ATP6V(0)d2 controls Leishmania parasitophorous vacuole biogenesis via cholesterol homeostasis

Author(s):Show less -

Pessoa, Carina Carraro ^[1] ; Reis, Luiza Campos ^[2] ; Ramos-Sanchez, Eduardo Milton ^[2] ; Orikaza, Cristina Mary ^[1] ; Cortez, Cristian ^[3] ; de Castro Levatti, Erica Valadares ^[4] ; Benites Badaro, Ana Carolina ^[1] ; da Silva Yamamoto, Joyce Umbelino ^[5] ; D'Almeida, Vania ^[5] ; Goto, Hiro ^{[6, 2]} ; Mortara, Renato Arruda ^[1] ; Real, Fernando ^{[1, 7]}

Affiliation

^[1] Univ Fed Sao Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, Sao Paulo - Brazil

^[2] Univ Sao Paulo, Inst Med Trop, Lab Soroepidemiol & Imunobiol, Inst Med Trop, Sao Paulo - Brazil

^[3] Univ Mayor, Fac Ciencias, Ctr Genom & Bioinformat, Santiago - Chile

^[4] Univ Fed Sao Paulo, Dept Bioquim, Escola Paulista Med, Sao Paulo - Brazil

^[5] Univ Fed Sao Paulo, Dept Psicobiol, Escola Paulista Med, Sao Paulo - Brazil

^[6] Univ Sao Paulo, Fac Med, Dept Med Prevent, Sao Paulo - Brazil

^[7] INSERM U1016, Inst Cochin, Dept Infect, Immun, Inflammat, Paris - France

• Document type: Journal article
• Source: PLOS PATHOGENS; v. 15, n. 6 JUN 2019.
• Web of Science Citations: 0

Abstract

V-ATPases are part of the membrane components of pathogen-containing vacuoles, although their function in intracellular infection remains elusive. In addition to organelle acidification, V-ATPases are alternatively implicated in membrane fusion and anti-inflammatory functions controlled by ATP6V(0)d2, the d subunit variant of the V-ATPase complex. Therefore, we evaluated the role of ATP6V(0)d2 in the biogenesis of pathogen-containing vacuoles using ATP6V(0)d2 knock-down macrophages infected with the protozoan parasite Leishmania amazonensis. These parasites survive within IFN gamma/LPS-activated inflammatory macrophages, multiplying in large/fusogenic parasitophorous vacuoles (PVs) and inducing ATP6V(0)d2 upregulation. ATP6V(0)d2 knock-down decreased macrophage cholesterol levels and inhibited PV enlargement without interfering with parasite multiplication. However, parasites required ATP6V(0)d2 to resist the influx of oxidized low-density lipoprotein (ox-LDL)-derived cholesterol, which restored PV enlargement in ATP6V(0)d2 knock-down macrophages by replenishing macrophage cholesterol pools. Thus, we reveal parasite-mediated subversion of host V-ATPase function toward cholesterol retention, which is required for establishing an inflammation-resistant intracellular parasite niche. Author summary V-ATPases control acidification and other processes at intracellular vesicles that bacteria and parasites exploit as compartments for replication and immune evasion. We report that the protozoan intracellular parasite Leishmania amazonensis resists inflammatory macrophage immune responses and upregulates an alternative isoform of subunit d of V-ATPase (ATP6V(0)d2). Leishmania are still sequestered within acidified parasitophorous vacuoles (PVs) in cells lacking ATP6V(0)d2, but these PVs do not enlarge in volume, a distinguishing feature of intracellular infection by these parasites. Cholesterol levels in ATP6V(0)d2-deficient cells are reduced and exogenous cholesterol repletion can restore vacuole size, leading to enhanced parasite killing. This study demonstrates the ATP6V(0)d2-mediated interplay of macrophage cholesterol retention and control of the biogenesis of large pathogen-containing vacuoles. The study provides grounds for the development of new therapeutic strategies for diseases caused by intracellular pathogens sheltered in host cell compartments. (AU)

• FAPESP's process: 15/14205-9 - Investigation of the role of vacuolar ATPase of d2 subunit ( ATP6V0d2 ) in the biogenese of parasitophorous vacuoles formed by Leishmania (Leishmania) amazonensis in RAW 264.7 macrophages
• Grantee: Carina Carraro Pessoa
• Support type: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/15000-4 - Trypanosoma cruzi: intra and interspecific genomic variability and mechanisms of cell invasion/egress
• Grantee: Renato Arruda Mortara
• Support type: Research Projects - Thematic Grants