7c55dad048e87e3300c8253d51dcc775 265554.pdf 3a99fc5d586dcdb773fc02a036b85850ba558049 265554.pdf ade0e6f19b155f7a51eb9faf5c56b2484341f98b9d50202300e0d770d458e14d 265554.pdf Title: Novel Endometrial Cancer Models Using Sensitive Metastasis Tracing for CXCR4-Targeted Therapy in Advanced Disease Subject: Advanced endometrial cancer (EC) lacks therapy, thus, there is a need for novel treatment targets. CXCR4 overexpression is associated with a poor prognosis in several cancers, whereas its inhibition prevents metastases. We assessed CXCR4 expression in EC in women by using IHC. Orthotopic models were generated with transendometrial implantation of CXCR4-transduced EC cells. After in vitro evaluation of the CXCR4-targeted T22-GFP-H6 nanocarrier, subcutaneous EC models were used to study its uptake in tumor and normal organs. Of the women, 91% overexpressed CXCR4, making them candidates for CXCR4-targeted therapies. Thus, we developed CXCR4+ EC mouse models to improve metastagenesis compared to current models and to use them to develop novel CXCR4-targeted therapies for unresponsive EC. It showed enhanced dissemination, especially in the lungs and liver, and displayed 100% metastasis penetrance at all clinically relevant sites with anti-hVimentin IHC, improving detection sensitivity. Regarding the CXCR4-targeted nanocarrier, 60% accumulated in the SC tumor; therefore, selectively targeting CXCR4+ cancer cells, without toxicity in non-tumor organs. Our CXCR4+ EC models will allow testing of novel CXCR4-targeted drugs and development of nanomedicines derived from T22-GFP-H6 to deliver drugs to CXCR4+ cells in advanced EC. This novel approach provides a therapeutic option for women with metastatic, high risk or recurrent EC that have a dismal prognosis and lack effective therapies. Keywords: advanced endometrial cancer; orthotopic model; metastasis; CXCR4-targeted nanoparticles; animal model Author: Esperanza Medina-Gutiérrez, María Virtudes Céspedes, Alberto Gallardo, Elisa Rioja-Blanco, Miquel Àngel Pavón, Laura Asensio-Puig, Lourdes Farré, Lorena Alba-Castellón, Ugutz Unzueta, Antonio Villaverde, Esther Vázquez, Isolda Casanova and Ramon Mangues Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Jul 14 12:01:29 2022 CEST ModDate: Fri Jul 15 08:41:31 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 18 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3491485 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- XMESMT+VnURWPalladioL Type 1 Custom yes yes no 10 0 KCWTDO+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 FSMHAS+URWPalladioL-Roma Type 1 Custom yes yes no 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Endometrial Cancer Patient Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Cell Culture Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Cell Line Constructs and Stable Cell Line Generation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Lentiviral Transduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Flow Cytometry Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Cell Viability Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Cell Blocks Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: In Vivo Experiments Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Children: Item: Title: Orthotopic EC Models Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Subcutaneous CXCR4+ EC Models Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: In Vivo T22-GFP-H6 Biodistribution Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Necropsy and Histological Examination Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Bioluminescence Intensity Assessment Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Immunocytochemistry and Immunohistochemistry Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Children: Item: Title: Immunohistochemical Evaluation of CXCR4 Expression in EC Patient Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Generation of CXCR4+ Luciferase+ Human EC Cell Lines Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Development of a Subcutaneous Tumor Model Bearing Human EC Cells in Swiss Nude and Follow-Up Markers of Cancer Cell Growth Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Bioluminescent Follow-Up of Primary Tumor and Metastatic Dissemination in a Novel Orthotopic Model of Advanced EC in NSG Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Marker-Guided Comparison of Metastatic Yield in the EC Intrauterine Orthotopic Models Generated from CXCR4- or CXCR4+ EC Cells in NSG Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: In Vitro Uptake of the Fluorescent T22-GFP-H6 Nanocarrier and Its Cytotoxicity in Human EC Cell Lines Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Biodistribution of CXCR4-Targeted Nanocarrier T22-GFP-H6 in a CXCR4+ Subcutaneous EC Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Children: Item: Title: CXCR4 Expression Pattern in EC Patients Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Development of an Aggressive CXCR4+ Advanced EC Metastatic Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: CXCR4 Overexpression Is Associated with Enhanced Metastatic Dissemination in EC Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Item: Title: Use of Highly Sensitive Human-Vimentin as EC Tumor Cells Marker to Detect Metastatic Foci Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71 Item: Title: Development of a CXCR4 Subcutaneous Tumor Model and Its Use to Evaluate Targeting of Protein-Based Nanocarriers to CXCR4+ EC Cells Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1b26f7b2 Item: Title: Future Contribution of the Novel Models for the Development of Targeted Therapies in Advanced EC Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@491cc5c9 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@74ad1f1f Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6a1aab78 Info: Title: Novel Endometrial Cancer Models Using Sensitive Metastasis Tracing for CXCR4-Targeted Therapy in Advanced Disease Author: Esperanza Medina-Gutiérrez, María Virtudes Céspedes, Alberto Gallardo, Elisa Rioja-Blanco, Miquel Àngel Pavón, Laura Asensio-Puig, Lourdes Farré, Lorena Alba-Castellón, Ugutz Unzueta, Antonio Villaverde, Esther Vázquez, Isolda Casanova and Ramon Mangues Subject: Advanced endometrial cancer (EC) lacks therapy, thus, there is a need for novel treatment targets. CXCR4 overexpression is associated with a poor prognosis in several cancers, whereas its inhibition prevents metastases. We assessed CXCR4 expression in EC in women by using IHC. Orthotopic models were generated with transendometrial implantation of CXCR4-transduced EC cells. After in vitro evaluation of the CXCR4-targeted T22-GFP-H6 nanocarrier, subcutaneous EC models were used to study its uptake in tumor and normal organs. Of the women, 91% overexpressed CXCR4, making them candidates for CXCR4-targeted therapies. Thus, we developed CXCR4+ EC mouse models to improve metastagenesis compared to current models and to use them to develop novel CXCR4-targeted therapies for unresponsive EC. It showed enhanced dissemination, especially in the lungs and liver, and displayed 100% metastasis penetrance at all clinically relevant sites with anti-hVimentin IHC, improving detection sensitivity. Regarding the CXCR4-targeted nanocarrier, 60% accumulated in the SC tumor; therefore, selectively targeting CXCR4+ cancer cells, without toxicity in non-tumor organs. Our CXCR4+ EC models will allow testing of novel CXCR4-targeted drugs and development of nanomedicines derived from T22-GFP-H6 to deliver drugs to CXCR4+ cells in advanced EC. This novel approach provides a therapeutic option for women with metastatic, high risk or recurrent EC that have a dismal prognosis and lack effective therapies. 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