DOCSLIB.ORG

Genomic Expression Profiles in Cumulus Cells Derived from Germinal Vesicle and MII Mouse Oocytes

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Genomic Expression Profiles in Cumulus Cells Derived from Germinal Vesicle and MII Mouse Oocytes CSIRO PUBLISHING Reproduction, Fertility and Development http://dx.doi.org/10.1071/RD15077 Genomic expression profiles in cumulus cells derived from germinal vesicle and MII mouse oocytes Li ShaoA, Ri-Cheng ChianA,B,C, Yixin XuA, Zhengjie YanA, Yihui ZhangA, Chao GaoA, Li GaoA, Jiayin LiuA and Yugui CuiA,C AState Key Laboratory of Reproductive Medicine, Center for Clinical Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, 140 Hanzhong Road, Nanjing 210029, China. BDepartment of Obstetrics and Gynecology, McGill University, 687 Pine Avenue West, Montreal H3A 1A1, Canada. CCorresponding authors. Emails: [email protected]; [email protected] Abstract. Cumulus cells (CCs) are distinct from other granulosa cells and the mutual communication between CCs and oocytes is essential for the establishment of oocyte competence. In the present study we assessed genomic expression profiles in mouse CCs before and after oocyte maturation in vitro. Microarray analysis revealed significant changes in gene expression in CCs between the germinal vesicle (GV) and metaphase II (MII) stages, with 2615 upregulated and 2808 downregulated genes. Genes related to epidermal growth factor, extracellular matrix (Ptgs2, Ereg, Tnfaip6 and Efemp1), mitochondrial metabolism (Fdx1 and Aifm2), gap junctions and the cell cycle (Gja1, Gja4, Ccnd2, Ccna2 and Ccnb2) were highlighted as being differentially expressed between the two development stages. Real-time polymerase chain reaction confirmed the validity and reproducibility of the results for the selected differentially expressed genes. Similar expression patterns were identified by western blot analysis for some functional proteins, including EFEMP1, FDX1, GJA1 and CCND2, followed by immunofluorescence localisation. These genes may be potential biomarkers for oocyte develop- mental competence following fertilisation and will be investigated further in future studies. Additional keywords: biomarkers, developmental competence, in vitro maturation. Received 25 February 2015, accepted 21 April 2015, published online 20 May 2015 Introduction potential on the basis of gene expression profiles of human CCs, The cumulus–oocyte complex (COC) comprises a secondary Assou et al. (2008) found that there was a correlation between oocyte surrounded by a subgroup of distinctive somatic cells, the expression of some genes in CCs and embryo potential or namely cumulus cells (CCs). It has been widely accepted that pregnancy outcome, and proposed BCL2 L11, PCK1 and NFIB CCs maintain an intimate relationship with the oocyte through as biomarkers for predicting pregnancy. Interestingly, a recent gap junctions and paracrine signalling during folliculogenesis report indicated that there were significant differences in the (Su et al. 2004; Gilchrist et al. 2008). Following the LH surge, expression of SPSB2 and TP5313 between CCs of normal and communication between the oocyte and CCs induces oocyte chromosomally abnormal oocytes (Fragouli et al. 2012). maturation, because CCs compensate for oocyte-deficient It has been reported that gene expression differs between CCs metabolic molecules, such as pyruvate, alanine and cholesterol, associated with oocytes at the metaphase II (MII) stage com- to support oocyte development. Meanwhile, the oocyte secrets a pared with those at the germinal vesicle (GV) and metaphase I series of paracrine factors, such as growth differentiation factor (MI) stages, suggesting that the gene expression profile of CCs 9 (GDF9), bone morphogenetic protein 15 (BMP15) and of the mature MII oocyte may be used as a predictor of oocyte fibroblast growth factor 8 (FGF8), which act on adjacent