Carlos Logullo

Upon subsequent blood meals Aedes aegypti can transmit pathogens like dengue, Zika, and chikungunya. The hematophagic behavior does not start right after transition from pupae to adult mosquito, and it is deeply influenced by the metabolic balance after emergence, which regulates the biochemical and molecular changes necessary to seek the vertebrate host. The reproductive success, biting rate, number of eggs laid, and adult longevity are also affected by the metabolic balance. The study of the Insulin/insulin-like growth factor-1 signaling (IIS), is an important source of information that may aid in the development of vector control strategies. The protein tyrosine phosphatases (PTPs) family regulates the IIS, however, they are poorly explored in invertebrates. In the present work, we explore the PTP Leukocyte Common Antigen-Related (LAR) in the mosquito Ae. aegypti (AeLAR), silencing it after emergence to demonstrate that this enzyme also modulates the mosquito IIS pathway. We found that AeLAR knockdown induced glycogen accumulation, even on a restricted diet. The consequence of this accumulation was associated with increased tolerance to starvation and reduced locomotor activity. We also observed changes in the expression of clock genes, host-seeking behavior, blood feeding, and in the number of eggs laid. These results show the importance of AeLAR in insect metabolism and physiology. Disrupting mosquito metabolism during the onset of the adult stage, as demonstrated here, can be used as an additional resource to existing control methods before the mosquitoes become infected

Epigenetic modifications are long-lasting changes to the genome that influence a cell’s transcriptional potential, thereby altering its function. These modifications can trigger adaptive responses that impact protein expression and various cellular processes, including differentiation and growth. The primary epigenetic mechanisms identified to date include DNA and RNA methylation, histone modifications, and microRNA-mediated regulation of gene expression. The intricate crosstalk among these mechanisms makes epigenetics a compelling field for the development of novel control strategies, particularly through the use of epigenetic drugs targeting arthropod vectors such as ticks. In this study, we identified the Rhipicephalus microplus orthologs of canonical histone-modifying enzymes, along with components of the machinery responsible for m⁵C and ⁶mA-DNA, and m⁶A-RNA methylations. We further characterized their transcriptional profiles and enzymatic activities during embryonic development. To explore the functional consequences of epigenetic regulation in R. microplus, we evaluated the effects of various epigenetic inhibitors on the BME26 tick embryonic cell line. Molecular docking simulations were performed to predict the binding modes of these inhibitors to tick enzymes, followed by in vitro assessment of their effects on cell viability and morphology. Tick cells exposed to these inhibitors presented phenotypic and molecular alterations. Notably, we observed high levels of DNA methylation in the nuclear genome. Importantly, inhibition of DNA methylation using 5′-azacytidine (5′-AZA) was associated with increased activity of the mitochondrial electron transport chain and ATP synthesis but reduced cellular proliferation. Our findings highlight the importance of epigenetic regulation during tick embryogenesis and suggest that targeting these pathways may constitute a novel and promising strategy for tick control.

The advancement of multi-omics technologies is crucial to deepen knowledge on tick biology. These approaches, used to study diverse phenomena, are applied to experiments that aim to understand changes in gene transcription, protein function, cellular processes, and prediction of systems at global biological levels. This review addressed the application of omics data to investigate and elucidate tick physiological processes, such as feeding, digestion, reproduction, neuronal, endocrine systems, understanding population dynamics, transmitted pathogens, control, and identifying new vaccine targets. Furthermore, new therapeutic perspectives using tick bioactive molecules, such as anti-inflammatory, analgesic, and antitumor, were summarized. Taken together, the application of omics technologies can help to understand the protein functions and biological behavior of ticks, as well as the identification of potential new antigens influencing the development of alternative control strategies and, consequently, the tick-borne disease prevention in veterinary and public health contexts. Finally, tick population dynamics have been determined through a combination of environmental factors, host availability, and genetic adaptations, and recent advances in omics technologies have improved our understanding of their ecological resilience and resistance mechanisms. Future directions point to the integration of spatial omics and artificial intelligence to further unravel tick biology and improve control strategies.

