DOCSLIB.ORG

Adipose Stem Cells from Patients with Crohn's Disease Show a Distinctive

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Adipose Stem Cells from Patients with Crohn's Disease Show a Distinctive Serena et al. Clinical Epigenetics (2020) 12:53 https://doi.org/10.1186/s13148-020-00843-3 RESEARCH Open Access Adipose stem cells from patients with Crohn’s disease show a distinctive DNA methylation pattern Carolina Serena1,2, Monica Millan3,4, Miriam Ejarque1,2, Alfonso Saera-Vila1, Elsa Maymó-Masip1,2, Catalina Núñez-Roa1,2, Diandra Monfort-Ferré1, Margarida Terrón-Puig1,2, Michelle Bautista5, Margarita Menacho5, Marc Martí6, Eloy Espin6, Joan Vendrell1,2,7* and Sonia Fernández-Veledo1,2* Abstract Background: Crohn’s disease (CD) is characterized by persistent inflammation and ulceration of the small or large bowel, and expansion of mesenteric adipose tissue, termed creeping fat (CF). We previously demonstrated that human adipose-derived stem cells (hASCs) from CF of patients with CD exhibit dysfunctional phenotypes, including a pro-inflammatory profile, high phagocytic capacity, and weak immunosuppressive properties. Importantly, these phenotypes persist in patients in remission and are found in all adipose depots explored including subcutaneous fat. We hypothesized that changes in hASCs are a consequence of epigenetic modifications. Methods: We applied epigenome-wide profiling with a methylation array (Illumina EPIC/850k array) and gene expression analysis to explore the impact of CD on the methylation signature of hASCs isolated from the subcutaneous fat of patients with CD and healthy controls (n = 7 and 5, respectively; cohort I). Differentially methylated positions (p value cutoff < 1 × 10−4 and ten or more DMPs per gene) and regions (inclusion threshold 0.2, p value cutoff < 1 × 10−2 and more than 2 DMRs per gene) were identified using dmpfinder and Bumphunter (minfi), respectively. Changes in the expression of differentially methylated genes in hASCs were validated in a second cohort (n = 10/10 inactive and active CD and 10 controls; including patients from cohort I) and also in peripheral blood mononuclear cells (PBMCs) of patients with active/inactive CD and of healthy controls (cohort III; n = 30 independent subjects). Results: We found a distinct DNA methylation landscape in hASCs from patients with CD, leading to changes in the expression of differentially methylated genes involved in immune response, metabolic, cell differentiation, and development processes. Notably, the expression of several of these genes in hASCs and PBMCs such as tumor necrosis factor alpha (TNFA) and PR domain zinc finger protein 16 (PRDM16) were not restored to normal (healthy) levels after disease remission. Conclusions: hASCs of patients with CD exhibit a unique DNA methylation and gene expression profile, but the expression of several genes are only partially restored in patients with inactive CD, both in hASCs and PBMCs. Understanding how CD shapes the functionality of hASCs is critical for investigating the complex pathophysiology of this disease, as well as for the success of cell-based therapies. Keywords: Methylome, Inflammatory bowel disease, Adipose tissue, Epigenetics, Gene expression * Correspondence: [email protected]; [email protected] 1Institut d´Investigació Sanitària Pere Virgili, Hospital Universitari Joan XXIII, Dr Mallafré Guasch, 4, 43007 Tarragona, Spain Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Serena et al. Clinical Epigenetics (2020) 12:53 Page 2 of 15 Background and third, we assessed the reproducibility of the changes The prevalence of inflammatory bowel disease (IBD), in- in gene expression of candidates genes in an more easily cluding Crohn’s disease (CD) and ulcerative colitis, is in- accessible sample, by testing for a specific blood signature. creasing, especially in western countries, and it is likely to represent an important social and economic burden Results in the coming years [1, 2]. CD is characterized by per- Methylation signature of adipose stem cells from sistent inflammation and ulcerations at the small or subcutaneous fat in patients with Crohn’s large bowel, provoking chronic low-grade systemic in- We first assessed genome-wide patterns of DNA methy- flammation and an undulating course of activity, with lation in hASCs isolated from subcutaneous fat depots relapsing cycles occurring after periods of remission. of patients with CD and compared them with those from Adipose tissue (AT) is believed to play an active role in hASCs isolated from healthy controls (n = 7 patients the pathogenesis of CD, as the expansion of mesenteric with CD and 5 healthy controls; cohort I). No differ- fat attached to the inflamed segments of intestine, also ences in overall mean DNA methylation (calculated by known as “creeping fat,” is a hallmark of the disease that averaging across all probes on the Illumina EPIC/850k seems to be directly related to disease activity [3–6]. In array) were observed between the two groups (p = 0.33; fact, it has recently been demonstrated that inclusion of Student’s t test) (data not shown), indicating that CD is the mesentery in ileocolic resection reduces the rate of not associated with global changes to DNA methylation surgical recurrence [7]. in this cell type. By contrast, DNA methylation at indi- AT is now recognized as a highly active endocrine vidual CpG sites showed