Project description:Mycetoma is a neglected, chronic granulomatous infection of the subcutaneous tissue, most often caused by the fungal pathogen Madurella mycetomatis. Characteristic of the infection is the formation of grains.
Project description:Mycetoma is a neglected, chronic granulomatous infection of the subcutaneous tissue, most often caused by the fungal pathogen Madurella mycetomatis. Characteristic of the infection is the formation of grains.
Project description:Mycetoma is a neglected, chronic granulomatous infection of the subcutaneous tissue, most often caused by the fungal pathogen Madurella mycetomatis. Characteristic of the infection is the formation of grains. However, knowledge of the function and formation of the grain is limited. To map the processes leading to M. mycetomatis grain formation, we used a Galleria mellonella larvae infection model and time-course transcriptomic profiling. In the infected G. mellonella 88.0% of the RNA sequence reads mapped to G. mellonella, only 0.01% mapped to M. mycetomatis. Differential Gene Expression analysis revealed that 3.498 G. mellonella and 137 M. mycetomatis genes were differentially expressed during infection. Most of the enriched GO terms of both host and pathogen are linked to energy pathways, nucleobase metabolic process, and cation and iron transport. Genes related to iron transport were highly expressed by both G. mellonella (transferrin and ferritin) and M. mycetomatis (SidA, SidD and SidI). A protein-protein interaction network analysis of D. melanogaster homologues genes in M. mycetomatis revealed the expression of the entire siderophore biosynthesis pathway throughout infection. Many host and pathogen genes were differentially expressed during infection. The identification of the importance of iron acquisition during grain formation can be exploited as a potential novel diagnostic and therapeutic strategy for mycetoma.
Project description:Mycetoma is a neglected, chronic granulomatous infection of the subcutaneous tissue, most often caused by the fungal pathogen Madurella mycetomatis. Characteristic of the infection is the formation of grains. However, knowledge of the function and formation of the grain is limited. To map the processes leading to M. mycetomatis grain formation, we used a Galleria mellonella larvae infection model and time-course transcriptomic profiling. In the infected G. mellonella 88.0% of the RNA sequence reads mapped to G. mellonella, only 0.01% mapped to M. mycetomatis. Differential Gene Expression analysis revealed that 3.498 G. mellonella and 137 M. mycetomatis genes were differentially expressed during infection. Most of the enriched GO terms of both host and pathogen are linked to energy pathways, nucleobase metabolic process, and cation and iron transport. Genes related to iron transport were highly expressed by both G. mellonella (transferrin and ferritin) and M. mycetomatis (SidA, SidD and SidI). A protein-protein interaction network analysis of D. melanogaster homologues genes in M. mycetomatis revealed the expression of the entire siderophore biosynthesis pathway throughout infection. Many host and pathogen genes were differentially expressed during infection. The identification of the importance of iron acquisition during grain formation can be exploited as a potential novel diagnostic and therapeutic strategy for mycetoma.
Project description:Mycetoma is a chronic granulomatous infection of the subcutaneous tissue, most often caused by the fungal pathogen Madurella mycetomatis. Characteristic of the infection is the formation of grains. However, knowledge of the function and formation of the grain is limited. To map the processes leading to M. mycetomatis grain formation, we used a Galleria mellonella larvae infection model and transcriptomic profiling. G. mellonella larvae were infected with M. mycetomatis and after 4h, 24h, 72h and 168h post-inoculation, RNA was extracted from larval content and sequenced. Differential Gene Expression analysis revealed that 3,498 G. mellonella and 136 M. mycetomatis genes were differentially expressed during infection. Among these genes, genes related to iron transport were highly expressed by both G. mellonella (transferrin and ferritin) and M. mycetomatis (SidA, SidD and SidI). LC-MS/MS analysis of M. mycetomatis cultured under iron-limiting conditions revealed the presence of SidA and SidD orthologs and concurrent RP-HPLC and LC-MS identified a singly charged, putative siderophore in culture supernatant. M. mycetomatis could also obtain iron from holoferritin. The identification of the importance of iron acquisition during grain formation can be exploited as a potential novel diagnostic and therapeutic strategy for mycetoma.
Project description:Using Low Quantity single strand CAGE (LQ-ssCAGE) we mapped the transcription start sites (TSS) and annotated the 5' end of the invertebrate Galleria mellonella which is upcoming and booming in the last years as an experimental model in infection disease and immunology research. The current genome annotation of this model lacks the annotation of the 5' end and TSS information. To map TSS under healthy and infection conditions, the G. mellonella larva was infected with the fungal pathogen Madurella mycetomatis After 4, 30 and 52 hours of the infection, larvae were treated with itraconazole or ravuconazole. RNA-seq and LQ-ssCAGE libraries prepared and sequenced. The LQ-ssCAGE data was processed to identify CAGE transcription start site (CTSS), uni- and bi-directional clusters. LQ-ssCAGE enabled us to precisely identify (39,410) TSS and (249) active enhancers. We assigned genomic features to the resulting TSSs and enhancers. The majority of the TSS peaks are annotated as promoter regions while the enhancers were annotated as intergenic and genic. Furthermore, we confirmed the quality of TSS by promoter shapes and GC bias. We identified a set of super-enhancers and predicted de-novo motifs. In this study we reported the first atlas of TSS and active enhancers of the G. mellonella.