DOCSLIB.ORG

Connective Tissue Growth Factor Production by Activated Pancreatic

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Connective Tissue Growth Factor Production by Activated Pancreatic Laboratory Investigation (2010) 90, 1179–1188 & 2010 USCAP, Inc All rights reserved 0023-6837/10 $32.00 Connective tissue growth factor production by activated pancreatic stellate cells in mouse alcoholic chronic pancreatitis Alyssa L Charrier1,2 and David R Brigstock1,2,3 Alcoholic chronic pancreatitis (ACP) is characterized by pancreatic necrosis, inflammation, and scarring, the latter of which is due to excessive collagen deposition by activated pancreatic stellate cells (PSC). The aim of this study was to establish a model of ACP in mice, a species that is usually resistant to the toxic effects of alcohol, and to identify the cell type(s) responsible for production of connective tissue growth factor (CTGF), a pro-fibrotic molecule. C57Bl/6 male mice received intraperitoneal ethanol injections for 3 weeks against a background of cerulein-induced acute pancreatitis. Peak blood alcohol levels remained consistently high in ethanol-treated mice as compared with control mice. In mice receiving ethanol plus cerulein, there was increased collagen deposition as compared with other treatment groups as well as increased frequency of a-smooth muscle actin and desmin-positive PSC, which also showed significantly enhanced CTGF protein production. Expression of mRNA for collagen a1(I), a-smooth muscle actin or CTGF were all increased and co- localized exclusively to activated PSC in ACP. Pancreatic expression of mRNA for key profibrotic markers were all increased in ACP. In conclusion, a mouse model of ACP has been developed that mimics key pathophysiological features of the disease in humans and which shows that activated PSC are the principal producers of collagen and CTGF. PSC-derived CTGF is thus a candidate therapeutic target in anti-fibrotic strategies for ACP. Laboratory Investigation (2010) 90, 1179–1188; doi:10.1038/labinvest.2010.82; published online 5 April 2010 KEYWORDS: connective tissue growth factor; fibrosis; pancreas Chronic pancreatitis stems from relapsing episodes of acute come activated and transition to a myofibroblast-like, alpha- pancreatitis,1 and is characterized by destruction of acinar smooth muscle actin (a-SMA)-positive cell type that is cap- tissue, a sustained pancreatic inflammatory response, and able of depositing large quantities of fibrillar collagen in the fibrosis.2,3 Although development of chronic pancreatitis is interstitial spaces.11,12 The effects of alcohol exposure on PSC strongly associated with heavy alcohol consumption, only a function have been largely deduced from in vitro studies and small percentage of heavy drinkers actually develop the dis- from analysis of human pathological specimens. Studies of ease4–7 suggesting that other predisposing factors (for ex- the mechanisms of alcohol on PSC function in vivo remain ample, genetics, environment, injury) must also be present ambiguous and are confounded by the fact that a good an- for disease progression. Pancreatitis is responsible for ap- imal model of alcoholic chronic pancreatitis (ACP) does not proximately 3500 deaths each year in the USA, 16% of which yet exist.9,13 are due to chronic disease with about half of those deaths due Connective tissue growth factor (CTGF; also known as to alcohol.8 CCN2) is a member of the CCN family of proteins14,15 which Although the pathophysiology of chronic pancreatitis re- associates with components of the extracellular matrix or cell mains poorly understood,9 pancreatic stellate cells (PSC) are surface integrins16 and regulates cellular processes such believed to have a central role in the initiation and progres- as, adhesion, migration, mitogenesis and differentiation.15 sion of the fibrotic response.10 Under normal circumstances Although CTGF has an important role in vertebrate