PMID- 37729272
OWN - NLM
STAT- MEDLINE
DCOM- 20231003
LR  - 20240214
IS  - 1935-2735 (Electronic)
IS  - 1935-2727 (Print)
IS  - 1935-2727 (Linking)
VI  - 17
IP  - 9
DP  - 2023 Sep
TI  - Disruption of the inositol phosphorylceramide synthase gene affects Trypanosoma 
      cruzi differentiation and infection capacity.
PG  - e0011646
LID - 10.1371/journal.pntd.0011646 [doi]
LID - e0011646
AB  - Sphingolipids (SLs) are essential components of all eukaryotic cellular 
      membranes. In fungi, plants and many protozoa, the primary SL is 
      inositol-phosphorylceramide (IPC). Trypanosoma cruzi is a protozoan parasite that 
      causes Chagas disease (CD), a chronic illness for which no vaccines or effective 
      treatments are available. IPC synthase (IPCS) has been considered an ideal target 
      enzyme for drug development because phosphoinositol-containing SL is absent in 
      mammalian cells and the enzyme activity has been described in all parasite forms 
      of T. cruzi. Furthermore, IPCS is an integral membrane protein conserved amongst 
      other kinetoplastids, including Leishmania major, for which specific inhibitors 
      have been identified. Using a CRISPR-Cas9 protocol, we generated T. cruzi 
      knockout (KO) mutants in which both alleles of the IPCS gene were disrupted. We 
      demonstrated that the lack of IPCS activity does not affect epimastigote 
      proliferation or its susceptibility to compounds that have been identified as 
      inhibitors of the L. major IPCS. However, disruption of the T. cruzi IPCS gene 
      negatively affected epimastigote differentiation into metacyclic trypomastigotes 
      as well as proliferation of intracellular amastigotes and differentiation of 
      amastigotes into tissue culture-derived trypomastigotes. In accordance with 
      previous studies suggesting that IPC is a membrane component essential for 
      parasite survival in the mammalian host, we showed that T. cruzi IPCS null 
      mutants are unable to establish an infection in vivo, even in immune deficient 
      mice.
CI  - Copyright: (c) 2023 Dos Santos et al. This is an open access article distributed 
      under the terms of the Creative Commons Attribution License, which permits 
      unrestricted use, distribution, and reproduction in any medium, provided the 
      original author and source are credited.
FAU - Dos Santos, Nailma S A
AU  - Dos Santos NSA
AD  - Departamento de Bioquimica e Imunologia, Universidade Federal de Minas Gerais, 
      Belo Horizonte, Brazil.
FAU - Estevez-Castro, Carlos F
AU  - Estevez-Castro CF
AD  - Departamento de Bioquimica e Imunologia, Universidade Federal de Minas Gerais, 
      Belo Horizonte, Brazil.
FAU - Macedo, Juan P
AU  - Macedo JP
AD  - Departamento de Bioquimica e Imunologia, Universidade Federal de Minas Gerais, 
      Belo Horizonte, Brazil.
FAU - Chame, Daniela F
AU  - Chame DF
AD  - Departamento de Bioquimica e Imunologia, Universidade Federal de Minas Gerais, 
      Belo Horizonte, Brazil.
FAU - Castro-Gomes, Thiago
AU  - Castro-Gomes T
AD  - Departamento de Parasitologia, Universidade Federal de Minas, Belo Horizonte, 
      Brazil.
FAU - Santos-Cardoso, Mariana
AU  - Santos-Cardoso M
AD  - Departamento de Bioquimica e Imunologia, Universidade Federal de Minas Gerais, 
      Belo Horizonte, Brazil.
FAU - Burle-Caldas, Gabriela A
AU  - Burle-Caldas GA
AD  - Departamento de Bioquimica e Imunologia, Universidade Federal de Minas Gerais, 
      Belo Horizonte, Brazil.
FAU - Covington, Courtney N
AU  - Covington CN
AD  - Department of Chemistry and Centre for Global Infectious Disease, Durham 
      University, Durham, United Kingdom.
FAU - Steel, Patrick G
AU  - Steel PG
AD  - Department of Chemistry and Centre for Global Infectious Disease, Durham 
      University, Durham, United Kingdom.
FAU - Smith, Terry K
AU  - Smith TK
AD  - BSRC School of Biology, Biomolecular Science Building, St Andrews, United 
      Kingdom.
FAU - Denny, Paul W
AU  - Denny PW
AD  - Department of Biosciences and Centre for Global Infectious Diseases, Durham 
      University, Durham, United Kingdom.
FAU - Teixeira, Santuza M R
AU  - Teixeira SMR
AUID- ORCID: 0000-0002-2588-2842
AD  - Departamento de Bioquimica e Imunologia, Universidade Federal de Minas Gerais, 
      Belo Horizonte, Brazil.
LA  - eng
GR  - MR/P027989/1/MRC_/Medical Research Council/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20230920
PL  - United States
TA  - PLoS Negl Trop Dis
JT  - PLoS neglected tropical diseases
JID - 101291488
RN  - 4L6452S749 (Inositol)
SB  - IM
MH  - Mice
MH  - Animals
MH  - *Trypanosoma cruzi
MH  - *Chagas Disease
MH  - *Leishmania major/genetics
MH  - Cell Differentiation
MH  - Inositol/metabolism/pharmacology
MH  - Mammals
PMC - PMC10545103
COIS- The authors have declared that no competing interests exist.
EDAT- 2023/09/20 18:42
MHDA- 2023/10/03 06:47
PMCR- 2023/09/20
CRDT- 2023/09/20 13:44
PHST- 2022/09/30 00:00 [received]
PHST- 2023/09/07 00:00 [accepted]
PHST- 2023/10/02 00:00 [revised]
PHST- 2023/10/03 06:47 [medline]
PHST- 2023/09/20 18:42 [pubmed]
PHST- 2023/09/20 13:44 [entrez]
PHST- 2023/09/20 00:00 [pmc-release]
AID - PNTD-D-22-01249 [pii]
AID - 10.1371/journal.pntd.0011646 [doi]
PST - epublish
SO  - PLoS Negl Trop Dis. 2023 Sep 20;17(9):e0011646. doi: 
      10.1371/journal.pntd.0011646. eCollection 2023 Sep.