DOCSLIB.ORG

Glycosylation of Recombinant Anticancer Therapeutics in Different Expression Systems with Emerging Technologies

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Glycosylation of Recombinant Anticancer Therapeutics in Different Expression Systems with Emerging Technologies Published OnlineFirst May 22, 2018; DOI: 10.1158/0008-5472.CAN-18-0032 Cancer Review Research Glycosylation of Recombinant Anticancer Therapeutics in Different Expression Systems with Emerging Technologies Tariq Nadeem1, Mohsin Ahmad Khan1, Bushra Ijaz1, Nadeem Ahmed1, Zia ur Rahman1, Muhammad Shahzad Latif1, Qurban Ali1,2, and Muhammad Adeel Rana3 Abstract Glycosylation, a posttranslational modification, has a major system to obtain structurally and functionally identical glycans, role in recombinant anticancer therapeutic proteins, as most of as in humans. In many expression systems, the N-glycosylation the approved recombinant therapeutics are glycoproteins. The pathway remains conserved in the endoplasmic reticulum, constant amino acid sequence of therapeutics determines the but divergence is observed when the protein enters the Golgi enzymatic activity, while the presence of glycans influences complex. Hence, in recent decades, numerous approaches their pharmacokinetics, solubility, distribution, serum half-life, have been adopted to engineer the Golgi's N-glycosylation effector function, and binding to receptors. Glycoproteins pathway to attain human-like glycans. Several researchers have expressed in different expression systems acquire their own tried to engineer the N-glycosylation pathway of expression oligosaccharides, which increases the protein diversity. The systems. In this review, we examine the glycosylation pattern heterogeneity of glycans creates hurdles in downstream proces- in various expression systems, along with emerging technologies sing, ultimately leading to variable anticancer therapeutic effi- for glycosylation engineering of anticancer therapeutic drugs. cacy. Therefore, glycoproteins require an appropriate expression Cancer Res; 78(11); 1–12. Ó2018 AACR. Introduction ket value of protein-based drugs is growing, with a compounded annual growth rate of 16% compared with the pharmaceutical Cancer is the second leading cause of death in humans, devour- market growth rate of 8% (9). Among the total approved bio- ing the lives of 8.8 million people in 2015 (1, 2). In 2025, 19.3 pharmaceuticals, almost 70% are glycoproteins, which contain million new cases are predicted (3). This disease is characterized carbohydrate moieties gained as a posttranslational modification by abnormal and uncontrolled growth of cells, which have the in the process of glycosylation (10–13). This glycosylation diver- potential to invade other parts of the body through metastasis sifies the class of biopharmaceuticals. Many functions of antican- (4, 5). Currently, most common cancer treatments include cer glycoproteins are associated with glycan attachments, such as radiotherapy, surgery, and chemotherapy. With the advancement solubility, pharmacodistribution, pharmacokinetics, proper of technologies, efforts are being made in clinical treatment to structural folding, binding to receptors, and serum half-life (14). identify effective state-of-the-art therapies to replace conventional The most significant anticancer therapeutic recombinant pro- methods (6, 7). teins are mAbs, which are glycosylated in their Fc region (15). Recent advances have paved the way for the development of Alteration of the composition and structure of glycans causes recombinant anticancer therapeutics through engineered cell conformational changes in the Fc domain of antibodies, affecting lines. As anticancerous agents, these drugs improve the delivery their binding affinity to Fcg receptors (16, 17). This process leads of immune cells to tumor tissues, altering the tumor microenvi- to a change in immune effector functions, including complement- ronment, enhancing antigen priming, and facilitating effector cell dependent cytotoxicity, antibody-dependent cell-mediated cyto- activation and maturation (6, 7). Production of anticancer ther- toxicity (ADCC), and antibody-dependent cell-mediated phago- apeutic proteins as a class of drugs is dominating the drug cytosis (18). Deglycosylation of antibodies reduces their binding industry, partly because of the high demand and partly because affinity and hence their effector functions (19, 20). Changes in the of advancements in recombinant DNA technology (8). The mar- glycoforms of therapeutic mAbs or Fc-fusion proteins can impact the pharmacokinetics of proteins; for example, the negative impact of hypermannosylation on pharmacokinetics can trigger 1Center of Excellence in Molecular Biology, University of the Punjab, Lahore, the C-type lectin clearance mechanism (15, 18, 21). In many cases, Pakistan. 2Institute of Molecular Biology and Biotechnology, University of the terminal sugars in the glycans can affect the pharmacokinetics 3 Lahore, Lahore, Pakistan. Department of Microbiology, Quaid-I-Azam Univer- of an antibody due to glycan binding to receptors on tissues, sity, Islamabad, Pakistan. ultimately leading to its removal from circulation. The major Corresponding Authors: Qurban Ali, Center of Excellence in Molecular Biology, glycan receptors that remove glycoproteins are the mannose University of the Punjab, Lahore 57300, Pakistan. Phone: 321-962-1929; E-mail: receptor and the asialoglycoprotein receptor (22, 23). As both [email protected]; and Tariq Nadeem, [email protected] these receptors are abundantly expressed in the liver, it is likely doi: 10.1158/0008-5472.CAN-18-0032 that glycoproteins with terminal mannose or galactose residues Ó2018 American Association for Cancer Research. will be distributed predominantly in the liver and be catabolized