scholarly journals Single-cell resolution analysis of the human pancreatic ductal progenitor cell niche

2020 ◽  
Vol 117 (20) ◽  
pp. 10876-10887 ◽  
Author(s):  
Mirza Muhammad Fahd Qadir ◽  
Silvia Álvarez-Cubela ◽  
Dagmar Klein ◽  
Jasmijn van Dijk ◽  
Rocío Muñiz-Anquela ◽  
...  
Keyword(s):  
Single Cell ◽  
Pancreatic Ducts ◽  
Human Pancreas ◽  
Immunodeficient Mice ◽  
Ductal Cells ◽  
Surrogate Surface ◽  
Morphogenetic Protein ◽  
Resolution Analysis ◽  
Cell Niche ◽  
Acinar Tissue

We have described multipotent progenitor-like cells within the major pancreatic ducts (MPDs) of the human pancreas. They express PDX1, its surrogate surface marker P2RY1, and the bone morphogenetic protein (BMP) receptor 1A (BMPR1A)/activin-like kinase 3 (ALK3), but not carbonic anhydrase II (CAII). Here we report the single-cell RNA sequencing (scRNA-seq) of ALK3bright+-sorted ductal cells, a fraction that harbors BMP-responsive progenitor-like cells. Our analysis unveiled the existence of multiple subpopulations along two major axes, one that encompasses a gradient of ductal cell differentiation stages, and another featuring cells with transitional phenotypes toward acinar tissue. A third potential ducto-endocrine axis is revealed upon integration of the ALK3bright+ dataset with a single-cell whole-pancreas transcriptome. When transplanted into immunodeficient mice, P2RY1+/ALK3bright+ populations (enriched in PDX1+/ALK3+/CAII− cells) differentiate into all pancreatic lineages, including functional β-cells. This process is accelerated when hosts are treated systemically with an ALK3 agonist. We found PDX1+/ALK3+/CAII− progenitor-like cells in the MPDs of types 1 and 2 diabetes donors, regardless of the duration of the disease. Our findings open the door to the pharmacological activation of progenitor cells in situ.

scholarly journals Human pluripotent stem cell-derived acinar/ductal organoids generate human pancreas upon orthotopic transplantation and allow disease modelling

Gut ◽  
2016 ◽  
Vol 66 (3) ◽  
pp. 473-486 ◽  
Author(s):  
Meike Hohwieler ◽  
Anett Illing ◽  
Patrick C Hermann ◽  
Tobias Mayer ◽  
Marianne Stockmann ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Global Gene Expression ◽  
Pancreatic Disease ◽  
Pancreatic Ducts ◽  
Human Pancreas ◽  
Orthotopic Transplantation ◽  
Immunodeficient Mice ◽  
Gene Therapy Approach ◽  
Ductal Cells

ObjectiveThe generation of acinar and ductal cells from human pluripotent stem cells (PSCs) is a poorly studied process, although various diseases arise from this compartment.DesignWe designed a straightforward approach to direct human PSCs towards pancreatic organoids resembling acinar and ductal progeny.ResultsExtensive phenotyping of the organoids not only shows the appropriate marker profile but also ultrastructural, global gene expression and functional hallmarks of the human pancreas in the dish. Upon orthotopic transplantation into immunodeficient mice, these organoids form normal pancreatic ducts and acinar tissue resembling fetal human pancreas without evidence of tumour formation or transformation. Finally, we implemented this unique phenotyping tool as a model to study the pancreatic facets of cystic fibrosis (CF). For the first time, we provide evidence that in vitro,but also in our xenograft transplantation assay, pancreatic commitment occurs generally unhindered in CF. Importantly, cystic fibrosis transmembrane conductance regulator (CFTR) activation in mutated pancreatic organoids not only mirrors the CF phenotype in functional assays but also at a global expression level. We also conducted a scalable proof-of-concept screen in CF pancreatic organoids using a set of CFTR correctors and activators, and established an mRNA-mediated gene therapy approach in CF organoids.ConclusionsTaken together, our platform provides novel opportunities to model pancreatic disease and development, screen for disease-rescuing agents and to test therapeutic procedures.

