Integrated Analysis of Transcriptomic and Proteomic Analyses Reveals Different Metabolic Patterns in the Livers of Tibetan and Yorkshire Pigs

Total Page:16

File Type:pdf, Size:1020Kb

Integrated Analysis of Transcriptomic and Proteomic Analyses Reveals Different Metabolic Patterns in the Livers of Tibetan and Yorkshire Pigs Open Access Anim Biosci Vol. 34, No. 5:922-930 May 2021 https://doi.org/10.5713/ajas.20.0342 pISSN 2765-0189 eISSN 2765-0235 Integrated analysis of transcriptomic and proteomic analyses reveals different metabolic patterns in the livers of Tibetan and Yorkshire pigs Mengqi Duan1,a, Zhenmei Wang1,a, Xinying Guo1, Kejun Wang2, Siyuan Liu1, Bo Zhang3,*, and Peng Shang1,* * Corresponding Authors: Objective: Tibetan pigs, predominantly originating from the Tibetan Plateau, have been Bo Zhang Tel: +86-010-62734852, subjected to long-term natural selection in an extreme environment. To characterize the Fax: +86-010-62734852, metabolic adaptations to hypoxic conditions, transcriptomic and proteomic expression E-mail: [email protected] patterns in the livers of Tibetan and Yorkshire pigs were compared. Peng Shang Tel: +86-0894-5822924, Methods: RNA and protein were extracted from liver tissue of Tibetan and Yorkshire pigs Fax: +86-0894-5822924, (n = 3, each). Differentially expressed genes and proteins were subjected to gene ontology E-mail: [email protected] and Kyoto encyclopedia of genes and genomes functional enrichment analyses. 1 College of Animal Science, Tibet Agriculture Results: In the RNA-Seq and isobaric tags for relative and absolute quantitation analyses, a and Animal Husbandry University, Linzhi, total of 18,791 genes and 3,390 proteins were detected and compared. Of these, 273 and Xizang 86000, China 257 differentially expressed genes and proteins were identified. Evidence from functional 2 College of Animal Sciences and Veterinary Medicine, Henan Agricultural University, enrichment analysis showed that many genes were involved in metabolic processes. The Zhengzhou, Henan 450046, China combined transcriptomic and proteomic analyses revealed that small molecular biosynthesis, 3 National Engineering Laboratory for Animal metabolic processes, and organic hydroxyl compound metabolic processes were the major Breeding/Beijing Key Laboratory for Animal processes operating differently in the two breeds. The important genes include retinol Genetic Improvement, China Agricultural University, Beijing 100193, China dehydrogenase 16, adenine phosphoribosyltransferase, prenylcysteine oxidase 1, sorbin and SH3 domain containing 2, ENSSSCG00000036224, perilipin 2, ladinin 1, kynurenine a These authors contributed equally to this aminotransferase 1, and dimethylarginine dimethylaminohydrolase 1. work. Conclusion: The findings of this study provide novel insight into the high-altitude metabolic ORCID adaptation of Tibetan pigs. Mengqi Duan https://orcid.org/0000-0002-9792-129X Zhenmei Wang Keywords: Hypoxia; Metabolic; Proteome; Tibetan Pig; Transcriptome https://orcid.org/0000-0002-0696-3276 Xinying Guo https://orcid.org/0000-0003-1405-4483 Kejun Wang https://orcid.org/0000-0003-4337-980X Siyuan Liu INTRODUCTION https://orcid.org/0000-0003-0418-5762 Bo Zhang https://orcid.org/0000-0002-7331-1550 Tibetan pigs, mainly originating from the Tibetan Plateau, have endured long-term natural Peng Shang selection in an extreme environment, such as hypoxia, low temperature, and high solar https://orcid.org/0000-0001-5427-7986 radiation [1,2]. Adaptation to the extreme environment is complex and reflected in several Submitted May 16, 2020; Revised Jun 23, 2020; aspects, including cardiopulmonary function, fat metabolism, immunity, blood pressure, and Accepted Sept 13, 2020 coat color [3-5]. Some studies have been conducted on hypoxia adaptation, physiological features, meat quality, and animal origin [6-8] of Tibetan pigs. Liver metabolic processes are also an important adaptive feature, as they play a key role in catalytic decomposition, biosynthesis, and energy delivery. Characterization of mRNA and protein expression in the liver tissues of Tibetan pigs remains to be performed. Recently, high-throughput sequenc- ing of DNA, RNA, and proteins, has been widely used to study physiological processes and traits [9,10] in pigs. In the present study, we combined proteomic and transcriptomic analyses to characterize liver function in Tibetan and Yorkshire pigs, to better under- Copyright © 2021 by Animal Bioscience This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, 922 and reproduction in any medium, provided the original work is properly cited. www.animbiosci.org Duan et al (2021) Anim Biosci 34:922-930 stand the high-altitude adaptations of Tibetan pigs. according to the manufacturer’s instructions (Thermo Fisher Scientific, Waltham, MA, USA). The cDNA synthesis was MATERIALS AND METHODS performed with 1 μg of total RNA, following the protocol accompanying the FastQuant RT Kit (Tiangen Biotech (Bei- Ethics statement and sample collection jing) Co., LTD, Beijing, China). The qPCR amplification was The procedures for animal care were approved by the Ani- performed as described previously [1]. Relative expression is mal Welfare Committee of the Tibet Agriculture and Animal presented as mean±standard error of the mean. Differences Husbandry College, and all experiments were conducted in were tested for statistical significance using the Student’s t- accordance with approved relevant guidelines and regula- test. A p-value <0.05 was considered as the threshold for tions. Tibetan and Yorkshire pigs were raised under identical statistical significance (* p<0.05; ** p<0.01). conditions with same diet at the farm of the Tibet Agricul- ture and Animal Husbandry College, located in Linzhi city Protein extraction and labeling (Tibet, 2,900 m, above mean sea level). All pigs were housed Liver tissue protein was extracted using a mammalian tissue in standard conditions with natural, uncontrolled room tem- total protein extraction kit (AP0601-50, Beijing Bangfei perature and light. Complete formula meal feed was fed Biotechnology Co., Ltd., Beijing, China). Quantification three times per day and pigs had ad libitum access to water. was investigated using a protein quantification kit (Dingguo At the age of 6 months, three Tibetan pigs and three York- Changsheng, Beijing, China). Protein peptides from each shire pigs were randomly selected to slaughter and sample. group were labeled using the 8-plex isobaric tags for rela- Liver tissues from each individual were collected, immedi- tive and absolute quantitation (iTRAQ) reagents multiplex ately frozen in liquid nitrogen, and stored at –80°C until use. kit (ABI, Foster City, CA, USA). For each sample, approxi- mately 200 μg protein was precipitated with 10 volumes of RNA isolation and library construction acetone at –20°C overnight. After centrifugation, the pro- The total RNA from liver tissues was extracted following the tein pellet was dissolved in 60 μL of dissolution buffer. The phenol-chloroform method. RNA concentration and integ- iTRAQ labeling reagents were added to the peptide samples rity were determined using an Agilent 2100 Bioanalyzer and and reacted at room temperature for 1 h, before investiga- the Agilent RNA 6000 Nano Kit (Agilent Technologies, Palo tion of the labeling and extraction efficiency. Samples were Alto, CA, USA). RNA-seq libraries were constructed using pooled, vacuum-dried, and dissolved in solution. The re- the NEBNext Ultra RNA Library Prep Kit for Illumina (# constituted peptides were analyzed with a Q-Exactive mass E7530L, NEB; San Diego, CA, USA) according to the manu- spectrometer (Thermo Fisher Scientific, USA), coupled with facturer’s instructions. Paired-end 150 bp reads were generated a nano high-performance liquid chromatography system using the HiSeq Xten platform. (Thermo Fisher Scientific, USA). The detailed steps for en- zymolysis and iTRAQ labeling have been described previously Data analysis of RNA-Seq [5,9]. Data analysis included quality control of raw reads, filtering, alignment, assembly, expression count, and annotation. Con- Retrieval of database and protein data analysis taminated reads were filtered using a Perl script in-house. Data analysis was performed using Proteome Discover v2.1 Clean reads were mapped to the reference genome (Sus scrofa (Thermo Fisher Scientific, USA) software. Peptide identifi- genome v11.1, downloaded from ENSEMBL web server) cation was performed using the SEQUEST search engine, using HISAT2 v2.1.0 [11]. Reads for each gene in each sample using the Uniprot database (Sus scrofa 49003 entries, 20190102). were counted by HTSeq v0.6.0 (https://github.com/simon- Proteins were identified according to the following criteria: anders/htseq). Fragments per kilobase per million mapped mass error was set to 15 ppm for precursor ions and 0.02 Da reads