The role of hookworm glycoproteins in parasite biology and disease pathogenesis

The National Institute of Allergy and Infectious Diseases awarded Yale University $100,228 on October 9, 2026, under the Allergy and Infectious Diseases Research program (CFDA 93.855) to investigate the role of hookworm glycoproteins in parasite biology and disease pathogenesis.

The research characterizes glycosylated antigens in human hookworm species and their roles in immunopathogenesis. The project will identify glycoproteins present throughout hookworm development in two species of human hookworm and define the glycan structures present in each. The work includes generating native glycan microarrays from hookworm proteins and probing them with serum and mucosal reagents using a permissive animal model. The research will identify excretory/secretory glycoproteins recognized by host immunoglobulin G and mucosal immunoglobulin A to investigate their contribution to hookworm immunopathogenesis.

Prior work in this research area identified 83 glycoproteins (326 glycopeptides) in larvae of the human hookworm Ancylostoma ceylanicum, with 38 glycopeptides modified with phosphorylcholine, a known immunogenic glycan substitution.

Performance occurs in Connecticut. The period of performance runs through June 30, 2029. This is a Project Grant award classified as assistance funding.

Abstract

Hookworms are parasitic worms (HELMINTHS) that attach to the intestinal mucosa, where they ingest host blood from lacerated blood vessels, leading to severe anemia and growth delay. Elucidating mechanisms of hookworm pathogenesis is critical to the development of novel interventions that can be used to combat these and related helminthiases. Molecular interactions at the host-parasite interface are driven by the release of excretory/secretory proteins (ES) by adult hookworms in the small intestine. Previous studies conducted in our lab have demonstrated robust serum IGG and mucosal IGA responses to hookworm ES post-infection. Intriguingly, literature from related helminth systems has reported roles for antigen glycosylation in mediating host antibody responses to infection. However, specific glycan moieties on hookworm ES proteins have yet to be identified, and their roles in immunopathogenesis remain unstudied. We compiled a library of nematode glycan structures and developed a glycoproteomic workflow that identified 83 glycoproteins (326 glycopeptides) in the larvae of the human hookworm ancylostoma ceylanicum. Thirty-eight of these glycopeptides were modified with phosphorylcholine, a known immunogenic glycan substitution, suggesting a role for protein glycosylation in hookworm pathogenesis. This research will leverage advances in glycobiology to identify glycosylated hookworm antigens and interrogate their roles in hookworm immunopathogenesis. First, we will characterize the glycoproteins present throughout development in two species of human hookworm and define the glycan structures present in each. Next, we will generate native glycan microarrays from hookworm proteins and probe them with serum and mucosal reagents using a permissive animal model. Finally, we propose experiments to identify ES glycoproteins recognized by host IGG and mucosal IGA and investigate their role in mediating resistance to infection. This research will provide novel insight into glycosylation-mediated mechanisms underlying hookworm immunopathogenesis, ultimately leading to future work aimed at developing novel vaccines, drugs, and diagnostics to combat this globally important parasite. The combined expertise of the sponsor and a team of internal/external collaborators constitute a robust training environment that will support development as an independent scientist committed to advancing research in parasitic diseases.