devel- PSC remain quiescent but after pancreatic injury they be- opment17–19 it is weakly expressed in adult connective tissues 1Center for Clinical and Translational Research, The Research Institute at Nationwide Children’s Hospital, Columbus, OH, USA; 2Molecular, Cellular and Developmental Biology Program, The Ohio State University, Columbus, OH, USA and 3Department of Surgery, The Ohio State University, Columbus, OH, USA Correspondence: Dr DR Brigstock, PhD, Room WA2022, Center for Clinical and Translational Research, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus OH 43205, USA. E-mail: [email protected] Received 23 October 2009; revised 18 December 2009; accepted 25 January 2010 www.laboratoryinvestigation.org | Laboratory Investigation | Volume 90 August 2010 1179 CTGF in alcoholic chronic pancreatitis in mice AL Charrier and DR Brigstock except during wound healing, tissue regeneration, cancer or and AEC chromogenic substrate (all from ScyTek Labora- fibrosis.16 A role for CTGF in pancreatic fibrosis was first tories, Logan, UT, USA). Double immunofluorescent detec- proposed from studies that showed CTGF overexpression in tion of a-SMA and CTGF or a-SMA and desmin acute necrotizing pancreatitis20,21 and in desmoplastic re- was achieved by incubation of sections with anti-a-SMA gions of pancreatic cancer.22 In in vitro studies, activated PSC and NH1 anti-CTGF IgY (5 mg/ml; an in-house antibody have been shown to co-express CTGF, transforming growth raised in chickens against recombinant human CTGF) or factor b-1 (TGF-b1), collagen 1 and other extracellular ma- a-SMA and desmin (H:76) (1:50, Santa Cruz Laboratories, trix proteins23,24 and to synthesize CTGF after exposure to Santa Cruz, CA, USA) followed by incubation with ethanol or acetaldehyde.25 However, data showing that CTGF Alexa Fluors 647 goat-anti mouse IgG (1:1000, is expressed by activated PSC in ACP in vivo are lacking. Invitrogen, Carlsbad, CA, USA) and goat-anti chicken Ig- We have produced a rapid and efficient model of ACP in Biotin (1:400, Santa Cruz Laboratories) or Alexa Flour 647 mice that mimics key pathophysiological features of the goat-anti mouse IgG and Alexa Fluor 488 goat-anti rabbit human form of the disease and which shows that activated IgG (1:1000, Invitrogen) for 1 h at room temperature. Slides PSC are a principal source of CTGF in ethanol-induced were incubated with Fluorescent NeutrAvidin (1:200, pancreatic fibrosis. Invitrogen), mounted with Vectashield Mounting Medium with DAPI (Vector Laboratories, Burlingame, CA, USA), and examined by confocal laser microscopy (LSM510, Carl Zeiss MATERIALS AND METHODS Co. Ltd, Jena, Germany) using an oil-immersion objective Animal Model lens (Plan-Apochromat  63/NA ¼ 1.4). All animal procedures were approved by the Institutional Animal Care and Use Committee of The Research Institute at Nationwide Children’s Hospital (Columbus, OH). Male In Situ Hybridization C57Bl/6 mice 6–8 weeks old were injected with ethanol Total RNA was extracted from cultured murine PSC with (3.2 g/kg; administered in a 33.3% ethanol: 67.7% water so- RNA STAT-60 (Tel-Test Inc., Friendswood, TX, USA) fol- lution) i.p. one time per day, six times per week, for 3 weeks. lowing the manufacturer’s instructions as previously de- On one day each week, some mice also received an i.p. in- scribed.25 Aliquots of 4 mg of total RNA were reverse jection of cerulein every hour for 6 hours. (50 mg/kg; Sigma transcribed using a SuperScript II Reverse Transcriptase kit Chemical Co., St Louis, MI, USA). Control mice received (Invitrogen). The cDNA was then amplified by polymerase either ethanol alone, cerulein alone, or water alone (n ¼ 6 per chain reaction using primers specific for a-SMA: 50- group). Mice were housed three to a cage and fed a low-fat CTACTGCCGAGCGTGAGATTGTCC-30 (sense) and 50-AGG diet ad libitum. Twice weekly, serum samples were collected GCCCAGCTTCGTCGTATTC-30 (antisense) (497 bp); col- 1 hour