A Plasmodium falciparum genetic cross reveals the contributions of pfcrt and plasmepsin II/III to piperaquine drug resistance
Piperaquine (PPQ) is widely used in combination with dihydroartemisinin (DHA) as a first-line treatment against malaria parasites. Multiple genetic drivers of PPQ resistance have been reported, including mutations in the Plasmodium falciparum chloroquine resistance transporter (pfcrt) and increased copies of plasmepsin II/III (pm2/3). We generated a cross between a Cambodia-derived multi-drug resistant KEL1/PLA1 lineage isolate (KH004) and a drug susceptible parasite isolated in Malawi (Mal31). Mal31 harbors a wild-type (3D7-like) pfcrt allele and a single copy of pm2/3, while KH004 has a chloroquine-resistant (Dd2-like) pfcrt allele with an additional G367C substitution and four copies of pm2/3. We recovered 104 unique recombinant progeny and examined a targeted set of progeny representing all possible combinations of variants at pfcrt and pm2/3 for detailed analysis of competitive fitness and a range of PPQ susceptibility phenotypes, including PPQ survival assay (PSA), area under the dose-response curve (AUC), and a limited point IC50 (LP-IC50). We find that inheritance of the KH004 pfcrt allele is required for PPQ resistance, whereas copy number variation in pm2/3 further enhances resistance but does not confer resistance in the absence of PPQ-R-associated mutations in pfcrt. Deeper investigation of genotype-phenotype relationships demonstrates that progeny clones from experimental crosses can be used to understand the relative contributions of pfcrt, pm2/3, and parasite genetic background, to a range of PPQ-related traits and confirm the critical role of the PfCRT G367C substitution in PPQ resistance.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - year:2023 |
|---|---|
| Enthalten in: |
bioRxiv : the preprint server for biology - (2023) vom: 17. Sept. |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Kane, John [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
|---|
| Anmerkungen: |
Date Revised 27.02.2024 published: Electronic Citation Status PubMed-not-MEDLINE |
|---|
| doi: |
10.1101/2023.06.06.543862 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM362419582 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM362419582 | ||
| 003 | DE-627 | ||
| 005 | 20240229163628.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1101/2023.06.06.543862 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1308.xml |
| 035 | |a (DE-627)NLM362419582 | ||
| 035 | |a (NLM)37745488 | ||
| 035 | |a (PII)2023.06.06.543862 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Kane, John |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A Plasmodium falciparum genetic cross reveals the contributions of pfcrt and plasmepsin II/III to piperaquine drug resistance |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Revised 27.02.2024 | ||
| 500 | |a published: Electronic | ||
| 500 | |a Citation Status PubMed-not-MEDLINE | ||
| 520 | |a Piperaquine (PPQ) is widely used in combination with dihydroartemisinin (DHA) as a first-line treatment against malaria parasites. Multiple genetic drivers of PPQ resistance have been reported, including mutations in the Plasmodium falciparum chloroquine resistance transporter (pfcrt) and increased copies of plasmepsin II/III (pm2/3). We generated a cross between a Cambodia-derived multi-drug resistant KEL1/PLA1 lineage isolate (KH004) and a drug susceptible parasite isolated in Malawi (Mal31). Mal31 harbors a wild-type (3D7-like) pfcrt allele and a single copy of pm2/3, while KH004 has a chloroquine-resistant (Dd2-like) pfcrt allele with an additional G367C substitution and four copies of pm2/3. We recovered 104 unique recombinant progeny and examined a targeted set of progeny representing all possible combinations of variants at pfcrt and pm2/3 for detailed analysis of competitive fitness and a range of PPQ susceptibility phenotypes, including PPQ survival assay (PSA), area under the dose-response curve (AUC), and a limited point IC50 (LP-IC50). We find that inheritance of the KH004 pfcrt allele is required for PPQ resistance, whereas copy number variation in pm2/3 further enhances resistance but does not confer resistance in the absence of PPQ-R-associated mutations in pfcrt. Deeper investigation of genotype-phenotype relationships demonstrates that progeny clones from experimental crosses can be used to understand the relative contributions of pfcrt, pm2/3, and parasite genetic background, to a range of PPQ-related traits and confirm the critical role of the PfCRT G367C substitution in PPQ resistance | ||
| 650 | 4 | |a Preprint | |
| 700 | 1 | |a Li, Xue |e verfasserin |4 aut | |
| 700 | 1 | |a Kumar, Sudhir |e verfasserin |4 aut | |
| 700 | 1 | |a Button-Simons, Katrina A |e verfasserin |4 aut | |
| 700 | 1 | |a Brenneman, Katelyn M Vendrely |e verfasserin |4 aut | |
| 700 | 1 | |a Dahlhoff, Haley |e verfasserin |4 aut | |
| 700 | 1 | |a Sievert, Mackenzie A C |e verfasserin |4 aut | |
| 700 | 1 | |a Checkley, Lisa A |e verfasserin |4 aut | |
| 700 | 1 | |a Shoue, Douglas A |e verfasserin |4 aut | |
| 700 | 1 | |a Singh, Puspendra P |e verfasserin |4 aut | |
| 700 | 1 | |a Abatiyow, Biley A |e verfasserin |4 aut | |
| 700 | 1 | |a Haile, Meseret T |e verfasserin |4 aut | |
| 700 | 1 | |a Nair, Shalini |e verfasserin |4 aut | |
| 700 | 1 | |a Reyes, Ann |e verfasserin |4 aut | |
| 700 | 1 | |a Tripura, Rupam |e verfasserin |4 aut | |
| 700 | 1 | |a Peto, Tom |e verfasserin |4 aut | |
| 700 | 1 | |a Lek, Dysoley |e verfasserin |4 aut | |
| 700 | 1 | |a Kappe, Stefan H I |e verfasserin |4 aut | |
| 700 | 1 | |a Dhorda, Mehul |e verfasserin |4 aut | |
| 700 | 1 | |a Nkhoma, Standwell C |e verfasserin |4 aut | |
| 700 | 1 | |a Cheeseman, Ian H |e verfasserin |4 aut | |
| 700 | 1 | |a Vaughan, Ashley M |e verfasserin |4 aut | |
| 700 | 1 | |a Anderson, Timothy J C |e verfasserin |4 aut | |
| 700 | 1 | |a Ferdig, Michael T |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t bioRxiv : the preprint server for biology |d 2020 |g (2023) vom: 17. Sept. |w (DE-627)NLM31090014X |7 nnns |
| 773 | 1 | 8 | |g year:2023 |g day:17 |g month:09 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1101/2023.06.06.543862 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |j 2023 |b 17 |c 09 | ||