considerable variability between organ that controls metabolic homeostasis. Emerging CD and controls. Principal component analysis (PCA) of evidence points to the immune system as a major con- the first two principal components (PC1 and PC2) of the tributor to AT homeostasis and function [8]. Indeed, AT methylation data revealed a difference between hASCs contributes importantly to low-grade systemic inflamma- of CD and those of controls (Fig. 1a) with PC1 explain- tion in obesity and its comorbidities [9–11]. AT contains ing 20.56% of the variance. We also performed a correl- an abundant population of multipotent human adipose- ation analysis between the confounding variables, derived stem cells (hASCs) that differentiate into several hospital, sex, age, and body mass index (BMI) and the specialized cells, including adipocytes and endothelial principal components (Fig. 1B; data is represented by a cells, and play key roles in immune response [12, 13]. heat map). No correlation was found between the six For example, hASCs administered to the circulation principal components and hospital, sex, age, or BMI, have the ability to home to inflamed/injured tissues [14, suggesting that these confounding variables did not 15]. Of note, some metabolic stress-induced conditions significantly affect the methylome pattern. Notably, we including CD alter the anti-inflammatory properties of found a significant correlation between the presence or these cells [16–18]. Remarkably, the altered properties of absence of the disease and PC1 (Fig. 1b). hASCs persist even when patients with CD are in remis- It is now recognized that, in addition to the expected sion, in all AT depots explored including subcutaneous binary behavior of CpG methylation (either hypo- or fat [18]. The constancy of this dysfunctional phenotype hypermethylated), some cytosines show intermediate, yet might reflect changes at the epigenetic level, which ul- low, levels of methylation [22]. Thus, we explored poten- timately hamper the resolution of the inflammatory tial differences in the segmented methylomes as fully events in the sinus of the affected mesenteric AT. In methylated regions (FMRs; > 50% methylated CpGs), support of this proposition, recent DNA methylation stud- low-methylated regions (LMRs; 3.9–50%), and unmethy- ies in peripheral blood mononuclear cells (PBMCs) revealed lated regions (UMRs; 1–3.9%), as described [22–26]. The that patients with CD have a distinct DNA methylome FMR class comprised over 50% of the methylome in all linked to the expression of differentially methylated genes samples (Fig. 1c), and the global percentage of methyla- associated with immune response and inflammation [19– tion was stable between control- and CD-hASCs. Never- 21]. We hypothesized that hASCs from patients with CD theless, there was a trend for greater methylation in the are conditioned by a chronic inflammatory milieu, which LMR region in hASCs isolated from patients with CD may alter their DNA methylome and influence their anti- relative to those from healthy controls. inflammatory and regenerative capacity. To address this issue, we designed a three-step study: Differentially methylated positions in adipose stem cells first, we evaluated whether
Load more

Recommended publications
  • Multifactorial Erβ and NOTCH1 Control of Squamous Differentiation and Cancer
    Multifactorial ERβ and NOTCH1 control of squamous differentiation and cancer Yang Sui Brooks, … , Karine Lefort, G. Paolo Dotto J Clin Invest. 2014;124(5):2260-2276. https://doi.org/10.1172/JCI72718. Research Article Oncology Downmodulation or loss-of-function mutations of the gene encoding NOTCH1 are associated with dysfunctional squamous cell differentiation and development of squamous cell carcinoma (SCC) in skin and internal organs. While NOTCH1 receptor activation has been well characterized, little is known about how NOTCH1 gene transcription is regulated. Using bioinformatics and functional screening approaches, we identified several regulators of the NOTCH1 gene in keratinocytes, with the transcription factors DLX5 and EGR3 and estrogen receptor β (ERβ) directly controlling its expression in differentiation. DLX5 and ERG3 are required for RNA polymerase II (PolII) recruitment to the NOTCH1 locus, while ERβ controls NOTCH1 transcription through RNA PolII pause release. Expression of several identified NOTCH1 regulators, including ERβ, is frequently compromised in skin, head and neck, and lung SCCs and SCC-derived cell lines. Furthermore, a keratinocyte ERβ–dependent program of gene expression is subverted in SCCs from various body sites, and there are consistent differences in mutation and gene-expression signatures of head and neck and lung SCCs in female versus male patients. Experimentally increased ERβ expression or treatment with ERβ agonists inhibited proliferation of SCC cells and promoted NOTCH1 expression and squamous differentiation both in vitro and in mouse xenotransplants. Our data identify a link between transcriptional control of NOTCH1 expression and the estrogen response in keratinocytes, with implications for differentiation therapy of squamous cancer. Find the latest version: https://jci.me/72718/pdf Research article Multifactorial ERβ and NOTCH1 control of squamous differentiation and cancer Yang Sui Brooks,1,2 Paola Ostano,3 Seung-Hee Jo,1,2 Jun Dai,1,2 Spiro Getsios,4 Piotr Dziunycz,5 Günther F.L.
