DOCSLIB.ORG

Srebp1c-PAX4 Axis Mediates Pancreatic Β-Cell Compensatory

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Srebp1c-PAX4 Axis Mediates Pancreatic Β-Cell Compensatory Diabetes Volume 68, January 2019 81 SREBP1c-PAX4 Axis Mediates Pancreatic b-Cell Compensatory Responses Upon Metabolic Stress Gung Lee,1 Hagoon Jang,1 Ye Young Kim,1 Sung Sik Choe,1 Jinuk Kong,1 Injae Hwang,1 Jeu Park,1 Seung-Soon Im,2 and Jae Bum Kim1 Diabetes 2019;68:81–94 | https://doi.org/10.2337/db18-0556 SREBP1c is a key transcription factor for de novo lipo- or hyperglycemia in obesity, pancreatic islets adaptively genesis. Although SREBP1c is expressed in pancreatic elevate the level of serum insulin through a b-cell com- islets, its physiological roles in pancreatic b-cells are pensatory mechanism. This process includes the expansion largely unknown. In this study, we demonstrate that of b-cell mass and the augmentation of insulin production b SREBP1c regulates -cell compensation under meta- (3,4). Although b-cell neogenesis and protection from bolic stress. SREBP1c expression level was augmented apoptosis can contribute to b-cell mass expansion, accu- in pancreatic islets from obese and diabetic animals. In mulating evidence suggests that an increase in b-cell pro- pancreatic b-cells, SREBP1c activation promoted the b liferation would be the principal mechanism of postnatal ISLET STUDIES expression of cell cycle genes and stimulated -cell b b proliferation through its novel target gene, PAX4. Com- -cell growth (5,6). Thus, the potentiation of -cell pro- pared with SREBP1c+/+ mice, SREBP1c2/2 mice showed liferation is important to maintain glucose homeostasis glucose intolerance with low insulin levels. Moreover, and to protect against the onset of diabetes. b-cells from SREBP1c2/2 mice exhibited reduced SREBP1c is a basic helix-loop-helix leucine zipper tran- capacity to proliferate and secrete insulin. Conversely, scription factor that governs de novo lipogenesis (7–10). transplantation of SREBP1c-overexpressing islets re- During the postprandial state, insulin activates SREBP1c stored insulin levels and relieved hyperglycemia in strep- via AKT (protein kinase B) and mTORC1 (mammalian tozotocin-induced diabetic animals. Collectively, these target of rapamycin C1) (11,12). SREBP1c stimulates de data suggest that pancreatic SREBP1c is a key player novo lipogenesis by elevating the expression of lipogenic in mediating b-cell compensatory responses in obesity. target genes including fatty acid synthase, stearoyl-CoA desaturase 1, and acetyl-CoA carboxylase (12–14). More- over, SREBP1c has been implicated in lipid metabolisms of Pancreatic islets regulate glucose homeostasis through the pancreatic islets (15,16). For instance, it has been reported secretion of several hormones, particularly glucagon and that the activation of SREBP1c by liver X receptor agonist insulin. Under hypoglycemic conditions, glucagon is re- increases intracellular lipid accumulation in pancreatic leased from pancreatic a-cells to promote hepatic glyco- islets (16). SREBP1 is associated with cell growth through genolysis and gluconeogenesis to increase blood glucose lipid metabolism and/or cell cycle progression in cer- levels (1). In contrast, insulin secretion from pancreatic tain cell types (17,18). However, it remains unclear b-cells is stimulated by hyperglycemia. Insulin effectively whether SREBP1c might be associated with pancreatic b-cell lowers the level of blood glucose via glucose uptake into proliferation. peripheral tissues and hinders hepatic glucose production. Paired box 4 (PAX4) belongs to the PAX gene family, On the other hand, metabolic stresses such as obesity and a group of transcription factors that carry out essential insulin resistance increase insulin demand and disrupt roles in embryogenesis as well as in cellular plasticity glucose homeostasis (2). In response to chronic fuel surfeit in adults (19). PAX4 is mainly expressed in endocrine 1National Creative Research Initiatives Center for Adipose Tissue Remodeling, This article contains Supplementary Data online at http://diabetes Institute of Molecular Biology and Genetics, Department of Biological Sciences, .diabetesjournals.org/lookup/suppl/doi:10.2337/db18-0556/-/DC1. Seoul National University, Seoul, South Korea © 2018 by the American Diabetes Association. Readers may use this article as 2 Department of Physiology and Medical Research Center, Keimyung University long as the work is properly cited, the