@article{fdi:010077350,
  title = {{A} single amino acid substitution ({H}451{Y}) in {L}eishmania calcium-dependent kinase {SCAMK} confers high tolerance and resistance to antimony},
  author = {{V}ergnes, {B}aptiste and {G}azanion, {E}lodie and {M}ariac, {C}{\'e}dric and {D}u {M}anoir, {M}. and {S}ollelis, {L}. and {L}opez-{R}ubio, {J}. {J}. and {S}terkers, {Y}. and {B}anuls, {A}nne-{L}aure},
  editor = {},
  language = {{ENG}},
  abstract = {{B}ackground: {F}or almost a century, antimonials have remained the first-line drugs for the treatment of leishmaniasis. {H}owever, little is known about their mode of action and clinical resistance mechanisms. {O}bjectives: {W}e have previously shown that {L}eishmania nicotinamidase ({PNC}1) is an essential enzyme for parasite {NAD}+ homeostasis and virulence in vivo. {H}ere, we found that parasites lacking the pnc1 gene ({D}elta pnc1) are hypersusceptible to the active form of antimony ({S}b{III}) and used these mutant parasites to better understand antimony's mode of action and the mechanisms leading to resistance. {M}ethods: {S}b{III}-resistant {WT} and {D}elta pnc1 parasites were selected in vitro by a stepwise selection method. {NAD}({H})/{NADP}({H}) dosages and quantitative {RT}-{PCR} experiments were performed to explain the susceptibility differences observed between strains. {WGS} and a marker-free {CRISPR}/{C}as9 base-editing approach were used to identify and validate the role of a new resistance mutation. {R}esults: {NAD}+-depleted {D}elta pnc1 parasites were highly susceptible to {S}b{III} and this phenotype could be rescued by {NAD}+ precursor or trypanothione precursor supplementation. {D}elta pnc1 parasites could become resistant to {S}b{III} by an unknown mechanism. {WGS} revealed a unique amino acid substitution ({H}451{Y}) in an {EF}-hand domain of an orphan calcium-dependent kinase, recently named {SCAMK}. {W}hen introduced into a {WT} reference strain by base editing, the {H}451{Y} mutation allowed {L}eishmania parasites to survive at extreme concentrations of {S}b{III}, potentiating the rapid emergence of resistant parasites. {C}onclusions: {T}hese results establish that {L}eishmania {SCAMK} is a new central hub of antimony's mode of action and resistance development, and uncover the importance of drug tolerance mutations in the evolution of parasite drug resistance.},
  keywords = {},
  booktitle = {},
  journal = {{J}ournal of {A}ntimicrobial {C}hemotherapy},
  volume = {74},
  numero = {11},
  pages = {3231--3239},
  ISSN = {0305-7453},
  year = {2019},
  DOI = {10.1093/jac/dkz334},
  URL = {https://www.documentation.ird.fr/hor/fdi:010077350},
}