    [Show full text]
  • ¬LACZ-REPORTER MAPPING of Dlx5/6 EXPRESSION AND
    Received: 26 December 2019 Revised: 10 May 2020 Accepted: 11 May 2020 DOI: 10.1002/cne.24952 RESEARCH ARTICLE LacZ-reporter mapping of Dlx5/6 expression and genoarchitectural analysis of the postnatal mouse prethalamus Luis Puelles1 | Carmen Diaz2 | Thorsten Stühmer3 | José L. Ferran1 | Margaret Martínez-de la Torre1 | John L. R. Rubenstein3 1Department of Human Anatomy and Psychobiology and IMIB-Arrixaca Institute, Abstract University of Murcia, Murcia, Spain We present here a thorough and complete analysis of mouse P0-P140 prethalamic 2 Department of Medical Sciences, School of histogenetic subdivisions and corresponding nuclear derivatives, in the context of Medicine and Institute for Research in Neurological Disabilities, University of Castilla- local tract landmarks. The study used as fundamental material brains from a transgenic La Mancha, Albacete, Spain mouse line that expresses LacZ under the control of an intragenic enhancer of Dlx5 3Nina Ireland Laboratory of Developmental Neurobiology, Department of Psychiatry, and Dlx6 (Dlx5/6-LacZ). Subtle shadings of LacZ signal, jointly with pan-DLX immu- UCSF Medical School, San Francisco, noreaction, and several other ancillary protein or RNA markers, including Calb2 and California Nkx2.2 ISH (for the prethalamic eminence, and derivatives of the rostral zona limitans Correspondence shell domain, respectively) were mapped across the prethalamus. The resulting model Luis Puelles, Department of Human Anatomy and Psychobiology, School of Medicine, of the prethalamic region postulates tetrapartite rostrocaudal and dorsoventral subdi- University of Murcia, Murcia 30071, Spain. visions, as well as a tripartite radial stratification, each cell population showing a char- Email: [email protected] acteristic molecular profile. Some novel nuclei are proposed, and some instances of Carmen Díaz, Department of Medical potential tangential cell migration were noted.
    [Show full text]
  • Exclusion of Dlx5/6 Expression from the Distal-Most Mandibular Arches Enables BMP-Mediated Specification of the Distal Cap
    Exclusion of Dlx5/6 expression from the distal-most mandibular arches enables BMP-mediated specification of the distal cap Joshua W. Vincentza, Jose J. Casasnovasa, Ralston M. Barnesb, Jianwen Quec, David E. Clouthierd, Jun Wange, and Anthony B. Firullia,1 aRiley Heart Research Center, Herman B. Wells Center for Pediatric Research Division of Pediatric Cardiology, Departments of Anatomy, Biochemistry, and Medical and Molecular Genetics, Indiana University Medical School, Indianapolis, IN 46202; bBiologics Discovery California, Bristol-Myers Squibb, Redwood City, CA 94063; cDepartment of Medicine, Columbia University Medical Center, New York, NY 10032; dDepartment of Craniofacial Biology, University of Colorado Anschutz Medical Campus, Aurora, CO 80108; and eDepartment of Molecular Physiology and Biophysics, Texas Heart Institute, Baylor College of Medicine, Houston, TX 77030 Edited by Clifford J. Tabin, Harvard Medical School, Boston, MA, and approved May 26, 2016 (received for review March 8, 2016) Cranial neural crest cells (crNCCs) migrate from the neural tube to Smads, H2, and GATA2/3 provide positive transcriptional inputs the pharyngeal arches (PAs) of the developing embryo and, sub- that serve to counteract the activity of Dlx proteins, thereby re- sequently, differentiate into bone and connective tissue to form the lieving EDN1-meditated repression. Together, these findings in- mandible. Within the PAs, crNCCs respond to local signaling cues to tegrate the communication between BMP and EDN1 signaling partition into the proximo-distally oriented subdomains that convey that establishes the distal cap of the mandible. positional information to these developing tissues. Here, we show that the distal-most of these subdomains, the distal cap, is marked Results by expression of the transcription factor Hand1 (H1) and gives rise to The H1Cre Lineage Gives Rise to the Lower Incisors, in a HAND Factor- the ectomesenchymal derivatives of the lower incisors.