use is educational and not for profit, and the School of Medicine, Daegu, Republic of Korea work is not altered. More information is available at http://www.diabetesjournals Corresponding author: Jae Bum Kim, [email protected] .org/content/license. Received 18 May 2018 and accepted 3 October 2018 82 SREBP1c-PAX4 Axis in b-Cell Compensation Diabetes Volume 68, January 2019 pancreas where it plays an essential role to induce differ- using the sensitive colorimetric assay for viable cells entiation toward b-cells (20). Also, PAX4 has been impli- according to the manufacturer protocol (#CK04-11; cated in b-cell plasticity of adult pancreatic islets. For Dojindo Molecular Technologies). example, it has been revealed that ectopic expression of PAX4 in human or murine islets enhances b-cell prolifer- Quantitative Real-time PCR ation by stimulating the transcription of cell cycle genes Total RNAs were isolated from MIN6 and aTC1–6 cells (21–23). Moreover, the overexpression of PAX4 protects using TRIzol reagent (Invitrogen). Islet RNAs were pre- b-cells from apoptosis induced by streptozotocin (STZ) pared using the RNeasy Mini Kit (Qiagen). Subsequently, (24). Recently, it has been reported that mutations of the equal amounts of RNA were synthesized to cDNA using PAX4 gene are associated with type 1 and 2 diabetes as well RevertAid reverse transcriptase (#EP0441; Thermo Fisher as ketosis-prone diabetes in various ethnic groups (25). Scientific). Relative amounts of mRNA were calculated by Although the correlation between PAX4 dysfunction and using a CFX Real-Time Quantitative PCR Detection System diabetes has been demonstrated, the regulatory mecha- (Bio-Rad) after normalization to cyclophilin mRNA. The nism of PAX4 expression remains elusive. primer sequences used are listed in Supplementary Table 1. +/+ In this study, we investigated SREBP1c and 2/2 SREBP1c mice under normal chow diet (NCD) or Preparation of Recombinant Adenovirus a high-fat diet (HFD) to understand the roles of SREBP1c Adenoviral plasmid were constructed as previously de- in pancreatic islets. We demonstrate that pancreatic scribed (26). cDNAs for SREBP1c and PAX4 were incor- SREBP1c plays an important role in b-cell compensatory porated into an AdTrack-CMV shuttle vector and an responses upon metabolic stress, accompanied with regu- Ad-Easy vector. Adenoviruses were amplified in human lating b-cell proliferation and survival. In pancreatic islets, embryonic kidney 293A (HEK293A) cells and isolated by we identified that PAX4 could act as a downstream medi- CsCl density centrifugation. Green fluorescent protein ator of SREBP1c to control b-cell growth. In addition, was coexpressed from an independent promoter with transplantation of SREBP1c-overexpressing primary islets inserted cDNA. Empty virus expressing only the gene ameliorated glucose intolerance in diabetic animals, im- for green fluorescent protein served as control (MOCK). plying that elevated expression of SREBP1c would poten- b tiate -cell function. Collectively, our data suggest that the Cell Culture and siRNA SREBP1c-PAX4 axis would play a pivotal role in compen- MIN6 cells were cultured in DMEM medium containing satory responses of pancreatic islets under metabolic 15% FBS (HyClone). HEK293T, aTC1–6 cells were cultured stress. in DMEM containing 10% FBS. Harvested primary islets were cultured in RPMI 1640 medium (HyClone) with 10% RESEARCH DESIGN AND METHODS FBS. Two kinds of siRNAs were designed and produced Animals and Treatment by Bioneer (Daejeon, South Korea). The antisense siPAX4 2/2 SREBP1c mice were provided by Dr. Jay D. Horton at sequences were as follows: antisense, UGGUACUCCUCA- the University of Texas Southwestern Medical Center CAGAAGG and ACUGUCAAAUAGAGGCCUC. siRNA was (Dallas, TX). db/+ and db/db male mice were obtained transfected using lipofectamine-iMAX (Thermo Fisher 2/2 from Daehan Bio (Seoul, Korea). SREBP1c and Scientific) into cell lines. +/+ SREBP1 littermates were maintained on a NCD (Zeigler) for 8 weeks. They were fed with NCD or 60% HFD (Re- Immunohistochemistry search Diets Inc.) for 12 weeks. For fasted/refed experi- The pancreata were isolated from mice, fixed in 4% para- ments, blood samples were collected after fasting for 16 h formaldehyde, and embedded in paraffin block. Paraffin and refeeding for 2 h. Mouse tissue specimens were blocks were cut into 5-mm-thick sections with 30-mm immediately stored at 280°C. For the glucose tolerance intervals. Pancreas sections from NCD- or HFD-fed