www.aacrjournals.org OF1 Downloaded from cancerres.aacrjournals.org on September 27, 2021. © 2018 American Association for Cancer Research. Published OnlineFirst May 22, 2018; DOI: 10.1158/0008-5472.CAN-18-0032 Nadeem et al. there, as shown by Wright and colleagues in the case of an IgG with their effector functions. This article reviews N-glycosylation occur- terminal mannose or galactose residues (24). ring in different expression systems, and we have summarized the Development of recombinant anticancer therapeutics has various strategies adopted in different glycosylation engineering made substantial progress for the treatment of various solid and technologies. hematologic tumors over the past decade (25–27). Naked mAbs are most commonly used to treat cancer. Rituximab was the first N-glycosylation in the Endoplasmic fi recombinant mAb approved by FDA in 1997 and nds its ther- Reticulum apeutic applications in variety of hematologic cancers including lymphocytic leukemia and non-Hodgkin lymphoma. It is an Eukaryotic N-glycosylation occurs in two organelles, the endo- anti-CD20 humanized recombinant drug that plays a pivotal role plasmic reticulum (ER) and the Golgi complex. Dolichol-linked in B-cell malignancies (28–30). On the other hand, trastuzumab glycan precursor formation and transfer to a nascent protein with is one of the mAbs used for the treatment of solid tumor that is a little bit of processing occurs in the ER, while complete matu- þ capable of ADCC through interactions with Fcg/R immune cell ration and processing of N-linked glycans occurs in the Golgi subsets. It has transformed the treatment of HER-2–positive breast apparatus. N-glycosylation occurs at asparagine residue in the cancer (31–33). Tumor targeted recombinant mAbs can also be consensus amino acid sequence (Asn-X-Ser/Thr), where X can be conjugated to other forms of anticancer therapy that enhances any amino acid, but not proline. The process is initially started and their efficacy by lessening the systemic toxicities to normal cells. processed in the ER, where oligosaccharide transferase (OT) There are three types conjugated mAbs: radiolabeled that are catalyzes the transfer of glycan onto the nascent protein (48). attached to radionuclide moieties, chemolabeled that are linked Secretory proteins containing signal peptides are directed by to antineoplastic drugs, and immunotoxin mAbs that are associ- signal recognition particles across the membrane into the lumen ated with bacterial and plant toxins (34–36). Revolution in of the ER, followed by its movement into the OT-mediated recombinant DNA technology has facilitated the progress toward glycosylation machinery. Glycosylation is not dependent on more specific and less toxic anticancer therapy (29). protein folding or tertiary structure. However, some evidence has In eukaryotic organisms, N-glycosylation is the most prevalent shown that secondary structures on both sides of the Asn con- type of glycosylation, in which a preassembled oligosaccharide is sensus sequences may help in this enzymatic reaction (48). The transferred onto asparagine (Asn) in the consensus sequence of whole process of glycosylation is completed in the ER and Golgi the nascent protein. This oligosaccharide processing and matu- bodies. Glycoproteins processed in the ER usually show homol- ration occurs regardless of the protein template (12, 37–40). ogy and remain conserved in higher eukaryotes and yeast (49). Hence, cancer glycoproteins expressed in different expression Tetradecasaccharide (Glc3Man9GlcNAc2b1) attached to Asn- systems acquire glycans depending on their own glycosylation amide groups is derived from the dolichol pathway (50). The machinery. Glycoproteins expressed in yeast show hypermanno- synthesis of tetradecasaccharide starts on the cytosolic face of the sylated glycans, which compromise their therapeutic efficacy ER by transferring GlcNAc onto the membrane-anchored Dol-P, (41–45). Similarly, glycoproteins expressed in plant cells acquire
Load more

Recommended publications
  • A Systematic Review on the Implications of O-Linked Glycan Branching and Truncating Enzymes on Cancer Progression and Metastasis
    cells Review A Systematic Review on the Implications of O-linked Glycan Branching and Truncating Enzymes on Cancer Progression and Metastasis 1, 1, 1 1,2,3, Rohitesh Gupta y, Frank Leon y, Sanchita Rauth , Surinder K. Batra * and Moorthy P. Ponnusamy 1,2,* 1 Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE 68105, USA; [email protected] (R.G.); [email protected] (F.L.); [email protected] (S.R.) 2 Fred and Pamela Buffett Cancer Center, Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE 681980-5900, USA 3 Department of Pathology and Microbiology, UNMC, Omaha, NE 68198-5900, USA * Correspondence: [email protected] (S.K.B.); [email protected] (M.P.P.); Tel.: +402-559-5455 (S.K.B.); +402-559-1170 (M.P.P.); Fax: +402-559-6650 (S.K.B. & M.P.P.) Equal contribution. y Received: 21 January 2020; Accepted: 12 February 2020; Published: 14 February 2020 Abstract: Glycosylation is the most commonly occurring post-translational modifications, and is believed to modify over 50% of all proteins. The process of glycan modification is directed by different glycosyltransferases, depending on the cell in which it is expressed. These small carbohydrate molecules consist of multiple glycan families that facilitate cell–cell interactions, protein interactions, and downstream signaling. An alteration of several types of O-glycan core structures have been implicated in multiple cancers, largely due to differential glycosyltransferase expression or activity. Consequently, aberrant O-linked glycosylation has been extensively demonstrated to affect biological function and protein integrity that directly result in cancer growth and progression of several diseases.
