Identificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncer

ilustraciones, diagramas, gráficas. tablas

Autores:
Martínez Enríquez, Laura Camila
Tipo de recurso:
Fecha de publicación:
2022
Institución:
Universidad Nacional de Colombia
Repositorio:
Universidad Nacional de Colombia
Idioma:
spa
OAI Identifier:
oai:repositorio.unal.edu.co:unal/82934
Acceso en línea:
https://repositorio.unal.edu.co/handle/unal/82934
https://repositorio.unal.edu.co/
Palabra clave:
610 - Medicina y salud::616 - Enfermedades
Prevención del cáncer
Sistema Inmunológico
Cancer Prevention
Immune System
Neoantígenos
inmunogenicidad
donantes sanos
Linfocitos T CD8
tetrámero
Sistemas de cultivo
Neoantigens
CD8 T cell
healthy donor
Immugenicity
Rights
openAccess
License
Reconocimiento 4.0 Internacional
id UNACIONAL2_5cef99512c9c3bd55dad20dc2c345a36
oai_identifier_str oai:repositorio.unal.edu.co:unal/82934
network_acronym_str UNACIONAL2
network_name_str Universidad Nacional de Colombia
repository_id_str
dc.title.spa.fl_str_mv Identificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncer
dc.title.translated.eng.fl_str_mv Identification and characterization of antigen specific t cells from healthy donors for cancer immunotherapy
title Identificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncer
spellingShingle Identificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncer
610 - Medicina y salud::616 - Enfermedades
Prevención del cáncer
Sistema Inmunológico
Cancer Prevention
Immune System
Neoantígenos
inmunogenicidad
donantes sanos
Linfocitos T CD8
tetrámero
Sistemas de cultivo
Neoantigens
CD8 T cell
healthy donor
Immugenicity
title_short Identificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncer
title_full Identificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncer
title_fullStr Identificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncer
title_full_unstemmed Identificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncer
title_sort Identificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncer
dc.creator.fl_str_mv Martínez Enríquez, Laura Camila
dc.contributor.advisor.none.fl_str_mv Parra López, Carlos Alberto
dc.contributor.author.none.fl_str_mv Martínez Enríquez, Laura Camila
dc.contributor.researchgroup.spa.fl_str_mv Inmunología y Medicina Traslacional
dc.contributor.orcid.spa.fl_str_mv Martinez Enriquez, Laura Camila [0000-0003-0799-942X]
dc.contributor.cvlac.spa.fl_str_mv Martínez Enríquez, Laura Camila [0001705413]
dc.subject.ddc.spa.fl_str_mv 610 - Medicina y salud::616 - Enfermedades
topic 610 - Medicina y salud::616 - Enfermedades
Prevención del cáncer
Sistema Inmunológico
Cancer Prevention
Immune System
Neoantígenos
inmunogenicidad
donantes sanos
Linfocitos T CD8
tetrámero
Sistemas de cultivo
Neoantigens
CD8 T cell
healthy donor
Immugenicity
dc.subject.decs.spa.fl_str_mv Prevención del cáncer
Sistema Inmunológico
dc.subject.decs.eng.fl_str_mv Cancer Prevention
Immune System
dc.subject.proposal.spa.fl_str_mv Neoantígenos
inmunogenicidad
donantes sanos
Linfocitos T CD8
tetrámero
Sistemas de cultivo
dc.subject.proposal.eng.fl_str_mv Neoantigens
CD8 T cell
healthy donor
Immugenicity
description ilustraciones, diagramas, gráficas. tablas
publishDate 2022
dc.date.issued.none.fl_str_mv 2022-11-15
dc.date.accessioned.none.fl_str_mv 2023-01-16T12:57:53Z
dc.date.available.none.fl_str_mv 2023-01-16T12:57:53Z
dc.type.spa.fl_str_mv Trabajo de grado - Maestría
dc.type.driver.spa.fl_str_mv info:eu-repo/semantics/masterThesis
dc.type.version.spa.fl_str_mv info:eu-repo/semantics/acceptedVersion
dc.type.content.spa.fl_str_mv Text
dc.type.redcol.spa.fl_str_mv http://purl.org/redcol/resource_type/TM
status_str acceptedVersion
dc.identifier.uri.none.fl_str_mv https://repositorio.unal.edu.co/handle/unal/82934
dc.identifier.instname.spa.fl_str_mv Universidad Nacional de Colombia
dc.identifier.reponame.spa.fl_str_mv Repositorio Institucional Universidad Nacional de Colombia
dc.identifier.repourl.spa.fl_str_mv https://repositorio.unal.edu.co/
url https://repositorio.unal.edu.co/handle/unal/82934
https://repositorio.unal.edu.co/
identifier_str_mv Universidad Nacional de Colombia
Repositorio Institucional Universidad Nacional de Colombia
