Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transfer

IL-12 is a potent cytokine for cancer immunotherapy. However, its systemic delivery as a recombinant protein has shown unacceptable toxicity in early clinical trials, mostly related to the secondary increase in IFN- levels in circulation. In the last two decades, a wide plethora of methodologies has...

Full description

Bibliographic Details
Author: Di-Trani, C.A. (Claudia Augusta)|||/items/5d899126-779d-4c97-8232-95bfe18f9f26
Format: doctoral thesis
Publication Date:2023
Country:España
Institution:Universidad de Navarra
Repository:Dadun. Depósito Académico Digital de la Universidad de Navarra
Language:English
OAI Identifier:oai:dadun.unav.edu:10171/67876
Online Access:https://hdl.handle.net/10171/67876
Access Level:Open access
Keyword:Materias Investigacion::Ciencias de la Salud::Inmunología
Cancer immunotherapy
mRNA-based technology
Immunocytokines
Armored T cell transfer
IL-12
Locoregional immunotherapy
Cytokine therapy
id ES_2d3bb7f55259b72da65a7a0fa3e663a7
oai_identifier_str oai:dadun.unav.edu:10171/67876
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transfer
title Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transfer
spellingShingle Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transfer
Di-Trani, C.A. (Claudia Augusta)|||/items/5d899126-779d-4c97-8232-95bfe18f9f26
Materias Investigacion::Ciencias de la Salud::Inmunología
Cancer immunotherapy
mRNA-based technology
Immunocytokines
Armored T cell transfer
IL-12
Locoregional immunotherapy
Cytokine therapy
title_short Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transfer
title_full Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transfer
title_fullStr Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transfer
title_full_unstemmed Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transfer
title_sort Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transfer
dc.creator.none.fl_str_mv Di-Trani, C.A. (Claudia Augusta)|||/items/5d899126-779d-4c97-8232-95bfe18f9f26
author Di-Trani, C.A. (Claudia Augusta)|||/items/5d899126-779d-4c97-8232-95bfe18f9f26
author_facet Di-Trani, C.A. (Claudia Augusta)|||/items/5d899126-779d-4c97-8232-95bfe18f9f26
author_role author
dc.contributor.none.fl_str_mv Berraondo-López, P. (Pedro)
Melero, I. (Ignacio)
Dadun. Depósito Académico Digital Universidad de Navarra
dc.subject.none.fl_str_mv Materias Investigacion::Ciencias de la Salud::Inmunología
Cancer immunotherapy
mRNA-based technology
Immunocytokines
Armored T cell transfer
IL-12
Locoregional immunotherapy
Cytokine therapy
topic Materias Investigacion::Ciencias de la Salud::Inmunología
Cancer immunotherapy
mRNA-based technology
Immunocytokines
Armored T cell transfer
IL-12
Locoregional immunotherapy
Cytokine therapy
description IL-12 is a potent cytokine for cancer immunotherapy. However, its systemic delivery as a recombinant protein has shown unacceptable toxicity in early clinical trials, mostly related to the secondary increase in IFN- levels in circulation. In the last two decades, a wide plethora of methodologies has been employed to harness the desired anti-tumor features of IL-12 whilst limiting its side effects. Pro-cytokines and immunocytokines are some of the examples used to favor IL-12 bioactivity selectively in the TME. Also, the intra-tumoral delivery of IL-12-encoding mRNA or DNA is being evaluated in clinical trials. The first chapter of the thesis provides evidence of a viable platform to generate mRNAs encoding for chimeric constructs designed to customize the desired properties of the therapeutic cytokines. Upon intra-tumor injection of naked mRNA in murine models, our results demonstrated that the in vivo translated and secreted molecules encompassing single-chain IL-12 fused to diabodies targeting CSF1R, PD-L1 or both tether this potent cytokine onto the surface of tumor-infiltrating myeloid cells, preventing its systemic leakage, when compared to non-chimerized IL-12. To our surprise, the IL-12-diabody targeting PD-L1 is sufficient to achieve high tethering capacity, refuting our prediction that the double targeting would be more effective than the single ones. Furthermore, the anti-tumor efficacy of the IL-12-diabodies was comparable to that achieved by non-targeted IL-12, demonstrating that the fusion of the diabody to the cytokine does not hinder its potent bioactivity in vivo. The transient expression of IL-12 in the tumor niche, as well as the prevented leakage to the periphery mediated by the mRNA-encoded immunocytokines, holds promise for expanding the methodologies aimed at making the most of IL-12-anti-tumor properties whilst limiting its associated side effects. In the second chapter of the thesis we evaluated the effectiveness of tumor-specific CD8+ T cells, engineered with IL-12-encoding mRNA, against murine models of peritoneal metastasis. In order to recapitulate the human disease where the omentum represents a preferential location of metastasis, we employed intraperitoneal injection of tumor cells that preferentially localized to the omentum. We compared the intravenous and locoregional routes of administration of engineered OT-I T cell therapy in terms of anti-tumoral efficacy and reprogramming of the tumor microenvironment. Our findings clearly demonstrated the superiority of intracavitary immunotherapy: local delivery rapidly localized the armored T cell therapy to the omentum, where early encounter with tumor cell antigens and localized exposure of the tumor niche to secreted IL-12 resulted in a significantly higher anti-tumor response and in a more favorable transcriptional reprogramming of the omentum. Transient IL-12 expression was responsible for higher activation status of the immune cells in the TME, longer persistence of transferred T lymphocytes in vivo, and the development of immunity to endogenous antigens following primary tumor eradication. The anti-tumor efficacy of T cells with lower affinity TCRs was also improved by IL-12-mRNA-engineering, but to a lesser extent than T cells bearing higher affinity TCRs. Our data point to an alternative and safer route for the intracavitary delivery of adoptive T-cell therapies engineered to express IL-12 transiently.
