Abstract
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.