dc.language.iso.spa.fl_str_mv spa
language spa
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rights_invalid_str_mv Reconocimiento 4.0 Internacional
Derechos reservados al autor, 2016
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dc.format.extent.spa.fl_str_mv 144 páginas
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dc.publisher.spa.fl_str_mv Universidad Nacional de Colombia
dc.publisher.program.spa.fl_str_mv Bogotá - Medicina - Maestría en Inmunología
dc.publisher.faculty.spa.fl_str_mv Facultad de Medicina
dc.publisher.place.spa.fl_str_mv Bogotá, Colombia
dc.publisher.branch.spa.fl_str_mv Universidad Nacional de Colombia - Sede Bogotá
institution Universidad Nacional de Colombia
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spelling Reconocimiento 4.0 InternacionalDerechos reservados al autor, 2016http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Parra López, Carlos Alberto29924f3507fcedbca9501300464d5b61Martínez Enríquez, Laura Camilad5ebd3344dcfeec087b3f188a87a136cInmunología y Medicina TraslacionalMartinez Enriquez, Laura Camila [0000-0003-0799-942X]Martínez Enríquez, Laura Camila [0001705413]2023-01-16T12:57:53Z2023-01-16T12:57:53Z2022-11-15https://repositorio.unal.edu.co/handle/unal/82934Universidad Nacional de ColombiaRepositorio Institucional Universidad Nacional de Colombiahttps://repositorio.unal.edu.co/ilustraciones, diagramas, gráficas. tablasLa inmunoterapia basada en neoantígenos permite estimular el sistema inmune del paciente con cáncer al inducir una respuesta antitumoral dirigida mediada por Linfocitos T (LT). Los neoantígenos son generados por mutaciones somáticas en el ADN que producen cambios en la secuencia de aminoácidos y que son exclusivas de las células tumorales. La selección de neoantígenos inmunogénicos se realiza por medio de herramientas in-silico que predicen la afinidad y tiempo de unión del neoantígeno a la molécula de HLA, luego estos son evaluados en sistemas de cultivo in-vitro con las células de los pacientes, sin embargo, la frecuencia reportada de respuestas a neoantígenos es aún baja. Por lo tanto, este estudio planteó dos acercamientos diferentes para identificar y caracterizar neoantígenos inmunogénicos. Por un lado, se propuso la implementación de sistemas de cultivo con células de donantes sanos para evaluar la inmunogenicidad de los neoantígenos de manera in-vitro. Por otra parte, se propuso el uso del docking y la dinámica molecular para identificar características moleculares asociadas a la inmunogenicidad de los neoantígenos. Para el primer enfoque se utilizaron células mononucleares de sangre periférica (PBMCs) de donantes sanos HLA-A*02:01. Se evaluaron cuatro tipos de cultivos diferentes, manteniendo el uso de las citoquinas IL-21, IL-15 e IL-7 pero modificando las células de partida: i) PBMCs totales, ii) cocultivo acelerado con células dendríticas a partir de PBMCs, iii) cocultivo de células dendríticas in-situ (DCs in-situ) con LT CD8+ vírgenes enriquecidos y iv) cocultivo de células dendríticas por adherencia de monocitos (moDCs) con LT CD8+ vírgenes enriquecidos. Los neoantígenos restringidos a HLA-A*02:01 fueron seleccionados a partir de una búsqueda en la literatura y se evaluaron en forma de pool. El reconocimiento de los LT CD8+ a neoantígenos se evaluó mediante la producción de las citoquinas IFN-γ y TNF-a y por la marcación de tetrámeros. Como resultados se pudo observar que es necesaria la presencia de células presentadoras profesionales, como lo son las DC, y un enriquecimiento de los LT CD8 vírgenes, pues fue en este cultivo que se logró detectar, aunque en baja proporción, LT específicos contra los neoantígenos. No obstante, estos resultados solo se observaron en 2 de 4 donantes evaluados, lo cual indica que es necesario realizar ensayos adicionales para poder determinar que este sistema de cultivo es el indicado. Para el segundo enfoque se realizó una prueba de concepto con dos neoantígenos (uno inmunogénico y otro no inmunogénico) para evaluar el uso del docking y la dinámica molecular como herramientas de tamizaje para la identificación de neoantígenos inmunogénicos, permitiendo determinar que un neoantígeno inmunogénico debe formar un complejo