were then calculated to estimate the expression level of for fragment ions. Trypsin was used as a cleavage enzyme, each gene. Deseq (http://bioconductor.org/packages/release/ and two missing cleavages were allowed. The false discovery bioc/html/DESeq.html) was used to identify the differentially rate was adjusted to 0.01. Differentially expressed proteins expressed genes (DEGs), with the criteria of fold change >2 (DEPs) were identified as having a fold change >1.2 and p- and adjusted p-value <0.05 [12]. value <0.05. Expression level validation by quantitative-polymerase Functional enrichment analysis chain reaction Gene ontology (GO) and Kyoto encyclopedia of genes and Differentially expressed mRNAs were selected for validation genomes (KEGG) functional enrichment analyses were per- by quantitative-polymerase chain reaction (qPCR). Total formed, using the
Recommended publications
  • Screening and Identification of Key Biomarkers in Clear Cell Renal Cell Carcinoma Based on Bioinformatics Analysis
    bioRxiv preprint doi: https://doi.org/10.1101/2020.12.21.423889; this version posted December 23, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Screening and identification of key biomarkers in clear cell renal cell carcinoma based on bioinformatics analysis Basavaraj Vastrad1, Chanabasayya Vastrad*2 , Iranna Kotturshetti 1. Department of Biochemistry, Basaveshwar College of Pharmacy, Gadag, Karnataka 582103, India. 2. Biostatistics and Bioinformatics, Chanabasava Nilaya, Bharthinagar, Dharwad 580001, Karanataka, India. 3. Department of Ayurveda, Rajiv Gandhi Education Society`s Ayurvedic Medical College, Ron, Karnataka 562209, India. * Chanabasayya Vastrad [email protected] Ph: +919480073398 Chanabasava Nilaya, Bharthinagar, Dharwad 580001 , Karanataka, India bioRxiv preprint doi: https://doi.org/10.1101/2020.12.21.423889; this version posted December 23, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract Clear cell renal cell carcinoma (ccRCC) is one of the most common types of malignancy of the urinary system. The pathogenesis and effective diagnosis of ccRCC have become popular topics for research in the previous decade. In the current study, an integrated bioinformatics analysis was performed to identify core genes associated in ccRCC. An expression dataset (GSE105261) was downloaded from the Gene Expression Omnibus database, and included 26 ccRCC and 9 normal kideny samples. Assessment of the microarray dataset led to the recognition of differentially expressed genes (DEGs), which was subsequently used for pathway and gene ontology (GO) enrichment analysis.
    [Show full text]
  • 1 Metabolic Dysfunction Is Restricted to the Sciatic Nerve in Experimental
    Page 1 of 255 Diabetes Metabolic dysfunction is restricted to the sciatic nerve in experimental diabetic neuropathy Oliver J. Freeman1,2, Richard D. Unwin2,3, Andrew W. Dowsey2,3, Paul Begley2,3, Sumia Ali1, Katherine A. Hollywood2,3, Nitin Rustogi2,3, Rasmus S. Petersen1, Warwick B. Dunn2,3†, Garth J.S. Cooper2,3,4,5* & Natalie J. Gardiner1* 1 Faculty of Life Sciences, University of Manchester, UK 2 Centre for Advanced Discovery and Experimental Therapeutics (CADET), Central Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Sciences Centre, Manchester, UK 3 Centre for Endocrinology and Diabetes, Institute of Human Development, Faculty of Medical and Human Sciences, University of Manchester, UK 4 School of Biological Sciences, University of Auckland, New Zealand 5 Department of Pharmacology, Medical Sciences Division, University of Oxford, UK † Present address: School of Biosciences, University of Birmingham, UK *Joint corresponding authors: Natalie J. Gardiner and Garth J.S. Cooper Email: [email protected]; [email protected] Address: University of Manchester, AV Hill Building, Oxford Road, Manchester, M13 9PT, United Kingdom Telephone: +44 161 275 5768; +44 161 701 0240 Word count: 4,490 Number of tables: 1, Number of figures: 6 Running title: Metabolic dysfunction in diabetic neuropathy 1 Diabetes Publish Ahead of Print, published online October 15, 2015 Diabetes Page 2 of 255 Abstract High glucose levels in the peripheral nervous system (PNS) have been implicated in the pathogenesis of diabetic neuropathy (DN). However our understanding of the molecular mechanisms which cause the marked distal pathology is incomplete. Here we performed a comprehensive, system-wide analysis of the PNS of a rodent model of DN.