after ethanol administration for assessment of peak lagen a(I): 50-GACGTCCCTGGTGAAGTTGGT-30 (sense) blood alcohol levels (BAL). Mice were killed 2 days after the and 50-AGCCACGATGACCCTTTATG-30 (antisense) last ethanol treatment and pancreata were removed before (587 bp); or CTGF: 50-AGTGGAGATGCCAGGAGAAA-30 fixation in 4% paraformaldehyde (pH 7.2–7.4) or for RNA (sense) and 50-ACCTGCACAGCATTTGTTTG-30 (antisense) extraction, as described below. (540 bp). Following initial denaturation at 941C for 2 min, the samples were amplified by 35 cycles of denaturation at Blood Alcohol Levels (BAL) 941C for 30 s, annealing at 601C (621C for CTGF or 651C for Peak BAL were measured using an Ethanol-L3K assay (Di- a-SMA) for 40 s, extension at 721C for 90 s, and ended by agnostic Chemicals Limited, Oxford, CT, USA). NADH extension at 721C for 10 min. The PCR products were se- produced in the assay reaction was quantified by measuring parated on 1.5% agarose gels and then gel extraction was absorption at 340 nm with a SpectraMax M2 Microplate performed using QIAquick Gel Extraction (Qiagen, Valencia, Reader (Molecular Devices, Sunnyvale, CA, USA). CA, USA). Gel extraction products were subcloned into the pCRII-TOPO transcription vector (Invitrogen) according to Histology the manufacturer’s protocol and sequenced using the M13R Fixed pancreatic tissues were embedded in paraffin, cut into sequencing primer. 4 mm sections and stained with haemotoxylin-eosin for Antisense and sense RNA probes labeled with digoxignein- standard histological examination. Collagen staining was UTP or Biotin-UTP were generated by in vitro transcription performed using acidified 0.1% Sirius red F3B (Pfalz & of linearized plasmids with SP6 and T7 RNA polymerases Bauer, Waterbury, CT) as described.26,27 Immunohisto- (Roche Molecular Biochemicals, Mannheim, Germany) chemical staining for a-SMA or F4/80 was performed using according to the manufacturer’s protocol. The digoxigenin- monoclonal mouse-anti human a-SMA (1:100, Dako Cyto- labeled RNA was localized using Anti-digoxignenin-fluor- mation, Denmark) or anti-mouse F4/80 antigen respectively escein, Fab fragments (Roche Molecular Biochemicals). (1:100, eBioscience, San Diego, CA, USA), followed by Biotin-labeled RNA was localized using NeutrAvidin or development with UltraTek Anti-Polyvalent, UltraTek HRP Texas Red (Invitrogen).
Load more

Recommended publications
  • Do Humans Possess the Capability to Regenerate?
    The Science Journal of the Lander College of Arts and Sciences Volume 12 Number 2 Spring 2019 - 2019 Do Humans Possess the Capability to Regenerate? Chasha Wuensch Touro College Follow this and additional works at: https://touroscholar.touro.edu/sjlcas Part of the Biology Commons, and the Pharmacology, Toxicology and Environmental Health Commons Recommended Citation Wuensch, C. (2019). Do Humans Possess the Capability to Regenerate?. The Science Journal of the Lander College of Arts and Sciences, 12(2). Retrieved from https://touroscholar.touro.edu/sjlcas/vol12/ iss2/2 This Article is brought to you for free and open access by the Lander College of Arts and Sciences at Touro Scholar. It has been accepted for inclusion in The Science Journal of the Lander College of Arts and Sciences by an authorized editor of Touro Scholar. For more information, please contact [email protected]. Do Humans Possess the Capability to Regenerate? Chasha Wuensch A Chasha Wuensch graduated in May 2018 with a Bachelor of Science degree in Biology and will be attending pharmacy school. Abstract Urodele amphibians, including newts and salamanders, are amongst the most commonly studied research models for regenera- tion .The ability to regenerate, however, is not limited to amphibians, and the regenerative process has been observed in mammals as well .This paper discusses methods by which amphibians and mammals regenerate to lend insights into human regenerative mechanisms and regenerative potential .A focus is placed on the urodele and murine digit tip models,
