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Supplemental Figure 2. Calcipotriol does not alter LRP6 expression in Low-BAR/Low-VDR cell lines. (A) Western blot for LRP6 or tubulin (loading control) following treatment with vehicle or 10 μM calcipotriol. Representative experiments from 2-3 biologic replicates are shown.

Supplemental Figure 1. Calcipotriol does not inhibit Wnt signaling through VDR sequestration of β-catenin or induction of DKK1. (A) AsPC-1 cells were treated with 100 nM calcipotriol for 4 hours. Whole cell lysates were collected and immunoprecipitated with anti- β-catenin or isotype control antibodies followed by western blots for β-catenin, E-cadherin or VDR. (B) Western blot for DKK-1 and tubulin (loading control) and (C) BAR reporter activity for indicated conditions. AsPC-1 cells were transfected with control or DKK1 siRNA for 48 hours and treated with vehicle or 100 nM calcipotriol for an additional 24 hours. BAR was normalized to vehicle and siRNA control and is reported as mean {plus minus} SD, with one representative experiment of 3 biological repeats shown.

Supplemental Figure 3. Calcipotriol reduces VDR message while paradoxically increasing VDR protein levels. (A) VDR expression in AsPC-1 by qPCR after treatments described in Figure 3E. (B) Western blots for VDR or tubulin (loading control) for AsPC-1 lysates at indicated timepoints after calcipotriol treatment. Arrowhead indicates specific band representing VDR protein. qPCR reported as mean {plus minus} SEM. Representative experiments from 2-3 biologic replicates are shown. **P

Supplemental Table 1. QPCR PRIMERS

Supplemental Table 2. RNA-Seq analysis showing significant gene expression changes following 6 hour calcipotriol treatment of AsPC-1 cell line

Die Corona-Pandemie hat kommunale Verwaltungen vor große Herausforderungen gestellt. Gleichzeitig bietet die anhaltende Krise Chancen für digitale Wandlungsprozesse innerhalb der öffentlichen Verwaltungen. Die vorliegende Studie untersucht Digitalisierungsprozesse in den Kommunalverwaltungen Nordrhein-Westfalens (NRW) im Kontext der Corona-Pandemie. Empirisch stützt sich die Studie auf eine Online-Befragung von 118 Kommunen. Die erhobenen Daten lassen Rückschlüsse auf den Stand der Verwaltungsdigitalisierung vor, die Entwicklung während und potenzielle Veränderungen nach der Corona-Pandemie zu. Die Ergebnisse bestätigen, dass die Corona-Pandemie Digitalisierungsprozesse in der Kommunalverwaltung in NRW insgesamt vorangetrieben hat und die Befragten erwarten, dass die neuen digitalen Arbeitsmodi auch nach der Pandemie genutzt werden. Als Digitalisierungshemmnisse werden vor allem die finanzielle und technische Ausstattung der Kommunalverwaltungen, unsichere Rechtslagen und zurückhaltende Führungskräfte genannt.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy in need of more effective treatment approaches. One potential therapeutic target is Wnt/β-catenin signaling, which plays important roles in PDAC tumor initiation and progression. Among Wnt inhibitors with suitable in vivo biologic activity is vitamin D, which is known to antagonize Wnt/β-catenin signaling in colorectal cancer and have antitumor activity in PDAC. For this study, the relationship between vitamin D signaling, Wnt/β-catenin activity, and tumor cell growth in PDAC was investigated through the use of calcipotriol, a potent non-hypercalcemic vitamin D analogue. PDAC tumor cell growth inhibition by calcipotriol was positively correlated with vitamin D receptor expression and Wnt/β-catenin activity. Furthermore, vitamin D and Wnt signaling activity were found to be reciprocally linked through feedback regulation. Calcipotriol inhibited autocrine Wnt/β-catenin signaling in PDAC cell lines in parallel with decreased protein levels of the low-density lipoprotein receptor-related protein 6 (LRP6), a requisite coreceptor for ligand-dependent canonical Wnt signaling. Decrease in LRP6 protein seen with calcipotriol was mediated through a novel mechanism involving transcriptional upregulation of low-density lipoprotein receptor adaptor protein 1 (LDLRAP1). Finally, changes in LRP6 or LDLRAP1 expression directly altered Wnt reporter activity, supporting their roles as regulators of ligand-dependent Wnt/β-catenin signaling. Implications: This study provides a novel biochemical target through which vitamin D signaling exerts inhibitory effects on Wnt/β-catenin signaling, as well as potential biomarkers for predicting and following tumor response to vitamin D–based therapy. Mol Cancer Res; 13(11); 1509–19. ©2015 AACR.

Brown adipose tissue (BAT) thermogenesis is critical to maintain homoeothermia and is centrally controlled via sympathetic outputs. Body temperature and BAT activity also impact energy expenditure, and obesity is commonly associated with decreased BAT capacity and sympathetic tone. Severely obese mice that lack leptin or its receptor (LepRb) show decreased BAT capacity, sympathetic tone, and body temperature and thus are unable to adapt to acute cold exposure (Trayhurn et al., 1976). LepRb-expressing neurons are found in several hypothalamic sites, including the dorsomedial hypothalamus (DMH) and median preoptic area (mPOA), both critical sites to regulate sympathetic, thermoregulatory BAT circuits. Specifically, a subpopulation in the DMH/dorsal hypothalamic area (DHA) is stimulated by fever-inducing endotoxins or cold exposure (Dimicco and Zaretsky, 2007; Morrison et al., 2008). Using the retrograde, transsynaptic tracer pseudorabies virus (PRV) injected into the BAT of mice, we identified PRV-labeled LepRb neurons in the DMH/DHA and mPOA (and other sites), thus indicating their involvement in the regulation of sympathetic BAT circuits. Indeed, acute cold exposure induced c-Fos (as a surrogate for neuronal activity) in DMH/DHA LepRb neurons, and a large number of mPOA LepRb neurons project to the DMH/DHA. Furthermore, DMH/DHA LepRb neurons (and a subpopulation of LepRb mPOA neurons) project and synaptically couple to rostral raphe pallidus neurons, consistent with the current understanding of BAT thermoregulatory circuits from the DMH/DHA and mPOA (Dimicco and Zaretsky, 2007; Morrison et al., 2008). Thus, these data present strong evidence that LepRb neurons in the DMH/DHA and mPOA mediate thermoregulatory leptin action.