    [Show full text]
  • Supplementary Table S4. FGA Co-Expressed Gene List in LUAD
    Supplementary Table S4. FGA co-expressed gene list in LUAD tumors Symbol R Locus Description FGG 0.919 4q28 fibrinogen gamma chain FGL1 0.635 8p22 fibrinogen-like 1 SLC7A2 0.536 8p22 solute carrier family 7 (cationic amino acid transporter, y+ system), member 2 DUSP4 0.521 8p12-p11 dual specificity phosphatase 4 HAL 0.51 12q22-q24.1histidine ammonia-lyase PDE4D 0.499 5q12 phosphodiesterase 4D, cAMP-specific FURIN 0.497 15q26.1 furin (paired basic amino acid cleaving enzyme) CPS1 0.49 2q35 carbamoyl-phosphate synthase 1, mitochondrial TESC 0.478 12q24.22 tescalcin INHA 0.465 2q35 inhibin, alpha S100P 0.461 4p16 S100 calcium binding protein P VPS37A 0.447 8p22 vacuolar protein sorting 37 homolog A (S. cerevisiae) SLC16A14 0.447 2q36.3 solute carrier family 16, member 14 PPARGC1A 0.443 4p15.1 peroxisome proliferator-activated receptor gamma, coactivator 1 alpha SIK1 0.435 21q22.3 salt-inducible kinase 1 IRS2 0.434 13q34 insulin receptor substrate 2 RND1 0.433 12q12 Rho family GTPase 1 HGD 0.433 3q13.33 homogentisate 1,2-dioxygenase PTP4A1 0.432 6q12 protein tyrosine phosphatase type IVA, member 1 C8orf4 0.428 8p11.2 chromosome 8 open reading frame 4 DDC 0.427 7p12.2 dopa decarboxylase (aromatic L-amino acid decarboxylase) TACC2 0.427 10q26 transforming, acidic coiled-coil containing protein 2 MUC13 0.422 3q21.2 mucin 13, cell surface associated C5 0.412 9q33-q34 complement component 5 NR4A2 0.412 2q22-q23 nuclear receptor subfamily 4, group A, member 2 EYS 0.411 6q12 eyes shut homolog (Drosophila) GPX2 0.406 14q24.1 glutathione peroxidase
    [Show full text]
  • Growth and Gene Expression Profile Analyses of Endometrial Cancer Cells Expressing Exogenous PTEN
    [CANCER RESEARCH 61, 3741–3749, May 1, 2001] Growth and Gene Expression Profile Analyses of Endometrial Cancer Cells Expressing Exogenous PTEN Mieko Matsushima-Nishiu, Motoko Unoki, Kenji Ono, Tatsuhiko Tsunoda, Takeo Minaguchi, Hiroyuki Kuramoto, Masato Nishida, Toyomi Satoh, Toshihiro Tanaka, and Yusuke Nakamura1 Laboratories of Molecular Medicine [M. M-N., M. U., K. O., T. M., T. Ta., Y. N.] and Genome Database [T. Ts.], Human Genome Center, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan; Department of Obstetrics and Gynecology, School of Medicine, Kitasato University, Sagamihara 228-8555, Japan [H. K.]; Department of Obstetrics and Gynecology, Institute of Clinical Medicine, University of Tsukuba, Tsukuba 305-8576, Japan [M. N.]; and Department of Obstetrics and Gynecology, Ibaraki Seinan Central Hospital, Tsukuba 306-0433, Japan [T. S.] ABSTRACT Akt/protein kinase B, cell survival, and cell proliferation (8). Over- expression of PTEN can decrease cell proliferation and tumorigenicity The PTEN tumor suppressor gene encodes a multifunctional phospha- (9, 10), an observation attributed to the ability of PTEN to induce cell tase that plays an important role in inhibiting the phosphatidylinositol-3- cycle arrest and apoptosis (11, 12). kinase pathway and downstream functions that include activation of Akt/protein kinase B, cell survival, and cell proliferation. Enforced ex- Thus, lack of PTEN expression may affect a complex set of pression of PTEN in various cancer cell lines decreases cell proliferation transcriptional targets. However, no systematic assessment of PTEN- through arrest of the cell cycle, accompanied in some cases by induction regulated targets in cancer cells has been reported to date.
