Tumor Marker B7-H6 Bound to the Coiled Coil Peptide-Polymer Conjugate Enables Targeted Therapy by Activating Human Natural Killer Cells
Targeted cancer immunotherapy is a promising tool for restoring immune surveillance and eradicating cancer cells. Hydrophilic polymers modified with coiled coil peptide tags can be used as universal carriers designed for cell-specific delivery of such biologically active proteins. Here, we describe...
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| Format: | Article |
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MDPI AG
2021-11-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/9/11/1597 |
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| author | Barbora Kalousková Ondřej Skořepa Denis Cmunt Celeste Abreu Kateřina Krejčová Jan Bláha Irena Sieglová Vlastimil Král Milan Fábry Robert Pola Michal Pechar Ondřej Vaněk |
| author_facet | Barbora Kalousková Ondřej Skořepa Denis Cmunt Celeste Abreu Kateřina Krejčová Jan Bláha Irena Sieglová Vlastimil Král Milan Fábry Robert Pola Michal Pechar Ondřej Vaněk |
| author_sort | Barbora Kalousková |
| collection | DOAJ |
| description | Targeted cancer immunotherapy is a promising tool for restoring immune surveillance and eradicating cancer cells. Hydrophilic polymers modified with coiled coil peptide tags can be used as universal carriers designed for cell-specific delivery of such biologically active proteins. Here, we describe the preparation of pHPMA-based copolymer conjugated with immunologically active protein B7-H6 via complementary coiled coil VAALEKE (peptide E) and VAALKEK (peptide K) sequences. Receptor B7-H6 was described as a binding partner of NKp30, and its expression has been proven for various tumor cell lines. The binding of B7-H6 to NKp30 activates NK cells and results in Fas ligand or granzyme-mediated apoptosis of target tumor cells. In this work, we optimized the expression of coiled coil tagged B7-H6, its ability to bind activating receptor NKp30 has been confirmed by isothermal titration calorimetry, and the binding stoichiometry of prepared chimeric biopolymer has been characterized by analytical ultracentrifugation. Furthermore, this coiled coil B7-H6-loaded polymer conjugate activates NK cells <i>in vitro</i> and, in combination with coiled coil scFv, enables their targeting towards a model tumor cell line. Prepared chimeric biopolymer represents a promising precursor for targeted cancer immunotherapy by activating the cytotoxic activity of natural killer cells. |
| first_indexed | 2024-03-10T05:40:59Z |
| format | Article |
| id | doaj.art-2c9043d3a1e7484f806ef4bdcff6e30e |
| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-10T05:40:59Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomedicines |
| spelling | doaj.art-2c9043d3a1e7484f806ef4bdcff6e30e2023-11-22T22:30:37ZengMDPI AGBiomedicines2227-90592021-11-01911159710.3390/biomedicines9111597Tumor Marker B7-H6 Bound to the Coiled Coil Peptide-Polymer Conjugate Enables Targeted Therapy by Activating Human Natural Killer CellsBarbora Kalousková0Ondřej Skořepa1Denis Cmunt2Celeste Abreu3Kateřina Krejčová4Jan Bláha5Irena Sieglová6Vlastimil Král7Milan Fábry8Robert Pola9Michal Pechar10Ondřej Vaněk11Department of Biochemistry, Faculty of Science, Charles University, Hlavova 2030, 12840 Prague, Czech RepublicDepartment of Biochemistry, Faculty of Science, Charles University, Hlavova 2030, 12840 Prague, Czech RepublicDepartment of Biochemistry, Faculty of Science, Charles University, Hlavova 2030, 12840 Prague, Czech RepublicDepartment of Biochemistry, Faculty of Science, Charles University, Hlavova 2030, 12840 Prague, Czech RepublicDepartment of Biochemistry, Faculty of Science, Charles University, Hlavova 