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| Status |
Public on Jun 27, 2013 |
| Title |
Leishmania donovani amastigotes: Nelfinavir (NFV)-sensitive (control) vs Nelfinavir-resistant [Comparative Genomic Hybridization (DNA) profiling] |
| Platform organism |
Leishmania |
| Sample organism |
Leishmania donovani |
| Experiment type |
Genome variation profiling by array
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| Summary |
Drug resistance is a major public health challenge in Leishmaniasis chemotherapy, particularly in the case of emerging Leishmania/HIV-1 co-infections. Recently, we have delineated the mechanism of cell death induced by the HIV-1 protease inhibitor, Nelfinavir, in the Leishmania parasite. In order to investigate the underlying molecular mechanism involved in Nelfinavir resistance, in vitro Nelfinavir resistant amastigotes were developed by direct drug pressure in culture. RNA expression profiling analyses of closely related Leishmania species were used as a screening tool to compare Nelfinavir-resistant and -sensitive parasites in order to identify candidate genes involved in drug resistance, and several genes were found to be differentially expressed. Comparative gene hybridization (CGH) analyses of Nelfinavir-resistant and -sensitive Leishmania using whole-genome 60-mer oligonucleotide microarrays were also carried out. RNA expression profiles and the CGH of Nelfinavir resistant vs sensitive Leishmania amastigotes suggest that parasites regulate mRNA levels either by modulating gene copy numbers through chromosome aneuploidy, or gene deletion/duplication by homologous recombination. Interestingly, supernumerary chromosomes 6 and 11 in the resistant parasites lead to upregulation of the ABC class of transporters, which are involved in vesicular trafficking. Transporter assays using radiolabeled Nelfinavir suggest that the drug accumulates in greater amounts in the resistant parasites and in a time dependent manner. Furthermore, high-resolution electron microscopy showed an increased number of vacuoles in Nelfinavir-resistant parasites. Together these results suggest that Nelfinavir is rapidly and dramatically sequestered in these intracellular vesicles.
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| Overall design |
Two condition experiment: NFV-sensitive vs resistant. Biological replicates: Three. One dye swap.
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| Contributor(s) |
Kumar P, Raymond F, Lodge R, Ouellette M, Tremblay MJ |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
Nov 09, 2012 |
| Last update date |
Jun 29, 2013 |
| Contact name |
Frederic Raymond |
| Organization name |
Université Laval
|
| Department |
École de nutrition
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| Lab |
INAF
|
| Street address |
2440 Bd Hochelaga Suite 1710
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| City |
Quebec City |
| State/province |
Quebec |
| ZIP/Postal code |
G1V 0A6 |
| Country |
Canada |
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| Platforms (1) |
| GPL11330 |
Agilent custom Leishmania microarray v2 |
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| Samples (3) |
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| This SubSeries is part of SuperSeries: |
| GSE42197 |
Leishmania donovani amastigotes: Nelfinavir (NFV)-sensitive (control) vs Nelfinavir-resistant |
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| Relations |
| BioProject |
PRJNA179277 |
