RNA interference-mediated hookworm control by gut-dwelling lactic acid bacteria

Area of Science:

• Parasitology
• Molecular Biology
• Probiotics
• RNA Interference (RNAi)

Background:

• Geohelminth infections pose challenges due to complex life cycles and resistance to traditional anthelmintics.
• Hookworm infections (Ancylostoma sp. and Necator sp.) are significant global health issues requiring novel therapeutic strategies.
• Existing treatments are becoming less effective, necessitating the development of sustainable parasite management solutions.

Purpose of the Study:

• To develop a novel probiotic-based therapy utilizing RNA interference (RNAi) against essential hookworm enzymes.
• To engineer Lactococcus lactis to produce and deliver double-stranded RNA (dsRNA) targeting astacin-like metalloprotease (Ac-MTP-1) in Ancylostoma sp.
• To evaluate the efficacy of this engineered probiotic in controlling hookworm infection and associated pathology.

Main Methods:

• Genetic engineering of Lactococcus lactis to produce dsRNA targeting the hookworm enzyme Ac-MTP-1.
• Development of a gut delivery RNAi vector for oral administration of engineered probiotics.
• In vivo testing in mouse models to assess the impact of engineered probiotics on hookworm infection and organ damage.

Main Results:

• Engineered probiotics successfully delivered dsRNA, leading to RNAi-mediated knockdown of the target enzyme in hookworms.
• A 70% reduction in larval penetration of the gut lining and a 50% decrease in migration to vital organs were observed.
• Significant reversal of liver (30%-40%) and kidney (50%-60%) damage in mouse models pre-treated with engineered L. lactis.

Conclusions:

• This study demonstrates the first successful oral administration of genetically engineered probiotics for RNAi-based gene knockdown in hookworms.
• The novel 'RNAi food' approach shows potential for interrupting the hookworm life cycle and managing infections.
• This strategy can complement existing anthelmintics and be extended to other hookworm species like A. duodenalis and A. ceylanicum.