    [Show full text]
  • Supplemental Material.Pdf
    Supplemental Material ZNF750 Interacts with KLF4 and RCOR1, KDM1A, and CTBP1/2 Chromatin Regulators to Repress Epidermal Progenitor Genes and Induce Differentiation Genes Lisa D. Boxer, Brook Barajas, Shiying Tao, Jiajing Zhang, and Paul Khavari Supplemental Inventory Figure S1. This figure supports Figure 1 and shows the Gene Ontology terms for ZNF750-bound but unaffected genes, the genomic enrichment of ZNF750 ChIP-seq peaks, and the percentage of peaks that contain the ZNF750 motif. Figure S2. This figure supports Figure 2 and shows gene expression changes with depletion of ZNF750-interacting proteins, confirms the knock-down of ZNF750-interacting proteins, and shows the quantification of Ki67 in ZNF750-interacting protein depleted organotypic tissue. Figure S3. This figure supports Figure 3 and shows the quantification of ZNF750 and interacting protein co-IPs, and IPs and Far western blots demonstrating competition between KLF4 and KDM1A for binding to ZNF750. Figure S4. This figure supports Figure 4 and shows the expression of ZNF750 mutant proteins and the effects of full-length or mutant ZNF750 on differentiation in organotypic tissue and on clonogenic growth. Figure S5. This figure supports Figure 5 and shows the effects of mutagenesis of ZNF750 and KLF4 motifs on reporter activity, and the changes in histone marks with depletion of ZNF750 and interacting proteins. Figure S6. This figure supports Figure 6 and shows the changes in mRNA and protein expression of ZNF750-interacting proteins during keratinocyte differentiation, and the expression of ZNF750-interacting proteins with ZNF750 depletion. Table S1. Supports Figure 1 and shows the genomic coordinates of ZNF750 ChIP-seq peaks.
    [Show full text]
  • The Tumor Suppressor HHEX Inhibits Axon Growth When Prematurely Expressed in Developing Central Nervous System Neurons
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by epublications@Marquette Marquette University e-Publications@Marquette Biological Sciences Faculty Research and Biological Sciences, Department of Publications 9-1-2015 The umorT Suppressor HHEX Inhibits Axon Growth when Prematurely Expressed in Developing Central Nervous System Neurons Matthew .T Simpson Marquette University Ishwariya Venkatesh Marquette University Ben L. Callif Marquette University Laura K. Thiel Marquette University Denise M. Coley Marquette University See next page for additional authors Accepted version. Molecular and Cellular Neuroscience, Vol 68 )September 2015): 272-283. DOI. © 2015 Elsevier Inc. Used with permission. NOTICE: this is the author’s version of a work that was accepted for publication in Molecular and Cellular Neuroscience. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Molecular and Cellular Neuroscience, Vol 68 )September 2015): 272-283. DOI. Authors Matthew T. Simpson, Ishwariya Venkatesh, Ben L. Callif, Laura K. Thiel, Denise M. Coley, Kristen N. Winsor, Zimei Wang, Audra A. Kramer, Jessica K. Lerch, and Murray G. Blackmore This article is available at e-Publications@Marquette: https://epublications.marquette.edu/bio_fac/515 NOT THE PUBLISHED VERSION; this is the author’s final, peer-reviewed manuscript. The published version may be accessed by following the link in the citation at the bottom of the page. The Tumor Suppressor HHEX Inhibits Axon Growth When Prematurely Expressed in Developing Central Nervous System Neurons Matthew T.