    [Show full text]
  • Early Growth Response 1 Regulates Hematopoietic Support and Proliferation in Human Primary Bone Marrow Stromal Cells
    Hematopoiesis SUPPLEMENTARY APPENDIX Early growth response 1 regulates hematopoietic support and proliferation in human primary bone marrow stromal cells Hongzhe Li, 1,2 Hooi-Ching Lim, 1,2 Dimitra Zacharaki, 1,2 Xiaojie Xian, 2,3 Keane J.G. Kenswil, 4 Sandro Bräunig, 1,2 Marc H.G.P. Raaijmakers, 4 Niels-Bjarne Woods, 2,3 Jenny Hansson, 1,2 and Stefan Scheding 1,2,5 1Division of Molecular Hematology, Department of Laboratory Medicine, Lund University, Lund, Sweden; 2Lund Stem Cell Center, Depart - ment of Laboratory Medicine, Lund University, Lund, Sweden; 3Division of Molecular Medicine and Gene Therapy, Department of Labora - tory Medicine, Lund University, Lund, Sweden; 4Department of Hematology, Erasmus MC Cancer Institute, Rotterdam, the Netherlands and 5Department of Hematology, Skåne University Hospital Lund, Skåne, Sweden ©2020 Ferrata Storti Foundation. This is an open-access paper. doi:10.3324/haematol. 2019.216648 Received: January 14, 2019. Accepted: July 19, 2019. Pre-published: August 1, 2019. Correspondence: STEFAN SCHEDING - [email protected] Li et al.: Supplemental data 1. Supplemental Materials and Methods BM-MNC isolation Bone marrow mononuclear cells (BM-MNC) from BM aspiration samples were isolated by density gradient centrifugation (LSM 1077 Lymphocyte, PAA, Pasching, Austria) either with or without prior incubation with RosetteSep Human Mesenchymal Stem Cell Enrichment Cocktail (STEMCELL Technologies, Vancouver, Canada) for lineage depletion (CD3, CD14, CD19, CD38, CD66b, glycophorin A). BM-MNCs from fetal long bones and adult hip bones were isolated as reported previously 1 by gently crushing bones (femora, tibiae, fibulae, humeri, radii and ulna) in PBS+0.5% FCS subsequent passing of the cell suspension through a 40-µm filter.