mice test (GTT), mice were fasted for 16 h and intraperitoneally and db/db mice were stained with anti-SREBP1 (SC-367; 2 injected with glucose (1 g $ kg 1 body weight for mice). Santa Cruz Biotechnology), anti-insulin (ab7842; Abcam), Blood glucose levels were measured in tail vein blood anti-pax4 (ab42450; Abcam), anti-glucagon (ab10983; samples by using a Contour TS Blood Glucose Meter Abcam), and anti-Ki67-FITC (11-5698-80; eBioscience) anti- (Bayer). For the insulin tolerance test (ITT), ad libitum bodies. Species-specific secondary antibodies staining or mice were intraperitoneally injected with insulin (1 unit/kg diaminobenzidine staining was followed. For the islet mor- body weight for mice). All experiments with mice were phology
Load more

Recommended publications
  • PAX4 R192H and P321H Polymorphisms in Type 2 Diabetes and Their Functional Defects
    Journal of Human Genetics (2016) 61, 943–949 & 2016 The Japan Society of Human Genetics All rights reserved 1434-5161/16 www.nature.com/jhg ORIGINAL ARTICLE PAX4 R192H and P321H polymorphisms in type 2 diabetes and their functional defects Jatuporn Sujjitjoon1,6, Suwattanee Kooptiwut2,6, Nalinee Chongjaroen3, Namoiy Semprasert2, Wanthanee Hanchang2, Kanjana Chanprasert3,4, Watip Tangjittipokin3, Pa-thai Yenchitsomanus1 and Nattachet Plengvidhya5 We have previously identified PAX4 mutations causing MODY9 and a recent genome-wide association study reported a susceptibility locus of type 2 diabetes (T2D) near PAX4. In this study, we aim to investigate the association between PAX4 polymorphisms and T2D in Thai patients and examine functions of PAX4 variant proteins. PAX4 rs2233580 (R192H) and rs712701 (P321H) were genotyped in 746 patients with T2D and 562 healthy normal control subjects by PCR and restriction- fragment length polymorphism method. PAX4 variant proteins were investigated for repressor function on human insulin and glucagon promoters and for cell viability and apoptosis upon high glucose exposure. Genotype and allele frequencies of PAX4 rs2233580 were more frequent in patients with T2D than in control subjects (P = 0.001 and 0.0006, respectively) with odds ratio of 1.66 (P = 0.001; 95% confidence interval, 1.22–2.27). PAX4 rs712701 was not associated with T2D but it was in linkage disequilibrium with rs2233580. The 192H/321H (A/A) haplotype was more frequent in T2D patients than in controls (9.5% vs 6.6%; P = 0.009). PAX4 R192H, but not PAX4 P321H, impaired repression activities on insulin and glucagon promoters and decreased transcript levels of genes required to maintain β-cell function, proliferation and survival.
    [Show full text]
  • Context-Specific A-To-B-Cell Reprogramming by Forced Pdx1
    Downloaded from genesdev.cshlp.org on September 30, 2021 - Published by Cold Spring Harbor Laboratory Press RESEARCH COMMUNICATION from a cells via a bihormonal glucagon+insulin+ (Gcg+Ins+) Context-specific a-to-b-cell transitional state (Thorel et al. 2010). The interconversion reprogramming by forced presumably occurs in response to a combination of the physiological need to replenish b cells and regeneration- Pdx1 expression induced stress, raising questions as to the local or systemic 1 2 1 signals triggered by such lesions. Direct superimposition of Yu-Ping Yang, Fabrizio Thorel, Daniel F. Boyer, apro-b-lineage condition was reported when Pax4 expres- Pedro L. Herrera,2 and Christopher V.E. Wright1,3 sion was forced in pancreatic or endocrine progenitors or in 1 embryonic a cells to redirect endocrine differentiation or Vanderbilt University Program in Developmental Biology, coax pre-existing a cells into b cells. The converted cells Department of Cell and Developmental Biology, Vanderbilt seemed similar to normal b cells and temporarily im- University Medical Center, Nashville, Tennessee 37232, USA; 2 proved glycemia under induced diabetes, although the Department of Cell Physiology and Metabolism, University effect was superseded by uncontrolled a-cell neogenesis of Geneva Faculty of Medicine, CH-1211 Geneva 4, Switzerland and fatality caused by extreme hyperglycemia (Collombat et al. 2009). These studies on the ability of a single Using single transcription factors to reprogram cells could lineage-allocating transcription factor to sustain com- produce important insights into the epigenetic mechanisms plete cell fate conversion suggest that similar analyses that direct normal differentiation, or counter inappropriate for other transcription factors could be insightful.
