Single-cell transcriptomics reveals a conserved metaplasia program in pancreatic injury

Ma, Z.; Lytle, N. K.; Chen, B.; Jyotsana, N.; Weiser Novak, S.; Cho, C. J.; Caplan, L.; Ben-Levy, O.; Neininger, A. C.; Burnette, D. T.; Trinh, V. Q.; Tan, M. C. B.; Patterson, E. A.; Arrojo e Drigo, R.; Giraddi, R. R.; Ramos, C.; Means, A. L.; Manor, U.; Mills, J. C.; Goldenring, J. R.; Lau, K.; Wahl, G.; DelGiorno, K. E.

doi:10.1101/2021.04.09.439243

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bioRxiv · 10.1101/2021.04.09.439243

Single-cell transcriptomics reveals a conserved metaplasia program in pancreatic injury

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Abstract

BACKGROUND & AIMSAcinar to ductal metaplasia (ADM) occurs in the pancreas in response to tissue injury and is a potential precursor for adenocarcinoma. The goal of these studies was to define the populations arising from ADM, the associated transcriptional changes, and markers of disease progression. METHODSAcinar cells were lineage-traced with enhanced yellow fluorescent protein (EYFP) to follow their fate upon injury. Transcripts of over 13,000 EYFP+ cells were determined using single-cell RNA sequencing (scRNA-seq). Developmental trajectories were generated. Data were compared to gastric metaplasia, KrasG12D-induced neoplasia, and human pancreatitis. Results were confirmed by immunostaining and electron microscopy. KrasG12D was expressed in injury-induced ADM using several inducible Cre drivers. Surgical specimens of chronic pancreatitis from 15 patients were evaluated by immunostaining. RESULTSscRNA-seq of ADM revealed emergence of a mucin/ductal population resembling gastric pyloric metaplasia. Lineage trajectories suggest that some pyloric metaplasia cells can generate tuft and enteroendocrine cells (EECs). Comparison to KrasG12D-induced ADM identifies populations associated with disease progression. Activation of KrasG12D expression in HNF1B+ or POU2F3+ ADM populations leads to neoplastic transformation and formation of MUC5AC+ gastric-pit-like cells. Human pancreatitis samples also harbor pyloric metaplasia with a similar transcriptional phenotype. CONCLUSIONSUnder conditions of chronic injury, acinar cells undergo a pyloric-type metaplasia to mucinous progenitor-like populations, which seed disparate tuft cell and EEC lineages. ADM-derived EEC subtypes are diverse. KrasG12D expression is sufficient to drive neoplasia from injury-induced ADM and offers an alternative origin for tumorigenesis. This program is conserved in human pancreatitis, providing insight into early events in pancreas diseases.

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Ma, Z., Lytle, N. K., Chen, B., Jyotsana, N., Weiser Novak, S., Cho, C. J., Caplan, L., Ben-Levy, O., Neininger, A. C., Burnette, D. T., Trinh, V. Q., Tan, M. C. B., Patterson, E. A., Arrojo e Drigo, R., Giraddi, R. R., Ramos, C., Means, A. L., Manor, U., Mills, J. C., Goldenring, J. R., Lau, K., Wahl, G., DelGiorno, K. E.. 2021-04-11. Single-cell transcriptomics reveals a conserved metaplasia program in pancreatic injury. https://doi.org/10.1101/2021.04.09.439243

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