{"id":{"repo_id":"dialnet","oai_identifier":"oai:dialnet.unirioja.es:TES0000023211"},"canonical_url":"https://search.dev.ndltd.org/etd/dialnet/oai:dialnet.unirioja.es:TES0000023211","repository":{"repo_id":"dialnet","name":"Dialnet","base_url":"https://dialnet.unirioja.es/oaites/OAIHandler"},"display":{"title":"Design and development of an RNA vaccine against tumor-mediated angiogenesis and metastasis targeting a KLH-Adrenomedullin fusion protein","abstract":"Adrenomedullin (AM) is an autocrine/paracrine growth factor as well as a crucial regulator of angiogenesis and immune response. AM is overexpressed by most solid tumors, which makes it a good target for anti-tumor therapy. In this study, we designed and tested a mRNA vaccine directed against a fusion antigen composed by a small piece of keyhole limpet hemocyanin (KLH), as a hapten, and mouse AM. The in vitro-synthesized mRNA was encapsulated in lipid nanoparticles (LNPs) and injected in C57BL/6J mice. Empty LNPs were used as a negative control. After five immunizations, B16- F10 melanoma cells were injected through the tail vein to induce lung metastases. In addition, transgenic mice expressing green fluorescent protein in the blood vessels (SMAA-GFP) were also immunized with both LNP types to assess the effect of the vaccine in subcutaneous tumors and potential side-effects of the vaccine on normal blood vessel formation. Antibody titers against AM and the number of CD8+ T cells were significantly higher in AMimmunized mice than in negative controls. Furthermore, the number and size of the lung metastases, as well as the number of blood vessels, were significantly reduced in the AMimmunized group. Furthermore, the volume of subcutaneous tumors was significantly reduced with significant differences observed in the number of blood vessels. In addition, no significant differences were observed in the number and distribution of blood vessels in normal tissues and organs after immunization of the SMAA-GFP animals. In summary, we have shown that a mRNA vaccine against the fusion KLH-AM peptide was able to break peripheral immunotolerance and induce a specific response against the angiogenic factor AM thus reducing tumor burden in two melanoma tumor models in the absence of disturbances to normal blood vessels. The study results indicate a great translational potential and possibility to use it in combinational anti-tumor treatment settings, as well as possibility to overcome challenges of existing therapies targeting angiogenesis and AM itself.","abstract_html":"Adrenomedullin (AM) is an autocrine/paracrine growth factor as well as a crucial regulator of angiogenesis and immune response. AM is overexpressed by most solid tumors, which makes it a good target for anti-tumor therapy. In this study, we designed and tested a mRNA vaccine directed against a fusion antigen composed by a small piece of keyhole limpet hemocyanin (KLH), as a hapten, and mouse AM. The in vitro-synthesized mRNA was encapsulated in lipid nanoparticles (LNPs) and injected in C57BL/6J mice. Empty LNPs were used as a negative control. After five immunizations, B16- F10 melanoma cells were injected through the tail vein to induce lung metastases. In addition, transgenic mice expressing green fluorescent protein in the blood vessels (SMAA-GFP) were also immunized with both LNP types to assess the effect of the vaccine in subcutaneous tumors and potential side-effects of the vaccine on normal blood vessel formation. Antibody titers against AM and the number of CD8+ T cells were significantly higher in AMimmunized mice than in negative controls. Furthermore, the number and size of the lung metastases, as well as the number of blood vessels, were significantly reduced in the AMimmunized group. Furthermore, the volume of subcutaneous tumors was significantly reduced with significant differences observed in the number of blood vessels. In addition, no significant differences were observed in the number and distribution of blood vessels in normal tissues and organs after immunization of the SMAA-GFP animals. In summary, we have shown that a mRNA vaccine against the fusion KLH-AM peptide was able to break peripheral immunotolerance and induce a specific response against the angiogenic factor AM thus reducing tumor burden in two melanoma tumor models in the absence of disturbances to normal blood vessels. The study results indicate a great translational potential and possibility to use it in combinational anti-tumor treatment settings, as well as possibility to overcome challenges of existing therapies targeting angiogenesis and AM itself.","abstract_has_math":false,"creators":["Tadic, Srdan"],"institution":"Universidad de La Rioja (España)","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Martínez Ramírez, Alfredo (Universidad de La Rioja)"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025","date_published":"2025","updated_at":"2026-07-24T06:27:11Z","subjects":[],"languages":["eng"],"rights":["LICENCIA DE USO: Los documentos a texto completo incluidos en Dialnet son de acceso libre y propiedad de sus autores y/o editores. Por tanto, cualquier acto de reproducción, distribución, comunicación pública y/o transformación total o parcial requiere el consentimiento expreso y escrito de aquéllos. Cualquier enlace al texto completo de estos documentos deberá hacerse a través de la URL oficial de éstos en Dialnet. Más información: https://dialnet.unirioja.es/info/derechosOAI | INTELLECTUAL PROPERTY RIGHTS STATEMENT: Full text documents hosted by Dialnet are protected by copyright and/or related rights. This digital object is accessible without charge, but its use is subject to the licensing conditions set by its authors or editors. Unless expressly stated otherwise in the licensing conditions, you are free to linking, browsing, printing and making a copy for your own personal purposes. All other acts of reproduction and communication to the public are subject to the licensing conditions expressed by editors and authors and require consent from them. Any link to this document should be made using its official URL in Dialnet. More info: https://dialnet.unirioja.es/info/derechosOAI"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://dialnet.unirioja.es/servlet/oaites?codigo=396432","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Martínez Ramírez, Alfredo (Universidad de La Rioja)"]},{"key":"dc:creator","label":"Author","values":["Tadic, Srdan"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2025"]},{"key":"dc:publisher","label":"Institution","values":["Universidad de La Rioja (España)"]},{"key":"dc:type","label":"Dc Type","values":["text (thesis)"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["LICENCIA DE USO: Los documentos a texto completo incluidos en Dialnet son de acceso libre y propiedad de sus autores y/o editores. Por tanto, cualquier acto de reproducción, distribución, comunicación pública y/o transformación total o parcial requiere el consentimiento expreso y escrito de aquéllos. Cualquier enlace al texto completo de estos documentos deberá hacerse a través de la URL oficial de éstos en Dialnet. Más información: https://dialnet.unirioja.es/info/derechosOAI | INTELLECTUAL PROPERTY RIGHTS STATEMENT: Full text documents hosted by Dialnet are protected by copyright and/or related rights. This digital object is accessible without charge, but its use is subject to the licensing conditions set by its authors or editors. Unless expressly stated otherwise in the licensing conditions, you are free to linking, browsing, printing and making a copy for your own personal purposes. All other acts of reproduction and communication to the public are subject to the licensing conditions expressed by editors and authors and require consent from them. Any link to this document should be made using its official URL in Dialnet. More info: https://dialnet.unirioja.es/info/derechosOAI"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://dialnet.unirioja.es/servlet/oaites?codigo=396432"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Adrenomedullin (AM) is an autocrine/paracrine growth factor as well as a crucial regulator of angiogenesis and immune response. AM is overexpressed by most solid tumors, which makes it a good target for anti-tumor therapy. In this study, we designed and tested a mRNA vaccine directed against a fusion antigen composed by a small piece of keyhole limpet hemocyanin (KLH), as a hapten, and mouse AM. The in vitro-synthesized mRNA was encapsulated in lipid nanoparticles (LNPs) and injected in C57BL/6J mice. Empty LNPs were used as a negative control. After five immunizations, B16- F10 melanoma cells were injected through the tail vein to induce lung metastases. In addition, transgenic mice expressing green fluorescent protein in the blood vessels (SMAA-GFP) were also immunized with both LNP types to assess the effect of the vaccine in subcutaneous tumors and potential side-effects of the vaccine on normal blood vessel formation. Antibody titers against AM and the number of CD8+ T cells were significantly higher in AMimmunized mice than in negative controls. Furthermore, the number and size of the lung metastases, as well as the number of blood vessels, were significantly reduced in the AMimmunized group. Furthermore, the volume of subcutaneous tumors was significantly reduced with significant differences observed in the number of blood vessels. In