scholarly journals SARS-CoV-2 Cell Entry Factors ACE2 and TMPRSS2 are Expressed in the Pancreas but are Not Enriched in Islet Endocrine Cells

2020 ◽  
Author(s):  
Katie C. Coate ◽  
Jeeyeon Cha ◽  
Shristi Shrestha ◽  
Wenliang Wang ◽  
Luciana Mateus Gonçalves ◽  
...  
Keyword(s):  
Pancreatic Islet ◽  
Endocrine Cells ◽  
Islet Cells ◽  
Cell Entry ◽  
Pancreatic Ducts ◽  
Human Pancreas ◽  
Β Cells ◽  
Ductal Cells ◽  
Putative Receptor ◽  
New Onset

Summary/AbstractReports of new-onset diabetes and diabetic ketoacidosis in individuals with COVID-19 have led to the hypothesis that SARS-CoV-2, the virus that causes COVID-19, is directly cytotoxic to pancreatic islet β cells. This would require binding and entry of SARS-CoV-2 into host β cells via cell surface co-expression of ACE2 and TMPRSS2, the putative receptor and effector protease, respectively. To define ACE2 and TMPRSS2 expression in the human pancreas, we examined six transcriptional datasets from primary human islet cells and assessed protein expression by immunofluorescence in pancreata from donors with and without diabetes. ACE2 and TMPRSS2 transcripts were low or undetectable in pancreatic islet endocrine cells as determined by bulk or single cell RNA sequencing, and neither protein was detected in α or β cells from these donors. Instead, ACE2 protein was expressed in the islet and exocrine tissue microvasculature and also found in a subset of pancreatic ducts, whereas TMPRSS2 protein was restricted to ductal cells. The absence of significant ACE2 and TMPRSS2 co-expression in islet endocrine cells reduces the likelihood that SARS-CoV-2 directly infects pancreatic islet β cells through these cell entry proteins.

scholarly journals Multiomics single-cell analysis of human pancreatic islets reveals novel cellular states in health and type 1 diabetes

2021 ◽  
Author(s):  
Maria Fasolino ◽  
Gregory W. Schwartz ◽  
Maria L. Golson ◽  
Yue J. Wang ◽  
Ashleigh Morgan ◽  
...  
Keyword(s):  
Type 1 Diabetes ◽  
Single Cell ◽  
Pancreatic Islets ◽  
Mhc Class Ii ◽  
Single Cell Analysis ◽  
Cell Types ◽  
Human Pancreas ◽  
Analytical Strategy ◽  
Ductal Cells

AbstractType 1 diabetes (T1D) is an autoimmune disease of only partially defined etiology in which immune cells destroy insulin-producing beta cells. Using single-cell transcriptomics and an advanced analytical strategy to assess pancreatic islets of T1D, autoantibody-positive, and non-diabetic organ donors, we identified both canonical cell types and rare insulin-expressing cells with a hybrid mixture of endocrine and exocrine gene signatures within all donors. We further found elevated expression of MHC Class II pathway genes in exocrine ductal cells of T1D donors, which we confirmed through CyTOF, in situ imaging mass cytometry, and immunofluorescence analysis. Taken together, our multimodal analyses identify novel cell types and processes that may contribute to T1D immunopathogenesis and provide new cellular and molecular insights into human pancreas function.

scholarly journals EPEN-31. SINGLE-CELL RNAseq OF CHILDHOOD EPENDYMOMA REVEALS DISTINCT NEOPLASTIC CELL SUBPOPULATIONS THAT IMPACT ETIOLOGY, MOLECULAR CLASSIFICATION AND OUTCOME

2020 ◽  
Vol 22 (Supplement_3) ◽  
pp. iii314-iii314
Author(s):  
Andrew Donson ◽  
Austin Gillen ◽  
Riemondy Kent ◽  
Ahmed Gilani ◽  
Sujatha Venkataraman ◽  
...  
Keyword(s):  
Single Cell ◽  
Histological Analysis ◽  
Molecular Classification ◽  
Neoplastic Cell ◽  
Good Prognosis ◽  
Cellular Heterogeneity ◽  
Mesenchymal Phenotype ◽  
Ependymal Differentiation ◽  
Cell Subpopulations ◽  
Cell Niche

