Charles University in Prague teamed up with Chang Gung University to study the role of protozoan exosome on virus transmission supported by the MOST-GACR Joint Research Cooperation Funding. Trichomonas vaginalis is the causative agent of trichomoniasis, the most common non-viral sexually transmitted infection (STI) with a worldwide incidence of over 220 million cases. Trichomonads often harbor viral (TVV) or Mycoplasma hominis endosymbionts that can amplify the severity of infection. However, the mechanisms of communication between endosymbiont, parasite, and host cell are unknown. This cooperation project is based on mutual sharing of data, personnel, and complementary expertise between two leading groups that provide a synergic effect towards investigation of a common aim: to elucidate how presence of endosymbionts (TVV and M. hominis) modify exosomal cargo of T. vaginalis and may stimulate the parasite virulence.

Rada P, Hrdý I, Zdrhal A, Narayanasamy RK, Smutna T, Horáčková J, Harant K, Benes V, Ong SC, Tsai CY, Luo HW, Chiu CH, Tang P*, Tachezy J*. (2022) Double-stranded RNA viruses are released from Trichomonas vaginalis inside small extracellular vesicles and modulate the exosomal cargo. Front. Microbiol. 13:893692

Trichomonas vaginalis is a flagellated parasitic protozoan of the human genitourinary tract. Previous studies showed that a double-stranded ribonucleic acid (dsRNA) virus, Trichomonasvirus (TVV), belongs to the Totiviridae family and can infect T. vaginalis. The TVV family has five subspecies; TVV1~TVV5. Accumulating reports demonstrated that a single T. vaginalis isolate could harbor up to five TVV subspecies. By mining the next-generation ribonucleic acid sequencing (RNA-seq) datasets, we elucidated the distribution of five TVV subspecies in twenty-four T. vaginalis isolates. Our results showed that only three isolates are TVV-free, while fifteen isolates are co-infected by more than one TVV subspecies. To clarify whether multiple TVV subspecies infected all the cells in the same isolate, we will use single-cell RNA sequencing technology to estimate the distribution of TVV subspecies in T. vaginalis isolates. In addition, we will isolated colonies derived from the parent isolate that contains ONLY specifc TVV subspecies or co-infected by combination of TVV2 subspecies. These sub-isolates derived from the same parent isolate and have identical genetic backgrounds except with or without TVV. They are potent tools for future studies.to pinpoint individual or co-infected TVV subspecies' role in the pathogenicity and drug susceptibility of T. vaginalis

Ong SC, Cheng WH, Ku FM, Tsai CY, Huang PJ, Lee CC, Yeh YM, Rada P, Hrdý I, Narayanasamy RK, Smutna T, Lin R. Luo HW, Chiu CH. Tachezy J*. Tang P* (2022) Identification of Endosymbiotic Virus in Small Extracellular Vesicles Derived from Trichomonas vaginalis. Genes (13)531.

Microprotein (MP) is defined as a protein composed of less than 100 amino acids. Accumulated experimental data showed that microproteins play important regulatory roles in various biological processes, including tumor progression, stress response, and signaling transduction. However, microprotein has been under-studied because classical genome annotation methods do not consider open reading frames (ORF) less than 100 amino acids as putative genes. Recent developments in new-generation sequencing techniques allowed us to identify novel MPs and obtain a more comprehensive view of these novel MP's expression profiles. Up to date, MP research had not been explored in any protozoan. This three-year research proposal will use Trichomonas vaginals as a model system to identify and characterize the putative functional roles of MP under iron stress. T. vaginalis is the causing agent of the most common sexually transmitted infection (STI) of nonviral origin in humans trichomoniasis. Iron is an essential element for the survival of T. vaginalis, which sustains hydrogenosomal quality, energy metabolism, and redox homeostasis.

Our preliminary study based on an integrated genome-wide ORF screening, transcriptomic and exosome proteomic data analysis showed that MP did express in the T. vaginalis transcriptome and proteome. These exciting results prompted us to initiate studies on this unexplored field. We aimed to investigate the functional roles of iron-dependent microproteins in T. vaginalis and to establish an experimental and analytical platform for studying microproteins.

Millions of people throughout the world suffer from parasite infections. Traditionally, technicians use manual eye inspection of microscopic specimens to perform a parasite examination. However, manual operations have limitations that hinder the ability to obtain precise egg counts and cause inefficient identification of infected parasites on co-infections. The technician requirements for handling a large number of microscopic examinations in countries that have limited medical resources are substantial. We developed the helminth egg analysis platform (HEAP) as a user-friendly microscopic helminth eggs identification and quantification platform to assist medical technicians during parasite infection examination. (http://heap.cgu.edu.tw)

Lee CC, Huang PJ, Yeh YM, Li PH, Chiu CH, Cheng WH, Tang P*. (2022) Helminth egg analysis platform (HEAP): An opened platform for microscopic helminth egg identification and quantification based on the integration of deep learning architectures. J Microbiol Immunol Infect. (55)395-404.