péptido-MHC estable en el tiempo. Este estudio demuestra la alta complejidad que representa el uso de células de donantes sanos y de las herramientas computacionales de docking y dinámica molecular, sin embargo, estos dos enfoques son prometedores ya que no solo permitirían mejorar la selección de neoantígenos inmunogénicos sino también tienen el potencial de identificar TCR específicos contra estos antígenos con fines de terapia adoptiva celular basada en modificación del TCR. (Texto tomado de la fuente)Immunotherapy based on neoantigens allows to stimulate the immune system of cancer patients by inducing a directed antitumor response mediated by T cells. Neoantigens are generated by somatic mutations in DNA that produce changes in the amino acid sequence and are exclusive to tumor cells. The selection of immunogenic neoantigens to predict the affinity and binding of the neoantigen to the HLA is performed in silico and then evaluated with in vitro culture assays with patient cells, however, the reported frequency of responses to neoantigens is low. Therefore, this study proposed two different approaches to identify and characterize immunogenic neoantigens. On the one hand, the implementation of culture systems with cells from healthy donors to evaluate the immunogenicity of neoantigens in vitro. On the other hand, the use of docking and molecular dynamics to identify in silico molecular characteristics associated with the immunogenicity of neoantigens. To evaluate the first approach, peripheral blood mononuclear cells (PBMCs) from healthy HLA-A*02:01 donors were used as a model. HLA-A*02:01-restricted neoantigens were selected from a literature search and evaluated as a pool. Four different types of cultures were evaluated, maintaining the use of the cytokines IL-21, IL-15 and IL-7 but modifying the starting cells: i) total PBMCs, ii) accelerated co-culture with dendritic cells (acDCs) from PBMCs and iii ) co-culture of dendritic cells in situ with Naïve CD8+ T cells and iv) co-culture of monocyte derived dendritic cells with Naïve CD8+ T cells. Recognition of CD8+ T cells to neoantigens was assessed by cytokine production and by tetramer labeling. It was possible to observe that the presence of professional antigen presenting cells, such as DCs, and an enrichment of Naïve CD8+ T cells is necessary to detect, although in low proportion, specific LTs against neoantigens. However, these results were only observed in 2 of 4 donors evaluated, which indicates that additional tests are necessary to determine if this culture system work. For the second approach, a proof of concept was carried out with two neoantigens (one immunogenic and one non-immunogenic) to evaluate the use of docking and molecular dynamics as screening tools for the identification of immunogenic neoantigens, allowing to determine that an immunogenic neoantigen should form a peptide-MHC complex stable over time. This study demonstrates the high complexity of using healthy donor cells and tools such as molecular dynamics and docking, however, these two approaches are promising since they would not only allow to improve the selection of immunogenic neoantigens but also the identification of TCRs specific to neoantigens for adoptive cell therapy with cell modification of the TCRMaestríaMagíster en InmunologíaVacunas contra el cáncer144 páginasapplication/pdfspaUniversidad Nacional de ColombiaBogotá - Medicina - Maestría en InmunologíaFacultad de MedicinaBogotá, ColombiaUniversidad Nacional de Colombia - Sede Bogotá610 - Medicina y salud::616 - EnfermedadesPrevención del cáncerSistema InmunológicoCancer PreventionImmune SystemNeoantígenosinmunogenicidaddonantes sanosLinfocitos T CD8tetrámeroSistemas de cultivoNeoantigensCD8 T cellhealthy donorImmugenicityIdentificación y caracterización de linfocitos T neoantígeno específicos de donantes sanos con fines de inmunoterapia en cáncerIdentification and characterization of antigen specific t cells from healthy donors for cancer immunotherapyTrabajo de grado - Maestríainfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/acceptedVersionTexthttp://purl.org/redcol/resource_type/TM1. 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