Uncovering the role of a Cryptosporidium parvum thrombospondin protein (CpTSP8) in host cell attachment

Lain, Joshua Aaron

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https://hdl.handle.net/2142/127424 Copy

Description

Title

Uncovering the role of a Cryptosporidium parvum thrombospondin protein (CpTSP8) in host cell attachment

Author(s)

Lain, Joshua Aaron

Issue Date

2024-07-25

Director of Research (if dissertation) or Advisor (if thesis)

Vinayak Alam, Sumiti

Department of Study

Pathobiology

Discipline

VMS - Pathobiology

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

M.S.

Degree Level

Thesis

Keyword(s)

• Pathobiology
• Parasitology
• Cryptosporidium
• Cryptosporidium Parvum
• Expansion Microscopy
• Thrombospondin
• Tsp8

Language

eng

Abstract

The protozoan parasite Cryptosporidium parvum is a leading cause of diarrheal disease and mortality in children and immunocompromised individuals. There is no vaccine or effective therapy to prevent or treat cryptosporidiosis. Very little is known about how Cryptosporidium’s invasive life stage, the sporozoite, recognizes and attaches to hosts’ intestinal epithelium. In related apicomplexan parasites such as Toxoplasma and Plasmodium, motility, attachment, and invasion are regulated by genes containing thrombospondin repeat (TSP) domains (Sultan et al., 1997; Brossier et al., 2003). Developing an understanding of how Cryptosporidium sporozoites attach to host cells is important for the future development of much needed vaccines and therapeutics. We have identified a C. parvum gene (cgd6_780) that is expressed only in the sporozoite stage and encodes for a TSP domain protein (CpTSP8). Recent hyperplexed localization of organelle proteins by isotope tagging (hyperLOPIT) analysis, a technique in which successive rounds of centrifugation followed by isotope tagging of proteins allows for localization of proteins to specific organelles, clusters this protein with other apical secretory organelle proteins. Based on the gene expression profile and secreted proteome data, we hypothesize the role of this parasite TSP in host cell attachment. Using CRISPR/Cas9 editing and immunocompromised mouse infection model, we created a transgenic C. parvum line that expresses this TSP amended with an epitope tag. Super-resolution and expansion microscopy of sporozoites revealed this protein’s localization on the apical end of the parasite’s surface and during early stages of infection. Immunoprecipitation was performed on this TSP to uncover interacting partners. Future studies are required to further investigate the molecular function of CpTS8, and its mechanistic role in host attachment and infection.

Graduation Semester

2024-12

Type of Resource

Thesis

Handle URL

https://hdl.handle.net/2142/127424

Copyright and License Information

Copyright 2024 Joshua Lain

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