    [Show full text]
  • Insulin/Glucose-Responsive Cells Derived from Induced Pluripotent Stem Cells: Disease Modeling and Treatment of Diabetes
    Insulin/Glucose-Responsive Cells Derived from Induced Pluripotent Stem Cells: Disease Modeling and Treatment of Diabetes Sevda Gheibi *, Tania Singh, Joao Paulo M.C.M. da Cunha, Malin Fex and Hindrik Mulder * Unit of Molecular Metabolism, Lund University Diabetes Centre, Jan Waldenströms gata 35; Box 50332, SE-202 13 Malmö, Sweden; [email protected] (T.S.); [email protected] (J.P.M.C.M.d.C.); [email protected] (M.F.) * Correspondence: [email protected] (S.G.); [email protected] (H.M.) Supplementary Table 1. Transcription factors associated with development of the pancreas. Developmental Gene Aliases Function Ref. Stage SRY-Box Transcription Factor Directs the primitive endoderm SOX17 DE, PFE, PSE [1] 17 specification. Establishes lineage-specific transcriptional programs which leads DE, PFE, PSE, Forkhead Box A2; Hepatocyte to proper differentiation of stem cells FOXA2 PMPs, EPs, [2] Nuclear Factor 3-β into pancreatic progenitors. Regulates Mature β-cells expression of PDX1 gene and aids in maturation of β-cells A pleiotropic developmental gene which regulates growth, and Sonic Hedgehog Signaling differentiation of several organs. SHH DE [3] Molecule Repression of SHH expression is vital for pancreas differentiation and development Promotes cell differentiation, proliferation, and survival. Controls C-X-C Motif Chemokine the spatiotemporal migration of the Receptor 4; Stromal Cell- CXCR4 DE angioblasts towards pre-pancreatic [4] Derived Factor 1 Receptor; endodermal region which aids the Neuropeptide Y3 Receptor induction of PDX1 expression giving rise to common pancreatic progenitors Crucial for generation of pancreatic HNF1 Homeobox B HNF1B PFE, PSE, PMPs multipotent progenitor cells and [5] Hepatocyte Nuclear Factor 1-β NGN3+ endocrine progenitors Master regulator of pancreatic organogenesis.
    [Show full text]
  • Combined Ectopic Expression of Pdx1 and Ptf1a/P48 Results in the Stable Conversion of Posterior Endoderm Into Endocrine and Exocrine Pancreatic Tissue
    Downloaded from genesdev.cshlp.org on October 2, 2021 - Published by Cold Spring Harbor Laboratory Press RESEARCH COMMUNICATION results reported in this communication, ectopic expres- Combined ectopic expression sion of Pdx1 in nonpancreatic chicken endoderm re- of Pdx1 and Ptf1a/p48 results sulted in the initiation of pancreatic budding, but it was not sufficient to promote differentiation of either exo- in the stable conversion crine or endocrine cells (Grapin-Botton et al. 2001). of posterior endoderm into Ptf1a/p48 is a bHLH transcription factor, which was originally identified as a part of a heterotrimeric pan- endocrine and exocrine creas transcription factor complex, referred to as PTF1 that activates transcription of exocrine specific pancre- pancreatic tissue atic genes in the mature pancreas (Cockell et al. 1989; Beres et al. 2006). Mice bearing a null mutation of Ptf1a/ Solomon Afelik, Yonglong Chen, p48 are completely devoid of exocrine pancreas, while 1 and Tomas Pieler endocrine pancreatic cells still form, but are found to be translocated to the spleen (Krapp et al. 1998). More re- Georg-August-Universität Göttingen, Zentrum Biochemie und cent studies have revealed that Ptf1a/p48 is already ex- Molekular Zellbiologie, Abteilung Entwicklungsbiochemie, pressed in pancreatic precursor cells which contribute to 37077 Göttingen, Germany all pancreatic cell types, and that, in the absence of Patterning of the embryonic endoderm into distinct sets Ptf1a/p48, pancreatic precursor cells adopt a duodenal fate (Kawaguchi et al. 2002). These findings suggest a of precursor cells involves the precisely regulated activi- role for Ptf1a/p48 that is not solely in exocrine differen- ties of key transcription regulators.