    [Show full text]
  • DLX Genes: Roles in Development and Cancer
    cancers Review DLX Genes: Roles in Development and Cancer Yinfei Tan 1,* and Joseph R. Testa 1,2,* 1 Genomics Facility, Fox Chase Cancer Center, Philadelphia, PA 19111, USA 2 Cancer Signaling and Epigenetics Program, Fox Chase Cancer Center, Philadelphia, PA 19111, USA * Correspondence: [email protected] (Y.T.); [email protected] (J.R.T.) Simple Summary: DLX homeobox family genes encode transcription factors that have indispensable roles in embryonic and postnatal development. These genes are critically linked to the morphogene- sis of craniofacial structures, branchial arches, forebrain, and sensory organs. DLX genes are also involved in postnatal homeostasis, particularly hematopoiesis and, when dysregulated, oncogen- esis. DLX1/2, DLX3/4, and DLX5/6 exist as bigenes on different chromosomes, sharing intergenic enhancers between gene pairs, which allows orchestrated spatiotemporal expression. Genomic alterations of human DLX gene enhancers or coding sequences result in congenital disorders such as split-hand/foot malformation. Aberrant postnatal expression of DLX genes is associated with hematological malignancies, including leukemias and lymphomas. In several mouse models of T-cell lymphoma, Dlx5 has been shown to act as an oncogene by cooperating with activated Akt, Notch1/3, and/or Wnt to drive tumor formation. In humans, DLX5 is aberrantly expressed in lung and ovarian carcinomas and holds promise as a therapeutic target. Abstract: Homeobox genes control body patterning and cell-fate decisions during development. The homeobox genes consist of many families, only some of which have been investigated regarding a possible role in tumorigenesis. Dysregulation of HOX family genes have been widely implicated in cancer etiology.
    [Show full text]
  • Supplementary Files:To “Allelic Switching of DLX5, GRB10 and SVOPL During Colorectal
    Supplementary files:to “Allelic switching of DLX5, GRB10 and SVOPL during colorectal cancer tumorigenesis” Arnoud Boot, Jan Oosting, Saskia Doorn, Sarah Ouahoud, Marina Ventayol Garcia, Dina Ruano, Hans Morreau, Tom van Wezel* Supplementary figure 1: ASE scores for selected chromosome 7 genes as determined by cSNP arrays. ASE scores were only calculated for cell lines with a heterozygous cSNP in the DNA. Supplementary figure 2: Gene expression of SVOPL and DLX5. Supplementary table 1: ASE results per cell line for all genes with at least 1 heterozygous sample Per sample, each gene was assigned an ASE score, based on the data from the heterozygous cSNPs, as described in the Supplementary methods. Samples displaying ASE for that gene are marked 1, samples not showing ASE are marked 0. For samples where no heterozygous cSNPs were identified, the field was left empty. Genes for which none of the samples showed a heterozygous cSNP were removed, as ASE could not be calculated. Supplementary table 2: DLX5 ASE results in paired normal and cancer samples. ASE scores for paired normal mucosa and colorectal cancer samples. The 4th column states the case- level conclusion concerning allelic switching during tumorigenesis. Supplementary table 3: PRPS1L1 KASPar genotyping results in paired normal and cancer samples. Supplementary table 4: Samples used in this study. Supplementary methods: ASE detection method and array analysis QC Figure S1: ASE scores for selected chromosome 7 genes Figure S2: Gene expression of SVOPL and DLX5 Supplementary methods: ASE detection method and array analysis QC Detection of ASE was performed by hybridizing both DNA and cDNA to the Infinium HumanExome- 12v1 beadchips.