    [Show full text]
  • Glycosylation: Rising Potential for Prostate Cancer Evaluation
    cancers Review Glycosylation: Rising Potential for Prostate Cancer Evaluation Anna Kałuza˙ * , Justyna Szczykutowicz and Mirosława Ferens-Sieczkowska Department of Chemistry and Immunochemistry, Wroclaw Medical University, Sklodowskiej-Curie 48/50, 50-369 Wroclaw, Poland; [email protected] (J.S.); [email protected] (M.F.-S.) * Correspondence: [email protected]; Tel.: +48-71-770-30-66 Simple Summary: Aberrant protein glycosylation is a well-known hallmark of cancer and is as- sociated with differential expression of enzymes such as glycosyltransferases and glycosidases. The altered expression of the enzymes triggers cancer cells to produce glycoproteins with specific cancer-related aberrations in glycan structures. Increasing number of data indicate that glycosylation patterns of PSA and other prostate-originated proteins exert a potential to distinguish between benign prostate disease and cancer as well as among different stages of prostate cancer development and aggressiveness. This review summarizes the alterations in glycan sialylation, fucosylation, truncated O-glycans, and LacdiNAc groups outlining their potential applications in non-invasive diagnostic procedures of prostate diseases. Further research is desired to develop more general algorithms exploiting glycobiology data for the improvement of prostate diseases evaluation. Abstract: Prostate cancer is the second most commonly diagnosed cancer among men. Alterations in protein glycosylation are confirmed to be a reliable hallmark of cancer. Prostate-specific antigen is the biomarker that is used most frequently for prostate cancer detection, although its lack of sensitivity and specificity results in many unnecessary biopsies. A wide range of glycosylation alterations in Citation: Kałuza,˙ A.; Szczykutowicz, prostate cancer cells, including increased sialylation and fucosylation, can modify protein function J.; Ferens-Sieczkowska, M.
    [Show full text]
  • Yeast Genome Gazetteer P35-65
    gazetteer Metabolism 35 tRNA modification mitochondrial transport amino-acid metabolism other tRNA-transcription activities vesicular transport (Golgi network, etc.) nitrogen and sulphur metabolism mRNA synthesis peroxisomal transport nucleotide metabolism mRNA processing (splicing) vacuolar transport phosphate metabolism mRNA processing (5’-end, 3’-end processing extracellular transport carbohydrate metabolism and mRNA degradation) cellular import lipid, fatty-acid and sterol metabolism other mRNA-transcription activities other intracellular-transport activities biosynthesis of vitamins, cofactors and RNA transport prosthetic groups other transcription activities Cellular organization and biogenesis 54 ionic homeostasis organization and biogenesis of cell wall and Protein synthesis 48 plasma membrane Energy 40 ribosomal proteins organization and biogenesis of glycolysis translation (initiation,elongation and cytoskeleton gluconeogenesis termination) organization and biogenesis of endoplasmic pentose-phosphate pathway translational control reticulum and Golgi tricarboxylic-acid pathway tRNA synthetases organization and biogenesis of chromosome respiration other protein-synthesis activities structure fermentation mitochondrial organization and biogenesis metabolism of energy reserves (glycogen Protein destination 49 peroxisomal organization and biogenesis and trehalose) protein folding and stabilization endosomal organization and biogenesis other energy-generation activities protein targeting, sorting and translocation vacuolar and lysosomal
    [Show full text]
  • Congenital Disorders of Glycosylation from a Neurological Perspective