    [Show full text]
  • Transcription Factors That Govern Development and Disease: an Achilles Heel in Cancer
    G C A T T A C G G C A T genes Review Transcription Factors That Govern Development and Disease: An Achilles Heel in Cancer Dhananjay Huilgol , Prabhadevi Venkataramani , Saikat Nandi * and Sonali Bhattacharjee * Bungtown Road, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, NY 11724, USA; [email protected] (D.H.); [email protected] (P.V.) * Correspondence: [email protected] (S.N.); [email protected] (S.B.) Received: 15 September 2019; Accepted: 8 October 2019; Published: 12 October 2019 Abstract: Development requires the careful orchestration of several biological events in order to create any structure and, eventually, to build an entire organism. On the other hand, the fate transformation of terminally differentiated cells is a consequence of erroneous development, and ultimately leads to cancer. In this review, we elaborate how development and cancer share several biological processes, including molecular controls. Transcription factors (TF) are at the helm of both these processes, among many others, and are evolutionarily conserved, ranging from yeast to humans. Here, we discuss four families of TFs that play a pivotal role and have been studied extensively in both embryonic development and cancer—high mobility group box (HMG), GATA, paired box (PAX) and basic helix-loop-helix (bHLH) in the context of their role in development, cancer, and their conservation across several species. Finally, we review TFs as possible therapeutic targets for cancer and reflect on the importance of natural resistance against cancer in certain organisms, yielding knowledge regarding TF function and cancer biology. Keywords: transcription factors; embryonic development; evolution; cancer; clinical trials; high mobility group box (HMG); basic helix loop helix (bHLH); paired box (Pax); GATA 1.
    [Show full text]
  • Xo GENE PANEL
    xO GENE PANEL Targeted panel of 1714 genes | Tumor DNA Coverage: 500x | RNA reads: 50 million Onco-seq panel includes clinically relevant genes and a wide array of biologically relevant genes Genes A-C Genes D-F Genes G-I Genes J-L AATK ATAD2B BTG1 CDH7 CREM DACH1 EPHA1 FES G6PC3 HGF IL18RAP JADE1 LMO1 ABCA1 ATF1 BTG2 CDK1 CRHR1 DACH2 EPHA2 FEV G6PD HIF1A IL1R1 JAK1 LMO2 ABCB1 ATM BTG3 CDK10 CRK DAXX EPHA3 FGF1 GAB1 HIF1AN IL1R2 JAK2 LMO7 ABCB11 ATR BTK CDK11A CRKL DBH EPHA4 FGF10 GAB2 HIST1H1E IL1RAP JAK3 LMTK2 ABCB4 ATRX BTRC CDK11B CRLF2 DCC EPHA5 FGF11 GABPA HIST1H3B IL20RA JARID2 LMTK3 ABCC1 AURKA BUB1 CDK12 CRTC1 DCUN1D1 EPHA6 FGF12 GALNT12 HIST1H4E IL20RB JAZF1 LPHN2 ABCC2 AURKB BUB1B CDK13 CRTC2 DCUN1D2 EPHA7 FGF13 GATA1 HLA-A IL21R JMJD1C LPHN3 ABCG1 AURKC BUB3 CDK14 CRTC3 DDB2 EPHA8 FGF14 GATA2 HLA-B IL22RA1 JMJD4 LPP ABCG2 AXIN1 C11orf30 CDK15 CSF1 DDIT3 EPHB1 FGF16 GATA3 HLF IL22RA2 JMJD6 LRP1B ABI1 AXIN2 CACNA1C CDK16 CSF1R DDR1 EPHB2 FGF17 GATA5 HLTF IL23R JMJD7 LRP5 ABL1 AXL CACNA1S CDK17 CSF2RA DDR2 EPHB3 FGF18 GATA6 HMGA1 IL2RA JMJD8 LRP6 ABL2 B2M CACNB2 CDK18 CSF2RB DDX3X EPHB4 FGF19 GDNF HMGA2 IL2RB JUN LRRK2 ACE BABAM1 CADM2 CDK19 CSF3R DDX5 EPHB6 FGF2 GFI1 HMGCR IL2RG JUNB LSM1 ACSL6 BACH1 CALR CDK2 CSK DDX6 EPOR FGF20 GFI1B HNF1A IL3 JUND LTK ACTA2 BACH2 CAMTA1 CDK20 CSNK1D DEK ERBB2 FGF21 GFRA4 HNF1B IL3RA JUP LYL1 ACTC1 BAG4 CAPRIN2 CDK3 CSNK1E DHFR ERBB3 FGF22 GGCX HNRNPA3 IL4R KAT2A LYN ACVR1 BAI3 CARD10 CDK4 CTCF DHH ERBB4 FGF23 GHR HOXA10 IL5RA KAT2B LZTR1 ACVR1B BAP1 CARD11 CDK5 CTCFL DIAPH1 ERCC1 FGF3 GID4
    [Show full text]
