86b7ad12b47be4d976a3640f0e4f6556 pmc_34208189.pdf 1ebb1c6919d3177d23e56e4cadaf31805550a7a3 pmc_34208189.pdf 36c8177ba6bc5422e60e5db6980d92cd37cd78562cb36aee39cad47e1743d348 pmc_34208189.pdf Title: Biparatopic Protein Nanoparticles for the Precision Therapy of CXCR4+ Cancers Subject: The accumulated molecular knowledge about human cancer enables the identification of multiple cell surface markers as highly specific therapeutic targets. A proper tumor targeting could significantly avoid drug exposure of healthy cells, minimizing side effects, but it is also expected to increase the therapeutic index. Specifically, colorectal cancer has a particularly poor prognosis in late stages, being drug targeting an appropriate strategy to substantially improve the therapeutic efficacy. In this study, we have explored the potential of the human albumin-derived peptide, EPI-X4, as a suitable ligand to target colorectal cancer via the cell surface protein CXCR4, a chemokine receptor overexpressed in cancer stem cells. To explore the potential use of this ligand, self-assembling protein nanoparticles have been generated displaying an engineered EPI-X4 version, which conferred a modest CXCR4 targeting and fast and high level of cell apoptosis in tumor CXCR4+ cells, in vitro and in vivo. In addition, when EPI-X4-based building blocks are combined with biologically inert polypeptides containing the CXCR4 ligand T22, the resulting biparatopic nanoparticles show a dramatically improved biodistribution in mouse models of CXCR4+ human cancer, faster cell internalization and enhanced target cell death when compared to the version based on a single ligand. The generation of biparatopic materials opens exciting possibilities in oncotherapies based on high precision drug delivery based on the receptor CXCR4. Keywords: tumor homing; EPI-X4; CXCR4; biparatopic nanoparticles; tumor targeting; drug delivery Author: Olivia Cano-Garrido, Patricia Álamo, Laura Sánchez-García, Aïda Falgàs, Alejandro Sánchez-Chardi, Naroa Serna, Eloi Parladé, Ugutz Unzueta, Mònica Roldán, Eric Voltà-Durán, Isolda Casanova, Antonio Villaverde, Ramón Mangues and Esther Vázquez Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Sat Jun 12 10:25:59 2021 CEST ModDate: Sat Jun 12 11:09:55 2021 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 8330902 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- WHZOOC+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 IYDCBB+VnURWPalladioL Type 1 Custom yes yes no 15 0 TPWTEJ+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 YOIBLS+URWPalladioL-Bold Type 1 Custom yes yes no 24 0 MGUDHE+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 KVEXVN+CMSY10 Type 1 Builtin yes yes no 32 0 LCJEVJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 41 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 63 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 67 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 71 0 NXQZWG+EURM10 Type 1 Builtin yes yes no 86 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 118 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 122 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 126 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 130 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 150 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 154 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 158 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 162 0 BCDJEE+PalatinoLinotype-BoldItalic TrueType WinAnsi yes yes no 169 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-10-02 03:12:02 CEST RepresentationInformation: pmc_34208189.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 12:14:23 CEST Size: 8330902 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 456 FreeObjects: 3 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Protein Design, Production and Purification Destination: subsection.2.1 Item: Title: Protein Characterization Destination: subsection.2.2 Item: Title: Electron Microscopy Destination: subsection.2.3 Item: Title: Cell Culture, Flow Cytometry and Cytotoxicity Assay Destination: subsection.2.4 Item: Title: Production and Characterization of Biparatopic Nanoparticles Destination: subsection.2.5 Item: Title: Cytotoxicity Studies Destination: subsection.2.6 Item: Title: Analysis of Apoptosis Destination: subsection.2.7 Item: Title: Confocal Assay Destination: subsection.2.8 Item: Title: Evaluation of EPIX4-(RK)-GFP-H6 and Biparatopic Nanoparticles Biodistribution in a Colorectal Subcutaneous Cancer Mouse Destination: subsection.2.9 Item: Title: Histopathology and Detection of Apoptotic Bodies and Mitotic Figures Destination: subsection.2.10 Item: Title: Statistical Analyses Destination: subsection.2.11 Item: Title: Results Destination: section.3 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: Patents Destination: section.6 Item: Title: References Destination: section.6 Info: Title: Biparatopic Protein Nanoparticles for the Precision Therapy of CXCR4+ Cancers Author: Olivia Cano-Garrido, Patricia Álamo, Laura Sánchez-García, Aïda Falgàs, Alejandro Sánchez-Chardi, Naroa Serna, Eloi Parladé, Ugutz Unzueta, Mònica Roldán, Eric Voltà-Durán, Isolda Casanova, Antonio Villaverde, Ramón Mangues and Esther Vázquez Subject: The accumulated molecular knowledge about human cancer enables the identification of multiple cell surface markers as highly specific therapeutic targets. A proper tumor targeting could significantly avoid drug exposure of healthy cells, minimizing side effects, but it is also expected to increase the therapeutic index. Specifically, colorectal cancer has a particularly poor prognosis in late stages, being drug targeting an appropriate strategy to substantially improve the therapeutic efficacy. In this study, we have explored the potential of the human albumin-derived peptide, EPI-X4, as a suitable ligand to target colorectal cancer via the cell surface protein CXCR4, a chemokine receptor overexpressed in cancer stem cells. To explore the potential use of this ligand, self-assembling protein nanoparticles have been generated displaying an engineered EPI-X4 version, which conferred a modest CXCR4 targeting and fast and high level of cell apoptosis in tumor CXCR4+ cells, in vitro and in vivo. In addition, when EPI-X4-based building blocks are combined with biologically inert polypeptides containing the CXCR4 ligand T22, the resulting biparatopic nanoparticles show a dramatically improved biodistribution in mouse models of CXCR4+ human cancer, faster cell internalization and enhanced target cell death when compared to the version based on a single ligand. The generation of biparatopic materials opens exciting possibilities in oncotherapies based on high precision drug delivery based on the receptor CXCR4. Keywords: tumor homing; EPI-X4; CXCR4; biparatopic nanoparticles; tumor targeting; drug delivery Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Sat Jun 12 10:25:59 CEST 2021 ModDate: Sat Jun 12 11:09:55 CEST 2021 ID: 0x289f2144f876ef741974d3fb0af47140, 0x289f2144f876ef741974d3fb0af47140 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: KVEXVN+CMSY10 FontSubset: true FirstChar: 0 LastChar: 20 FontDescriptor: FontName: KVEXVN+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true Font: BaseFont: TPWTEJ+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 122 FontDescriptor: FontName: TPWTEJ+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: NXQZWG+EURM10 FontSubset: true FirstChar: 11 LastChar: 22 FontDescriptor: FontName: NXQZWG+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: YOIBLS+URWPalladioL-Bold FontSubset: 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