publishDate 2023
dc.date.none.fl_str_mv 2023
2023-11-16
2023
2023-11-16
2023
2023-11-16
2023
2023-07-04
dc.type.none.fl_str_mv doctoral thesis
http://purl.org/coar/resource_type/c_db06
dc.type.openaire.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
dc.identifier.none.fl_str_mv https://hdl.handle.net/10171/67876
url https://hdl.handle.net/10171/67876
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidad de Navarra
publisher.none.fl_str_mv Universidad de Navarra
dc.source.none.fl_str_mv reponame:Dadun. Depósito Académico Digital de la Universidad de Navarra
instname:Universidad de Navarra
instname_str Universidad de Navarra
reponame_str Dadun. Depósito Académico Digital de la Universidad de Navarra
collection Dadun. Depósito Académico Digital de la Universidad de Navarra
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869405301137997824
spelling Tailoring the bioactivities of IL-12 for cancer immunotherapy with mRNA-based technology: from immunocytokines to armored T cell transferDi-Trani, C.A. (Claudia Augusta)|||/items/5d899126-779d-4c97-8232-95bfe18f9f26Materias Investigacion::Ciencias de la Salud::InmunologíaCancer immunotherapymRNA-based technologyImmunocytokinesArmored T cell transferIL-12Locoregional immunotherapyCytokine therapyIL-12 is a potent cytokine for cancer immunotherapy. However, its systemic delivery as a recombinant protein has shown unacceptable toxicity in early clinical trials, mostly related to the secondary increase in IFN- levels in circulation. In the last two decades, a wide plethora of methodologies has been employed to harness the desired anti-tumor features of IL-12 whilst limiting its side effects. Pro-cytokines and immunocytokines are some of the examples used to favor IL-12 bioactivity selectively in the TME. Also, the intra-tumoral delivery of IL-12-encoding mRNA or DNA is being evaluated in clinical trials. The first chapter of the thesis provides evidence of a viable platform to generate mRNAs encoding for chimeric constructs designed to customize the desired properties of the therapeutic cytokines. Upon intra-tumor injection of naked mRNA in murine models, our results demonstrated that the in vivo translated and secreted molecules encompassing single-chain IL-12 fused to diabodies targeting CSF1R, PD-L1 or both tether this potent cytokine onto the surface of tumor-infiltrating myeloid cells, preventing its systemic leakage, when compared to non-chimerized IL-12. To our surprise, the IL-12-diabody targeting PD-L1 is sufficient to achieve high tethering capacity, refuting our prediction that the double targeting would be more effective than the single ones. Furthermore, the anti-tumor efficacy of the IL-12-diabodies was comparable to that achieved by non-targeted IL-12, demonstrating that the fusion of the diabody to the cytokine does not hinder its potent bioactivity in vivo. The transient expression of IL-12 in the tumor niche, as well as the prevented leakage to the periphery mediated by the mRNA-encoded immunocytokines, holds promise for expanding the methodologies aimed at making the most of IL-12-anti-tumor properties whilst limiting its associated side effects. In the second chapter of the thesis we evaluated the effectiveness of tumor-specific CD8+ T cells, engineered with IL-12-encoding mRNA, against murine models of peritoneal metastasis. In order to recapitulate the human disease where the omentum represents a preferential location of metastasis, we employed intraperitoneal injection of tumor cells that preferentially localized to the omentum. We compared the intravenous and locoregional routes of administration of engineered OT-I T cell therapy in terms of anti-tumoral efficacy and reprogramming of the tumor microenvironment. Our findings clearly demonstrated the superiority of intracavitary immunotherapy: local delivery rapidly localized the armored T cell therapy to the omentum, where early encounter with tumor cell antigens and localized exposure of the tumor niche to secreted IL-12 resulted in a significantly higher anti-tumor response and in a more favorable transcriptional reprogramming of the omentum. Transient IL-12 expression was responsible for higher activation status of the immune cells in the TME, longer persistence of transferred T lymphocytes in vivo, and the development of immunity to endogenous antigens following primary tumor eradication. The anti-tumor efficacy of T cells with lower affinity TCRs was also improved by IL-12-mRNA-engineering, but to a lesser extent than T cells bearing higher affinity TCRs. Our data point to an alternative and safer route for the intracavitary delivery of adoptive T-cell therapies engineered to express IL-12 transiently.Universidad de NavarraBerraondo-López, P. (Pedro)Melero, I. (Ignacio)Dadun. Depósito Académico Digital Universidad de Navarra20232023-11-1620232023-11-1620232023-11-1620232023-07-04doctoral thesishttp://purl.org/coar/resource_type/c_db06info:eu-repo/semantics/doctoralThesisapplication/pdfhttps://hdl.handle.net/10171/67876reponame:Dadun. Depósito Académico Digital de la Universidad de Navarrainstname:Universidad de NavarraInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:dadun.unav.edu:10171/678762026-06-21T12:47:57Z
score 15,301603

Similar Items