# Experimental genetic crosses for malaria research

> **NIH NIH P01** · UNIVERSITY OF NOTRE DAME · 2024 · $404,950

## Abstract

PROJECT SUMMARY
A concrete way to map genotypes that cause Plasmodium falciparum drug resistance phenotypes is by
performing well thought out experimental genetic crosses between drug susceptible and resistant strains,
isolating recombinant progeny and then using quantitative trait loci mapping to link genotype to phenotype.
As an alternative, bulk segregant analysis under drug pressure can be used to pinpoint loci of interest. P.
falciparum genetic crosses were initially carried out in splenectomized chimpanzees, but NIH has banned
chimpanzee research for financial and ethical reasons. To overcome this, we have developed and improved
on a human-liver chimeric mouse model (the FRG NOD huHep mouse) to replace the chimpanzee for the
generation of recombinant progeny from P. falciparum genetic crosses.
The FRG NOD mouse lacks the fumaryl acetoacetate hydrolase gene (F designation) and this causes
hepatocyte cell death. However, hepatocyte death is controlled with the drug nitisinone. Since only mouse
hepatocytes lack fumaryl acetoacetate hydrolase, this enables repopulation of the mouse with human
hepatocytes over time with on-off drug use to control the death of mouse hepatocytes and their replacement
with human hepatocytes. In close collaboration with the Yecuris Corporation, who creates the FRG NOD
huHep mouse, we ensure that the mice we use for our studies have maximal human hepatocyte chimerism
and are susceptible to P. falciparum sporozoite infection. Additionally, the mice are able to maintain a human
red blood cell (huRBC) population after huRBC infusion and this allows for P. falciparum liver stage-to-blood
stage transition in the mouse. Following blood removal, the in vitro expansion of asexual P. falciparum blood
stages allows for bulk segregant analysis as well as downstream cloning and then –omics analyses and
phenotypic analyses of recombinant progeny.
We have clearly demonstrated our ability to use the FRG NOD huHep/huRBC mouse for the generation of
recombinant progeny from experimental crosses and Core A will continue to produce recombinant progeny
from a further ten well-conceived experimental genetic crosses between P. falciparum drug resistant and drug
susceptible strains as part of this P01 renewal. The phenotyping of bulk and isolated progeny supplied by
Core A and downstream mapping of genetic loci responsible for observed phenotypes are integral parts of
RP01, RP02 & RP03 and, as such, Core A is the linchpin of this P01 renewal. Successful creation of progeny
for this P01 will further our understanding of the spread of artemisinin drug resistance, which has now been
documented to have occurred independently in Africa and is of grave concern to malaria control and
elimination efforts.

## Key facts

- **NIH application ID:** 10863149
- **Project number:** 2P01AI127338-06A1
- **Recipient organization:** UNIVERSITY OF NOTRE DAME
- **Principal Investigator:** Ashley M Vaughan
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $404,950
- **Award type:** 2
- **Project period:** 2017-08-01 → 2029-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10863149

## Citation

> US National Institutes of Health, RePORTER application 10863149, Experimental genetic crosses for malaria research (2P01AI127338-06A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10863149. Licensed CC0.

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