    [Show full text]
  • Dysregulation at Multiple Points of the Kynurenine Pathway Is a Ubiquitous Feature of Renal Cancer: Implications for Tumour Immune Evasion
    This is a repository copy of Dysregulation at multiple points of the kynurenine pathway is a ubiquitous feature of renal cancer: implications for tumour immune evasion. White Rose Research Online URL for this paper: http://eprints.whiterose.ac.uk/158462/ Version: Accepted Version Article: Hornigold, N, Dunn, KR, Craven, RA et al. (22 more authors) (2020) Dysregulation at multiple points of the kynurenine pathway is a ubiquitous feature of renal cancer: implications for tumour immune evasion. British Journal of Cancer. ISSN 0007-0920 https://doi.org/10.1038/s41416-020-0874-y © The Author(s), under exclusive licence to Cancer Research UK 2020. This is an author produced version of an article published in British Journal of Cancer. Uploaded in accordance with the publisher's self-archiving policy. Reuse Items deposited in White Rose Research Online are protected by copyright, with all rights reserved unless indicated otherwise. They may be downloaded and/or printed for private study, or other acts as permitted by national copyright laws. The publisher or other rights holders may allow further reproduction and re-use of the full text version. This is indicated by the licence information on the White Rose Research Online record for the item. Takedown If you consider content in White Rose Research Online to be in breach of UK law, please notify us by emailing [email protected] including the URL of the record and the reason for the withdrawal request. [email protected] https://eprints.whiterose.ac.uk/ 1 Dysregulation at multiple points
    [Show full text]
  • Protein Network Analyses of Pulmonary Endothelial Cells In
    www.nature.com/scientificreports OPEN Protein network analyses of pulmonary endothelial cells in chronic thromboembolic pulmonary hypertension Sarath Babu Nukala1,8,9*, Olga Tura‑Ceide3,4,5,9, Giancarlo Aldini1, Valérie F. E. D. Smolders2,3, Isabel Blanco3,4, Victor I. Peinado3,4, Manuel Castell6, Joan Albert Barber3,4, Alessandra Altomare1, Giovanna Baron1, Marina Carini1, Marta Cascante2,7,9 & Alfonsina D’Amato1,9* Chronic thromboembolic pulmonary hypertension (CTEPH) is a vascular disease characterized by the presence of organized thromboembolic material in pulmonary arteries leading to increased vascular resistance, heart failure and death. Dysfunction of endothelial cells is involved in CTEPH. The present study describes for the frst time the molecular processes underlying endothelial dysfunction in the development of the CTEPH. The advanced analytical approach and the protein network analyses of patient derived CTEPH endothelial cells allowed the quantitation of 3258 proteins. The 673 diferentially regulated proteins were associated with functional and disease protein network modules. The protein network analyses resulted in the characterization of dysregulated pathways associated with endothelial dysfunction, such as mitochondrial dysfunction, oxidative phosphorylation, sirtuin signaling, infammatory response, oxidative stress and fatty acid metabolism related pathways. In addition, the quantifcation of advanced oxidation protein products, total protein carbonyl content, and intracellular reactive oxygen species resulted increased
    [Show full text]
  • Prenylcysteine Oxidase 1, a Pro-Oxidant Enzyme of Low Density Lipoproteins
    Prenylcysteine oxidase 1, a pro-oxidant enzyme of low density lipoproteins Luis V. Herrera-Marcos,1,2 Jose M. Lou-Bonafonte,2,3,4 Maria V. Martinez-Gracia,4 Carmen Arnal,2,4,5, Maria A. Navarro1,2,4, Jesus Osada1,2,4 1Departamento de Bioquimica y Biologia Molecular y Celular, Facultad de Veterinaria, Instituto de Investigacion Sanitaria de Aragon-Universidad de Zaragoza, Zaragoza, E-50013, Spain, 2Instituto Agroalimentario de Aragon, CITA-Universidad de Zaragoza, Spain, 3Departamento de Farmacologia y Fisiologia, Facultad de Ciencias de la Salud y del Deporte, Instituto de Investigacion Sanitaria de Aragon-Universidad de Zaragoza, Huesca, E-22002, Spain, 4CIBER de Fisiopatologia de la Obesidad y Nutricion, Instituto de Salud Carlos III, Madrid, E-28029, Spain, 5Departamento de Patologia Animal, Facultad de Veterinaria, Instituto de Investigacion Sanitaria de Aragon-Universidad de Zaragoza, Zaragoza, E-50013, Spain, TABLE OF CONTENTS 1. Abstract 2. Introduction 3. Human PCYOX1 gene 3.1. Gene polymorphisms 3.2. Hepatic PCYOX1 transcripts 3.3. Hepatic PCYOX1 transcriptional regulation 3.4. Hepatic PCYOX1 post-transcriptional regulation 4. PCYOX1 Protein 4.1. Post-translational modifications (PTMs) 4.2. Protein structure and function 4.3. Protein interactions 5. PCYOX1 in physiology and disease 5.1. Lipoproteins 5.2. Pcyox1-deficient mice 5.3. PCYOX1 and the liver 5.4. PCYOX1 gene and cancer 5.5. PCYOX1 and neurodegenerative diseases 5.6. PCYOX1 and obesity 6. Conclusions 7. Acknowledgments 8. References 1. ABSTRACT Elevated levels of low density lipoproteins (LDLs) cause atherosclerotic disease, and proteomic analyses have found that these lipoproteins are endowed with prenylcysteine lyase.
