Exploring pathogenicity and tumorigenic potential of the nematode Anisakis using human intestinal organoids and extracellular vesicles

Anisakiasis is an extremely underestimated accidental zoonosis both epidemiologically and medically. Its chronic form can potentially lead to erosive ulcers, granuloma formation and chronic inflammation at the human gastro-intestinal tract; features potentially involved in the onset of a carcinogenic microenvironment. In this regard, case reports of gastric or intestinal tumors in co-occurrence with anisakiasis are increasing from endemic countries. Giving that, investigations on tumorigenic potential of anisakiasis and its zoonotic agent Anisakis need to be exploited. Our work centers on the study of Anisakis-human interactions through a comparative transcriptomic approach using: i) the human intestinal organoids (HIO), a bi or three-dimensional cutting-edge model capable of self-organization, self-renewal, and exhibiting the architecture and functionality of the organ of origin, in this case the intestine; and ii) a newly discovered Anisakis messenger of pathogenicity, the extracellular vesiscles (EVs). EVs are nano-scale particles of 100-1000 nm involved in cell-cell and inter-species communication, having a new role in host-pathogen interface. EVs content is characterized by key elements such as lipids, nucleic acids, proteins and miRNAs, which are able to affect pivotal host pathways, being involved in a complex regulatory network and modulating host’s genes expression. Analysis of data revealed top 100 abundant transcripts in the HIO and 7 differentially expressed genes in HIO treated with Anisakis EVs, among them, transcripts showing potential link to cancer processes have been detected. In particular, a downregulation of EPHB2 and a upregulation of NUPR1 emerged and, interestingly, these alterations have been associated to colorectal cancer. In addition, qRT-PCR carried out on inflammatory products (Il33, Il8, Il1β) showed a decreasing trend in Il33 gene expression, an increasing trend in Il1β and no alteration in Il8, a dynamic that were previous described as involved in helminth infection chronicity. This project represent the first attempt aimed to investigate Anisakis tumorigenic potential using HIO and EVs, revealing interesting outcomes that we hope will open new ways for the study of anisakiasis and its potential consequences in humans, a field to be explored.