addition, no significant differences were observed in the number and distribution of blood vessels in normal tissues and organs after immunization of the SMAA-GFP animals. In summary, we have shown that a mRNA vaccine against the fusion KLH-AM peptide was able to break peripheral immunotolerance and induce a specific response against the angiogenic factor AM thus reducing tumor burden in two melanoma tumor models in the absence of disturbances to normal blood vessels. The study results indicate a great translational potential and possibility to use it in combinational anti-tumor treatment settings, as well as possibility to overcome challenges of existing therapies targeting angiogenesis and AM itself.","La adrenomedulina (AM) es un factor de crecimiento autocrino/paracrino, además de un regulador crucial de la angiogénesis y la respuesta inmunitaria. La AM se sobreexpresa en la mayoría de los tumores sólidos, lo que la convierte en un buen objetivo para la terapia antitumoral. En este estudio, diseñamos y probamos una vacuna de mRNA dirigida contra un antígeno de fusión compuesto por un pequeño fragmento de hemocianina de lapa (KLH), como hapteno, y AM de ratón. El mRNA sintetizado in vitro se encapsuló en nanopartículas lipídicas (LNP) y se inyectó en ratones C57BL/6J. Se utilizaron LNPs vacías como control negativo. Tras cinco inmunizaciones, se inyectaron células de melanoma B16-F10 a través de la vena de la cola para inducir metástasis pulmonares. Además, se inmunizaron ratones transgénicos que expresaban la proteína verde fluorescente en los vasos sanguíneos (SMAA-GFP) con ambos tipos de LNPs para evaluar el efecto de la vacuna en tumores subcutáneos y sus posibles efectos secundarios sobre la formación de vasos sanguíneos no tumorales. Los títulos de anticuerpos contra la AM y el número de linfocitos T CD8+ fueron significativamente mayores en los ratones inmunizados con KLH-AM que en los controles negativos. Además, el número y el tamaño de las metástasis pulmonares, así como el número de vasos sanguíneos intratumorales, se redujeron significativamente en el grupo inmunizado con KLH-AM. Asimismo, el volumen de los tumores subcutáneos se redujo significativamente, observándose diferencias significativas en el número de vasos sanguíneos dentro de ellos. Además, no se observaron diferencias significativas en el número y la distribución de vasos sanguíneos en tejidos y órganos normales tras la inmunización de los animales SMAA-GFP. En resumen, hemos demostrado que una vacuna de mRNA contra la proteína de fusión KLH-AM logró romper la inmunotolerancia periférica e inducir una respuesta específica contra el factor angiogénico AM, reduciendo así la carga tumoral en dos modelos de melanoma, sin alteraciones en los vasos sanguíneos normales. Los resultados del estudio indican un gran potencial traslacional y la posibilidad de utilizar esta vacuna en tratamientos antitumorales combinados, así como la posibilidad de superar los desafíos de las terapias existentes dirigidas a la angiogénesis y a la propia AM."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Design and development of an RNA vaccine against tumor-mediated angiogenesis and metastasis targeting a KLH-Adrenomedullin fusion protein"]}]}],"canonical_facts":{"dc:contributor":["Martínez Ramírez, Alfredo (Universidad de La Rioja)"],"dc:creator":["Tadic, Srdan"],"dc:date":["2025"],"dc:description":["Adrenomedullin (AM) is an autocrine/paracrine growth factor as well as a crucial regulator of angiogenesis and immune response. AM is overexpressed by most solid tumors, which makes it a good target for anti-tumor therapy. In this study, we designed and tested a mRNA vaccine directed against a fusion antigen composed by a small piece of keyhole limpet hemocyanin (KLH), as a hapten, and mouse AM. The in vitro-synthesized mRNA was encapsulated in lipid nanoparticles (LNPs) and injected in C57BL/6J mice. Empty LNPs were used as a negative control. After five immunizations, B16- F10 melanoma cells were injected through the tail vein to induce lung metastases. In addition, transgenic mice expressing green fluorescent protein in the blood vessels (SMAA-GFP) were also immunized with both LNP types to assess the effect of the vaccine in subcutaneous tumors and potential side-effects of the vaccine on normal blood vessel formation. Antibody titers against AM and the number of CD8+ T cells were significantly higher in AMimmunized mice than in negative controls. Furthermore, the number and size of the lung metastases, as well as the number of blood vessels, were significantly reduced in the AMimmunized group. Furthermore, the volume of subcutaneous tumors was significantly reduced with significant