The essential host genome for Cryptosporidium intracellular survival exposes metabolic dependencies that can be leveraged for treatment

Mapping how pathogens interact with their host cells can reveal unexpected pathogen and host cell biology, paving the way for new treatments. Cryptosporidium is an intracellular parasite of intestinal epithelial cells, and a leading cause of diarrheal death and disease in infants worldwide. Despite...

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Bibliographic Details
Published in:bioRxiv 2024-12
Main Authors: N Bishara Marzook, Song, Ok-Ryul, Baumgärtel, Lotta, Bernitz, Netanya, Mkandawire, Tapoka T, Watson, Lucy C, Nunes, Vanessa, Warchal, Scott, Macrae, James I, Howell, Michael, Sateriale, Adam
Format: Article
Language:English
Subjects:
Actin
Cell death
Cell survival
Cholesterol
CRISPR
Cryptosporidium
Diarrhea
Epithelial cells
Gene mapping
Glutathione
Infections
Intracellular
Parasites
Pathogens
Peptide mapping
Protein arrays
Squalene
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Summary:Mapping how pathogens interact with their host cells can reveal unexpected pathogen and host cell biology, paving the way for new treatments. Cryptosporidium is an intracellular parasite of intestinal epithelial cells, and a leading cause of diarrheal death and disease in infants worldwide. Despite this, very little is known about the cell biology of infection of this eukaryotic pathogen. Here, we designed and implemented a unique microscopy-based arrayed CRISPR-Cas9 screen to interrogate the effects of the loss of every protein-coding human gene on a Cryptosporidium infection. As the experimental readout is image-based, we extracted multiple phenotypic features of infection, including parasite growth, progression of the parasite to its sexual life stage, and recruitment of host actin to pedestals beneath the parasite vacuole. Using this dataset, we discovered a tipping point in the host cholesterol biosynthesis pathway that controls Cryptosporidium infection. Parasite growth can either be inhibited or promoted by the intermediary metabolite squalene. A build-up of squalene in epithelial cells creates a reducing environment, with more reduced host glutathione available for uptake by the parasite. Because Cryptosporidium has lost the ability to synthesize glutathione, this uptake from the host cell is required for growth and progression through its life cycle. We demonstrate that this dependency can be leveraged for treatment with the abandoned drug lapaquistat, an inhibitor of host squalene synthase that has efficacy against Cryptosporidium in vitro and in vivo.Competing Interest StatementThe authors have declared no competing interest.
DOI:10.1101/2024.12.04.626561