  • The Transcription Factor PAX4 Acts As a Survival Gene in INS-1E Insulinoma Cells
    Oncogene (2007) 26, 4261–4271 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc ORIGINAL ARTICLE The transcription factor PAX4 acts as a survival gene in INS-1E insulinoma cells T Brun, DL Duhamel, KH Hu He, CB Wollheim and BR Gauthier Department of Cell Physiology and Metabolism, University Medical Center, Geneva 4, Switzerland The paired/homeodomain transcription factor Pax4 is domain nuclear factor Pax4 is essential for b-cell essential for islet b-cell generation during pancreas maturation. Indeed, Pax4 mRNA is detected in the development and their survival in adulthood. High Pax4 mouse pancreatic bud at E9.5, becoming maximal at expression was reported in human insulinomas indicating E13.5–15.5 during the period of massive b-cell expan- that deregulation of the gene may be associated with sion and thereafter declining. In mature islets, low levels tumorigenesis. We report that rat insulinoma INS-1E of Pax4 expression are detected in human, mouse and cells express 25-fold higher Pax4 mRNA levels than rat rat b-cells (Zhang et al., 2001; Heremans et al., 2002; islets. In contrast to primary b-cells, activin A but not Kojima et al., 2003; Zalzman et al., 2003; Brun et al., betacellulin or glucose induced Pax4 mRNA levels 2004, 2005; Theis et al., 2004; Gunton et al., 2005). indicating dissociation of Pax4 expression from insulino- Consistent with its tissue-specific expression pattern, ma cell proliferation. Short hairpin RNA adenoviral targeted disruption of the pax4 gene in mice results constructs targeted to the paired domain or homeodomain in the absence of mature pancreatic b- and d-cells with (viPax4PD and viPax4HD) were generated.
    [Show full text]
  • The Global Gene Expression Profile of the Secondary Transition During Pancreatic Development
    ÔØ ÅÒÙ×Ö ÔØ The global gene expression profile of the secondary transition during pancre- atic development Stefanie J. Willmann, Nikola S. Mueller, Silvia Engert, Michael Sterr, Ingo Burtscher, Aurelia Raducanu, Martin Irmler, Johannes Beckers, Steffen Sass, Fabian J. Theis, Heiko Lickert PII: S0925-4773(15)30037-X DOI: doi: 10.1016/j.mod.2015.11.004 Reference: MOD 3386 To appear in: Received date: 19 June 2015 Revised date: 26 November 2015 Accepted date: 27 November 2015 Please cite this article as: Willmann, Stefanie J., Mueller, Nikola S., Engert, Sil- via, Sterr, Michael, Burtscher, Ingo, Raducanu, Aurelia, Irmler, Martin, Beckers, Johannes, Sass, Steffen, Theis, Fabian J., Lickert, Heiko, The global gene expres- sion profile of the secondary transition during pancreatic development, (2015), doi: 10.1016/j.mod.2015.11.004 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. ACCEPTED MANUSCRIPT The global gene expression profile of the secondary transition during pancreatic development Stefanie J. Willmann*1,5, Nikola S. Mueller*2, Silvia Engert1, Michael Sterr1, Ingo Burtscher1, Aurelia Raducanu1, Martin Irmler3, Johannes Beckers3,4,5,
    [Show full text]
  • Mafa Enables Pdx1 to Effectively Convert Pancreatic Islet Progenitors and Committed Islet A-Cells Into B-Cells in Vivo