    [Show full text]
  • Repair, Regeneration, and Fibrosis Gregory C
    91731_ch03 12/8/06 7:33 PM Page 71 3 Repair, Regeneration, and Fibrosis Gregory C. Sephel Stephen C. Woodward The Basic Processes of Healing Regeneration Migration of Cells Stem cells Extracellular Matrix Cell Proliferation Remodeling Conditions That Modify Repair Cell Proliferation Local Factors Repair Repair Patterns Repair and Regeneration Suboptimal Wound Repair Wound Healing bservations regarding the repair of wounds (i.e., wound architecture are unaltered. Thus, wounds that do not heal may re- healing) date to physicians in ancient Egypt and battle flect excess proteinase activity, decreased matrix accumulation, Osurgeons in classic Greece. The liver’s ability to regenerate or altered matrix assembly. Conversely, fibrosis and scarring forms the basis of the Greek myth involving Prometheus. The may result from reduced proteinase activity or increased matrix clotting of blood to prevent exsanguination was recognized as accumulation. Whereas the formation of new collagen during the first necessary event in wound healing. At the time of the repair is required for increased strength of the healing site, American Civil War, the development of “laudable pus” in chronic fibrosis is a major component of diseases that involve wounds was thought to be necessary, and its emergence was not chronic injury. appreciated as a symptom of infection but considered a positive sign in the healing process. Later studies of wound infection led The Basic Processes of Healing to the discovery that inflammatory cells are primary actors in the repair process. Although scurvy (see Chapter 8) was described in Many of the basic cellular and molecular mechanisms necessary the 16th century by the British navy, it was not until the 20th for wound healing are found in other processes involving dynamic century that vitamin C (ascorbic acid) was found to be necessary tissue changes, such as development and tumor growth.
    [Show full text]
  • Wound Healing: a Paradigm for Regeneration
    SYMPOSIUM ON REGENERATIVE MEDICINE Wound Healing: A Paradigm for Regeneration Victor W. Wong, MD; Geoffrey C. Gurtner, MD; and Michael T. Longaker, MD, MBA From the Hagey Laboratory for Pediatric Regenerative Medi- CME Activity cine, Department of Surgery, Stanford University, Stanford, Target Audience: The target audience for Mayo Clinic Proceedings is primar- relationships with any commercial interest related to the subject matter ily internal medicine physicians and other clinicians who wish to advance of the educational activity. Safeguards against commercial bias have been CA. their current knowledge of clinical medicine and who wish to stay abreast put in place. Faculty also will disclose any off-label and/or investigational of advances in medical research. use of pharmaceuticals or instruments discussed in their presentation. Statement of Need: General internists and primary care physicians must Disclosure of this information will be published in course materials so maintain an extensive knowledge base on a wide variety of topics covering that those participants in the activity may formulate their own judgments all body systems as well as common and uncommon disorders. Mayo Clinic regarding the presentation. Proceedings aims to leverage the expertise of its authors to help physicians In their editorial and administrative roles, William L. Lanier, Jr, MD, Terry L. understand best practices in diagnosis and management of conditions Jopke, Kimberly D. Sankey, and Nicki M. Smith, MPA, have control of the encountered in the clinical setting. content of this program but have no relevant financial relationship(s) with Accreditation: College of Medicine, Mayo Clinic is accredited by the Accred- industry. itation Council for Continuing Medical Education to provide continuing med- The authors report no competing interests.