differences observed in the number of blood vessels. In addition, no significant differences were observed in the number and distribution of blood vessels in normal tissues and organs after immunization of the SMAA-GFP animals. In summary, we have shown that a mRNA vaccine against the fusion KLH-AM peptide was able to break peripheral immunotolerance and induce a specific response against the angiogenic factor AM thus reducing tumor burden in two melanoma tumor models in the absence of disturbances to normal blood vessels. The study results indicate a great translational potential and possibility to use it in combinational anti-tumor treatment settings, as well as possibility to overcome challenges of existing therapies targeting angiogenesis and AM itself.","La adrenomedulina (AM) es un factor de crecimiento autocrino/paracrino, además de un regulador crucial de la angiogénesis y la respuesta inmunitaria. La AM se sobreexpresa en la mayoría de los tumores sólidos, lo que la convierte en un buen objetivo para la terapia antitumoral. En este estudio, diseñamos y probamos una vacuna de mRNA dirigida contra un antígeno de fusión compuesto por un pequeño fragmento de hemocianina de lapa (KLH), como hapteno, y AM de ratón. El mRNA sintetizado in vitro se encapsuló en nanopartículas lipídicas (LNP) y se inyectó en ratones C57BL/6J. Se utilizaron LNPs vacías como control negativo. Tras cinco inmunizaciones, se inyectaron células de melanoma B16-F10 a través de la vena de la cola para inducir metástasis pulmonares. Además, se inmunizaron ratones transgénicos que expresaban la proteína verde fluorescente en los vasos sanguíneos (SMAA-GFP) con ambos tipos de LNPs para evaluar el efecto de la vacuna en tumores subcutáneos y sus posibles efectos secundarios sobre la formación de vasos sanguíneos no tumorales. Los títulos de anticuerpos contra la AM y el número de linfocitos T CD8+ fueron significativamente mayores en los ratones inmunizados con KLH-AM que en los controles negativos. Además, el número y el tamaño de las metástasis pulmonares, así como el número de vasos sanguíneos intratumorales, se redujeron significativamente en el grupo inmunizado con KLH-AM. Asimismo, el volumen de los tumores subcutáneos se redujo significativamente, observándose diferencias significativas en el número de vasos sanguíneos dentro de ellos. Además, no se observaron diferencias significativas en el número y la distribución de vasos sanguíneos en tejidos y órganos normales tras la inmunización de los animales SMAA-GFP. En resumen, hemos demostrado que una vacuna de mRNA contra la proteína de fusión KLH-AM logró romper la inmunotolerancia periférica e inducir una respuesta específica contra el factor angiogénico AM, reduciendo así la carga tumoral en dos modelos de melanoma, sin alteraciones en los vasos sanguíneos normales. Los resultados del estudio indican un gran potencial traslacional y la posibilidad de utilizar esta vacuna en tratamientos antitumorales combinados, así como la posibilidad de superar los desafíos de las terapias existentes dirigidas a la angiogénesis y a la propia AM."],"dc:format":["application/pdf"],"dc:identifier":["https://dialnet.unirioja.es/servlet/oaites?codigo=396432"],"dc:language":["eng"],"dc:publisher":["Universidad de La Rioja (España)"],"dc:rights":["LICENCIA DE USO: Los documentos a texto completo incluidos en Dialnet son de acceso libre y propiedad de sus autores y/o editores. Por tanto, cualquier acto de reproducción, distribución, comunicación pública y/o transformación total o parcial requiere el consentimiento expreso y escrito de aquéllos. Cualquier enlace al texto completo de estos documentos deberá hacerse a través de la URL oficial de éstos en Dialnet. Más información: https://dialnet.unirioja.es/info/derechosOAI | INTELLECTUAL PROPERTY RIGHTS STATEMENT: Full text documents hosted by Dialnet are protected by copyright and/or related rights. This digital object is accessible without charge, but its use is subject to the licensing conditions set by its authors or editors. Unless expressly stated otherwise in the licensing conditions, you are free to linking, browsing, printing and making a copy for your own personal purposes. All other acts of reproduction and communication to the public are subject to the licensing conditions expressed by editors and authors and require consent from them. Any link to this document should be made using its official URL in Dialnet. More info: https://dialnet.unirioja.es/info/derechosOAI"],"dc:title":["Design and development of an RNA vaccine against tumor-mediated angiogenesis and metastasis targeting a KLH-Adrenomedullin fusion protein"],"dc:type":["text (thesis)"]},"updated_at":"2026-07-24T06:27:11Z"}