CCs to quality(Ouandaogo et al. 2011). Furthermore, when comparing promote proliferation and differentiation (Gilchrist et al. 2008; the transcriptome profiles of CCs from in vivo- and in vitro- Su et al. 2009). matured human COCs at different nuclear maturation stages, Previous studies have used microarray analysis and real-time Ouandaogo et al. (2012) found that the transcriptomic signature polymerase chain reaction (PCR) to focus on gene expression of the CCs varies according to both oocyte maturation stages and profiles in granulosa cells (GCs) or CCs in an effort to find the maturation conditions, and suggested that there was a delay potential biomarkers of oocyte quality (Assou et al. 2006, 2010; in the acquisition of the mature CC phenotype following in vitro Patrizio et al. 2007; Huang and Wells 2010; Assidi et al. 2011; maturation (IVM). Therefore, it seems likely that screening for Vigone et al. 2013). Using a non-invasive test to assess embryo changes in gene expression in CCs under different maturation Journal compilation Ó CSIRO 2015 www.publish.csiro.au/journals/rfd B Reproduction, Fertility and Development L. Shao et al. conditions would make it possible to optimise the in vitro culture combined with mechanical isolation by pipetting. CCs were system (Kind et al. 2013; Salhab et al. 2013). However, only grouped according to the relevant oocyte maturation stage, limited data are available regarding gene expression profiles of which were evaluated under a stereomicroscope. Oocytes with a CCs in association with oocyte maturation. GV in the ooplasm were defined as being at the GV stage, IVM can imitate in vivo progress of oocyte maturation, which whereas oocytes with a first polar body in the perivitelline space is a good model to find some biomarkers during oocyte matura- were defined as being at the MII stage. CCs were washed in cold tion in CCs, and to understand the molecular mechanisms of the phosphate-buffered saline (PBS) before being centrifuged at mutual communication between two kinds of cells. It is neces- 300g for 10 min at 378C. The supernatant was discarded and the sary to clarify the molecular mechanism of oocyte maturation pellet was resuspended in Trizol reagent (Invitrogen, Waltham, during IVM in order to optimise the culture system and to obtain MA, USA) before being stored at À808C until microarray high developmental potential oocytes for clinical outcomes. The analysis. CCs from more than 100 COCs were pooled together objective of this study was to compare the differences of as one sample; each group contained at least three independent genomic expression profiles in CCs before and after IVM and pooled samples for microarray analysis. was to try to find some potential target genes expressed in CCs for further study of oocyte developmental competence. Total RNA extraction and microarray analysis Materials and methods Total RNA was extracted from pooled CC samples (three each Animal preparation from the GV-CCs and MII-CCs groups) as described previ- ously (Dai et al. 2014) and was isolated using an RNeasy Mini Three-week-old ICR mice were obtained from the Medical Kit (7410; Qiagen, Valencia, CA, USA) according to the Experimental Animal Center of Nanjing Medical University and manufacturer’s instructions. The quality of the RNA was maintained at 20–228C under a 12-h light–dark cycle. Sterilised confirmed using a NanoDrop ND-1000 spectrophotometer food and water were provided ad libitum. Animal experiments (Thermo Scientific, Waltham, MA, USA). The microarray were conducted in accordance with the Animal Research analysis was performed on an Agilent Technologies (Santa Committee Guidelines of Nanjing Medical University, which Clara, CA, USA) array platform (Whole Mouse Genome Oligo were approved by the Animal and Human Ethics Board of Microarray). Briefly, total RNA from each sample was Nanjing Medical