Banca de DEFESA: DYONATAN FONSECA SILVA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
STUDENT : DYONATAN FONSECA SILVA
DATE: 26/06/2024
TIME: 13:00
LOCAL: remota
TITLE:

EVALUATION OF THE METALLOPROTEINASE PROFILE IN TISSUE SAMPLES FROM ANIMALS EXPERIMENTALLY INFECTED WITH Trypanosoma cruzi

KEY WORDS:

Zymography, Metalloproteinase, Trypanosoma cruzi, Chagas disease, Pathogenesis

PAGES: 83
BIG AREA: Ciências Biológicas
AREA: Bioquímica
SUBÁREA: Bioquímica dos Microorganismos
SUMMARY:

Chagas disease (DCh) is caused by the parasite Trypanosoma cruzi (T. cruzi) and is characterized by two clinical phases: acute and chronic. During the acute phase, the parasite targets specific tissues and dysregulates cytokines and matrix metalloproteinases (MMPs), which are major risk factors for the development of cardiopathies, common complications in both the acute and chronic phases of the disease. This study aimed to optimize gel-based methods, such as SDS-PAGE and zymography, to evaluate whether T. cruzi infection could alter the metalloproteinase profile of key organs during the acute phase of DCh. For this purpose, total protein extract from epimastigote forms of T. cruzi Y strain from axenic culture was obtained, and experimental infection with blood trypomastigote forms was performed in female mice, divided into three groups: uninfected (CN, negative control, n=3) and infected, euthanized at 17 (D17, n=3) and 32 (D32, n=3) days post-infection. Control animals were used as a baseline for comparison with infected animals. Heart, spleen, liver, and blood from all groups were collected to produce total extract and blood plasma. Optimization allowed the determination of the protein profile of T. cruzi, revealing bands with approximate molecular weights of 95, 85, 63, 57, 48-52, 43-45, 40-41, 29, and 18-20 kDa, which were discussed in terms of their possible identification and function in the infection. The results also demonstrated the protein profiles of cardiac, splenic, hepatic tissues, and plasma in the three analyzed groups. The T. cruzi zymogram revealed three activity bands with approximate molecular weights of 66 kDa, 55 kDa, and 35 kDa, along with a faint activity band between 66 and 35 kDa. In the infected animal group, clinical signs of hepatomegaly and splenomegaly characteristic of the acute phase of DCh were observed. Several gelatin-degrading metalloproteinases were detected in the zymogram of tissues and plasma, especially in splenic tissue. Differences in their expression during acute infection were observed only in splenic and cardiac tissues, with the alteration in cardiac tissue related to the increased expression of a 93 kDa protease, likely proMMP-9, and in the spleen, a 146 kDa unknown protease. In summary, the findings provide information on the protein and proteolytic expression patterns for T. cruzi and the analyzed organs plus plasma, revealing abundant proteins as potential immunogenic targets against T. cruzi. These data serve for comparison of the genetic and functional diversity of the parasite along with other studies, besides confirming changes in protease expression during the acute phase of DCh, opening avenues for diagnostics and treatment based on the necessity and feasibility of inhibiting these proteins.

COMMITTEE MEMBERS:
Externa à Instituição - ALINE RIMOLDI RIBEIRO
Externa à Instituição - CLÁUDIA JASSICA GONÇALVES MORENO
Presidente - 2275890 - MARCELO DE SOUSA DA SILVA
Externo à Instituição - TAFFAREL MELO TORRES - UFERSA