Impaired Regulation of PMCA Activity by Defective CFTR Expression Promotes Epithelial Cell Damage in Alcoholic Pancreatitis and Hepatitis

Background and aims. Alcoholic pancreatitis and hepatitis are frequent, potentially lethal diseases with limited treatment options. Our previous study reported that the expression of CFTR Cl- channel is impaired by ethanol in pancreatic ductal cells leading to more severe alcohol-induced pancreatitis. In addition to determining epithelial ion secretion, CFTR has multiple interactions with other proteins, which may influence intracellular Ca2+ signaling. Thus, we aimed to investigate the impact of ethanol-mediated CFTR damage on intracellular Ca2+ homeostasis in pancreatic ductal epithelial cells and cholangiocytes.Methods. Human and mouse pancreas and liver samples and ex vivo organoids were used to study ion secretion, intracellular signaling and protein expression and interaction. The effect of PMCA4 inhibition was analysed in a mouse model of alcohol-induced pancreatitis.Results. The decreased CFTR expression impaired PMCA function and resulted in sustained intracellular Ca2+ elevation in ethanol-treated and mouse and human pancreatic organoids. Liver samples derived from alcoholic hepatitis patients and ethanol-treated mouse liver organoids showed decreased CFTR expression and function, and impaired PMCA4 activity. PMCA4 co-localizes and physically interacts with CFTR on the apical membrane of polarized epithelial cells, where CFTR-dependent calmodulin recruitment determines PMCA4 activity. The sustained intracellular Ca2+ elevation in the absence of CFTR inhibited mitochondrial function and was accompanied with increased apoptosis in pancreatic epithelial cells and PMCA4 inhibition increased the severity of alcohol-induced AP in mice.Conclusion. Our results suggest that improving Ca2+ extrusion in epithelial cells may be a potential novel therapeutic approach to protect the exocrine pancreatic function in alcoholic pancreatitis and prevent the development of cholestasis in alcoholic hepatitis.

Pancreatic Duct Cells Isolated From Canines Differentiate Into Beta-Like Pancreatic Islet Cells

In this study, we isolated and cultured pancreatic ductal cells from canines and revealed the possibility for using them to differentiate into functional pancreatic beta cells in vitro. Passaged pancreatic ductal cells were induced to differentiate into beta-like pancreatic islet cells using a mixture of induced factors. Differentiated pancreatic ductal cells were analyzed based on intracellular insulin granules using transmission electron microscopy, the expression of insulin and glucagon using immunofluorescence, and glucose-stimulated insulin secretion using ELISA. Our data revealed that differentiated pancreatic ductal cells not only expressed insulin and glucagon but also synthesized insulin granules and secreted insulin at different glucose concentrations. Our study might assist in the development of effective cell therapies for the treatment of type 1 diabetes mellitus in dogs.

Differential expression of SPC25, NDC80 kinetochore complex component in triple negative breast cancer.

Women diagnosed with triple negative breast cancer can benefit neither from endocrine therapy nor from HER2-targeted therapies (1). We mined published microarray datasets (2, 3) to determine in an unbiased fashion and at the systems level genes most differentially expressed in the primary tumors of patients with breast cancer. We report here significant differential expression of the gene encoding SPC25, NDC80 kinetochore complex component, SPC25, when comparing the tumor cells of patients with triple negative breast cancer to normal mammary ductal cells (2). SPC25 was also differentially expressed in bulk tumor in human breast cancer (3). SPC25 mRNA was present at significantly increased quantities in TNBC tumor cells relative to normal mammary ductal cells. Analysis of human survival data revealed that expression of SPC25 in primary tumors of the breast was correlated with overall survival in patients with luminal A type cancer, while within triple negative breast cancer, primary tumor expression of SPC25 was correlated with overall survival in patients with luminal androgen receptor subtype disease. SPC25 may be of relevance to initiation, maintenance or progression of triple negative breast cancers.

Differential expression of kinesin family member 2C in triple negative breast cancer.

Women diagnosed with triple negative breast cancer can benefit neither from endocrine therapy nor from HER2-targeted therapies (1). We mined published microarray datasets (2, 3) to determine in an unbiased fashion and at the systems level genes most differentially expressed in the primary tumors of patients with breast cancer. We report here significant differential expression of the gene encoding kinesin family member 2C, KIF2C, when comparing the tumor cells of patients with triple negative breast cancer to normal mammary ductal cells (2). KIF2C was also differentially expressed in bulk tumor in human breast cancer (3). KIF2C mRNA was present at significantly increased quantities in TNBC tumor cells relative to normal mammary ductal cells. Analysis of human survival data revealed that expression of KIF2C in primary tumors of the breast was correlated with overall survival in patients with basal-like type cancer, while within triple negative breast cancer, primary tumor expression of KIF2C was correlated with overall survival in patients with mesenchymal and luminal androgen receptor subtype disease. KIF2C may be of relevance to initiation, maintenance or progression of triple negative breast cancers.

Differential expression of lamin B1 in triple negative breast cancer.

Women diagnosed with triple negative breast cancer can benefit neither from endocrine therapy nor from HER2-targeted therapies (1). We mined published microarray datasets (2, 3) to determine in an unbiased fashion and at the systems level genes most differentially expressed in the primary tumors of patients with breast cancer. We report here significant differential expression of the gene encoding lamin B1, LMNB1, when comparing the tumor cells of patients with triple negative breast cancer to normal mammary ductal cells (2). LMNB1 was also differentially expressed in bulk tumor in human breast cancer (3). LMNB1 mRNA was present at significantly increased quantities in TNBC tumor cells relative to normal mammary ductal cells. Analysis of human survival data revealed that expression of LMNB1 in primary tumors of the breast was correlated with overall survival in patients with basal-like and luminal A subtype cancer, while within triple negative breast cancer, primary tumor expression of LMNB1 was correlated with overall survival in patients with basal-like 2, immunomodulatory and mesenchymal subtype disease. LMNB1 may be of relevance to initiation, maintenance or progression of triple negative breast cancers.