University. amplifiedlinearlyandlabelledwithCy3-UTPusingthe Agilent One-Colour Quick Amp Labelling Kit, resulting in Ovarian stimulation, COC collection and IVM labelled cRNAs. The labelled cRNAs were then purified using Mice were injected with 5 IU, i.p., equine chorionic gonado- the RNasey Mini Kit and the corresponding concentration and tropin (eCG; Folligon; Intervet, Castle Hill, NSW, Australia) to specific activity (pmol Cy3 per mgcRNA)wereassessed stimulate follicle development. Then, 46–48 h after eCG injec- using the NanoDrop ND-1000 spectrophotometer. Next, a 1-mg tion, mice were killed by cervical dislocation and the ovaries aliquot of each labelled cRNA was fragmented, followed by dissected out and placed in M2 medium (M716; Sigma, St Louis, hybridisation to the Mouse 4x44K Genome Oligo Microarray MO, USA). Immature COCs were released by puncturing the (Agilent Technologies), which contained 39 000þ mouse ovarian surface with a sterile 27-G needle and collected using genes and transcripts, using Agilent’s 60-mer SurePrint tech- Pasteur micropipettes; only those with a compacted cumulus nology. After hybridisation, the arrays were washed, fixed and were selected for use in the present study. COCs from each scanned using an Agilent G2505C scanner. mouse were randomly divided into two groups, the first to be For analysis of microarray data, the array images were used immediately (GV-CCs group) and second to be used for extracted using Agilent Feature Extraction software (version IVM (MII-CCs group). COCs in the latter group were washed 11.0.1.1). Raw signal intensity data were quintile normalised twice with IVM medium and then groups of 10 COCs were place and subsequently processed using GeneSpring GX v12.0 m in 30- L droplets of IVM medium (see Zhang et al. 2014) (Agilent Technologies). After quintile normalisation, genes covered by mineral oil (MKBG7544V; Sigma) and then cul- for which at least three of six samples were flagged as ‘Detected’ 8 tured
Load more

Recommended publications
  • Mutant RAMP2 Causes Primary Open-Angle Glaucoma Via the CRLR-Camp Axis
    © American College of Medical Genetics and Genomics ARTICLE Mutant RAMP2 causes primary open-angle glaucoma via the CRLR-cAMP axis Bo Gong, PhD1,2, Houbin Zhang, PhD1, Lulin Huang, PhD1, Yuhong Chen, MD3, Yi Shi, PhD1, Pancy Oi-Sin Tam, MS4, Xianjun Zhu, PhD1, Yi Huang, MD1,5, Bo Lei, PhD6, Periasamy Sundaresan, PhD7, Xi Li, MS1, Linxin Jiang, MS1, Jialiang Yang, MS1, Ying Lin, MS1, Fang Lu, PhD1, Lijia Chen, MRCSEd (Ophth), PhD4, Yuanfeng Li, MS1, Christopher Kai-Shun Leung, MD4, Xiaoxin Guo, MS1, Shanshan Zhang, MS1, Guo Huang, MS1, Yaqi Wu, MS1, Tongdan Zhou, MS1, Ping Shuai, PhD1, Clement Chee-Yung Tham, FRCOphth4, Nicole Weisschuh, PhD8, Subbaiah Ramasamy Krishnadas, MD7, Christian Mardin, MD9, André Reis, PhD10, Jiyun Yang, MD1, Lin Zhang, PhD1,3, Yu Zhou, PhD1, Ziyan Wang, PhD1, Chao Qu, MD11, Peter X. Shaw, PhD12, Chi-Pui Pang, DPhil4, Xinghuai Sun, MD3, Weiquan Zhu, MD5, Dean Yaw Li, MD5, Francesca Pasutto, PhD10 and Zhenglin Yang, MD, PhD1,2 Purpose: Primary open-angle glaucoma (POAG) is the leading 4763 POAG patients, whereas no variants were detected in any cause of irreversible blindness worldwide and mutations in known exon of RAMP2 in 10,953 control individuals. Mutant RAMP2s genes can only explain 5–6% of POAG. This study was conducted aggregated in transfected cells and resulted in damage to the AM- to identify novel POAG-causing genes and explore the pathogenesis RAMP2/CRLR-cAMP signaling pathway. Ablation of one Ramp2 of this disease. allele led to cAMP reduction and retinal ganglion cell death in mice. Methods: Exome sequencing was performed in a Han Chinese Conclusion: This study demonstrated that disruption of RAMP2/ cohort comprising 398 sporadic cases with POAG and 2010 CRLR-cAMP axis could cause POAG and identified a potential controls, followed by replication studies by Sanger sequencing.