    brain sciences Review Congenital Disorders of Glycosylation from a Neurological Perspective Justyna Paprocka 1,* , Aleksandra Jezela-Stanek 2 , Anna Tylki-Szyma´nska 3 and Stephanie Grunewald 4 1 Department of Pediatric Neurology, Faculty of Medical Science in Katowice, Medical University of Silesia, 40-752 Katowice, Poland 2 Department of Genetics and Clinical Immunology, National Institute of Tuberculosis and Lung Diseases, 01-138 Warsaw, Poland; [email protected] 3 Department of Pediatrics, Nutrition and Metabolic Diseases, The Children’s Memorial Health Institute, W 04-730 Warsaw, Poland; [email protected] 4 NIHR Biomedical Research Center (BRC), Metabolic Unit, Great Ormond Street Hospital and Institute of Child Health, University College London, London SE1 9RT, UK; [email protected] * Correspondence: [email protected]; Tel.: +48-606-415-888 Abstract: Most plasma proteins, cell membrane proteins and other proteins are glycoproteins with sugar chains attached to the polypeptide-glycans. Glycosylation is the main element of the post- translational transformation of most human proteins. Since glycosylation processes are necessary for many different biological processes, patients present a diverse spectrum of phenotypes and severity of symptoms. The most frequently observed neurological symptoms in congenital disorders of glycosylation (CDG) are: epilepsy, intellectual disability, myopathies, neuropathies and stroke-like episodes. Epilepsy is seen in many CDG subtypes and particularly present in the case of mutations
    [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]
  • Molecular Diagnostic Requisition
    BAYLOR MIRACA GENETICS LABORATORIES SHIP TO: Baylor Miraca Genetics Laboratories 2450 Holcombe, Grand Blvd. -Receiving Dock PHONE: 800-411-GENE | FAX: 713-798-2787 | www.bmgl.com Houston, TX 77021-2024 Phone: 713-798-6555 MOLECULAR DIAGNOSTIC REQUISITION PATIENT INFORMATION SAMPLE INFORMATION NAME: DATE OF COLLECTION: / / LAST NAME FIRST NAME MI MM DD YY HOSPITAL#: ACCESSION#: DATE OF BIRTH: / / GENDER (Please select one): FEMALE MALE MM DD YY SAMPLE TYPE (Please select one): ETHNIC BACKGROUND (Select all that apply): UNKNOWN BLOOD AFRICAN AMERICAN CORD BLOOD ASIAN SKELETAL MUSCLE ASHKENAZIC JEWISH MUSCLE EUROPEAN CAUCASIAN -OR- DNA (Specify Source): HISPANIC NATIVE AMERICAN INDIAN PLACE PATIENT STICKER HERE OTHER JEWISH OTHER (Specify): OTHER (Please specify): REPORTING INFORMATION ADDITIONAL PROFESSIONAL REPORT RECIPIENTS PHYSICIAN: NAME: INSTITUTION: PHONE: FAX: PHONE: FAX: NAME: EMAIL (INTERNATIONAL CLIENT REQUIREMENT): PHONE: FAX: INDICATION FOR STUDY SYMPTOMATIC (Summarize below.): *FAMILIAL MUTATION/VARIANT ANALYSIS: COMPLETE ALL FIELDS BELOW AND ATTACH THE PROBAND'S REPORT. GENE NAME: ASYMPTOMATIC/POSITIVE FAMILY HISTORY: (ATTACH FAMILY HISTORY) MUTATION/UNCLASSIFIED VARIANT: RELATIONSHIP TO PROBAND: THIS INDIVIDUAL IS CURRENTLY: SYMPTOMATIC ASYMPTOMATIC *If family mutation is known, complete the FAMILIAL MUTATION/ VARIANT ANALYSIS section. NAME OF PROBAND: ASYMPTOMATIC/POPULATION SCREENING RELATIONSHIP TO PROBAND: OTHER (Specify clinical findings below): BMGL LAB#: A COPY OF ORIGINAL RESULTS ATTACHED IF PROBAND TESTING WAS PERFORMED AT ANOTHER LAB, CALL TO DISCUSS PRIOR TO SENDING SAMPLE. A POSITIVE CONTROL MAY BE REQUIRED IN SOME CASES. REQUIRED: NEW YORK STATE PHYSICIAN SIGNATURE OF CONSENT I certify that the patient specified above and/or their legal guardian has been informed of the benefits, risks, and limitations of the laboratory test(s) requested.
    [Show full text]
  • ALG6-CDG in South Africa: Genotype-Phenotype Description of Five Novel Patients
    JIMD Reports DOI 10.1007/8904_2012_150 CASE REPORT ALG6-CDG in South Africa: Genotype-Phenotype Description of Five Novel Patients M. Dercksen • A. C. