Biogênese do vacúolo de Leishmania: papel da subunidade ATP6V0d2 na homeostase de colesterol
Autor(a) principal: | |
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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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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 |
_version_ |
1814268433777295360 |