    [Show full text]
  • Human Induced Pluripotent Stem Cell–Derived Podocytes Mature Into Vascularized Glomeruli Upon Experimental Transplantation
    BASIC RESEARCH www.jasn.org Human Induced Pluripotent Stem Cell–Derived Podocytes Mature into Vascularized Glomeruli upon Experimental Transplantation † Sazia Sharmin,* Atsuhiro Taguchi,* Yusuke Kaku,* Yasuhiro Yoshimura,* Tomoko Ohmori,* ‡ † ‡ Tetsushi Sakuma, Masashi Mukoyama, Takashi Yamamoto, Hidetake Kurihara,§ and | Ryuichi Nishinakamura* *Department of Kidney Development, Institute of Molecular Embryology and Genetics, and †Department of Nephrology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan; ‡Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Hiroshima, Japan; §Division of Anatomy, Juntendo University School of Medicine, Tokyo, Japan; and |Japan Science and Technology Agency, CREST, Kumamoto, Japan ABSTRACT Glomerular podocytes express proteins, such as nephrin, that constitute the slit diaphragm, thereby contributing to the filtration process in the kidney. Glomerular development has been analyzed mainly in mice, whereas analysis of human kidney development has been minimal because of limited access to embryonic kidneys. We previously reported the induction of three-dimensional primordial glomeruli from human induced pluripotent stem (iPS) cells. Here, using transcription activator–like effector nuclease-mediated homologous recombination, we generated human iPS cell lines that express green fluorescent protein (GFP) in the NPHS1 locus, which encodes nephrin, and we show that GFP expression facilitated accurate visualization of nephrin-positive podocyte formation in
    [Show full text]
  • WNT16 Is a New Marker of Senescence
    Table S1. A. Complete list of 177 genes overexpressed in replicative senescence Value Gene Description UniGene RefSeq 2.440 WNT16 wingless-type MMTV integration site family, member 16 (WNT16), transcript variant 2, mRNA. Hs.272375 NM_016087 2.355 MMP10 matrix metallopeptidase 10 (stromelysin 2) (MMP10), mRNA. Hs.2258 NM_002425 2.344 MMP3 matrix metallopeptidase 3 (stromelysin 1, progelatinase) (MMP3), mRNA. Hs.375129 NM_002422 2.300 HIST1H2AC Histone cluster 1, H2ac Hs.484950 2.134 CLDN1 claudin 1 (CLDN1), mRNA. Hs.439060 NM_021101 2.119 TSPAN13 tetraspanin 13 (TSPAN13), mRNA. Hs.364544 NM_014399 2.112 HIST2H2BE histone cluster 2, H2be (HIST2H2BE), mRNA. Hs.2178 NM_003528 2.070 HIST2H2BE histone cluster 2, H2be (HIST2H2BE), mRNA. Hs.2178 NM_003528 2.026 DCBLD2 discoidin, CUB and LCCL domain containing 2 (DCBLD2), mRNA. Hs.203691 NM_080927 2.007 SERPINB2 serpin peptidase inhibitor, clade B (ovalbumin), member 2 (SERPINB2), mRNA. Hs.594481 NM_002575 2.004 HIST2H2BE histone cluster 2, H2be (HIST2H2BE), mRNA. Hs.2178 NM_003528 1.989 OBFC2A Oligonucleotide/oligosaccharide-binding fold containing 2A Hs.591610 1.962 HIST2H2BE histone cluster 2, H2be (HIST2H2BE), mRNA. Hs.2178 NM_003528 1.947 PLCB4 phospholipase C, beta 4 (PLCB4), transcript variant 2, mRNA. Hs.472101 NM_182797 1.934 PLCB4 phospholipase C, beta 4 (PLCB4), transcript variant 1, mRNA. Hs.472101 NM_000933 1.933 KRTAP1-5 keratin associated protein 1-5 (KRTAP1-5), mRNA. Hs.534499 NM_031957 1.894 HIST2H2BE histone cluster 2, H2be (HIST2H2BE), mRNA. Hs.2178 NM_003528 1.884 CYTL1 cytokine-like 1 (CYTL1), mRNA. Hs.13872 NM_018659 tumor necrosis factor receptor superfamily, member 10d, decoy with truncated death domain (TNFRSF10D), 1.848 TNFRSF10D Hs.213467 NM_003840 mRNA.
