eac9cb3 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-13T09:39:40Z |
| publishDate | 2015-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-37d38e1312b442c5a080a1e62eac9cb32022-12-22T02:51:58ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2015-06-01610.3389/fmicb.2015.00628152904Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteinsJames W. Gillespie0Amanda L. Gross1Anatoliy T. Puzyrev2Deepa eBedi3Valery A. Petrenko4Auburn UniversityAuburn UniversityAuburn UniversityAuburn UniversityAuburn UniversityActive tumor targeting of nanomedicines has recently shown significant improvements in the therapeutic activity of currently existing drug delivery systems, such as liposomal doxorubicin (Doxil/Caelyx/Lipodox). Previously, we have shown that isolated pVIII major coat proteins of the fd tet filamentous phage vector, containing cancer cell-specific peptide fusions at their N terminus, can be used as active targeting ligands in a liposomal doxorubicin delivery system in vitro and in vivo. Here, we show a novel major coat protein isolation procedure in 2-propanol that allows spontaneous incorporation of the hydrophobic protein core into preformed liposomal doxorubicin with minimal damage or drug loss while still retaining the targeting ligand exposed for cell-specific targeting. Using a panel of 12 structurally unique ligands with specificity towards breast, lung, and/or pancreatic cancer, we showed the feasibility of pVIII major coat proteins to significantly increase the throughput of targeting ligand screening in a common nanomedicine core. Phage protein-modified Lipodox samples showed an average doxorubicin recovery of 82.8% across all samples with 100% of protein incorporation in the correct orientation (N-terminus exposed). Following cytotoxicity screening in a doxorubicin-sensitive breast cancer line (MCF-7), three major groups of ligands were identified. Ligands showing the most improved cytotoxicity included: DMPGTVLP, ANGRPSMT, VNGRAEAP, and ANDVYLD showing a 25-fold improvement (p < 0.05) in toxicity. Similarly DGQYLGSQ, ETYNQPYL, and GSSEQLYL ligands with specificity towards a doxorubicin-insensitive pancreatic cancer line (PANC-1) showed significant increases in toxicity (2-fold; p < 0.05). Thus, we demonstrated proof-of-concept that pVIII major coat proteins can be screened in significantly higher throughput to identify novel ligands displaying improved therapeutic activity in a desired cancer phenotype.http://journal.frontiersin.org/Journal/10.3389/fmicb.2015.00628/fullDoxorubicinbreast cancerPancreatic Cancerphage displayLiposomal Drug DeliveryTargeted nanomedicines |
| spellingShingle | James W. Gillespie Amanda L. Gross Anatoliy T. Puzyrev Deepa eBedi Valery A. Petrenko Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteins Frontiers in Microbiology Doxorubicin breast cancer Pancreatic Cancer phage display Liposomal Drug Delivery Targeted nanomedicines |
| title | Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteins |
| title_full | Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteins |
| title_fullStr | Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteins |
| title_full_unstemmed | Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteins |
| title_short | Combinatorial synthesis and screening of cancer cell-specific nanomedicines targeted via phage fusion proteins |
| title_sort | combinatorial synthesis and screening of cancer cell specific nanomedicines targeted via phage fusion proteins |
| topic | Doxorubicin breast cancer Pancreatic Cancer phage display Liposomal Drug Delivery Targeted nanomedicines |
| url | http://journal.frontiersin.org/Journal/10.3389/fmicb.2015.00628/full |
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