Trypanosoma Cruzi Induces Phospho‐proteome Profile in Primary Colon Epithelial Cells During Early Infection

Abstract

Trypanosoma cruzi, an obligate intracellular protozoan parasite, is the etiological agent of Chagas Disease. Due to globalization, the disease is now present in all economically advanced countries of the world. About 30% of T. cruzi infected individuals will develop cardiac, neurological, and/or gastrointestinal pathology, particularly megacolon. Megacolon, one of Chagas disease major pathologies, is accompanied by gastrointestinal motility disorders that have been attributed to alterations in the number of interstitial cells of Cajal and enteric nervous system defects. The objective of this study is to begin to define early molecular mechanisms that contribute to chagasic megacolon that can be exploited for development of molecular intervention strategies. In order to do this, we challenged primary colon epithelial cells (HCoEpiC) with invasive T. cruzi trypomastigotes at different time points to decipher changes in levels of phosphorylated proteins. Using a proteome profiler Human phospho‐kinase array, we show that T. cruzi induced a unique phospho‐proteome profile. We validated the phospho‐kinase array data via immunoblotting and immunofluorescence. Modulators of host immune response such JAK2, STAT1, and NF‐kB showed increased activation. Increased phosphorylated levels of kinases such, as p‐Src and p‐Fyn and the Heat Shock protein (p‐HSP27) chaperone illustrate dysregulation of intracellular signaling. We found that p‐JNK and p‐AKT exhibited increased phosphorylation and their downstream effectors p‐CREB and p‐c‐Jun showed increased nuclear colocalization. Early infection also induced increase thrompsopondin‐1 expression, a known fibrogenic protein. We generated a biological interaction network using Ingenuity Pathway Analysis (IPA) to further characterize protein‐protein interactions within the phospho‐proteome profile. In conclusion, we demonstrate a unique phospho‐proteome profile induced by T. cruzi during early infection of HCoEpiC and illustrate biological networks and signal transduction pathways that are activated during infection.Support or Funding InformationThis work was supported by NIH grants SC1AI127352, 5U54MD007586 (The RCMI Program in Health Disparities Research), and 5R25GM059994‐19 (RISE).