    [Show full text]
  • Live Imaging and Computational Modelling of Tissue Growth and Tissue Regeneration in Drosophila
    Live Imaging and Computational Modelling of Tissue Growth and Tissue Regeneration in Drosophila Jamie Rickman 19th January 2015 Abstract Wound healing in the epithelium of the Drosophila wing imaginal disc is a regenerative process which is known to proceed via the assembly of a contractile actin cable that circumscribes the wound drawing the leading edge cells together in a characterstic rosette formation. Here we computation- ally explore various other biological and mechanical processes that are involved in tissue repair in Drosophila; the formation of dynamic actin protrusions that pull each other forward to close the wound; the active extrusion of dead cells from the wound site; and the effect of a global tension force operating on the tissue. In silico simulation results, using the vertex model of epithelial tissue, and in vivo data are compared. It is found that patterning of line tensions in wounded cells and cells neighbouring the wound can drive rosette formation and wound closure. Results from simulating a global tension force indicate this could recapitulate the initial expansion phase of wound healing seen in experiment. Contents 1 Introduction 2 1.1 Tissue regeneration . 2 1.2 Regenerative healing in Drosophila wing imaginal discs . 2 1.3 The mechanical drivers of epithelial wound healing in Drosophila . 3 2 The Vertex Model 4 3 Current work and results 5 3.1 Wound closure in vivo ...................................... 5 3.1.1 Wound closure rate . 5 3.1.2 Rosette formation . 5 3.1.3 Inital expansion of wound following ablation . 6 3.2 Wound Closure in silico ..................................... 7 3.2.1 Implementation of the vertex model .
    [Show full text]
  • Serotonin Signaling Regulates Insulin-Like Peptides for Growth, Reproduction, and Metabolism in the Disease Vector Aedes Aegypti
    Serotonin signaling regulates insulin-like peptides for growth, reproduction, and metabolism in the disease vector Aedes aegypti Lin Linga,b and Alexander S. Raikhela,b,1 aDepartment of Entomology, University of California, Riverside, CA 92521; and bInstitute for Integrative Genome Biology, University of California, Riverside, CA 92521 Contributed by Alexander S. Raikhel, September 6, 2018 (sent for review May 15, 2018; reviewed by Christen Mirth, Michael R. Strand, and Marc Tatar) Disease-transmitting female mosquitoes require a vertebrate individuals have completed adequate growth to enter the next de- blood meal to produce their eggs. An obligatory hematophagous velopmental stage (5, 6). Although other DILPs promote growth, lifestyle, rapid reproduction, and existence of a large number of their specific expression patterns suggest that they might carry out transmittable diseases make mosquitoes the world’s deadliest an- distinct physiological functions (2). In Drosophila larvae, dilps1,-2, imals. Attaining optimal body size and nutritional status is critical -3,and-5 are expressed predominantly in neurosecretory cells for mosquitoes to become reproductively competent and effective (IPCs, insulin producing cells) of the brain; ablation of larval IPCs disease vectors. We report that blood feeding boosts serotonin reduces body size with delayed metamorphosis (7). These DILPs concentration and elevates the serotonin receptor Aa5HT2B regulate growth by means of the canonical insulin/insulin-like (Aedes aegypti 5-hydroxytryptamine receptor, type 2B) transcript growth factor (IGF) pathway. Single gene mutations in insulin/ level in the fat-body, an insect analog of the vertebrate liver and IGF components have been shown to cause a reduction in growth adipose tissue.
    [Show full text]
  • Particular Reference to the Development of Theneural
    The role of embryology in teratological research, with particular reference to the development of the neural tube and heart M. H. Kaufman Department ofAnatomy, University of Cambridge, Cambridge CB2 3DY, U.K. Introduction There are many intriguing questions that the experimental embryologist can attempt to answer which could provide insight into the pathogenesis of many of the commoner types of congenital abnormalities that affect the human population. I should like to draw attention to certain areas of embryological interest, and make some general and personal observations on teratological research. I shall concentrate on two particular areas of interest, namely studies into the pathogenesis of neural tube closure defects and cardiovascular anomalies because these are the topics of my special interests. Two quite different, but complementary approaches may be employed in experimental teratological research to investigate how certain agents act on specific organs or tissues of the embryo or fetus to induce their abnormal development. The first approach takes cognizance of the nature and chemical structure of the agent under investigation, in order to determine at the biochemical level what the underlying mechanism and principal sites of action are likely to be. The primary effect of the agent may be to disrupt maternal metabolism, and the effect on the embryo or fetus may be secondary to the effect on the mother. In these latter circumstances it is not uncommon for an embryotoxic effect to be produced before any toxic manifestation becomes apparent in the mother. The second approach takes cognizance of the morphological abnormalities which may be induced by embryonic exposure to a teratogenic agent, in order to determine in the first instance at which stage of gestation the teratogenic insult is likely to have occurred, and, also, its site of action at the cellular and tissue level.