    [Show full text]
  • Clinical Impact of Copy Number Variation Changes in Bladder Cancer Samples
    EXPERIMENTAL AND THERAPEUTIC MEDICINE 22: 901, 2021 Clinical impact of copy number variation changes in bladder cancer samples VICTORIA SPASOVA1, BORIS MLADENOV2, SIMEON RANGELOV3, ZORA HAMMOUDEH1, DESISLAVA NESHEVA1, DIMITAR SERBEZOV1, RADA STANEVA1,4, SAVINA HADJIDEKOVA1,4, MIHAIL GANEV1, LUBOMIR BALABANSKI1,5, RADOSLAVA VAZHAROVA5,6, CHAVDAR SLAVOV3, DRAGA TONCHEVA1 and OLGA ANTONOVA1 1Department of Medical Genetics, Medical University‑Sofia, 1431 Sofia;2 Department of Urology, UMBALSM N.I. Pirogov, 1606 Sofia; 3Department of Urology, Tsaritsa Yoanna University Hospital, 1527 Sofia; 4Medical Genetics Laboratory, Nadezhda Women's Health Hospital, 1373 Sofia; 5Medical Genetics Laboratory, GARH Malinov, 1680 Sofia; 6Department of Biology, Medical Genetics and Microbiology, Faculty of Medicine, Sofia University St. Kliment Ohridski, 1407 Sofia, Bulgaria Received November 30, 2019; Accepted February 18, 2021 DOI: 10.3892/etm.2021.10333 Abstract. The aim of the present study was to detect copy uroepithelial tumours may lay a foundation for implementing number variations (CNVs) related to tumour progression and molecular CNV profiling of bladder tumours as part of a metastasis of urothelial carcinoma through whole‑genome routine progression risk estimation strategy, thus expanding scanning. A total of 30 bladder cancer samples staged from the personalized therapeutic approach. pTa to pT4 were included in the study. DNA was extracted from freshly frozen tissue via standard phenol‑chloroform extraction Introduction and CNV analysis was performed on two alternative platforms (CytoChip Oligo aCGH, 4x44K and Infinium OncoArray‑500K The most successful approach to treating a disease has BeadChip; Illumina, Inc.). Data were analysed with BlueFuse always been etiological therapy. In the case of bladder Multi software and Karyostudio, respectively.
    [Show full text]
  • Deciphering Pre-Leukaemic Dysregulation Caused by Mutant C/Ebpα at the Level of the Single Cell
    Deciphering pre-leukaemic dysregulation caused by mutant C/EBPa at the level of the single cell Moosa Rashid Qureshi Department of Haematology University of Cambridge This dissertation is submitted for the degree of Doctor of Philosophy Queens’ College August 2019 DeClaration This thesis is the result of my own work and inCludes nothing whiCh is the outCome of work done in Collaboration exCept as deClared in the Acknowledgments and speCified in the text. It is not substantially the same as any that I have submitted, or, is being ConCurrently submitted for a degree or diploma or other qualifiCation at the University of Cambridge or any other University or similar institution exCept as deClared in the Acknowledgments and speCified in the text. I further state that no substantial part of my thesis has already been submitted, or, is being ConCurrently submitted for any suCh degree, diploma or other qualification at the University of Cambridge or any other University or similar institution exCept as deClared in the Acknowledgments and speCified in the text. It does not exCeed the prescribed word limit for the relevant Degree Committee Moosa Qureshi August 2019 I AbstraCt DeCiphering pre-leukaemic dysregulation Caused by mutant C/EBPa at the level of the single cell Moosa Rashid Qureshi C/EBPa is an ideal Candidate to explore pre-leukaemiC dysregulation of transCriptional networks beCause it is a signifiCant player in normal myelopoiesis, it interaCts with other transCription faCtors (TFs) in lineage speCification, and the CEBPA gene has been identified as an early mutation in acute myeloid leukaemia (AML). N321D is a mutation affeCting the leuCine zipper domain of C/EBPa whiCh has previously generated an aggressive AML phenotype in a mouse model.
    [Show full text]