Heavy metal-laden particulate dust disrupts intestinal epithelial stem cells and lineage differentiation

Chronic heavy metals exposure has been associated with intestinal inflammation and higher incidences of colorectal cancer, however there is no molecular evidence that metal laden-particulates can cause injury to the gut epithelia. Communities that live adjacent to abandoned uranium mines have higher incidence of metabolic and digestive diseases; however, the cause is unknown. The goal of this study is to characterize particulate dust containing non-fissile uranium as an environmental toxicant that damages intestinal epithelia and characterize the subsequent transcriptomic changes. We hypothesized that non-fissile uranium will alter the differentiation pattern of progenitor cells in the crypt biasing them towards the secretory lineage. Human colonoids derived from healthy adult colonic biopsies (UNM IRB approved study 18-626 and 18-171) were established and shown to physiologically model the intestinal epithelia. Colonoids were acutely exposed (24h) to non-fissile uranium dust obtained near the Jackpile uranium mine in New Mexico, one of the largest open pit uranium mines in the US, located on the Laguna Pueblo. Control and dust-exposed colonoids (n=3 unique donors) were dissociated into single cells and processed for droplet-based single cell sequencing (scRNAseq). scRNAseq identified significant changes in the secretory lineage in uranium dust-exposed colonoids. Specifically, there was a four-fold expansion of enteroendocrine cells (EEC) with increases in EEC hormones serotonin and PYY in all colonoid lines. Proliferative pathways, such as Wnt signaling and negative regulation of apoptosis were upregulated in these EECs. These transcriptomic changes in EECs and proliferative cells and increased basolateral secretion of secretory granules and serotonin was validated in colonoids (˃5 biologically unique human donors) via RNA FISH, immunofluorescence and ELISAs. Our results demonstrate that acute uranium dust exposure directly induces transcriptomic and cellular changes in intestinal epithelia independently of microbiota, stroma, and immune cells. These results indicate that adult human colonoids are a relevant model for investigating environmental toxicants on intestinal epithelia and provide an improved understanding of uranium dust as a model of intestinal injury. This study was funded by NIDDK (K01DK106323), NIEHS (R56ES034400, P42ES025589), and NIGMS (P20GM121176, P20GM130422). This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.