2030, 12840 Prague, Czech RepublicDepartment of Biochemistry, Faculty of Science, Charles University, Hlavova 2030, 12840 Prague, Czech RepublicInstitute of Molecular Genetics, Czech Academy of Sciences, Vídeňská 1083, 14220 Prague, Czech RepublicInstitute of Molecular Genetics, Czech Academy of Sciences, Vídeňská 1083, 14220 Prague, Czech RepublicInstitute of Molecular Genetics, Czech Academy of Sciences, Vídeňská 1083, 14220 Prague, Czech RepublicInstitute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského nám. 2, 16206 Prague, Czech RepublicInstitute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského nám. 2, 16206 Prague, Czech RepublicDepartment of Biochemistry, Faculty of Science, Charles University, Hlavova 2030, 12840 Prague, Czech RepublicTargeted cancer immunotherapy is a promising tool for restoring immune surveillance and eradicating cancer cells. Hydrophilic polymers modified with coiled coil peptide tags can be used as universal carriers designed for cell-specific delivery of such biologically active proteins. Here, we describe the preparation of pHPMA-based copolymer conjugated with immunologically active protein B7-H6 via complementary coiled coil VAALEKE (peptide E) and VAALKEK (peptide K) sequences. Receptor B7-H6 was described as a binding partner of NKp30, and its expression has been proven for various tumor cell lines. The binding of B7-H6 to NKp30 activates NK cells and results in Fas ligand or granzyme-mediated apoptosis of target tumor cells. In this work, we optimized the expression of coiled coil tagged B7-H6, its ability to bind activating receptor NKp30 has been confirmed by isothermal titration calorimetry, and the binding stoichiometry of prepared chimeric biopolymer has been characterized by analytical ultracentrifugation. Furthermore, this coiled coil B7-H6-loaded polymer conjugate activates NK cells <i>in vitro</i> and, in combination with coiled coil scFv, enables their targeting towards a model tumor cell line. Prepared chimeric biopolymer represents a promising precursor for targeted cancer immunotherapy by activating the cytotoxic activity of natural killer cells.https://www.mdpi.com/2227-9059/9/11/1597coiled coilHPMA polymerNK cellNKp30B7-H6immunotherapy |
| spellingShingle | Barbora Kalousková Ondřej Skořepa Denis Cmunt Celeste Abreu Kateřina Krejčová Jan Bláha Irena Sieglová Vlastimil Král Milan Fábry Robert Pola Michal Pechar Ondřej Vaněk Tumor Marker B7-H6 Bound to the Coiled Coil Peptide-Polymer Conjugate Enables Targeted Therapy by Activating Human Natural Killer Cells Biomedicines coiled coil HPMA polymer NK cell NKp30 B7-H6 immunotherapy |
| title | Tumor Marker B7-H6 Bound to the Coiled Coil Peptide-Polymer Conjugate Enables Targeted Therapy by Activating Human Natural Killer Cells |
| title_full | Tumor Marker B7-H6 Bound to the Coiled Coil Peptide-Polymer Conjugate Enables Targeted Therapy by Activating Human Natural Killer Cells |
| title_fullStr | Tumor Marker B7-H6 Bound to the Coiled Coil Peptide-Polymer Conjugate Enables Targeted Therapy by Activating Human Natural Killer Cells |
| title_full_unstemmed | Tumor Marker B7-H6 Bound to the Coiled Coil Peptide-Polymer Conjugate Enables Targeted Therapy by Activating Human Natural Killer Cells |
| title_short | Tumor Marker B7-H6 Bound to the Coiled Coil Peptide-Polymer Conjugate Enables Targeted Therapy by Activating Human Natural Killer Cells |
| title_sort | tumor marker b7 h6 bound to the coiled coil peptide polymer conjugate enables targeted therapy by activating human natural killer cells |
| topic | coiled coil HPMA polymer NK cell NKp30 B7-H6 immunotherapy |
| url | https://www.mdpi.com/2227-9059/9/11/1597 |
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