    [Show full text]
  • Directing Human Embryonic Stem Cells to Generate Vascular Progenitor Cells
    Gene Therapy (2008) 15, 89–95 & 2008 Nature Publishing Group All rights reserved 0969-7128/08 $30.00 www.nature.com/gt REVIEW Directing human embryonic stem cells to generate vascular progenitor cells H Bai and ZZ Wang Center for Molecular Medicine, Maine Medical Center Research Institute, Scarborough, ME, USA Pluripotent human embryonic stem cells (hESCs) differenti- it is also possible that endothelial cells and SMCs derived ate into most of the cell types of the adult human body, from hESCs can be used to engineer artificial vessels to including vascular cells. Vascular cells, such as endothelial repair damaged vessels and form vessel networks in cells and vascular smooth muscle cells (SMCs) are engineered tissues. Here we review the current status of significant contributors to tissue repair and regeneration. In directing hESCs to differentiate to vascular cells. addition to their potential applications for treatment of Gene Therapy (2008) 15, 89–95; doi:10.1038/sj.gt.3303005; vascular diseases and stimulation of ischemic tissue growth, published online 16 August 2007 Keywords: human embryonic stem cells; endothelial cells; smooth muscle cells Introduction which give rise to hematopoietic stem cells and vascular endothelial cells.10–12 Even both of them give rise to Some degenerative diseases are caused by the loss of hematopoietic and endothelial cells, it is unclear whether tissue-specific cells in organs, examples include: (i) type I hemangioblasts are the same as hemogenic endothelial diabetes, in which insulin-producing b cells are de- cells. For easy to discuss, we will use the term stroyed by an autoimmune disorder; (ii) Parkinson’s hemangioblast for both of them.
    [Show full text]
  • Promoting Longevity by Maintaining Metabolic and Proliferative
    © 2014. Published by The Company of Biologists Ltd | The Journal of Experimental Biology (2014) 217, 109-118 doi:10.1242/jeb.089920 REVIEW Promoting longevity by maintaining metabolic and proliferative homeostasis Lifen Wang*, Jason Karpac* and Heinrich Jasper‡ ABSTRACT preventing or counteracting this decline have evolved. Aging is characterized by a widespread loss of homeostasis in Understanding the integration of these diverse processes, and thus biological systems. An important part of this decline is caused by age- gaining comprehensive insight into the causes and mechanisms of related deregulation of regulatory processes that coordinate cellular the aging process, remains a challenge. In recent years, however, the responses to changing environmental conditions, maintaining cell and study of aging has progressed from the identification of single-gene tissue function. Studies in genetically accessible model organisms mutations that influence lifespan toward more complex integrative have made significant progress in elucidating the function of such analysis of the aging process. This work has also been facilitated by regulatory processes and the consequences of their deregulation for genetically accessible model systems and includes detailed analysis tissue function and longevity. Here, we review such studies, focusing of the interaction between environmental factors and genetics in the on the characterization of processes that maintain metabolic and control of longevity, as well as assessment of tissue-specific proliferative homeostasis in the fruitfly Drosophila melanogaster. The contributions to lifespan regulation (Guarente and Kenyon, 2000; primary regulatory axis addressed in these studies is the interaction Tissenbaum and Guarente, 2002; Kenyon, 2005; Greer and Brunet, between signaling pathways that govern the response to oxidative 2008; Mair and Dillin, 2008; Broughton and Partridge, 2009; stress, and signaling pathways that regulate cellular metabolism and Panowski and Dillin, 2009; Toivonen and Partridge, 2009; Taylor growth.