    [Show full text]
  • Viewed Under 23 (B) Or 203 (C) fi M M Male Cko Mice, and Largely Unaffected Magni Cation; Scale Bars, 500 M (B) and 50 M (C)
    BRIEF COMMUNICATION www.jasn.org Renal Fanconi Syndrome and Hypophosphatemic Rickets in the Absence of Xenotropic and Polytropic Retroviral Receptor in the Nephron Camille Ansermet,* Matthias B. Moor,* Gabriel Centeno,* Muriel Auberson,* † † ‡ Dorothy Zhang Hu, Roland Baron, Svetlana Nikolaeva,* Barbara Haenzi,* | Natalya Katanaeva,* Ivan Gautschi,* Vladimir Katanaev,*§ Samuel Rotman, Robert Koesters,¶ †† Laurent Schild,* Sylvain Pradervand,** Olivier Bonny,* and Dmitri Firsov* BRIEF COMMUNICATION *Department of Pharmacology and Toxicology and **Genomic Technologies Facility, University of Lausanne, Lausanne, Switzerland; †Department of Oral Medicine, Infection, and Immunity, Harvard School of Dental Medicine, Boston, Massachusetts; ‡Institute of Evolutionary Physiology and Biochemistry, St. Petersburg, Russia; §School of Biomedicine, Far Eastern Federal University, Vladivostok, Russia; |Services of Pathology and ††Nephrology, Department of Medicine, University Hospital of Lausanne, Lausanne, Switzerland; and ¶Université Pierre et Marie Curie, Paris, France ABSTRACT Tight control of extracellular and intracellular inorganic phosphate (Pi) levels is crit- leaves.4 Most recently, Legati et al. have ical to most biochemical and physiologic processes. Urinary Pi is freely filtered at the shown an association between genetic kidney glomerulus and is reabsorbed in the renal tubule by the action of the apical polymorphisms in Xpr1 and primary fa- sodium-dependent phosphate transporters, NaPi-IIa/NaPi-IIc/Pit2. However, the milial brain calcification disorder.5 How- molecular identity of the protein(s) participating in the basolateral Pi efflux remains ever, the role of XPR1 in the maintenance unknown. Evidence has suggested that xenotropic and polytropic retroviral recep- of Pi homeostasis remains unknown. Here, tor 1 (XPR1) might be involved in this process. Here, we show that conditional in- we addressed this issue in mice deficient for activation of Xpr1 in the renal tubule in mice resulted in impaired renal Pi Xpr1 in the nephron.
    [Show full text]
  • Single-Cell RNA Sequencing Demonstrates the Molecular and Cellular Reprogramming of Metastatic Lung Adenocarcinoma
    ARTICLE https://doi.org/10.1038/s41467-020-16164-1 OPEN Single-cell RNA sequencing demonstrates the molecular and cellular reprogramming of metastatic lung adenocarcinoma Nayoung Kim 1,2,3,13, Hong Kwan Kim4,13, Kyungjong Lee 5,13, Yourae Hong 1,6, Jong Ho Cho4, Jung Won Choi7, Jung-Il Lee7, Yeon-Lim Suh8,BoMiKu9, Hye Hyeon Eum 1,2,3, Soyean Choi 1, Yoon-La Choi6,10,11, Je-Gun Joung1, Woong-Yang Park 1,2,6, Hyun Ae Jung12, Jong-Mu Sun12, Se-Hoon Lee12, ✉ ✉ Jin Seok Ahn12, Keunchil Park12, Myung-Ju Ahn 12 & Hae-Ock Lee 1,2,3,6 1234567890():,; Advanced metastatic cancer poses utmost clinical challenges and may present molecular and cellular features distinct from an early-stage cancer. Herein, we present single-cell tran- scriptome profiling of metastatic lung adenocarcinoma, the most prevalent histological lung cancer type diagnosed at stage IV in over 40% of all cases. From 208,506 cells populating the normal tissues or early to metastatic stage cancer in 44 patients, we identify a cancer cell subtype deviating from the normal differentiation trajectory and dominating the metastatic stage. In all stages, the stromal and immune cell dynamics reveal ontological and functional changes that create a pro-tumoral and immunosuppressive microenvironment. Normal resident myeloid cell populations are gradually replaced with monocyte-derived macrophages and dendritic cells, along with T-cell exhaustion. This extensive single-cell analysis enhances our understanding of molecular and cellular dynamics in metastatic lung cancer and reveals potential diagnostic and therapeutic targets in cancer-microenvironment interactions. 1 Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.