    Diabetes Volume 66, May 2017 1293 Mafa Enables Pdx1 to Effectively Convert Pancreatic Islet Progenitors and Committed Islet a-Cells Into b-Cells In Vivo Taka-aki Matsuoka,1 Satoshi Kawashima,1 Takeshi Miyatsuka,1,2 Shugo Sasaki,1 Naoki Shimo,1 Naoto Katakami,1 Dan Kawamori,1 Satomi Takebe,1 Pedro L. Herrera,3 Hideaki Kaneto,4 Roland Stein,5 and Iichiro Shimomura1 Diabetes 2017;66:1293–1300 | DOI: 10.2337/db16-0887 Among the therapeutic avenues being explored for re- including coronary and renal disease. A variety of innova- placement of the functional islet b-cell mass lost in tive approaches are being explored to produce b-cells from type 1 diabetes (T1D), reprogramming of adult cell types embryonic stem cells (1,2) and adult cell types (3–5). A into new b-cells has been actively pursued. Notably, supposition in these efforts involves producing conditions mouse islet a-cells will transdifferentiate into b-cells un- that correctly regulate the transcription factor networks der conditions of near b-cell loss, a condition similar to required in programming pancreatic progenitor cells into ISLET STUDIES a T1D. Moreover, human islet -cells also appear to poised b-cells and subsequently controlling mature islet cell func- for reprogramming into insulin-positive cells. Here we tion. These include transcription factors like Pdx1 (6–10), have generated transgenic mice conditionally expressing which is essential in the formation of early pancreatic ep- the islet b-cell–enriched Mafa and/or Pdx1 transcription ithelium, developing b-cells and adult islet b-cells, as well factors to examine their potential to transdifferentiate as neurogenin 3 (Ngn3) (11–13), which is required during embryonic pan–islet cell Ngn3-positive progenitors and embryogenesis for specification of all islet cell types (i.e., the later glucagon-positive a-cell population into b-cells.
    [Show full text]
  • Transcriptomic Analysis by RNA Sequencing Characterises Malignant Progression of Canine Insulinoma from Normal Tissue to Metastatic Disease Y
    www.nature.com/scientificreports OPEN Transcriptomic analysis by RNA sequencing characterises malignant progression of canine insulinoma from normal tissue to metastatic disease Y. Capodanno1,2*, F. O. Buishand 2,3, L. Y. Pang2, J. Kirpensteijn3,5, J. A. Mol3, R. Elders2,4 & D. J. Argyle2 Insulinomas (INS) are the most common human and canine functioning pancreatic neuroendocrine tumours. The long-term prognosis for malignant INS is poor, because micrometastases are frequently missed during surgery. As human and canine malignant INS share clinical and histopathological features, dogs have been proposed as models for INS research. Using RNA-sequencing, we conducted a pilot study to better understand the underlying molecular mechanisms of canine INS. Normal canine pancreas and lymph node control tissues were compared with primary INS and INS-metastatic lymph nodes, revealing more than 3,000 genes diferentially expressed in normal pancreas compared to primary INS. Only 164 genes were diferentially expressed between primary INS and INS-metastatic lymph nodes. Hierarchical clustering analysis demonstrated similar genetic profles in normal pancreas and early clinical stage primary INS, whereas late clinical stage primary INS resembled the genetic profle of INS-metastatic lymph nodes. These fndings suggest that markers of malignant behaviour could be identifed at the primary site of the disease. Finally, using the REACTOME pathways database, we revealed that an active collagen metabolism, extracellular matrix remodelling, beta- cell diferentiation and non-beta-cell trans-diferentiation might cause disease progression and hyperinsulinism in INS, identifying major pathways worthy of future research in this currently poorly controlled disease. Insulinomas (INS) are the most commonly diagnosed functioning pancreatic neuroendocrine tumours in humans and dogs1,2.
