Inhibition of Mist1 Homodimer Formation Induces Pancreatic Acinar-to-Ductal Metaplasia

Abstract

The pancreas consists of three main cell lineages (endocrine, exocrine, and duct) that develop from common primitive foregut precursors. The transcriptional network responsible for endocrine cell development has been studied extensively, but much less is known about the transcription factors that maintain the exocrine and duct cell lineages. One transcription factor that may be important to exocrine cell function is Mist1, a basic helix-loop-helix (bHLH) factor that is expressed in acinar cells. In order to perform a molecular characterization of this protein, we employed coimmunoprecipitation and bimolecular fluorescence complementation assays, coupled with electrophoretic mobility shift assay studies, to show that Mist1 exists in vivo as a homodimer complex. Analysis of transgenic mice expressing a dominant-negative Mist1 transgene (Mist1^{mutant basic} [Mist1^MB]) revealed the cell autonomous effect of inhibiting endogenous Mist1. Mist1^MB cells become disorganized, exhibit a severe depletion of intercellular gap junctions, and express high levels of the glycoprotein clusterin, which has been shown to demarcate immature acinar cells. Inhibition of Mist1 transcriptional activity also leads to activation of duct-specific genes, such as cytokeratin 19 and cytokeratin 20, suggesting that alterations in the bHLH network produce a direct acinar-to-ductal phenotypic switch in mature cells. We propose that Mist1 is a key transcriptional regulator of exocrine pancreatic cells and that in the absence of functional Mist1, acinar cells do not maintain their normal identity.

We thank Ray MacDonald, Galvin Swift, Rolf Kemler, Doris Stoffers, Chang-Deng Hu, and Tom Kerppola for providing valuable reagents used in this study and Judy Hallett and the Purdue Cancer Center Transgenic Mouse Core Facility for their assistance in generating the mice described in this report.

This work was supported in part by grants to S.F.K. from the National Institutes of Health (DK55489) and from the Purdue University Cancer Center. L.Z and J.M.R. were supported by Purdue Research Foundation graduate fellowships, and T.T. was supported by a GAANN graduate student training grant.