    [Show full text]
  • Amitriptyline-Mediated Cognitive Enhancement in Aged 36Tg Alzheimer’S Disease Mice Is Associated with Neurogenesis and Neurotrophic Activity
    Amitriptyline-Mediated Cognitive Enhancement in Aged 36Tg Alzheimer’s Disease Mice Is Associated with Neurogenesis and Neurotrophic Activity Wayne Chadwick1, Nick Mitchell2, Jenna Caroll3, Yu Zhou1, Sung-Soo Park1, Liyun Wang1, Kevin G. Becker4, Yongqing Zhang4, Elin Lehrmann4, William H. Wood III4, Bronwen Martin5, Stuart Maudsley1* 1 Receptor Pharmacology Unit, National Institute on Aging, National Institutes of Health, Baltimore, Maryland, United States of America, 2 Laboratory of Neurosciences, National Institute on Aging, National Institutes of Health, Baltimore, Maryland, United States of America, 3 Center for Neurodegenerative Disease Research, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America, 4 Genomics Unit, Research Resources Branch, National Institute on Aging, National Institutes of Health, Baltimore, Maryland, United States of America, 5 Metabolism Unit, National Institute on Aging, National Institutes of Health, Baltimore, Maryland, United States of America Abstract Approximately 35 million people worldwide suffer from Alzheimer’s disease (AD). Existing therapeutics, while moderately effective, are currently unable to stem the widespread rise in AD prevalence. AD is associated with an increase in amyloid beta (Ab) oligomers and hyperphosphorylated tau, along with cognitive impairment and neurodegeneration. Several antidepressants have shown promise in improving cognition and alleviating oxidative stress in AD but have failed as long- term therapeutics. In this study, amitriptyline, an FDA-approved tricyclic antidepressant, was administered orally to aged and cognitively impaired transgenic AD mice (36TgAD). After amitriptyline treatment, cognitive behavior testing demonstrated that there was a significant improvement in both long- and short-term memory retention. Amitriptyline treatment also caused a significant potentiation of non-toxic Ab monomer with a concomitant decrease in cytotoxic dimer Ab load, compared to vehicle-treated 36TgAD controls.
    [Show full text]
  • Characterization of Glomerular Extracellular Matrix in Iga Nephropathy by Proteomic Analysis of Laser-Captured Microdissected Gl
    Paunas et al. BMC Nephrology (2019) 20:410 https://doi.org/10.1186/s12882-019-1598-1 RESEARCH ARTICLE Open Access Characterization of glomerular extracellular matrix in IgA nephropathy by proteomic analysis of laser-captured microdissected glomeruli Flavia Teodora Ioana Paunas1,2* , Kenneth Finne2, Sabine Leh2,3, Tarig Al-Hadi Osman2, Hans-Peter Marti2,4, Frode Berven5 and Bjørn Egil Vikse1,2 Abstract Background: IgA nephropathy (IgAN) involves mesangial matrix expansion, but the proteomic composition of this matrix is unknown. The present study aimed to characterize changes in extracellular matrix in IgAN. Methods: In the present study we used mass spectrometry-based proteomics in order to quantitatively compare protein abundance between glomeruli of patients with IgAN (n = 25) and controls with normal biopsy findings (n = 15). Results: Using a previously published paper by Lennon et al. and cross-referencing with the Matrisome database we identified 179 extracellular matrix proteins. In the comparison between IgAN and controls, IgAN glomeruli showed significantly higher abundance of extracellular matrix structural proteins (e.g periostin, vitronectin, and extracellular matrix protein 1) and extracellular matrix associated proteins (e.g. azurocidin, myeloperoxidase, neutrophil elastase, matrix metalloproteinase-9 and matrix metalloproteinase 2). Periostin (fold change 3.3) and azurocidin (3.0) had the strongest fold change between IgAN and controls; periostin was also higher in IgAN patients who progressed to ESRD as compared to patients who did not. Conclusion: IgAN is associated with widespread changes of the glomerular extracellular matrix proteome. Proteins important in glomerular sclerosis or inflammation seem to be most strongly increased and periostin might be an important marker of glomerular damage in IgAN.
