Trypanosoma cruzi kDNA minicircle sequences integration into the vertebrate host genome: A biomarker for monitoring the efficacy of multidrug treatment of Chagas disease

Antonio Raimundo Lima Teixeira 1, * Clever Cardoso Gomes 2, Adriana Alves Sá 1, 3, Rubens José Nascimento 1, 4, Liana Lauria-Pires 1, 5, Ana Maria Castro 6, Francisco Ernesto Moreno Bernal 7 and Alessandro Oliveira de Sousa 1, 8

1 Department of Pathology, Faculty of Medicine of the University of Brasília. CEP 70.910-900, Federal District, Brazil.
2 Department of Histology, Embryology and Cell Biology, Federal University of Goiás. CEP 74605-90, Goiânia Brazil.
3 Sambaia Regional Hospital, QS 614, Cj C, Block 1/2, South Samambaia. CEP 72322-583, Federal District, Brazil.
4 National Technical Commission on Biosafety, Ministry of Science and Technology. South Police Sector, Area 5, Block 3, Block B, Rooms 10 to 14. CEP 70610-200, Brasília, Federal District, Brazil.
5 Catholic University of Brasilia, Department of Medicine, Taguatinga. CEP 71966-700, Federal District, Brazil.
6 Institute for Tropical Pathology and Public Health, Federal University of Goiás. CEP 74605-050, Goiânia, Brazil.
7 Animal Welfare and Avian Pathology Laboratory, Faculty of Agronomy and Veterinary Medicine, University of Brasília, CEP 70910-900, Federal District, Brazil.
8 Logistic Office for Health, Federal District Health Department, Building PO700, 2nd Floor, SRTV 702, W5 North. CEP 70723-040, Brasília, Brazil.
 
Research Article
International Journal of Scientific Research Updates, 2023, 05(01), 170–207.
Article DOI: 10.53430/ijsru.2023.5.1.0026
Publication history: 
Received on 01 February 2023; revised on 10 March 2023; accepted on 13 March 2023
 
Abstract: 
The protist Trypanosoma cruzi inserts kinetoplast DNA minicircle sequences into the host genome. Sensitive PCR with specific primer sets revealed protozoan nuclear DNA and kinetoplast DNA in agarose gels bands probed with radiolabel specific sequences for tissues of T. cruzi-infected rabbits and mice. Avian species are refractory to the infection, but chicks that hatched from T. cruzi-inoculated eggs retained the kinetoplast DNA in the embryonic germ line cells and developed parasite-free Chagas disease-like cardiomyopathy. A Target-primer TAIL-PCR with specific primer sets, Southern hybridization, cloning, and sequencing of the amplification products revealed kinetoplast minicircle sequences integration sites mainly in LINE-1 transposable elements and hitchhiking to several loci. This kinetoplast DNA biomarker was used to monitor the effect of multidrug treatment of T. cruzi-infected mice. A trypanocidal nitro heterocyclic compound in combination with an array of inhibitors of eukaryotic cell division prevented minicircle sequences transfer. Nine out of 12 inhibitors prevented the kinetoplast DNA integration into the macrophage genome. The multidrug treatment of T. cruzi-infected mice with benznidazole, azidothymidine and ofloxacin lowered the rate of minicircle sequences integrations into the mouse genome by 2.44-fold and reduced the rejection of the target heart cells.
 
Keywords: 
Trypanosoma cruzi; kDNA transfer; Biomarker; Multidrug treatment; Chagas disease
 
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