Abstract Ependymoma (EPN) is a brain tumor commonly presenting in childhood that remains fatal in the majority of children. Intra-tumoral cellular heterogeneity in bulk-tumor samples significantly confounds our understanding of EPN biology, impeding development of effective therapy. We therefore used single-cell RNA sequencing to catalog cellular heterogeneity of 26 childhood EPN, predominantly from ST-RELA, PFA1 and PFA2 subgroups. ST-RELA and PFA subgroups clustered separately, with ST-RELA clustering largely according to individual sample-of-origin. PFA1 and PFA2 subgroup EPNs cells were intermixed and revealed 4 major subpopulations – 2 with characteristics of ependymal differentiation (transporter and ciliated phenotype subpopulations), an undifferentiated subpopulation and a mesenchymal phenotype. Pseudotime analysis showed the undifferentiated progenitor subpopulation either differentiating into ependymal differentiation subpopulations or transitioning into the mesenchymal subpopulation. Histological analysis revealed that undifferentiated and mesenchymal subpopulations cells colocalized to perinecrotic/perivascular zones, the putative ependymoma stem cell niche. Deconvolution of PFA bulk transcriptome data showed that undifferentiated and mesenchymal subpopulations were associated with a poor prognosis; whereas the ciliated ependymal cell-differentiated subpopulation was associated with a good prognosis. In conflict with current distinct classification paradigms, the ratio of mesenchymal and ciliated subpopulations determined bulk-tumor subgroups assignment to PFA1 and PFA2 respectively. This atlas of EPN cellular heterogeneity provides an important advance in our understanding of EPN biology, identifying high-risk associated subpopulations for therapeutic targeting.

scholarly journals Regulation and expression of a renin-angiotensin system in human pancreas and pancreatic endocrine tumours

2002 ◽  
pp. 567-572 ◽  
Author(s):  
KY Lam ◽  
PS Leung
Keyword(s):  
Renin Angiotensin System ◽  
Receptor Protein ◽  
Pancreatic Ducts ◽  
Human Pancreas ◽  
Rt Pcr ◽  
Angiotensin System ◽  
Endocrine Tumours ◽  
Renin Angiotensin ◽  
Objective Evidence ◽  
Pancreatic Endocrine Tumours

OBJECTIVE: Evidence exists for the presence of a renin-angiotensin system (RAS) in the pancreas. The aims of this study were to prove the presence of an intrinsic RAS in the human pancreas and to analyse the role of such an RAS in pancreatic endocrine tumours (PETs). METHODS: Gene expression of key RAS components (angiotensinogen and angiotensin II receptors, namely AT1 and AT2) was investigated in human pancreas and in PETs by semi-quantitative RT-PCR and immunohistochemistry. RESULTS: Expression of mRNAs of RAS components was found in human pancreas and in PETs. Data from semi-quantitative RT-PCR analysis demonstrated an increase in the mRNA expression of angiotensinogen and AT2 receptor in PETs when compared with that in normal pancreas. By immunohistochemistry, angiotensinogen protein was predominantly localized in the pancreatic islets while AT1 receptor protein was in the pancreatic ducts. CONCLUSIONS: The data support the notion of the existence of an intrinsic RAS in the human pancreas. It also indicates, for the first time, that such a local pancreatic RAS is subject to regulation by PETs and its significant change may have pathophysiological relevance in patients with PETs.

scholarly journals A Novel Method to Identify the Differences Between Two Single Cell Groups at Single Gene, Gene Pair, and Gene Module Levels

2021 ◽  
Vol 12 ◽  
Author(s):  
Lingyu Cui ◽  
Bo Wang ◽  
Changjing Ren ◽  
Ailan Wang ◽  
Hong An ◽  
...  
Keyword(s):  
Single Cell ◽  
Molecular Mechanisms ◽  
Single Gene ◽  
Cell Types ◽  
Human Pancreas ◽  
Biological Difference ◽  
Cell Clusters ◽  
Cell Groups ◽  
Network Modules ◽  
Differential Gene