    [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]
  • A Genome-Wide Association Study Identifies Four Novel Susceptibility Loci Underlying Inguinal Hernia
    UCSF UC San Francisco Previously Published Works Title A genome-wide association study identifies four novel susceptibility loci underlying inguinal hernia. Permalink https://escholarship.org/uc/item/7g06z1k5 Journal Nature communications, 6(1) ISSN 2041-1723 Authors Jorgenson, Eric Makki, Nadja Shen, Ling et al. Publication Date 2015-12-21 DOI 10.1038/ncomms10130 Peer reviewed eScholarship.org Powered by the California Digital Library University of California ARTICLE Received 24 Aug 2015 | Accepted 6 Nov 2015 | Published 21 Dec 2015 DOI: 10.1038/ncomms10130 OPEN A genome-wide association study identifies four novel susceptibility loci underlying inguinal hernia Eric Jorgenson1,*, Nadja Makki2,3,*, Ling Shen1, David C. Chen4, Chao Tian5, Walter L. Eckalbar2,3, David Hinds5, Nadav Ahituv2,3 & Andrew Avins1 Inguinal hernia repair is one of the most commonly performed operations in the world, yet little is known about the genetic mechanisms that predispose individuals to develop inguinal hernias. We perform a genome-wide association analysis of surgically confirmed inguinal hernias in 72,805 subjects (5,295 cases and 67,510 controls) and confirm top associations in an independent cohort of 92,444 subjects with self-reported hernia repair surgeries (9,701 cases and 82,743 controls). We identify four novel inguinal hernia susceptibility loci in the regions of EFEMP1, WT1, EBF2 and ADAMTS6. Moreover, we observe expression of all four genes in mouse connective tissue and network analyses show an important role for two of these genes (EFEMP1 and WT1) in connective tissue maintenance/homoeostasis. Our findings provide insight into the aetiology of hernia development and highlight genetic pathways for studies of hernia development and its treatment.
    [Show full text]
  • Proteomics of Serum Extracellular Vesicles Identifies a Novel COPD Biomarker, Fibulin-3 from Elastic Fibres
    ORIGINAL ARTICLE COPD Proteomics of serum extracellular vesicles identifies a novel COPD biomarker, fibulin-3 from elastic fibres Taro Koba 1, Yoshito Takeda1, Ryohei Narumi2, Takashi Shiromizu2, Yosui Nojima 3, Mari Ito3, Muneyoshi Kuroyama1, Yu Futami1, Takayuki Takimoto4, Takanori Matsuki1, Ryuya Edahiro1, Satoshi Nojima5, Yoshitomo Hayama1, Kiyoharu Fukushima1, Haruhiko Hirata1, Shohei Koyama1, Kota Iwahori1, Izumi Nagatomo1, Mayumi Suzuki1, Yuya Shirai1, Teruaki Murakami1, Kaori Nakanishi 1, Takeshi Nakatani1, Yasuhiko Suga1, Kotaro Miyake1, Takayuki Shiroyama1, Hiroshi Kida 1, Takako Sasaki6, Koji Ueda7, Kenji Mizuguchi3, Jun Adachi2, Takeshi Tomonaga2 and Atsushi Kumanogoh1 ABSTRACT There is an unmet need for novel biomarkers in the diagnosis of multifactorial COPD. We applied next-generation proteomics to serum extracellular vesicles (EVs) to discover novel COPD biomarkers. EVs from 10 patients with COPD and six healthy controls were analysed by tandem mass tag-based non-targeted proteomics, and those from elastase-treated mouse models of emphysema were also analysed by non-targeted proteomics. For validation, EVs from 23 patients with COPD and 20 healthy controls were validated by targeted proteomics. Using non-targeted proteomics, we identified 406 proteins, 34 of which were significantly upregulated in patients with COPD. Of note, the EV protein signature from patients with COPD reflected inflammation and remodelling. We also identified 63 upregulated candidates from 1956 proteins by analysing EVs isolated from mouse models. Combining human and mouse biomarker candidates, we validated 45 proteins by targeted proteomics, selected reaction monitoring. Notably, levels of fibulin-3, tripeptidyl- peptidase 2, fibulin-1, and soluble scavenger receptor cysteine-rich domain-containing protein were significantly higher in patients with COPD.