    [Show full text]
  • Xo PANEL DNA GENE LIST
    xO PANEL DNA GENE LIST ~1700 gene comprehensive cancer panel enriched for clinically actionable genes with additional biologically relevant genes (at 400 -500x average coverage on tumor) Genes A-C Genes D-F Genes G-I Genes J-L AATK ATAD2B BTG1 CDH7 CREM DACH1 EPHA1 FES G6PC3 HGF IL18RAP JADE1 LMO1 ABCA1 ATF1 BTG2 CDK1 CRHR1 DACH2 EPHA2 FEV G6PD HIF1A IL1R1 JAK1 LMO2 ABCB1 ATM BTG3 CDK10 CRK DAXX EPHA3 FGF1 GAB1 HIF1AN IL1R2 JAK2 LMO7 ABCB11 ATR BTK CDK11A CRKL DBH EPHA4 FGF10 GAB2 HIST1H1E IL1RAP JAK3 LMTK2 ABCB4 ATRX BTRC CDK11B CRLF2 DCC EPHA5 FGF11 GABPA HIST1H3B IL20RA JARID2 LMTK3 ABCC1 AURKA BUB1 CDK12 CRTC1 DCUN1D1 EPHA6 FGF12 GALNT12 HIST1H4E IL20RB JAZF1 LPHN2 ABCC2 AURKB BUB1B CDK13 CRTC2 DCUN1D2 EPHA7 FGF13 GATA1 HLA-A IL21R JMJD1C LPHN3 ABCG1 AURKC BUB3 CDK14 CRTC3 DDB2 EPHA8 FGF14 GATA2 HLA-B IL22RA1 JMJD4 LPP ABCG2 AXIN1 C11orf30 CDK15 CSF1 DDIT3 EPHB1 FGF16 GATA3 HLF IL22RA2 JMJD6 LRP1B ABI1 AXIN2 CACNA1C CDK16 CSF1R DDR1 EPHB2 FGF17 GATA5 HLTF IL23R JMJD7 LRP5 ABL1 AXL CACNA1S CDK17 CSF2RA DDR2 EPHB3 FGF18 GATA6 HMGA1 IL2RA JMJD8 LRP6 ABL2 B2M CACNB2 CDK18 CSF2RB DDX3X EPHB4 FGF19 GDNF HMGA2 IL2RB JUN LRRK2 ACE BABAM1 CADM2 CDK19 CSF3R DDX5 EPHB6 FGF2 GFI1 HMGCR IL2RG JUNB LSM1 ACSL6 BACH1 CALR CDK2 CSK DDX6 EPOR FGF20 GFI1B HNF1A IL3 JUND LTK ACTA2 BACH2 CAMTA1 CDK20 CSNK1D DEK ERBB2 FGF21 GFRA4 HNF1B IL3RA JUP LYL1 ACTC1 BAG4 CAPRIN2 CDK3 CSNK1E DHFR ERBB3 FGF22 GGCX HNRNPA3 IL4R KAT2A LYN ACVR1 BAI3 CARD10 CDK4 CTCF DHH ERBB4 FGF23 GHR HOXA10 IL5RA KAT2B LZTR1 ACVR1B BAP1 CARD11 CDK5 CTCFL DIAPH1 ERCC1 FGF3 GID4 HOXA11
    [Show full text]
  • Molecular Basis for Intellectual Disability and Epilepsy: Role of the Human Homeobox Gene ARX
    I , I l¡ \-lN I ìo Molecular Basis for Intellectual Disability and Epilepsy: Role of the Human Homeobox Gene ARX A thesis submitted for the degree of Doctor of Philosophy to the University of Adelaide by Desiree Cloosterman BSc (Hons) school of Medicine, Department of Paediatrics, university of Adelaide November 2005 ll ,rlf the human brain were so simple that we coultl understønd it, we woukl be so simple thøt we couldn't" Emerson M. Pugh lll CONTENTS Summary IV Statement and Declaration v vl Acknowledgements..--- ------ --- List of Abbreviations vlll CHAPTER I Introduction-----------.-- 1 CHAPTER 2 Materials and Methods- 43 CHAPTER 3 Conservation of ARX. _.__-_._.1 10 CHAPTER 4 Yeast Two-Hybrid Screening __-_--_ I 33 CHAPTER 5 Confirmation of Y2H Interactions 176 CHAPTER 6 ZebrafishKnockdown Model------.- 203 CHAPTER 7 Conclusions 237 References lv SUMMARY Mental retardation (MR) is estimated to affect 2-3Yo of the population and is caused by both environmental and genetic factors. Mutations in Ihe Aristaless-related homeobox gene (AR$ have been found in numerous families with X-linked MR with and without other clinical features including infantile spasms, dystonia, lissencephaly, autism and dysarthria. The aim of this study was to investigate the normal function of the ARX protein within a cellular environment and in development. Discernment of ARX function will improve our understanding about the molecular pathology of intellectual disability and epilepsy, as well as improve our knowledge of the genes and mechanisms required for normal brain development. The first part of the thesis addressed the conservation of ARX domains and polyalanine regions by the identification and analysis of characterized and novel ARX orthologs.
    [Show full text]