19c730ae62b7dc40d72d46aabfc8c347 pmc_10054149.pdf 8d5d41b440e7554b51fe1fbf3e1c173ae013babf pmc_10054149.pdf ccfd6427f4f520742af465ad0578944d8fd46b0e5c8278165b94ef590dbede4a pmc_10054149.pdf Title: T22-PE24-H6 Nanotoxin Selectively Kills CXCR4-High Expressing AML Patient Cells In Vitro and Potently Blocks Dissemination In Vivo Subject: Despite advances in the development of targeted therapies for acute myeloid leukemia (AML), most patients relapse. For that reason, it is still necessary to develop novel therapies that improve treatment effectiveness and overcome drug resistance. We developed T22-PE24-H6, a protein nanoparticle that contains the exotoxin A from the bacterium Pseudomonas aeruginosa and is able to specifically deliver this cytotoxic domain to CXCR4+ leukemic cells. Next, we evaluated the selective delivery and antitumor activity of T22-PE24-H6 in CXCR4+ AML cell lines and BM samples from AML patients. Moreover, we assessed the in vivo antitumor effect of this nanotoxin in a disseminated mouse model generated from CXCR4+ AML cells. T22-PE24-H6 showed a potent, CXCR4-dependent antineoplastic effect in vitro in the MONO-MAC-6 AML cell line. In addition, mice treated with nanotoxins in daily doses reduced the dissemination of CXCR4+ AML cells compared to buffer-treated mice, as shown by the significant decrease in BLI signaling. Furthermore, we did not observe any sign of toxicity or changes in mouse body weight, biochemical parameters, or histopathology in normal tissues. Finally, T22-PE24-H6 exhibited a significant inhibition of cell viability in CXCR4high AML patient samples but showed no activity in CXCR4low samples. These data strongly support the use of T22-PE24-H6 therapy to benefit high-CXCR4-expressing AML patients. Keywords: acute myeloid leukemia; CXCR4; targeted nanoparticle; disseminated AML mouse model Author: Yáiza Núñez, Annabel Garcia-León, Aïda Falgàs, Naroa Serna, Laura Sánchez-García, Ana Garrido, Jorge Sierra, Alberto Gallardo, Ugutz Unzueta, Esther Vázquez, Antonio Villaverde, Ramon Mangues and Isolda Casanova Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Feb 22 08:52:56 2023 CET ModDate: Wed Feb 22 09:45:48 2023 CET 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: 7656966 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- OWNPHU+VnURWPalladioL Type 1 Custom yes yes no 10 0 UYQCRA+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 YTCYWJ+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 QJWGZP+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 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WinAnsi yes yes no 183 0 OJIMBG+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 186 0 OJIMBC+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 201 0 OJIMBE+PalatinoLinotype TrueType WinAnsi yes yes no 204 0 OJIMBG+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 207 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-10-20 02:26:26 CEST RepresentationInformation: pmc_10054149.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-07-12 02:23:26 CEST Size: 7656966 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 434 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Children: Item: Title: AML Cell Lines and Patient Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Lentiviral Luciferase Transduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Nanoparticle Design and Production Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Detection of CXCR4 Expression in AML Cell Lines Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Analysis of T22-PE24-H6 Antineoplastic Effect In Vitro Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Evaluation of T22-PE24-H6 Antineoplastic Activity in a Disseminated AML Mouse Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Immunohistochemical Analysis of CXCR4 and CD45 in AML Infiltrated Tissues Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Toxicity Analysis in Mouse Tissues and Blood Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Detection of CXCR4 Expression in AML Patients’ BM Cells Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Evaluation of T22-PE24-H6 Antineoplastic Activity in Cultured AML Patients’ BM Cells Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Children: Item: Title: T22-PE24-H6 Antineoplastic Effect in CXCR4+ AML Cell Lines Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: T22-PE24-H6 Induced Apoptosis in MONO-MAC-6 Cells Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Antineoplastic Effect of T22-PE24-H6 in a CXCR4+ AML Mouse Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Toxicity Evaluation in Mouse Tissues Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: T22-PE24-H6 In Vitro Effect in AML Patients’ BM Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Info: Title: T22-PE24-H6 Nanotoxin Selectively Kills CXCR4-High Expressing AML Patient Cells In Vitro and Potently Blocks Dissemination In Vivo Author: Yáiza Núñez, Annabel Garcia-León, Aïda Falgàs, Naroa Serna, Laura Sánchez-García, Ana Garrido, Jorge Sierra, Alberto Gallardo, Ugutz Unzueta, Esther Vázquez, Antonio Villaverde, Ramon Mangues and Isolda Casanova Subject: Despite advances in the development of targeted therapies for acute myeloid leukemia (AML), most patients relapse. For that reason, it is still necessary to develop novel therapies that improve treatment effectiveness and overcome drug resistance. We developed T22-PE24-H6, a protein nanoparticle that contains the exotoxin A from the bacterium Pseudomonas aeruginosa and is able to specifically deliver this cytotoxic domain to CXCR4+ leukemic cells. Next, we evaluated the selective delivery and antitumor activity of T22-PE24-H6 in CXCR4+ AML cell lines and BM samples from AML patients. Moreover, we assessed the in vivo antitumor effect of this nanotoxin in a disseminated mouse model generated from CXCR4+ AML cells. T22-PE24-H6 showed a potent, CXCR4-dependent antineoplastic effect in vitro in the MONO-MAC-6 AML cell line. In addition, mice treated with nanotoxins in daily doses reduced the dissemination of CXCR4+ AML cells compared to buffer-treated mice, as shown by the significant decrease in BLI signaling. Furthermore, we did not observe any sign of toxicity or changes in mouse body weight, biochemical parameters, or histopathology in normal tissues. Finally, T22-PE24-H6 exhibited a significant inhibition of cell viability in CXCR4high AML patient samples but showed no activity in CXCR4low samples. These data strongly support the use of T22-PE24-H6 therapy to benefit high-CXCR4-expressing AML patients. 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