    [Show full text]
  • BREAKTHROUGHS in BIOSCIENCE/ ADVISORY COMMITTEE REGENERATIVE MEDICINE CHAIR» AUTHOR» Paula H
    / FALL 2016 Regenerative Medicine Advances from the Convergence of Biology & Engineering WHAT'S INSIDE » EXCEPTIONAL REGENERATION IN NATURE 2 / NATURAL REGENERATION IN HUMAN TISSUES 3 TISSUE ENGINEERING 4 / CONSTRUCTING SKIN 7 / TUBULAR ORGANS 8 / BONE ENGINEERING 8 MENDING BROKEN HEARTS 8 / REAWAKENING THE HUMAN HEART 10 / THE ROAD AHEAD 11 BREAKTHROUGHS IN BIOSCIENCE/ ADVISORY COMMITTEE REGENERATIVE MEDICINE CHAIR» AUTHOR» Paula H. Stern, PhD Cathryn M. Delude, of Santa Fe, New Mexico, writes about Northwestern University Feinberg School of Medicine science and medicine for magazines, newspapers, and COMMITTEE MEMBERS» research institutes. Her articles have appeared in Nature Aditi Bhargava, PhD Outlook, The Journal of the National Cancer Association University of California, San Francisco (JNCI), AACR’s Cancer Discovery, Proto: Dispatches from David L. Brautigan, PhD the Frontiers of Medicine, Los Angeles Times, Boston Globe, University of Virginia School of Medicine New York Times, Scientific American, and The Scientist. She has also written for the Howard Hughes Medical Institute, David B. Burr, PhD Harvard Health Publications, Harvard School of Public Health, Indiana University School of Medicine Massachusetts General Hospital, Massachusetts Institute of Blanche Capel, PhD Technology, Dana Farber Cancer Center, Stowers Institute Duke University Medical Center for Medical Research, and the National Institutes of Health Rao L. Divi, PhD Office of Science Education. This is her fifth article in FASEB’s National Cancer Institute, National Institutes of Health Breakthroughs in Bioscience series. Marnie Halpern, PhD SCIENTIFIC ADVISOR» Carnegie Institution for Science Henry J. Donahue, PhD is the School of Engineering Foun- dation Professor and Chair of the Department of Biomedical Loraine Oman-Ganes, MD, FRCP(C), CCMG, FACMG Engineering at the Virginia Commonwealth University.
    [Show full text]
  • Stem-Cell-Based Embryo Models for Fundamental Research and Translation
    REVIEW ARTICLE https://doi.org/10.1038/s41563-020-00829-9 Stem-cell-based embryo models for fundamental research and translation Jianping Fu 1,2,3 ✉ , Aryeh Warmflash 4,5 ✉ and Matthias P. Lutolf 6,7 ✉ Despite its importance, understanding the early phases of human development has been limited by availability of human sam- ples. The recent emergence of stem-cell-derived embryo models, a new field aiming to use stem cells to construct in vitro models to recapitulate snapshots of the development of the mammalian conceptus, opens up exciting opportunities to promote fundamental understanding of human development and advance reproductive and regenerative medicine. This Review provides a summary of the current knowledge of early mammalian development, using mouse and human conceptuses as models, and emphasizes their similarities and critical differences. We then highlight existing embryo models that mimic different aspects of mouse and human development. We further discuss bioengineering tools used for controlling multicellular interactions and self-organization critical for the development of these models. We conclude with a discussion of the important next steps and exciting future opportunities of stem-cell-derived embryo models for fundamental discovery and translation. he development of a multicellular organism from a single notable progress in studying non-human primate (NHP) monkey fertilized egg is a brilliant triumph of evolution that has fas- embryos10–12, whose developments are similar to humans. However, Tcinated generations of scientists (Box 1). Understanding our NHP monkey models remain costly, are difficult to modify geneti- own development is of particular fundamental and practical inter- cally, and have their own ethical challenges.
