Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol

Detalhes bibliográficos
Autor(a) principal: Pessoa, Carina Carraro [UNIFESP]
Data de Publicação: 2019
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UNIFESP
Texto Completo: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=8044650
https://repositorio.unifesp.br/handle/11600/59272
Resumo: Vacuolar H+-ATPases (v-ATPases) are membrane-associated ATP-dependent multimeric enzymes responsible for pumping protons from the cytosol into the lumen of intracellular organelles, thus controlling the acidification of lysosomes, endosomes and other intracellular vesicles. It exhibit two functional domains, the V1 (cytosolic, composed of 8 subunits) and V0 (membranous, composed of 6 subunits). In addition to organelle acidification, v-ATPases are alternatively implicated in membrane fusion and anti-inflammatory functions controlled by a variant subunit, d2, of the V0 domain (ATP6V0d2). Leishmania spp. are parasites that cause cutaneous or visceral leishmaniasis in humans and other animals and are a major health problem in poor and developing countries. They are dimorphic parasites found extracellularly in the midgut of insect vectors as flagellated and elongated promastigotes and intracellularly in mammalian host macrophages as rounded amastigotes. Species such as Leishmania (Leishmania) amazonensis, L. mexicana and L. pifanoi are known to multiply in large and fusogenic vacuoles parasitoforos (VP), inducing the positive regulation of ATP6V0d2. In comparison to other Leishmania species, they also exhibit, at least in vitro, remarkable resistance to mechanisms of parasite elimination mediated by interferon-gamma (IFN-γ) and lipopolysaccharide (LPS) within macrophages or by direct treatment with species reactive oxygen species (ROS). However, a causal relationship between the development of large VPs and parasite resistance in inflammatory macrophages remains unclear, especially in vivo. Therefore, we evaluated the role of ATP6V0d2 in the biogenesis of pathogen-containing vacuoles using macrophages silenced to ATP6V0d2 infected with the protozoan parasite L. amazonensis. ATP6V0d2 knock-down decreased the cholesterol levels of the macrophages and inhibited the increase of the VP without interfering in the multiplication of the parasite. However, the parasites required ATP6V0d2 to resist the influx of oxidized low-density lipoprotein cholesterol (ox-LDL), which restored the increase of VP in silenced macrophages by replacing the cholesterol of the macrophages. Thus, we disclose the subversion of parasite-mediated V-ATPase function of the host cell relative to cholesterol retention, which is required to establish an inflammatory resistant intracellular parasite niche.
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spelling Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterolLeishmania vacuole biogenesis: role of the ATP6V0d2 subunit in cholesterol homeostasisv-ATPaseATP6V0d2LeishmaniamacrófagosoxLDLVacuolar H+-ATPases (v-ATPases) are membrane-associated ATP-dependent multimeric enzymes responsible for pumping protons from the cytosol into the lumen of intracellular organelles, thus controlling the acidification of lysosomes, endosomes and other intracellular vesicles. It exhibit two functional domains, the V1 (cytosolic, composed of 8 subunits) and V0 (membranous, composed of 6 subunits). In addition to organelle acidification, v-ATPases are alternatively implicated in membrane fusion and anti-inflammatory functions controlled by a variant subunit, d2, of the V0 domain (ATP6V0d2). Leishmania spp. are parasites that cause cutaneous or visceral leishmaniasis in humans and other animals and are a major health problem in poor and developing countries. They are dimorphic parasites found extracellularly in the midgut of insect vectors as flagellated and elongated promastigotes and intracellularly in mammalian host macrophages as rounded amastigotes. Species such as Leishmania (Leishmania) amazonensis, L. mexicana and L. pifanoi are known to multiply in large and fusogenic vacuoles parasitoforos (VP), inducing the positive regulation of ATP6V0d2. In comparison to other Leishmania species, they also exhibit, at least in vitro, remarkable resistance to mechanisms of parasite elimination mediated by interferon-gamma (IFN-γ) and lipopolysaccharide (LPS) within macrophages or by direct treatment with species reactive oxygen species (ROS). However, a causal relationship between the development of large VPs and parasite resistance in inflammatory macrophages remains unclear, especially in vivo. Therefore, we evaluated the role of ATP6V0d2 in the biogenesis of pathogen-containing vacuoles