Differential expression of pituitary tumor-transforming 1 in triple negative breast cancer.

Women diagnosed with triple negative breast cancer can benefit neither from endocrine therapy nor from HER2-targeted therapies (1). We mined published microarray datasets (2, 3) to determine in an unbiased fashion and at the systems level genes most differentially expressed in the primary tumors of patients with breast cancer. We report here significant differential expression of the gene encoding pituitary tumor-transforming 1, PTTG1, when comparing the tumor cells of patients with triple negative breast cancer to normal mammary ductal cells (2). PTTG1 was also differentially expressed in bulk tumor in human breast cancer (3). PTTG1 mRNA was present at significantly increased quantities in TNBC tumor cells relative to normal mammary ductal cells. Analysis of human survival data revealed that expression of PTTG1 in primary tumors of the breast was correlated with overall survival in patients with basal-like type cancer, while within triple negative breast cancer, primary tumor expression of PTTG1 was correlated with recurrence-free survival in patients with basal-like 1 and mesenchymal subtype disease. PTTG1 may be of relevance to initiation, maintenance or progression of triple negative breast cancers.

Differential expression of chromosome 20 open reading frame 24 in triple negative breast cancer.

Women diagnosed with triple negative breast cancer can benefit neither from endocrine therapy nor from HER2-targeted therapies (1). We mined published microarray datasets (2, 3) to determine in an unbiased fashion and at the systems level genes most differentially expressed in the primary tumors of patients with breast cancer. We report here significant differential expression of the gene encoding chromosome 20 open reading frame 24, C20orf24, when comparing the tumor cells of patients with triple negative breast cancer to normal mammary ductal cells (2). C20orf24 was also differentially expressed in bulk tumor in human breast cancer (3). C20orf24 mRNA was present at significantly increased quantities in TNBC tumor cells relative to normal mammary ductal cells. Analysis of human survival data revealed that expression of C20orf24 in primary tumors of the breast was correlated with overall survival in patients with luminal A type cancer, while within triple negative breast cancer, primary tumor expression of C20orf24 was correlated with overall survival in patients with basal-like 1 and immunomodulatory subtype disease. C20orf24 may be of relevance to initiation, maintenance or progression of triple negative breast cancers.

Cell Type of Pancreatic Ductal Adenocarcinoma Origin: Implications for Prognosis and Clinical Outcomes

<b><i>Background:</i></b> Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease that has no effective early detection method or treatment to date. <b><i>Summary:</i></b> The normal cell type that initiates PDAC, or its cellular origin, is still unknown. To investigate the contribution of distinct normal epithelial cell types to PDAC tumorigenesis, genetically engineered mouse models were used to show that both acinar and ductal cells are capable of giving rise to PDAC. These studies indicated that genetic mutations and pancreatic injury interact differently with each cellular origin to affect their predilection and process for forming PDAC. In this review, we summarize recent findings using various genetically engineered mouse models in the identification and characterization of the PDAC cell of origin. We also discuss potential implications for cellular origin on tumor development, PDAC transcriptional subtype, and disease prognosis of patients. <b><i>Key Message:</i></b> Although it is clear that both ductal and acinar cells have the potential to form PDAC, whether cellular origin can indeed influence patient prognosis and whether knowledge of cellular origin will aid in the diagnosis or treatment of patients in the future will need further study.

Differential expression of histone cluster 1, H2bh in triple negative breast cancer.

Women diagnosed with triple negative breast cancer can benefit neither from endocrine therapy nor from HER2-targeted therapies (1). We mined published microarray datasets (2, 3) to determine in an unbiased fashion and at the systems level genes most differentially expressed in the primary tumors of patients with breast cancer. We report here significant differential expression of the gene encoding histone cluster 1, H2bh, HIST1H2BH, when comparing the tumor cells of patients with triple negative breast cancer to normal mammary ductal cells (2). HIST1H2BH was also differentially expressed in the brain metastases of patients with metastastic breast cancer (3). HIST1H2BH mRNA was present at significantly increased quantities in TNBC tumor cells relative to normal mammary ductal cells. Analysis of human survival data revealed that expression of HIST1H2BH in primary tumors of the breast was correlated with overall survival in patients with luminal A type cancer, while within triple negative breast cancer, primary tumor expression of HIST1H2BH was correlated with distant metastasis-free survival in patients with immunomodulatory subtype disease. HIST1H2BH may be of relevance to initiation, maintenance or progression of triple negative breast cancers.

Differential expression of NDC80, kinetochore complex component in triple negative breast cancer.

Women diagnosed with triple negative breast cancer can benefit neither from endocrine therapy nor from HER2-targeted therapies (1). We mined published microarray datasets (2, 3) to determine in an unbiased fashion and at the systems level genes most differentially expressed in the primary tumors of patients with breast cancer. We report here significant differential expression of the gene encoding NDC80, kinetochore complex component, NDC80, when comparing the tumor cells of patients with triple negative breast cancer to normal mammary ductal cells (2). NDC80 was also differentially expressed in bulk tumor in human breast cancer (3). NDC80 mRNA was present at significantly increased quantities in TNBC tumor cells relative to normal mammary ductal cells. Analysis of human survival data revealed that expression of NDC80 in primary tumors of the breast was correlated with overall survival in patients with basal-like and luminal A subtype cancer, while within triple negative breast cancer, primary tumor expression of NDC80 was correlated with overall survival in patients with basal-like 1 subtype disease. NDC80 may be of relevance to initiation, maintenance or progression of triple negative breast cancers.