Single-cell sequencing technology can not only view the heterogeneity of cells from a molecular perspective, but also discover new cell types. Although there are many effective methods on dropout imputation, cell clustering, and lineage reconstruction based on single cell RNA sequencing (RNA-seq) data, there is no systemic pipeline on how to compare two single cell clusters at the molecular level. In the study, we present a novel pipeline on comparing two single cell clusters, including calling differential gene expression, coexpression network modules, and so on. The pipeline could reveal mechanisms behind the biological difference between cell clusters and cell types, and identify cell type specific molecular mechanisms. We applied the pipeline to two famous single-cell databases, Usoskin from mouse brain and Xin from human pancreas, which contained 622 and 1,600 cells, respectively, both of which were composed of four types of cells. As a result, we identified many significant differential genes, differential gene coexpression and network modules among the cell clusters, which confirmed that different cell clusters might perform different functions.

scholarly journals Uroguanylin and guanylate cyclase C in the human pancreas: expression and mutuality of ligand/receptor localization as indicators of intercellular paracrine signaling pathways

2001 ◽  
Vol 170 (1) ◽  
pp. 267-275 ◽  
Author(s):  
H Kulaksiz ◽  
Y Cetin
Keyword(s):  
Epithelial Cells ◽  
Guanylate Cyclase ◽  
Human Pancreas ◽  
Membrane Domain ◽  
Apical Cell Membrane ◽  
Ductal Cells ◽  
Pancreatic Ductal Cells ◽  
Cellular Expression ◽  
Guanylate Cyclase C ◽  
Electrolyte Secretion

The intestinal peptide hormone uroguanylin regulates electrolyte/fluid transport in the gastrointestinal epithelium by binding to its receptor, guanylate cyclase C (GC-C), and thus specifically coupling to activation of cystic fibrosis transmembrane conductance regulator (CFTR). Since CFTR is crucially involved in pancreatic electrolyte secretion, we investigated the human pancreas for expression and cell-specific localization of uroguanylin and guanylate cyclase C as potential regulatory components of pancreatic electrolyte secretion. RT-PCR analyses with specific primers revealed that uroguanylin and GC-C are expressed in the human pancreas (and in the duodenum, used as positive control); at the translational level, western blotting analyses with peptide- and region-specific antibodies identified the presence of 12.5 kDa uroguanylin and 130 kDa GC-C in both human pancreatic and intestinal extracts. At the cellular level, uroguanylin and GC-C immunoreactivities were absent from the islets of Langerhans but were exclusively confined to the exocrine parenchyma. Hence, uroguanylin was localized to the centroacinar cells typical of the pancreas, and also to epithelial cells of the intercalated, intralobular and interlobular ducts where the peptide was primarily concentrated adluminally to the apical portion of the respective cells. Coincidently, correlative studies localized the GC-C receptor to the epithelial cells of the ductal network, where it was confined exclusively to the apical cell membrane that evidently represents the functionally relevant target membrane domain for the regulatory peptide. In view of the fact that CFTR is highly expressed in pancreatic ductal cells where uroguanylin and its receptor are also localized, we assume that uroguanylin, an intrinsic pancreatic peptide, is involved in the regulation of electrolyte/water secretion in the ductal system via GC-C and CFTR. The particular cellular expression of uroguanylin in duct cells and the localization of GC-C to the duct cell apical membrane domain predict a novel route of intercellular signaling and luminal activation of GC-C via the pancreatic juice.

Expression of GRP78 protein in pancreatic ductal adenocarcinoma and intraductal papillary mucinous neoplasm.