    [Show full text]
  • Divergent Mechanisms for Regulating Growth and Development After Imaginal Disc Damage in the Tobacco Hornworm, Manduca Sexta Manuel A
    © 2019. Published by The Company of Biologists Ltd | Journal of Experimental Biology (2019) 222, jeb200352. doi:10.1242/jeb.200352 RESEARCH ARTICLE Divergent mechanisms for regulating growth and development after imaginal disc damage in the tobacco hornworm, Manduca sexta Manuel A. Rosero1, Benedict Abdon1, Nicholas J. Silva1, Brenda Cisneros Larios1, Jhony A. Zavaleta1, Tigran Makunts1, Ernest S. Chang2, S. Janna Bashar1, Louie S. Ramos1, Christopher A. Moffatt1 and Megumi Fuse1,* ABSTRACT ablation experiments have demonstrated the importance of these Holometabolous insects have been able to radiate to vast ecological discs to the coupling of growth and metamorphosis when injury niches as adults through the evolution of adult-specific structures such results in a developmental delay. These delays presumably provide as wings, antennae and eyes. These structures arise from imaginal sufficient time for tissue regeneration. Injury to imaginal discs discs that show regenerative capacity when damaged. During imaginal has also been shown to delay development in a variety of disc regeneration, development has been shown to be delayed in the holometabolous insects, whether through genetic tissue ablation fruit fly Drosophila melanogaster, but how conserved the delay- (Colombani et al., 2012; Hackney et al., 2012), surgical ablation or ’ inducing mechanisms are across holometabolous insects has not autotomy (Kunkel, 1977; O Farrell and Stock, 1953, 1954; Stock ’ been assessed. The goal of this research was to develop the hornworm and O Farrell, 1954), or X-ray-induced selective damage (Ely and ’ Manduca sexta as an alternative model organism to study such Jungreis, 1977; O Brien and Wolf, 1964; Poodry and Woods, 1990; damage-induced mechanisms, with the advantage of a larger Stieper et al., 2008).
    [Show full text]
  • The Evolving Roles of Canonical WNT Signaling in Stem Cells And
    Laboratory Investigation (2016) 96, 116–136 © 2016 USCAP, Inc All rights reserved 0023-6837/16 $32.00 PATHOBIOLOGY IN FOCUS The evolving roles of canonical WNT signaling in stem cells and tumorigenesis: implications in targeted cancer therapies Ke Yang1,2,3, Xin Wang2,4, Hongmei Zhang2,5, Zhongliang Wang1,2, Guoxin Nan1,2, Yasha Li1,2, Fugui Zhang2,5, Maryam K Mohammed2, Rex C Haydon2, Hue H Luu2, Yang Bi1,2 and Tong-Chuan He1,2,5 The canonical WNT/β-catenin signaling pathway governs a myriad of biological processes underlying the development and maintenance of adult tissue homeostasis, including regulation of stem cell self-renewal, cell proliferation, differentiation, and apoptosis. WNTs are secreted lipid-modified glycoproteins that act as short-range ligands to activate receptor-mediated signaling pathways. The hallmark of the canonical pathway is the activation of β-catenin-mediated transcriptional activity. Canonical WNTs control the β-catenin dynamics as the cytoplasmic level of β-catenin is tightly regulated via phosphorylation by the ‘destruction complex’, consisting of glycogen synthase kinase 3β (GSK3β), casein kinase 1α (CK1α), the scaffold protein AXIN, and the tumor suppressor adenomatous polyposis coli (APC). Aberrant regulation of this signaling cascade is associated with varieties of human diseases, especially cancers. Over the past decade, significant progress has been made in understanding the mechanisms of canonical WNT signaling. In this review, we focus on the current understanding of WNT signaling at the extracellular, cytoplasmic membrane, and intracellular/ nuclear levels, including the emerging knowledge of cross-talk with other pathways. Recent progresses in developing novel WNT pathway-targeted therapies will also be reviewed.
    [Show full text]