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Human enteroids as a tool to study conventional and ultra-high dose rate radiation

ABSTRACT: Human enteroids as a tool to study conventional and ultra-high dose rate radiation

PROVIDER: PRJNA1150164 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR30313500_1.fastq.gz	Fastqsanger.gz
	SRR30313500_2.fastq.gz	Fastqsanger.gz
	SRR30313501_1.fastq.gz	Fastqsanger.gz
	SRR30313501_2.fastq.gz	Fastqsanger.gz
	SRR30313502_1.fastq.gz	Fastqsanger.gz

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Human enteroids as a tool to study conventional and ultra-high dose rate radiation

Project description:Radiation therapy, one of the most effective therapies to treat cancer, is highly toxic to healthy tissue. The delivery of radiation at ultra-high dose rates, FLASH radiation therapy (FLASH), has been shown to maintain therapeutic anti-tumor efficacy while sparing normal tissues compared to conventional dose rate irradiation (CONV). Though promising, these studies have been limited mainly to murine models. Here, we leveraged enteroids, three-dimensional cell clusters that mimic the intestine, to study human-specific tissue response to radiation. We observed enteroids have a greater colony growth potential following FLASH compared with CONV. In addition, the enteroids that reformed following FLASH more frequently exhibited proper intestinal polarity. While we did not observe differences in enteroid damage across groups, we did see distinct transcriptomic changes. Specifically, the FLASH enteroids upregulated the expression of genes associated with the WNT-family, cell-cell adhesion, and hypoxia response. These studies validate human enteroids as a model to investigate FLASH and provide further evidence supporting clinical study of this therapy. Insight Box Promising work has been done to demonstrate the potential of ultra-high dose rate radiation (FLASH) to ablate cancerous tissue, while preserving healthy tissue. While encouraging, these findings have been primarily observed using pre-clinical murine and traditional two-dimensional cell culture. This study validates the use of human enteroids as a tool to investigate human-specific tissue response to FLASH. Specifically, the work described demonstrates the ability of enteroids to recapitulate previous in vivo findings, while also providing a lens through which to probe cellular and molecular-level responses to FLASH. The human enteroids described herein offer a powerful model that can be used to probe the underlying mechanisms of FLASH in future studies.

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Project description:Enteroendocrine cells are hormonal secreting cells in the gut. However, as they comprise <1% of the total epithelial cell population, studies on their response to stimuli have been limited. Chang-Graham, Danhof, Engevik, and Tomaro-Duchesneau et al (PMID 31029854) developed a model enteroid system to overcome this limitation. By driving expression of NGN3 in human jejunal enteroids, their system allows for analysis of hormonal secretion and transcriptional analysis in response to a stimulus. To further characterize these NGN3 enteroids, we performed RNASeq on these enteroids in the induced or uninduced state.

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INFANT AND ADULT HUMAN INTESTINAL ENTEROIDS ARE MORPHOLOGICALLY AND FUNCTIONALLY DISTINCT

Project description:Intestinal tissues from 3 infants and 3 adults were obtained and the stem cells were isolated to generate enteroids. Enteroids were differentiated for 5 days prior to RNA extraction and sequencing.

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Project description:Tuberculosis (TB) is a heterogeneous disease manifesting in a subset of individuals infected with aerosolized Mycobacterium tuberculosis (Mtb). Unlike human TB, murine infection results in uniformly high lung bacterial burdens and poorly organized granulomas. To develop a TB model that more closely resembles human disease, we infected mice with an ultra-low dose (ULD) of between 1-3 founding bacteria, reflecting a physiologic inoculum. ULD-infected mice exhibited highly heterogeneous bacterial burdens, well-circumscribed granulomas that shared features with human granulomas, and prolonged Mtb containment with unilateral pulmonary infection in some mice. We identified blood RNA signatures in mice infected with an ULD or a conventional Mtb dose (50-100 CFU) that correlated with lung bacterial burdens and predicted Mtb infection outcomes across species, including risk of progression to active TB in humans. Overall, these findings highlight the potential of the mouse TB model and show that ULD infection recapitulates key features of human TB.

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Expression data from human intestinal enteroids altered for Tgfbeta signaling

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