    [Show full text]
  • Identification of Shared and Unique Gene Families Associated with Oral
    International Journal of Oral Science (2017) 9, 104–109 OPEN www.nature.com/ijos ORIGINAL ARTICLE Identification of shared and unique gene families associated with oral clefts Noriko Funato and Masataka Nakamura Oral clefts, the most frequent congenital birth defects in humans, are multifactorial disorders caused by genetic and environmental factors. Epidemiological studies point to different etiologies underlying the oral cleft phenotypes, cleft lip (CL), CL and/or palate (CL/P) and cleft palate (CP). More than 350 genes have syndromic and/or nonsyndromic oral cleft associations in humans. Although genes related to genetic disorders associated with oral cleft phenotypes are known, a gap between detecting these associations and interpretation of their biological importance has remained. Here, using a gene ontology analysis approach, we grouped these candidate genes on the basis of different functional categories to gain insight into the genetic etiology of oral clefts. We identified different genetic profiles and found correlations between the functions of gene products and oral cleft phenotypes. Our results indicate inherent differences in the genetic etiologies that underlie oral cleft phenotypes and support epidemiological evidence that genes associated with CL/P are both developmentally and genetically different from CP only, incomplete CP, and submucous CP. The epidemiological differences among cleft phenotypes may reflect differences in the underlying genetic causes. Understanding the different causative etiologies of oral clefts is
    [Show full text]
  • The Transcriptome of Utricle Hair Cell Regeneration in the Avian Inner Ear
    The Journal of Neuroscience, March 5, 2014 • 34(10):3523–3535 • 3523 Development/Plasticity/Repair The Transcriptome of Utricle Hair Cell Regeneration in the Avian Inner Ear Yuan-Chieh Ku,1 Nicole A. Renaud,1 Rose A. Veile,1 Cynthia Helms,1 Courtney C.J. Voelker,2 Mark E. Warchol,2 and Michael Lovett1 1Department of Genetics and 2Department of Otolaryngology, Washington University School of Medicine, St Louis, Missouri 63110 Sensory hair cell loss is the major cause of hearing and balance disorders. Mammals are incapable of sustained hair cell regeneration, but lower vertebrates can regenerate these mechano-electrical transducers. We present the first comprehensive transcriptome (by mRNA- Seq) of hair cell regeneration in the chick utricle. We provide pathway and pattern annotations and correlate these with the phenotypic events that occur during regeneration. These patterns are surprisingly synchronous and highly punctuated. We show how these patterns are a new resource for identifying components of the hair cell transcriptome and identify 494 new putative hair-cell-specific genes and validate three of these (of three tested) by immunohistochemical staining. We describe many surprising new components and dynamic expression patterns, particularly within NOTCH signaling. For example, we show that HES7 is specifically expressed during utricle hair cell regeneration and closely parallels the expression of HES5. Likewise, the expression of ATOH1 is closely correlated with HEYL and the HLH inhibitory transcription factors ID1, ID2, and ID4.
    [Show full text]
  • Mis-Expression of a Cranial Neural Crest Cell-Specific Gene Program in Cardiac Neural Crest Cells Modulates HAND Factor Expressi
    Journal of Cardiovascular Development and Disease Article Mis-Expression of a Cranial Neural Crest Cell-Specific Gene Program in Cardiac Neural Crest Cells Modulates HAND Factor Expression, Causing Cardiac Outflow Tract Phenotypes Joshua W. Vincentz 1,*, David E. Clouthier 2 and Anthony B. Firulli 1,* 1 Herman B Wells Center for Pediatric Research, Departments of Pediatrics, Anatomy and Medical and Molecular Genetics, Indiana Medical School, Indianapolis, IN 46202, USA 2 Department of Craniofacial Biology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA; [email protected] * Correspondence: [email protected] (J.W.V.); tfi[email protected] (A.B.F.) Received: 30 March 2020; Accepted: 14 April 2020; Published: 20 April 2020 Abstract: Congenital heart defects (CHDs) occur with such a frequency that they constitute a significant cause of morbidity and mortality in both children and adults. A significant portion of CHDs can be attributed to aberrant development of the cardiac outflow tract (OFT), and of one of its cellular progenitors known as the cardiac neural crest cells (NCCs). The gene regulatory networks that identify cardiac NCCs as a distinct NCC population are not completely understood. Heart and neural crest derivatives (HAND) bHLH transcription factors play essential roles in NCC morphogenesis. The Hand1PA/OFT enhancer is dependent upon bone morphogenic protein (BMP) signaling in both cranial and cardiac NCCs. The Hand1PA/OFT enhancer is directly repressed by the endothelin-induced transcription factors DLX5 and DLX6 in cranial but not cardiac NCCs. This transcriptional distinction offers the unique opportunity to interrogate NCC specification, and to understand why, despite similarities, cranial NCC fate determination is so diverse.