using macrophages silenced to ATP6V0d2 infected with the protozoan parasite L. amazonensis. ATP6V0d2 knock-down decreased the cholesterol levels of the macrophages and inhibited the increase of the VP without interfering in the multiplication of the parasite. However, the parasites required ATP6V0d2 to resist the influx of oxidized low-density lipoprotein cholesterol (ox-LDL), which restored the increase of VP in silenced macrophages by replacing the cholesterol of the macrophages. Thus, we disclose the subversion of parasite-mediated V-ATPase function of the host cell relative to cholesterol retention, which is required to establish an inflammatory resistant intracellular parasite niche.As ATPases Vacuolares H + (v-ATPases) são enzimas multiméricas dependentes de ATP associadas à membrana, responsáveis por bombear prótons do citosol para o lúmen das organelas intracelulares, controlando assim a acidificação dos lisossomos, endossomos e outras vesículas intracelulares. Elas exibem dois domínios funcionais, o V1 (citosólico, composto por 8 subunidades) e V0 (membranar, composto por 6 subunidades). Além da acidificação de organelas, as v-ATPases são alternativamente implicadas na fusão de membrana e nas funções anti-inflamatórias controladas por uma subunidade variante, d2, do domínio V0 (ATP6V0d2). Leishmania spp. são parasitas que causam leishmaniose tegumentar ou visceral em humanos e outros animais e constituem um grande problema de saúde em países pobres e em desenvolvimento. São parasitas dimórficos encontrados extracelularmente no intestino médio dos insetos vetores como promastigotas flagelados e alongados e intracelularmente em macrófagos de hospedeiros mamíferos como amastigotas arredondadas. Espécies como Leishmania (Leishmania) amazonensis, L. mexicana e L. pifanoi são conhecidas por se multiplicarem em vacúolos parasitóforos (VP) grandes e fusogênicos, induzindo a regulação positiva de ATP6V0d2. Em comparação com outras espécies de Leishmania, elas também exibem, pelo menos in vitro, uma notável resistência a mecanismos de eliminação de parasitas mediados por interferon-gama (IFN-γ) e lipopolissacarídeo (LPS) dentro de macrófagos ou por tratamento direto com espécies reativas de oxigênio (ROS). Entretanto, uma relação causal entre o desenvolvimento de grandes VPs e a resistência do parasita em macrófagos inflamatórios permanece obscura, especialmente in vivo. Portanto, avaliamos o papel do ATP6V0d2 na biogênese de vacúolos contendo patógenos utilizando macrófagos silenciados para ATP6V0d2 infectados com o protozoário parasita L. amazonensis. O silenciamento de ATP6V0d2 diminuiu os níveis de colesterol dos macrófagos e inibiu o aumento do VP sem interferir na multiplicação do parasita. No entanto, os parasitas necessitaram de ATP6V0d2 para resistir ao influxo de colesterol derivado de lipoproteína de baixa densidade oxidada (ox-LDL), que restaurou o aumento do VP em macrófagos silenciados, repondo o colesterol dos macrófagos. Assim, revelamos a subversão da função da V-ATPase da célula hospedeira mediada por parasitas em relação à retenção de colesterol, que é necessária para estabelecer um nicho de parasita intracelular resistente à inflamação.Dados abertos - Sucupira - Teses e dissertações (2019)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).Universidade Federal de São Paulo (UNIFESP)Mortara, Renato Arruda [UNIFESP]http://lattes.cnpq.br/3754467086294573http://lattes.cnpq.br/4471823578370670Universidade Federal de São Paulo (UNIFESP)Pessoa, Carina Carraro [UNIFESP]2021-01-19T16:32:03Z2021-01-19T16:32:03Z2019-06-27info:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersion100 f.application/pdfhttps://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=8044650PESSOA, Carina Carraro. Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol. 2019. 100f. Tese (Doutorado em Microbiologia e Imunologia) – Escola Paulista de Medicina, Universidade Federal de São Paulo. São Paulo, 2019.https://repositorio.unifesp.br/handle/11600/59272porSão Pauloinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2024-08-10T21:03:46Zoai:repositorio.unifesp.br/:11600/59272Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-08-10T21:03:46Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false
dc.title.none.fl_str_mv Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol
Leishmania vacuole biogenesis: role of the ATP6V0d2 subunit in cholesterol homeostasis
title Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol
spellingShingle Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol
Pessoa, Carina Carraro [UNIFESP]
v-ATPase
ATP6V0d2
Leishmania
macrófagos
oxLDL
title_short Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol
title_full Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol
title_fullStr Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol
title_full_unstemmed Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol
title_sort Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol
author Pessoa, Carina Carraro [UNIFESP]
author_facet Pessoa, Carina Carraro [UNIFESP]
author_role author
dc.contributor.none.fl_str_mv Mortara, Renato Arruda [UNIFESP]
http://lattes.cnpq.br/3754467086294573
http://lattes.cnpq.br/4471823578370670
Universidade Federal de São Paulo (UNIFESP)
dc.contributor.author.fl_str_mv Pessoa, Carina Carraro [UNIFESP]
dc.subject.por.fl_str_mv v-ATPase
ATP6V0d2
Leishmania
macrófagos
oxLDL
topic v-ATPase
ATP6V0d2
Leishmania
macrófagos
oxLDL
description Vacuolar H+-ATPases (v-ATPases) are membrane-associated ATP-dependent multimeric enzymes responsible for pumping protons from the cytosol into the lumen of intracellular organelles, thus controlling the acidification of lysosomes, endosomes and other intracellular vesicles. It exhibit two functional domains, the V1 (cytosolic, composed of 8 subunits) and V0 (membranous, composed of 6 subunits). In addition to organelle acidification, v-ATPases are alternatively implicated in membrane fusion and anti-inflammatory functions controlled by a variant subunit, d2, of the V0 domain (ATP6V0d2). Leishmania spp. are parasites that cause cutaneous or visceral leishmaniasis in humans and other animals and are a major health problem in poor and developing countries. They are dimorphic parasites found extracellularly in the midgut of insect vectors as flagellated and elongated promastigotes and intracellularly in mammalian host macrophages as rounded amastigotes. Species such as Leishmania (Leishmania) amazonensis, L. mexicana and L. pifanoi are known to multiply in large and fusogenic vacuoles parasitoforos (VP), inducing the positive regulation of ATP6V0d2. In comparison to other Leishmania species, they also exhibit, at least in vitro, remarkable resistance to mechanisms of parasite elimination mediated by interferon-gamma (IFN-γ) and lipopolysaccharide (LPS) within macrophages or by direct treatment with species reactive oxygen species (ROS). However, a causal relationship between the development of large VPs and parasite resistance in inflammatory macrophages remains unclear, especially in vivo. Therefore, we evaluated the role of ATP6V0d2 in the biogenesis of pathogen-containing vacuoles using macrophages silenced to ATP6V0d2 infected with the protozoan parasite L. amazonensis. ATP6V0d2 knock-down decreased the cholesterol levels of the macrophages and inhibited the increase of the VP without interfering in the multiplication of the parasite. However, the parasites required ATP6V0d2 to resist the influx of oxidized low-density lipoprotein cholesterol (ox-LDL), which restored the increase of VP in silenced macrophages by replacing the cholesterol of the macrophages. Thus, we disclose the subversion of parasite-mediated V-ATPase function of the host cell relative to cholesterol retention, which is required to establish an inflammatory resistant intracellular parasite niche.
publishDate 2019
dc.date.none.fl_str_mv 2019-06-27
2021-01-19T16:32:03Z
2021-01-19T16:32:03Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=8044650
PESSOA, Carina Carraro. Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol. 2019. 100f. Tese (Doutorado em Microbiologia e Imunologia) – Escola Paulista de Medicina, Universidade Federal de São Paulo. São Paulo, 2019.
https://repositorio.unifesp.br/handle/11600/59272
url https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=8044650
https://repositorio.unifesp.br/handle/11600/59272
identifier_str_mv PESSOA, Carina Carraro. Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol. 2019. 100f. Tese (Doutorado em Microbiologia e Imunologia) – Escola Paulista de Medicina, Universidade Federal de São Paulo. São Paulo, 2019.
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 100 f.
application/pdf
dc.coverage.none.fl_str_mv São Paulo
dc.publisher.none.fl_str_mv Universidade Federal de São Paulo (UNIFESP)
publisher.none.fl_str_mv Universidade Federal de São Paulo (UNIFESP)
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNIFESP
instname:Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
instname_str Universidade Federal de São Paulo (UNIFESP)
instacron_str UNIFESP
institution UNIFESP
reponame_str Repositório Institucional da UNIFESP
collection Repositório Institucional da UNIFESP
repository.name.fl_str_mv Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)
repository.mail.fl_str_mv biblioteca.csp@unifesp.br
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