Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacy

[eng] Oncolytic adenoviruses that selectively replicate in cancer cells while sparing normal tissue have gained considerable attention as novel anticancer drugs. However, clinical trials with these viruses have identified the immune system as a major hurdle for their success in cancer patients. Desp...

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Autor: Fajardo Calderón, Carlos Alberto
Tipo de documento: tese
Estado:Versão publicada
Data de publicação:2017
País:España
Recursos:Universidad de Barcelona
Repositório:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/111644
Acesso em linha:https://hdl.handle.net/2445/111644
http://hdl.handle.net/10803/403492
Access Level:Acceso aberto
Palavra-chave:Virologia
Immunoteràpia
Adenovirus
Oncologia
Virology
Immunotheraphy
Adenoviruses
Oncology
id ES_2d0c501ce11e55553cfde3233d84df7d
oai_identifier_str oai:diposit.ub.edu:2445/111644
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacy
title Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacy
spellingShingle Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacy
Fajardo Calderón, Carlos Alberto
Virologia
Immunoteràpia
Adenovirus
Oncologia
Virology
Immunotheraphy
Adenoviruses
Oncology
title_short Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacy
title_full Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacy
title_fullStr Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacy
title_full_unstemmed Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacy
title_sort Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacy
dc.creator.none.fl_str_mv Fajardo Calderón, Carlos Alberto
author Fajardo Calderón, Carlos Alberto
author_facet Fajardo Calderón, Carlos Alberto
author_role author
dc.contributor.none.fl_str_mv Alemany Bonastre, Ramon
Viñals Canals, Francesc
Universitat de Barcelona. Departament de Bioquímica i Biologia Molecular (Farmàcia)
dc.subject.none.fl_str_mv Virologia
Immunoteràpia
Adenovirus
Oncologia
Virology
Immunotheraphy
Adenoviruses
Oncology
topic Virologia
Immunoteràpia
Adenovirus
Oncologia
Virology
Immunotheraphy
Adenoviruses
Oncology
description [eng] Oncolytic adenoviruses that selectively replicate in cancer cells while sparing normal tissue have gained considerable attention as novel anticancer drugs. However, clinical trials with these viruses have identified the immune system as a major hurdle for their success in cancer patients. Despite the existence of a highly immunosuppressive tumor environment, adenovirus-infected cells can nonetheless be efficiently cleared by virus-specific infiltrating cytotoxic T lymphocytes without compromising tumor burden. We hypothesize that arming oncolytic adenoviruses with bi-specific T-cell engagers (BiTEs), a new class of antibodies that re-direct T-cells to cancer cells, might favor antitumor rather than anti-viral immune responses. We have engineered the oncolytic adenovirus ICOVIR-15K to express EGFR-targeting BiTEs under the control of the major late promoter. ICOVIR-15K armed with a BiTE targeting human CD3 and EGFR (ICOVIR-15K-cBiTE) was successfully rescued and it showed similar oncolytic properties as the parental virus. cBiTE expression and secretion was detected in supernatants from ICOVIR-15K-cBiTE-infected cells, and the secreted BiTEs bound specifically to both CD3+ and EGFR+ cells. In cell co-culture assays, ICOVIR-15K-cBiTE-mediated oncolysis resulted in robust T-cell activation, proliferation, and bystander cell-mediated cytotoxicity. Notably, intratumoral injection of this cBiTE-expressing adenovirus increased the persistence and accumulation of tumor-infiltrating T cells in vivo. Moreover, in two distinct tumor xenograft models, combined delivery of ICOVIR-15K-cBiTE with peripheral blood mononuclear cells or T cells enhanced the antitumor efficacy achieved by the parental counterpart. We also demonstrate that another oncolytic adenovirus expressing a chimeric BiTE targeting human EGFR and mouse CD3 (ICOVIR-15K-mcBiTE) induce robust mouse T-cell activation, proliferation, and cell-mediated cytotoxicity of cancer cells in vitro. Thus, ICOVIR-15K-mcBiTE is a promising surrogate of ICOVIR-15K-cBiTE that will aid in future pharmacological and toxicological preclinical studies of BiTE-armed oncolytic adenoviruses. Finally, we show that the combination of ICOVIR-15K-cBiTE with chimeric antigen receptor T-cell therapy can overcome some of the limitations encountered by both agents as monotherapies. The results described in this thesis demonstrate that BiTE-armed oncolytic adenoviruses hold properties with the potential of solving key limitations in oncolytic virotherapy, and encourage their further evaluation and development.
publishDate 2017
dc.date.none.fl_str_mv 2017
dc.type.none.fl_str_mv info:eu-repo/semantics/doctoralThesis
info:eu-repo/semantics/publishedVersion
format doctoralThesis
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/111644
http://hdl.handle.net/10803/403492
url https://hdl.handle.net/2445/111644
http://hdl.handle.net/10803/403492
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv (c) Fajardo, 2017
info:eu-repo/semantics/openAccess
rights_invalid_str_mv (c) Fajardo, 2017
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universitat de Barcelona
publisher.none.fl_str_mv Universitat de Barcelona
dc.source.none.fl_str_mv Tesis Doctorals - Departament - Bioquímica i Biologia Molecular (Farmàcia)