The advancement of multi-omics technologies is crucial to deepen knowledge on tick biology. These approaches, used to study diverse phenomena, are applied to experiments that aim to understand changes in gene transcription, protein function, cellular processes, and prediction of systems at global biological levels. This review addressed the application of omics data to investigate and elucidate tick physiological processes, such as feeding, digestion, reproduction, neuronal, endocrine systems, understanding population dynamics, transmitted pathogens, control, and identifying new vaccine targets. Furthermore, new therapeutic perspectives using tick bioactive molecules, such as anti-inflammatory, analgesic, and antitumor were summarized. Taken together, the application of omics technologies can help to understand the protein functions and biological behavior of ticks, as well as the identification of potential new antigens influencing the development of alternative control strategies and consequently the tick borne-diseases prevention in veterinary and public health contexts. Finally, tick population dynamics have been determined by a combination of environmental factors, host availability and genetic adaptations, and recent advances in omics technologies have improved our understanding of their ecological resilience and resistance mechanisms. Future directions point to the integration of spatial omics and artificial intelligence to further unravel tick biology and improve control strategies.

Ticks are hematophagous ectoparasites that transmit pathogens and inflict significant economic losses on the cattle industry. Remarkably, they can survive extended periods of starvation in the absence of a host. The primary objective of this study was to investigate the metabolic adaptations that enable the tick Rhipicephalus microplus to endure starvation using the BME26 cell line as a model system. To simulate nutrient deprivation, cells were subjected to starvation conditions by replacing the L-15 culture medium with phosphate-buffered saline (PBS). Our findings show that these tick cells can endure experimental starvation for up to 48 h. The assessment of glycogen levels in starved cells shows a significant decrease, at both the 24 h and 48 h marks. Additionally, upregulation of phosphoenolpyruvate carboxykinase (PEPCK) gene expression, along with downregulation of hexokinase (HK) and pyruvate kinase (PK) gene expression, indicated that BME26 cells would prioritize the gluconeogenic pathway over the glycolytic pathway under starvation conditions. Moreover, the transcriptional levels of autophagy-related genes (ATG) were upregulated in response to starvation. Taken together, our findings suggest a potential role for autophagy in supplying substrates for the gluconeogenic pathway in nutrient-deprived tick cells. This work contributes to the understanding of metabolic regulation in R. microplus ticks and offers valuable insights for tick control strategies.

Aurora kinases (AURK) play a central role in controlling cell cycle in a wide range of organisms. They belong to the family of serine-threonine kinase proteins. Their role in the cell cycle includes, among others, the entry into mitosis, maturation of the centrosome and formation of the mitotic spindle. In mammals, 3 isoforms have been described: A, B and C, which are distinguished mainly by their function throughout the cell cycle. Two aurora kinase coding sequences have been identified in the transcriptome of the cattle tick Rhipicephalus microplus (Rm-AURKA and Rm-AURKB) containing the aurora kinase-specific domain. For both isoforms, the highest number of AURK coding transcripts is found in ovaries. Based on deduced amino acid sequences, it was possible to identify non-conserved threonine residues which are essential to AURK functions in vertebrates and which are not present in R. microplus sequences. A pan AURK inhibitor (CCT137690) caused cell viability decline in the BME26 tick embryonic cell line. In silico docking assay showed an interaction between Aurora kinase and CCT137690 with exclusive interaction sites in Rm-AURKA. The characterization of exclusive regions of the enzyme will enable new studies aimed at promoting species-specific enzymatic inhibition in ectoparasites.

The enzyme glucose-6-phosphate dehydrogenase (G6PDH) plays crucial roles in glucose homeostasis and the pentose phosphate pathway (PPP), being also involved in redox metabolism. The PPP is an important metabolic pathway that produces ribose and nicotinamide adenine dinucleotide phosphate (NADPH), which are essential for several physiologic and biochemical processes, such as the synthesis of fatty acids and nucleic acids. As a rate-limiting step in PPP, G6PDH is a highly conserved enzyme and its deficiency can lead to severe consequences for the organism, in particular for cell growth. Insufficient G6PDH activity can lead to cell growth arrest, impaired embryonic development, as well as a reduction in insulin sensitivity, inflammation, diabetes, and hypertension. While research on G6PDH and PPP has historically focused on mammalian models, particularly human disorders, recent studies have shed light on the regulation of this enzyme in arthropods, where new functions were discovered. This review will discuss the role of arthropod G6PDH in regulating redox homeostasis and immunometabolism and explore potential avenues for further research on this enzyme in various metabolic adaptations.