Chagas disease (CD) is a parasitic tropical pathology caused by the protozoan parasite Trypanosoma cruzi. It affects 6-7 million people worldwide, for which no vaccine or satisfactory treatment are available. Historically, CD has been considered endemic in Latin America, but more recently it has spread to Europe, North America, and the Pacific, due to global migrations. An estimated 300,000 people are currently infected in the United States, 45,000 of which exhibit cardiomyopathy, while most cases remain undiagnosed and untreated. There are only two accepted drugs to treat Chagas disease, but they are only available through the Centers for Disease Control and Prevention (CDC). Besides their adverse side effects, the efficacy of these drugs decreases the longer a person has been infected. Therefore, the development of new non-toxic chemotherapies to treat chronic patients of Chagas disease is highly demanded. We are interested in investigating proteins that can be used as chemotherapeutic targets for developing effective treatments against Chagas disease. Our approach is to identify metabolic pathways that are absent in human cells but necessary for parasite survival. Calcium ion (Ca2+) is an important second messenger in trypanosomatids that regulates a vast repertoire of cellular processes. In T. cruzi, Ca2+ signaling is particularly important for host cell invasion, differentiation, osmoregulation and cell bioenergetics. Several proteins involved in calcium homeostasis have been identified in trypanosomes. However, the T. cruzi genome contains a wide variety of calcium-interacting uncharacterized proteins. Using the CRISPR/Cas9 system, we have been able to perform knockout, complementation and in situ tagging of T. cruzi genes. Furthermore, we have established important differences between calcium signaling proteins in T. cruzi and their homologs in mammalian cells. The development of a CRISPR-based inducible silencing system is feasible in T. cruzi and we will use it to perform a large-scale screening of genes encoding calcium-interacting proteins in this organism. Then we will identify essential proteins involved in calcium signaling and perform their biological validation as drug targets, together with other essential proteins already identified. We finally expect to disclose relevant information on protein function, interacting partners, cellular localization and even inhibitors of essential calcium-related proteins. Taken together these results will eventually culminate in the chemical validation of them as target candidates for treatment development against Chagas disease. The mentored phase of this project is going to be developed in the laboratory of Dr. Roberto Docampo at the University of Georgia, which belongs to the Center for Tropical and Emerging Global Diseases. The academic environment, infrastructure and laboratory dynamics integrate the perfect conditions to complete my training plan, and will be determinant to achieve my actual goal: to become an independent researcher and a leader scientist, contributing to the generation of knowledge in the biomedical field.
Trypanosoma cruzi is the causative agent of Chagas disease, a tropical pathology endemic in Latin America considered one of the leading causes of heart disease in the world, for which there is no vaccine or satisfactory treatment available. We have been investigating metabolic differences between this parasite and human cells, with the ultimate goal of identifying new chemotherapeutic targets for the development of alternative treatments against Chagas disease. Understanding the strategies that T. cruzi has developed to survive the sharp environmental changes it faces throughout its life cycle will contribute to the identification of new drug targets.!
