TY - JOUR
T1 - The Plasmodium falciparum ABC transporter ABCI3 confers parasite strain-dependent pleiotropic antimalarial drug resistance
AU - Murithi, James M.
AU - Deni, Ioanna
AU - Pasaje, Charisse Flerida A.
AU - Okombo, John
AU - Bridgford, Jessica L.
AU - Gnädig, Nina F.
AU - Edwards, Rachel L.
AU - Yeo, Tomas
AU - Mok, Sachel
AU - Burkhard, Anna Y.
AU - Coburn-Flynn, Olivia
AU - Istvan, Eva S.
AU - Sakata-Kato, Tomoyo
AU - Gomez-Lorenzo, Maria G.
AU - Cowell, Annie N.
AU - Wicht, Kathryn J.
AU - Le Manach, Claire
AU - Kalantarov, Gavreel F.
AU - Dey, Sumanta
AU - Duffey, Maëlle
AU - Laleu, Benoît
AU - Lukens, Amanda K.
AU - Ottilie, Sabine
AU - Vanaerschot, Manu
AU - Trakht, Ilya N.
AU - Gamo, Francisco Javier
AU - Wirth, Dyann F.
AU - Goldberg, Daniel E.
AU - Odom John, Audrey R.
AU - Chibale, Kelly
AU - Winzeler, Elizabeth A.
AU - Niles, Jacquin C.
AU - Fidock, David A.
N1 - Funding Information:
D.A.F. gratefully acknowledges support from the Medicines for Malaria Venture (MMV) and the NIH ( R37 AI05234 , R01 AI124678 , R01 AI147628 ). J.C.N. acknowledges support from the Bill & Melinda Gates Foundation through the Grand Challenges Exploration initiative ( OPP1162467 ). K.C. gratefully acknowledges MMV, the South African Technology Innovation Agency (Project MMV09/0002 ), the South African Medical Research Council , and the South African Research Chairs Initiative of the Department of Science and Innovation, administered through the South African National Research Foundation. Funding to E.A.W., K.C., J.C.N., D.E.G., D.F.W., J.G., and D.A.F. is also provided by the Bill & Melinda Gates Foundation in support of the MalDA consortium (OPP1054480). S.M. is a recipient of a Long-Term Fellowship from the Human Frontiers of Science Program. We thank Lynn Wambua and Lauren Arendse (UCT) for kindly testing compounds against recombinant PKG. We also thank Wandy Beatty at Washington University for technical assistance with electron microscopy.
Funding Information:
D.A.F. gratefully acknowledges support from the Medicines for Malaria Venture (MMV) and the NIH (R37 AI05234, R01 AI124678, R01 AI147628). J.C.N. acknowledges support from the Bill & Melinda Gates Foundation through the Grand Challenges Exploration initiative (OPP1162467). K.C. gratefully acknowledges MMV, the South African Technology Innovation Agency (Project MMV09/0002), the South African Medical Research Council, and the South African Research Chairs Initiative of the Department of Science and Innovation, administered through the South African National Research Foundation. Funding to E.A.W. K.C. J.C.N. D.E.G. D.F.W. J.G. and D.A.F. is also provided by the Bill & Melinda Gates Foundation in support of the MalDA consortium (OPP1054480). S.M. is a recipient of a Long-Term Fellowship from the Human Frontiers of Science Program. We thank Lynn Wambua and Lauren Arendse (UCT) for kindly testing compounds against recombinant PKG. We also thank Wandy Beatty at Washington University for technical assistance with electron microscopy. K.J.W. and C.L.M. synthesized compounds 1 and 6. B.L. designed and optimized compound 5. I.D. O.C.-F. E.S.I. T.S.-K. and M.G.G.-L. performed resistance selections. J.M.M. and I.D. performed CRISPR/Cas9 SNP validation experiments. T.Y. S.M. and A.N.C. prepared and analyzed whole-genome sequencing data. J.M.M. I.D. J.L.B. and A.Y.B. performed asexual blood stage assays. C.F.A.P. and S.D. generated ABCI3 cKD parasites. J.M.M. and N.F.G. performed immunofluorescence assays. R.L.E. performed IEM assays. J.O. generated the heme fractionation data. G.F.K. produced PfCRT antibodies. A.K.L. S.O. M.V. I.N.T. F.-J.G. D.F.W. D.E.G. A.R.O.J. J.C.N. K.C. E.A.W. and D.A.F. supervised individual lab efforts and along with M.D. and B.L. provided funding and expertise for several. J.M.M. and D.A.F. wrote the manuscript with input from all authors. All authors approved the final manuscript. B.L. and M.D. are employees of MMV; M.G.G.-L. and F.-J.G. are employees of GSK.
Publisher Copyright:
© 2021 The Authors
PY - 2022/5/19
Y1 - 2022/5/19
N2 - Widespread Plasmodium falciparum resistance to first-line antimalarials underscores the vital need to develop compounds with novel modes of action and identify new druggable targets. Here, we profile five compounds that potently inhibit P. falciparum asexual blood stages. Resistance selection studies with three carboxamide-containing compounds, confirmed by gene editing and conditional knockdowns, identify point mutations in the parasite transporter ABCI3 as the primary mediator of resistance. Selection studies with imidazopyridine or quinoline-carboxamide compounds also yield changes in ABCI3, this time through gene amplification. Imidazopyridine mode of action is attributed to inhibition of heme detoxification, as evidenced by cellular accumulation and heme fractionation assays. For the copy-number variation-selecting imidazopyridine and quinoline-carboxamide compounds, we find that resistance, manifesting as a biphasic concentration-response curve, can independently be mediated by mutations in the chloroquine resistance transporter PfCRT. These studies reveal the interconnectedness of P. falciparum transporters in overcoming drug pressure in different parasite strains.
AB - Widespread Plasmodium falciparum resistance to first-line antimalarials underscores the vital need to develop compounds with novel modes of action and identify new druggable targets. Here, we profile five compounds that potently inhibit P. falciparum asexual blood stages. Resistance selection studies with three carboxamide-containing compounds, confirmed by gene editing and conditional knockdowns, identify point mutations in the parasite transporter ABCI3 as the primary mediator of resistance. Selection studies with imidazopyridine or quinoline-carboxamide compounds also yield changes in ABCI3, this time through gene amplification. Imidazopyridine mode of action is attributed to inhibition of heme detoxification, as evidenced by cellular accumulation and heme fractionation assays. For the copy-number variation-selecting imidazopyridine and quinoline-carboxamide compounds, we find that resistance, manifesting as a biphasic concentration-response curve, can independently be mediated by mutations in the chloroquine resistance transporter PfCRT. These studies reveal the interconnectedness of P. falciparum transporters in overcoming drug pressure in different parasite strains.
KW - ABCI3
KW - CRISPR/Cas9
KW - Plasmodium falciparum malaria
KW - biphasic dose-response curves
KW - cellular accumulation assays
KW - conditional knockdowns
KW - copy-number variations
KW - heme fractionation
KW - pfcrt
UR - http://www.scopus.com/inward/record.url?scp=85130525665&partnerID=8YFLogxK
U2 - 10.1016/j.chembiol.2021.06.006
DO - 10.1016/j.chembiol.2021.06.006
M3 - Article
C2 - 34233174
AN - SCOPUS:85130525665
SN - 2451-9456
VL - 29
SP - 824-839.e6
JO - Cell Chemical Biology
JF - Cell Chemical Biology
IS - 5
ER -