Expression and biological evaluation of rescombinant proteins from Simulium pertinax salivary gland that act in the hemostatic system

• Grant number: 20/08863-1
• Support Opportunities: Regular Research Grants
• Start date: July 01, 2021
• End date: June 30, 2023
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Geraldo Santana Magalhães
• Grantee: Geraldo Santana Magalhães
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated researchers: Andrea de Barros Pinto Viviani Cunha ; Eliana Faquim de Lima Mauro ; Keith Miller ; Peter Nicholas Strong

Abstract

Bloodsucking insects cause a wide range of problems in humans and other vertebrates, ranging from skin irritations to the transmission of a variety of pathogens that can result in acute illness, debilitation, or death. In this sense, the insects of the Simuliidae family, also known as black flies have medical and economic importance, especially in rural and coastal areas, since their bites cause intense itching and irritation and they are also vectors of diseases such as onchocerciasis (river blindness) and mansonellosis. The genus includes more than 2300 species and in the State of São Paulo, Simulium (Chirostilbia) pertinax stands out, which, due to its wide distribution, density, and voracious bloodsucking habits, has been subjected to control campaigns. Several pharmacologically active compounds are present in the salivary gland of these insects and play a crucial role in facilitating their blood supply, such as anticoagulants, antiplatelet factors, vasodilators, immunomodulators, anti-inflammatories, among others. Due to the particularity of acting with great efficiency in the hemostatic system, the study of the composition of the saliva of these insects can be beneficial for the discovery of new anti-hemostatic drugs. However, due to the small amount of these molecules in the saliva of these insects, their purification is very laborious and often impossible, which hinders their biochemical characterization. Thus, some works carry out the transcriptome of the salivary gland to unveil the profile of expressed genes, thus opening the possibility of obtaining several proteins in recombinant form. With this approach in mind, in this project, we intend to perform the transcriptome of S. (C.) pertinax to detail the profile of gene expression of the salivary gland of this species and we intend to analyze in-depth the transcripts involved in the hemostatic system and subsequently clone and express them in a heterologous system to assess their biological activities. This approach may allow the identification of new proteins that act in the hemostatic system and thus may contribute to the development of drugs with therapeutic activities for both humans and animals. (AU)