2017 ◽  
Vol 35 (4_suppl) ◽  
pp. 323-323
Author(s):  
Asha R. Dhanarajan ◽  
Reginald Hill ◽  
Sam G. Pappas ◽  
Gerard J. Abood ◽  
Jennifer Lynn Gnerlich ◽  
...  
Keyword(s):  
Pancreatic Ductal Adenocarcinoma ◽  
Intraductal Papillary Mucinous Neoplasm ◽  
Protein Quality ◽  
Expression Level ◽  
Ductal Adenocarcinoma ◽  
Pancreatic Ducts ◽  
Low Grade ◽  
Ductal Cells ◽  
Mucinous Neoplasm ◽  
Significant Difference

323 Background: Pancreatic ductal adenocarcinoma (PDAC) has a poor prognosis due to late detection, aggressive biology and resistance to chemotherapy. Markers for early detection of PDAC and mediators of chemoresistance are poorly understood. GRP78 is a major endoplasmic reticulum (ER) chaperone protein critical for protein quality control of the ER. Increased GRP78 expression promotes in vitro cancer cell survival, progression, and chemoresistance. This study aims to examine expression of GRP78 protein in PDAC and intraductal papillary mucinous neoplasm (IPMN) compared to normal pancreatic ducts using immunohistochemistry (IHC). Methods: The expression of GRP78 was assessed using IHC in 93 sections of formalin-fixed paraffin-embedded tissue: normal ducts (38), low grade IPMN (IPMN-LG, 26), high grade IPMN (IPMN-HG, 9), and PDAC (20). Immunostaining intensity of GRP78 protein was categorized as no or weak staining (0-1+) and strong staining (2-3+). Fisher’s exact test with two tails was performed using the GraphPad statistical software. Results: GRP78 expression was identified in the cytoplasm of normal pancreatic ducts, IPMN and PDAC with a fine granular pattern. All PDAC and IPMN-HG showed strong expression of GRP78 while the normal ductal cells showed only minimal expression of GRP78. 73% of IPMN-LG expresses GRP78 strongly. Statistical analysis revealed a significant difference in GRP78 expression level between normal ductal cells and all three pathological conditions including PDCA, IPMN-HG and IPMN-LG (all p < 0.0001). Conclusions: This study shows that the expression level of GRP78 is significantly increased in PDAC, IPMN-HG and IPMN-LG compared to normal ductal cells. There appears to be a progressive increase in GRP78 expression from IPMN-LG to IPMN-HG and invasive PDAC. Increased GRP78 expression may be a marker for those at risk of developing PDAC. Changes in GRP78 expression over time and its role in PDAC chemoresistance should be further assessed. [Table: see text]

scholarly journals Bone morphogenetic protein 2 induces pulmonary angiogenesis via Wnt–β-catenin and Wnt–RhoA–Rac1 pathways

2009 ◽  
Vol 184 (1) ◽  
pp. 83-99 ◽  
Author(s):  
Vinicio A. de Jesus Perez ◽  
Tero-Pekka Alastalo ◽  
Jenny C. Wu ◽  
Jeffrey D. Axelrod ◽  
John P. Cooke ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Bone Morphogenetic Protein ◽  
Bone Morphogenetic Protein 2 ◽  
Immunodeficient Mice ◽  
Vascular Growth ◽  
Small Vessels ◽  
Angiogenesis Assay ◽  
Morphogenetic Protein ◽  
Pulmonary Arterial

Mutations in bone morphogenetic protein (BMP) receptor II (BMPRII) are associated with pulmonary artery endothelial cell (PAEC) apoptosis and the loss of small vessels seen in idiopathic pulmonary arterial hypertension. Given the low penetrance of BMPRII mutations, abnormalities in other converging signaling pathways may be necessary for disease development. We hypothesized that BMPRII supports normal PAEC function by recruiting Wingless (Wnt) signaling pathways to promote proliferation, survival, and motility. In this study, we report that BMP-2, via BMPRII-mediated inhibition of GSK3-β, induces β-catenin (β-C) accumulation and transcriptional activity necessary for PAEC survival and proliferation. At the same time, BMP-2 mediates phosphorylated Smad1 (pSmad1) or, with loss of BMPRII, pSmad3-dependent recruitment of Disheveled (Dvl) to promote RhoA–Rac1 signaling necessary for motility. Finally, using an angiogenesis assay in severe combined immunodeficient mice, we demonstrate that both β-C– and Dvl-mediated RhoA–Rac1 activation are necessary for vascular growth in vivo. These findings suggest that the recruitment of both canonical and noncanonical Wnt pathways is required in BMP-2–mediated angiogenesis.

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