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ABSTRACT: RNA-seq for MTB infected mice BMDMs

PROVIDER: PRJNA723409 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14290429.fastq.gz	Fastqsanger.gz
	SRR14290430.fastq.gz	Fastqsanger.gz
	SRR14290431.fastq.gz	Fastqsanger.gz
	SRR14290432.fastq.gz	Fastqsanger.gz
	SRR14290433.fastq.gz	Fastqsanger.gz

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Effect of small molecule inhibitors on transcriptome of B6.Sst1S bone marrow derived macrophages stimulated with TNF and infected with virulent Mycobacterium tuberculosis

Project description:We investigated candidate host-directed therapies for tuberculosis in vitro. BMDMs from tuberculosis susceptible mice B6.Sst1S were stimulated with TNF and infected with Mtb. The infected macrophage were treated with Rocaglamide A, ISRIB and JNK inhibitor SP600125 alone or in combinations to asses the inhibitors' effectas on macrophage transcriptome.

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Next Generation Sequencing unveils the effect of caloric restriction on the splenic response to virulent H37rv Mycobacterium tubercolosis in DBA/2 mice

Project description:Purpose: The goal of this study was the transcriptome high-throughput data analysis of spleen cells from mice infected with a virulent strain of mycobacterium (H37rv) and either fed ad libitum (AL) or caloric restricted (CR) Methods: Spleen cell total RNA profile of 17 weeks old mice (1. MTB infected, Ad libitum fed; 2. MTB infected, Caloric restricted; 3. Not infected, Ad libitum fed; 4. Not infected, Caloric restricted; a pool of 5 animals per group) were generated by deep sequencing, in triplicate, using TruSeq Stranded Total RNA Illumina technology Results: Principal component analysis and unsupervised clustering showed that the RNA-seq profiles of spleen cells from these four groups of mice were clearly distinct and infection was the major force affecting gene expression, with 67% of variance coming from the infection status and another 23% variance being dependent on caloric intake . When directly comparing AL versus CR MTB infected spleens, 628 genes were found differentially expressed. Supervised hierarchical clustering based on the expression of these 628 genes was able to unveil the unique transcriptional profile of CR MTB infected mice compared to both AL MTB infected and Not infected mice. Coclusions: Caloric restriction is able to affect the expression of a significant number of genes in spleen cells isolated from mice infected with MTB. The study of biological process enrichements highlights relevant immune-related regulation in these animals, than may be at the base of the higher capacity of caloric restricted, compared to ad libitum fed, animals to resist MTB infection.

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Project description:Dual RNA-seq of Mtb infected macrophages

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Mycobacterium tuberculosis produces small noncoding RNAs that support mycobacterial survival in infected cells [miRNA discovery screen]

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Project description:Iron plays a critical role in the pathogenesis of many organisms including Mtb. It is the preferred redox cofactor in many basic cellular processes but due to its insolubility and potential toxicity under physiological conditions is a limiting nutrient in the host environment. Previously, we demonstrated that Mtb requires the iron storage protein ferritin (BfrB), for adaptation to both low and sufficient concentrations of environmental iron. We also showed that absence of bfrB compromises the ability of Mtb to overcome iron deficiency and prevent excess iron toxicity. In this study, we tested whether vaccination with ?bfrB could elicit host protective immune response against virulent Mtb infection. The results show that immunization of mice with the ?bfrB stimulates protective immunity associated with reduced immunopathology and better containment of the infection compared to vaccination with BCG. Genome-wide transcriptome analysis showed a distinct expression pattern of significantly differentially expressed genes (SDEG) between the ?bfrB and BCG-vaccinated, Mtb-infected mice lungs. Our network/pathway analysis of SDEG revealed significant inhibition of inflammatory response genes and activation of fibrosis genes in the ?bfrB, compared to BCG vaccinated, Mtb-infected mice lungs. The results provide a frame work for the study of mechanisms of protection relevant for the design of new and improved preventive strategies for TB. Female C57BL/6 mice were vaccinated subcutaneously with ?bfrB or BCG . At 8 weeks post-vaccination, mice were aerosol infected with Mtb H37Rv. At 4 weeks post infection, mice were sacrificed and lungs were removed. Lung total RNA from vaccinated and Mtb-infected mice was extracted, purified and were processed separately for microarray analysis.

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