    [Show full text]
  • Clonally Expanding Smooth Muscle Cells Promote Atherosclerosis by Escaping Efferocytosis and Activating the Complement Cascade
    Clonally expanding smooth muscle cells promote atherosclerosis by escaping efferocytosis and activating the complement cascade Ying Wanga,b, Vivek Nandaa,b,c, Daniel Direnzoa, Jianqin Yea, Sophia Xiaoa, Yoko Kojimaa, Kathryn L. Howea, Kai-Uwe Jarra, Alyssa M. Floresa, Pavlos Tsantilasa, Noah Tsaoa, Abhiram Raob,d, Alexandra A. C. Newmane, Anne V. Eberharda, James R. Priestf, Arno Ruusaleppg, Gerard Pasterkamph,i, Lars Maegdefesselj,k, Clint L. Millerl,m,n, Lars Lindo, Simon Koplevp, Johan L. M. Björkegrenp, Gary K. Owense, Erik Ingelssonb,o,q, Irving L. Weissmanr,1, and Nicholas J. Leepera,b,1 aDivision of Vascular Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, CA 94305; bStanford Cardiovascular Institute, Stanford University, Stanford, CA 94305; cDepartment of Molecular and Cellular Pathology, University of Alabama at Birmingham, Birmingham, AL 35294; dDepartment of Bioengineering, Stanford University, Stanford, CA 94305; eRobert M. Berne Cardiovascular Research Center, University of Virginia, Charlottesville, VA 22904; fDivision of Pediatric Cardiology, Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305; gDepartment of Cardiac Surgery, Tartu University Hospital, Tartu, Estonia 50406; hDepartment of Cardiology, University Medical Center Utrecht, 3584CX Utrecht, the Netherlands; iLaboratory of Clinical Chemistry, University Medical Center Utrecht, 3584CX Utrecht, the Netherlands; jDepartment for Vascular and Endovascular Surgery, Klinikum Rechts der Isar, Technical University
    [Show full text]
  • Investigating Developmental and Functional Deficits in Neurodegenerative
    UNIVERSITY OF CALIFORNIA, IRVINE Investigating developmental and functional deficits in neurodegenerative disease using transcriptomic analyses DISSERTATION submitted in partial satisfaction of the requirements for the degree of DOCTOR OF PHILOSOPHY in Biomedical Sciences by Ryan Gar-Lok Lim Dissertation Committee: Professor Leslie M. Thompson, Chair Assistant Professor Dritan Agalliu Professor Peter Donovan Professor Suzanne Sandmeyer 2016 Introduction, Figure 1.1 © 2014 Macmillan Publishers Limited. Appendix 1 © 2016 Elsevier Ltd. All other materials © 2016 Ryan Gar-Lok Lim DEDICATION This dissertation is dedicated to my parents, sister, and my wife. I love you all very much and could not have accomplished any of this without your love and support. Please take the time to reflect back on all of the moments we’ve shared, and know, that it is because of those moments I have been able to succeed. This accomplishment is as much yours as it is mine. ii TABLE OF CONTENTS Page LIST OF FIGURES vi LIST OF TABLES ix ACKNOWLEDGMENTS x CURRICULUM VITAE xiii ABSTRACT OF THE DISSERTATION xv Introduction Huntington’s disease, the neurovascular unit and the blood-brain barrier 1 1.1 Huntington’s Disease 1.2 HTT structure and function 1.2.1 Normal HTT function and possible loss-of-function contributions to HD 1.3 mHTT pathogenesis 1.3.1 The dominant pathological features of mHTT - a gain-of- toxic function? 1.3.2 Cellular pathologies and non-neuronal contributions to HD 1.4 The neurovascular unit and the blood-brain barrier 1.4.1 Structure and function
    [Show full text]
  • Transcriptomic Analysis of Neuregulin-1 Regulated Genes
