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Transcriptional control of the Cryptosporidium lifecycle

ABSTRACT: Transcriptional control of the Cryptosporidium lifecycle

PROVIDER: PRJNA971984 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24529970.fastq.gz	Fastqsanger.gz
	SRR24529971.fastq.gz	Fastqsanger.gz
	SRR24529972.fastq.gz	Fastqsanger.gz
	SRR24529973.fastq.gz	Fastqsanger.gz
	SRR24529974.fastq.gz	Fastqsanger.gz

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Transcriptional control of the Cryptosporidium lifecycle

Project description:The parasite Cryptosporidium is a leading agent of diarrheal disease in young children, and a cause and consequence of chronic malnutrition. There are no vaccines and only limited treatment options. The parasite infects enterocytes, where it engages in asexual and sexual replication, both of which are essential to continued infection and transmission. However, their molecular mechanisms remain largely unknown. Here we use single-cell RNA sequencing to reveal the gene expression program of the entire C. parvum life cycle in culture and infected animals. Diverging from the prevailing model, we find support for only three intracellular stages: asexual type-I meronts, male gamonts, and female gametes. We uncover a highly organized program for the assembly of components at each stage. Dissecting the underlying regulatory network, we identify the transcription factor Myb-M as the earliest determinant of male fate, in an organism that lacks genetic sex determination. Conditional expression of this factor overrides the developmental program and induces widespread maleness, while conditional deletion ablates male development. Both have a profound impact on the infection. A large set of stage-specific genes now provides the opportunity to understand, engineer, and disrupt parasite sex and life cycle progression to advance the development of vaccines and treatments.

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Lifecycle progression and sexual development of the apicomplexan parasite Cryptosporidium parvum

Project description:The apicomplexan parasite Cryptosporidium is a leading global cause of severe diarrheal disease and an important contributor to early childhood mortality. Currently there are no fully effective treatments or vaccines available. Transmission of the disease occurs through ingestion of oocysts, through direct contact or contaminated water or food. Oocysts are meiotic spores and the product of parasite sex. Cryptosporidium has a single host lifecycle where both asexual and sexual processes unfold in the intestine of infected hosts. Here we use the new-found ability to genetically engineer Cryptosporidium to make life cycle progression and parasite sex tractable. We derive reporter strains to follow parasite development in culture and infected mice and define the genes that orchestrate sex and oocyst formation through mRNA sequencing of sorted cells. After two days, parasites in cell culture show pronounced sexualization, but productive fertilization does not occur and infection falters. In contrast in infected mice, male gametes successfully fertilize females, leading to meiotic division and sporulation. To rigorously test for fertilization, we devised a two-component genetic crossing assay employing a Cre recombinase activated reporter. Our findings suggest obligate developmental progression towards sex in Cryptosporidium, which has important implications for the treatment and prevention of the infection.

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Project description:Cryptosporidium infects enterocytes, but their contribution to parasite control is not well understood. Early resistance to Cryptosporidium is dependent on the production of IFN gamma. Loss of STAT1 in enterocytes, but not dendritic cells or macrophages, antagonized early parasite control. Moreover, transcriptional profiling of enterocytes from infected mice revealed the induction of an IFN gamma signature that included multiple genes (IDO, GBP, IRG) associated with control of intracellular pathogens.

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Lifecycle progression and sexual development of the apicomplexan parasite Cryptosporidium parvum

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RNA-seq of intestinal epithelial cells transfected with an siRNA targeting the host long noncoding RNA, NR_033483, or a scrambled siRNA control, with or without Cryptosporidium infection

Project description:Cryptosporidium is a protozoan parasite that infects the gastrointestinal tract of humans and animals. It is an important opportunistic pathogen in children under the age of two and immunocompromised adults. There is currently no fully effective therapy or vaccine. Long noncoding RNAs are RNA transcripts, over 200 nt in length, that are capable of regulating gene expression through both transcriptional and translational methods. Our lab previously identified a panel of host long noncoding RNAs are that upregulated during Cryptosporidium infection, including NR_033483. Further experiments indicated this host long noncoding RNA may be playing a pro-parasitic role during infection. We hypothesized that NR_033483 is regulating the expression of host immune genes to aid the parasite. In this study, we treated intestinal epithelial cells with an siRNA targeting NR_033483 to knockdown its expression, or a scrambled siRNA control, and infected the samples with Cryptosporidium. We sought to determine changes in host gene expression when NR_033483 was knocked down verse control samples.

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MicroRNA expression profile of HCT-8 in the early phase of Cryptosporidium parvum infection

Project description:Cryptosporidium parvum is an important zoonotic parasitic disease worldwide, but the molecular mechanisms of the host–parasite interaction are not fully understood. Noncoding microRNAs (miRNAs) are considered key regulators of parasitic diseases. Therefore, we used microarray, qPCR, and bioinformatic analyses to investigate the intestinal epithelial miRNA expression profile after Cryptosporidium parvum infection.Twenty miRNAs were differentially expressed after infection (four upregulated and 16 downregulated). Further analysis of the differentially expressed miRNAs revealed that many important cellular responses were triggered by Cryptosporidium parvum infection, including cell apoptosis and the inflammatory and immune responses.This study demonstrates for the first time that the miRNA expression profile of human intestinal epithelium cells is altered by C. parvum infection. This dysregulation of miRNA expression may contribute to the regulation of host biological processes in response to C. parvum infection, including cell apoptosis and the immune responses. These results provide new insight into the regulatory mechanisms of host miRNAs during cryptosporidiosis, which may offer potential targets for future C. parvum control strategies.

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