    [Show full text]
  • Dlx5 Is a Positive Regulator of Chondrocyte Differentiation During Endochondral Ossification
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Developmental Biology 252, 257–270 (2002) doi:10.1006/dbio.2002.0862 Dlx5 Is a Positive Regulator of Chondrocyte Differentiation during Endochondral Ossification Deborah Ferrari and Robert A. Kosher1 Department of BioStructure and Function, University of Connecticut Health Center, Farmington, CT 06030 The process of endochondral ossification in which the bones of the limb are formed after generation of cartilage models is dependent on a precisely regulated program of chondrocyte maturation. Here, we show that the homeobox-containing gene Dlx5 is expressed at the onset of chondrocyte maturation during the conversion of immature proliferating chondrocytes into postmitotic hypertrophying chondrocytes, a critical step in the maturation process. Moreover, retroviral misexpression of Dlx5 during differentiation of the skeletal elements of the chick limb in vivo results in the formation of severely shortened skeletal elements that contain excessive numbers of hypertrophying chondrocytes which extend into ectopic regions, including sites normally occupied by immature chondrocytes. The expansion in the extent of hypertrophic maturation detectable histologically is accompanied by expanded and upregulated domains of expression of molecular markers of chondrocyte maturation, particularly type X collagen and osteopontin, and by expansion of mineralized cartilage matrix, which is characteristic of terminal hypertrophic differentiation. Furthermore, Dlx5 misexpression markedly reduces chondrocyte proliferation concomitant with promoting hypertrophic maturation. Taken together, these results indicate that Dlx5 is a positive regulator of chondrocyte maturation and suggest that it regulates the process at least in part by promoting conversion of immature proliferating chondrocytes into hypertrophying chondrocytes.
    [Show full text]
  • Supplementary File Table S1
    Supplementary file Table S1 Grouping of Homeobox genes according to their main known function. Anatomical Structure Morphogenesis EN1, HOXC10, HOXC13, HOXD3, LBX1, SIX2, SIX4 Organ Morphogenesis CDX1, CDX2, HOXA11, HOXA13, ISL1, LHX1, PAX3, PDHX, PITX2, PITX3, PROX1, SIX6 Body Pattern Formation ALX3, EMX2, HHEX, HOXA11, HOXA2, HOXA4, HOXA5, HOXA6, HOXB1, HOXB5, HOXB6, HOXC5, HOXD10, HOXD8, LMX1B, PITX2 Ectoderm Development PROX1, VAX2 Endoderm Development HOXC11 Brain & Nervous System Development Brain Development ALX1, DLX2, EMX2 Nervous System Development: ARX, DLX5, DLX6, HOXD10, LBX1, LHX1, OTP, PAX3, PHOX2A, PHOX2B Skeletal Development: ALX3, ALX4, DLX3, DLX5, DLX6, EN1, HOXA11, HOXA13, HOXA2, HOXB6, HOXD10, HOXD13, MSX2 Muscle Development: BARX2, MKX, SIRT1, SIRT2, SIX1 Other Homeobox Genes Involved In BARX1, CDX4, CUX1, DLX1, EMX1, EN2, Multicellular Organismal HOXA1, HOXA7, HOXA9, HOXB13, HOXB2, Development: HOXB3, HOXB4, HOXB7, HOXB8, HOXB9, HOXC12, HOXC8, HOXC9, HOXD1, HOXD11, HOXD12, HOXD9, ISL2, LBX2, LMX1A, MEIS1, NKX3-1, OTX1, TLX1, VAX1, VSX1, VSX2 Homeobox Genes Involved In Cell ARX, EMX2, HHEX, HLX, HOPX, LBX1, LHX1, Differentiation: LMX1B, MIXL1, OTP, PHOX2A, SIRT1, VSX2 Other Genes: PHTF1, SIRT3, SIRT6, SIRT7, ZHX1, ZHX2 Homeobox genes include two subsets of genes coding for transcription factors involved in multiple functions. The clustered HOX genes are indicated in bold. Supplementary file Figure S2 5’ Spatial collinearity 3’ HOXA Chr. 7p15.3 HOXB Chr. 17q21.3 HOXC Chr. 12q13.3 HOXD Chr. 2q31 13 12 11 10 9 8 7 6 5 4 3 2 1 Paralogous HOX groups Distribution of the 39 human HOX genes in four clusters located in different chromosomal regions*. Blue indicates anterior HOX genes. Yellow, paralogy group 3 Hox genes, green and purple indicatete central HOX genes and Red the posterior HOX genes.
    [Show full text]