    UC Riverside UC Riverside Previously Published Works Title Transcriptomic analysis of neuregulin-1 regulated genes following ischemic stroke by computational identification of promoter binding sites: A role for the ETS-1 transcription factor. Permalink https://escholarship.org/uc/item/2020r225 Journal PloS one, 13(6) ISSN 1932-6203 Authors Surles-Zeigler, Monique C Li, Yonggang Distel, Timothy J et al. Publication Date 2018 DOI 10.1371/journal.pone.0197092 Peer reviewed eScholarship.org Powered by the California Digital Library University of California RESEARCH ARTICLE Transcriptomic analysis of neuregulin-1 regulated genes following ischemic stroke by computational identification of promoter binding sites: A role for the ETS-1 transcription factor Monique C. Surles-Zeigler1, Yonggang Li2,3, Timothy J. Distel2, Hakeem Omotayo2, a1111111111 Shaokui Ge2, Byron D. Ford2* a1111111111 a1111111111 1 Department of Neurobiology, Morehouse School of Medicine, Atlanta, Georgia, United States of America, 2 Department of Biomedical Sciences, University of California±Riverside School of Medicine, Riverside, a1111111111 California, United States of America, 3 ICF, Atlanta, GA, United States of America a1111111111 * [email protected] Abstract OPEN ACCESS Citation: Surles-Zeigler MC, Li Y, Distel TJ, Ischemic stroke is a major cause of mortality in the United States. We previously showed Omotayo H, Ge S, Ford BD (2018) Transcriptomic that neuregulin-1 (NRG1) was neuroprotective in rat models of ischemic stroke. We used analysis of neuregulin-1 regulated genes following gene expression profiling to understand the early cellular and molecular mechanisms of ischemic stroke by computational identification of promoter binding sites: A role for the ETS-1 NRG1's effects after the induction of ischemia.
    [Show full text]
  • Coordination of Endothelial Cell Positioning and Fate Specification By
    ARTICLE https://doi.org/10.1038/s41467-021-24414-z OPEN Coordination of endothelial cell positioning and fate specification by the epicardium Pearl Quijada 1,8, Michael A. Trembley1, Adwiteeya Misra1,2, Jacquelyn A. Myers 3,4, Cameron D. Baker 3,4, Marta Pérez-Hernández 5, Jason R. Myers3,4, Ronald A. Dirkx Jr.1, ✉ Ethan David Cohen6, Mario Delmar5, John M. Ashton 3,4 & Eric M. Small 1,2,7 The organization of an integrated coronary vasculature requires the specification of immature 1234567890():,; endothelial cells (ECs) into arterial and venous fates based on their localization within the heart. It remains unclear how spatial information controls EC identity and behavior. Here we use single-cell RNA sequencing at key developmental timepoints to interrogate cellular contributions to coronary vessel patterning and maturation. We perform transcriptional profiling to define a heterogenous population of epicardium-derived cells (EPDCs) that express unique chemokine signatures. We identify a population of Slit2+ EPDCs that emerge following epithelial-to- mesenchymal transition (EMT), which we term vascular guidepost cells. We show that the expression of guidepost-derived chemokines such as Slit2 are induced in epicardial cells undergoing EMT, while mesothelium-derived chemokines are silenced. We demonstrate that epicardium-specific deletion of myocardin-related transcription factors in mouse embryos disrupts the expression of key guidance cues and alters EPDC-EC signaling, leading to the persistence of an immature angiogenic EC identity and inappropriate accumulation of ECs on the epicardial surface. Our study suggests that EC pathfinding and fate specification is controlled by a common mechanism and guided by paracrine signaling from EPDCs linking epicardial EMT to EC localization and fate specification in the developing heart.
    [Show full text]