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. 2023 Aug 12;13(1):13120.
doi: 10.1038/s41598-023-40353-9.

Insights into the microRNA landscape of Rhodnius prolixus, a vector of Chagas disease

Paula Beatriz Santiago  1 , Kaio Luís da Silva Bentes  1 , Waldeyr Mendes Cordeiro da Silva  2 , Yanna Reis Praça  1 , Sébastien Charneau  3 , Soraya Chaouch  4 , Philippe Grellier  4 , Marcos Antônio Dos Santos Silva Ferraz  5 , Izabela Marques Dourado Bastos  1 , Jaime Martins de Santana  1 , Carla Nunes de Araújo  6   7
Affiliations

Insights into the microRNA landscape of Rhodnius prolixus, a vector of Chagas disease

Paula Beatriz Santiago et al. Sci Rep. .

Abstract

The growing interest in microRNAs (miRNAs) over recent years has led to their characterization in numerous organisms. However, there is currently a lack of data available on miRNAs from triatomine bugs (Reduviidae: Triatominae), which are the vectors of the protozoan parasite Trypanosoma cruzi, the causative agent of Chagas disease. A comprehensive understanding of the molecular biology of vectors provides new insights into insect-host interactions and insect control approaches, which are key methods to prevent disease incidence in endemic areas. In this work, we describe the miRNome profiles from gut, hemolymph, and salivary gland tissues of the Rhodnius prolixus triatomine. Small RNA sequencing data revealed abundant expression of miRNAs, along with tRNA- and rRNA-derived fragments. Fifty-two mature miRNAs, previously reported in Ecdysozoa, were identified, including 39 ubiquitously expressed in the three tissues. Additionally, 112, 73, and 78 novel miRNAs were predicted in the gut, hemolymph, and salivary glands, respectively. In silico prediction showed that the top eight most highly expressed miRNAs from salivary glands potentially target human blood-expressed genes, suggesting that R. prolixus may modulate the host's gene expression at the bite site. This study provides the first characterization of miRNAs in a Triatominae species, shedding light on the role of these crucial regulatory molecules.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Read length distribution and abundance of small RNAs from Rhodnius prolixus. The graphs show the processed read numbers and size distribution (18–50 nt). The x-axis represents the length (nt) of the sequence. The y-axis represents the number of reads.
Figure 2
Figure 2
Abundance of Rhodnius prolixus miRNA reads aligned against known miRNAs database. The box plots illustrate hit species distribution results of R. prolixus genome-mapped reads aligned against a collection of known miRNAs from Ecdysozoa taxa. Features are based on sequence alignments and the abundance of reads from the extracted R. prolixus miRNAs. The box covers the interquartile interval, where 50% of the data is found. Each point represents a unique miRNA. Species are shown at the left for visualization.
Figure 3
Figure 3
Distribution profile of Rhodnius prolixus conserved miRNAs. Venn diagram depicts the degree of overlap of known miRNAs from the gut, hemolymph, and salivary glands.
Figure 4
Figure 4
Expression levels of Rhodnius prolixus miRNAs. The expression levels of miRNAs are based on normalization standards and AI levels, as described in the “Materials and methods” section. The blue color represents the highly expressed conserved miRNAs, the gray color represents the mild expressed, and yellow is the less expressed, all shown with color intensities degree. Colored circles indicate the presence of miRNA in the tissue as indicated: pink, gut; green, hemolymph; blue, salivary gland. The highlighted blue bars indicate the top expressed miRNAs. The miRNA sequences are shown.
Figure 5
Figure 5
Distribution profile of Rhodnius prolixus novel miRNAs. The Venn diagram depicts the degree of overlap of novel mature miRNAs from the gut, hemolymph, and salivary glands.
Figure 6
Figure 6
Small RNA abundance of Rhodnius prolixus samples. The bar graphs summarize reads abundance of genome-mapped reads aligned against a collection of sncRNAs from Triatominae downloaded from the RNAcentral database (https://rnacentral.org) and the extracted R. prolixus miRNAs sequences (conserved and novel). The horizontal axis represents the abundance (%) of aligned reads (log2 of the enrichment ratio).
Figure 7
Figure 7
Enriched pathways and Gene ontology Biological Process terms related to genes targeted by the top eight most highly expressed miRNAs from Rhodnius prolixus salivary glands. (A) Enriched pathways. The horizontal axis represents the Log2 of the enrichment ratio. The vertical axis represents the − Log10 value of the FDR rate. The size and color of the dots are proportional to the size of the category. The most significant pathways are towards the top. Especially meaningful categories for anti-hemostatic salivary function within the TOP 20 functional categories with FDR <  = 0.05 are labeled. (B) Enriched set of terms from Geneontology Biological Process. The horizontal axis represents the Log2 of the enrichment ratio. The vertical axis represents the − Log10 value of the FDR rate. The size and color of the dots are proportional to the size of the category. The most significant pathways are towards the top. The TOP 20 terms with FDR <  = 0.05 are labeled.
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