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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spelling Arming oncolytic adenoviruses with bi-specific T-cell engagers to improve antitumor efficacyFajardo Calderón, Carlos AlbertoVirologiaImmunoteràpiaAdenovirusOncologiaVirologyImmunotheraphyAdenovirusesOncology[eng] Oncolytic adenoviruses that selectively replicate in cancer cells while sparing normal tissue have gained considerable attention as novel anticancer drugs. However, clinical trials with these viruses have identified the immune system as a major hurdle for their success in cancer patients. Despite the existence of a highly immunosuppressive tumor environment, adenovirus-infected cells can nonetheless be efficiently cleared by virus-specific infiltrating cytotoxic T lymphocytes without compromising tumor burden. We hypothesize that arming oncolytic adenoviruses with bi-specific T-cell engagers (BiTEs), a new class of antibodies that re-direct T-cells to cancer cells, might favor antitumor rather than anti-viral immune responses. We have engineered the oncolytic adenovirus ICOVIR-15K to express EGFR-targeting BiTEs under the control of the major late promoter. ICOVIR-15K armed with a BiTE targeting human CD3 and EGFR (ICOVIR-15K-cBiTE) was successfully rescued and it showed similar oncolytic properties as the parental virus. cBiTE expression and secretion was detected in supernatants from ICOVIR-15K-cBiTE-infected cells, and the secreted BiTEs bound specifically to both CD3+ and EGFR+ cells. In cell co-culture assays, ICOVIR-15K-cBiTE-mediated oncolysis resulted in robust T-cell activation, proliferation, and bystander cell-mediated cytotoxicity. Notably, intratumoral injection of this cBiTE-expressing adenovirus increased the persistence and accumulation of tumor-infiltrating T cells in vivo. Moreover, in two distinct tumor xenograft models, combined delivery of ICOVIR-15K-cBiTE with peripheral blood mononuclear cells or T cells enhanced the antitumor efficacy achieved by the parental counterpart. We also demonstrate that another oncolytic adenovirus expressing a chimeric BiTE targeting human EGFR and mouse CD3 (ICOVIR-15K-mcBiTE) induce robust mouse T-cell activation, proliferation, and cell-mediated cytotoxicity of cancer cells in vitro. Thus, ICOVIR-15K-mcBiTE is a promising surrogate of ICOVIR-15K-cBiTE that will aid in future pharmacological and toxicological preclinical studies of BiTE-armed oncolytic adenoviruses. Finally, we show that the combination of ICOVIR-15K-cBiTE with chimeric antigen receptor T-cell therapy can overcome some of the limitations encountered by both agents as monotherapies. The results described in this thesis demonstrate that BiTE-armed oncolytic adenoviruses hold properties with the potential of solving key limitations in oncolytic virotherapy, and encourage their further evaluation and development.[spa] Los virus oncolitos, capaces de infectar selectivamente células cancerosas sin afectar aquellas sanas, han despertado interés en los últimos años como nueva terapia contra el cáncer. Sin embargo, los ensayos clínicos con estos virus han demostrado que el sistema inmune supone un obstáculo para el éxito de los mismos en pacientes con cáncer. A pesar de la inmunosupresión que se observa en el ambiente tumoral, las células cancerosas infectadas por el adenovirus pueden ser eliminadas eficientemente por los linfocitos T anti-adenovirales sin comprometer la carga tumoral. La hipótesis de esta tesis es que adenovirus oncoliticos expresando bi-specific T-cell engagers (BiTEs por sus siglas en inglés) capaces de redirgirir los linfocitos T para atacar las células cancerosas, puede favorecer la respuesta inmune antitumoral sobre la antiviral. El genoma del adenovirus oncolitico ICOVIR-15K fue modificado genéticamente para expresar BiTEs contra el receptor del factor de crecimiento epidérmico (EGFR por sus siglas en inglés) bajo el control del promotor mayor tardío. El virus ICOVIR-15K expresando un BiTE que reconoce el EGFR y el CD3 humanos (ICOVIR-15K-cBiTE) fue generado y retuvo propiedades oncoliticas similares a la del virus parental in vitro. La expresión y secreción del cBiTE fue detectada en los sobrenadantes de células infectadas ICOVIR-15K-cBiTE, y sus propiedades de unión a células CD3+ o EGFR+ fueron confirmadas in vitro. En experimentos de cocultivos, la oncolisis generada por ICOVIR-15K-cBiTE indujo la activación y proliferación de los linfocitos T, y aumentó la citotoxicidad de células cancerosas. La inyección de este adenovirus aumentó la persistencia y la acumulación de linfocitos infiltrantes de tumor in vivo. Adicionalmente, experimentos en modelos murinos de cáncer basados en la administración combinada de ICOVIR-15K-cBiTE y linfocitos humanos demostraron un aumento en la eficacia antitumoral comparado con el virus parental. Por último, hemos demostrado que la combinación de ICOVIR-15K-cBiTE y linfocitos T con receptores de antígeno quiméricos (CAR por sus siglas en inglés) pueden superar muchas de las carencias que tienen ambas terapias. Los resultados de esta tesis demuestran que los adenovirus oncoliticos expresando BiTEs tienen propiedades que puede superar muchas de las limitaciones de la viroterapia del cáncer, y alienta a continuar su evaluación y desarrollo a nivel clínico.Universitat de BarcelonaAlemany Bonastre, RamonViñals Canals, FrancescUniversitat de Barcelona. Departament de Bioquímica i Biologia Molecular (Farmàcia)2017info:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/111644http://hdl.handle.net/10803/403492Tesis Doctorals - Departament - Bioquímica i Biologia Molecular (Farmàcia)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglés(c) Fajardo, 2017info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/1116442026-05-27T06:46:51Z
score 15,300724

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