d26d420da8d8551193b8b0a39e073bfd cancers-15-00085.pdf d494c81820a2face39c0c7fa387bb38965bb0d85 cancers-15-00085.pdf a849745a2ab81ea69cda9194422566b57274a40263d7112d4a6669d86f7a3946 cancers-15-00085.pdf Title: Potent Anticancer Activity of CXCR4-Targeted Nanostructured Toxins in Aggressive Endometrial Cancer Models Subject: Patients with advanced endometrial cancer (EC) show poor outcomes. Thus, the development of new therapeutic approaches to prevent metastasis development in high-risk patients is an unmet need. CXCR4 is overexpressed in EC tumor tissue, epitomizing an unexploited therapeutic target for this malignancy. The in vitro antitumor activity of two CXCR4-targeted nanoparticles, including either the C. diphtheriae (T22-DITOX-H6) or P. aeruginosa (T22-PE24-H6) toxin, was evaluated using viability assays. Apoptotic activation was assessed by DAPI and caspase-3 and PARP cleavage in cell blocks. Both nanotoxins were repeatedly administrated to a subcutaneous EC mouse model, whereas T22-DITOX-H6 was also used in a highly metastatic EC orthotopic model. Tumor burden was assessed through bioluminescence, while metastatic foci and toxicity were studied using histological or immunohistochemical analysis. We found that both nanotoxins exerted a potent antitumor effect both in vitro and in vivo via apoptosis and extended the survival of nanotoxin-treated mice without inducing any off-target toxicity. Repeated T22-DITOX-H6 administration in the metastatic model induced a dramatic reduction in tumor burden while significantly blocking peritoneal, lung and liver metastasis without systemic toxicity. Both nanotoxins, but especially T22-DITOX-H6, represent a promising therapeutic alternative for EC patients that have a dismal prognosis and lack effective therapies. Keywords: advanced endometrial cancer; metastasis; protein nanoparticles; bacterial toxins; targeted drug delivery; CXCR4 Author: Esperanza Medina-Gutiérrez, Annabel García-León, Alberto Gallardo, Patricia Álamo, Lorena Alba-Castellón, Ugutz Unzueta, Antonio Villaverde, Esther Vázquez, Isolda Casanova and Ramon Mangues Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Dec 23 09:53:54 2022 CET ModDate: Fri Dec 23 10:06:31 2022 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 20 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4751714 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- MEXBTU+VnURWPalladioL Type 1 Custom yes yes no 10 0 PUGOCA+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 YGBEIL+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 ZBCMAM+URWPalladioL-Ital Type 1 Custom yes yes no 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BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 292 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 296 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 300 0 BCDHEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 304 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 321 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 325 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 329 0 BCDHEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 333 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 350 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 354 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 358 0 BCDHEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 362 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-07-04 02:30:17 CEST RepresentationInformation: cancers-15-00085.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-07-03 16:14:46 CEST Size: 4751714 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 533 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Cell Culture Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Cytotoxic Effect Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Apoptosis Assessment by Flow Cytometry Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Paraffin-Embedded Cell Blocks Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: DAPI Fluorescent Staining Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Immunocytochemistry Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Animals Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Children: Item: Title: Antitumor Effect of Nanotoxins in a Subcutaneous CXCR4+ EC Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Antimetastatic Effect of T22-DITOX-H6 Using a Highly Metastatic Orthotopic CXCR4+ EC Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Histological Examination Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Immunohistochemistry Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Children: Item: Title: Potent CXCR4-Dependent Cytotoxic Effect Induced by T22-DITOX-H6 and T22-PE24-H6 in CE Cells In Vitro Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: T22-DITOX-H6 and T22-PE24-H6-Induced EC Cell Death In Vitro Is Mediated by Apoptosis Induction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Repeated T22-DITOX-H6 or T22-PE24-H6 Treatment Inhibit Tumor Growth and Increases Survival without Systemic Toxicity in a CXCR4+ AN3CA Subcutaneous CE Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: T22-DITOX-H6 Repeated Administration Reduces Total Tumor Burden as Measured by Bioluminescence in a CXCR4+ AN3CA EC Orthotopic Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Potent Inhibition of the Development of Liver and Lung Metastases by T22-DITOX-H6 Repeated Dosage in a CXCR4+ AN3CA EC Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Inhibition of Lymph Node Metastases and Carcinomatotic Foci and Metastases after T22-DITOX-H6 Repeated Dosage in the Disseminated EC Model in the Absence of Systemic Toxicity Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Info: Title: Potent Anticancer Activity of CXCR4-Targeted Nanostructured Toxins in Aggressive Endometrial Cancer Models Author: Esperanza Medina-Gutiérrez, Annabel García-León, Alberto Gallardo, Patricia Álamo, Lorena Alba-Castellón, Ugutz Unzueta, Antonio Villaverde, Esther Vázquez, Isolda Casanova and Ramon Mangues Subject: Patients with advanced endometrial cancer (EC) show poor outcomes. Thus, the development of new therapeutic approaches to prevent metastasis development in high-risk patients is an unmet need. CXCR4 is overexpressed in EC tumor tissue, epitomizing an unexploited therapeutic target for this malignancy. The in vitro antitumor activity of two CXCR4-targeted nanoparticles, including either the C. diphtheriae (T22-DITOX-H6) or P. aeruginosa (T22-PE24-H6) toxin, was evaluated using viability assays. Apoptotic activation was assessed by DAPI and caspase-3 and PARP cleavage in cell blocks. Both nanotoxins were repeatedly administrated to a subcutaneous EC mouse model, whereas T22-DITOX-H6 was also used in a highly metastatic EC orthotopic model. Tumor burden was assessed through bioluminescence, while metastatic foci and toxicity were studied using histological or immunohistochemical analysis. We found that both nanotoxins exerted a potent antitumor effect both in vitro and in vivo via apoptosis and extended the survival of nanotoxin-treated mice without inducing any off-target toxicity. Repeated T22-DITOX-H6 administration in the metastatic model induced a dramatic reduction in tumor burden while significantly blocking peritoneal, lung and liver metastasis without systemic toxicity. Both nanotoxins, but especially T22-DITOX-H6, represent a promising therapeutic alternative for EC patients that have a dismal prognosis and lack effective therapies. 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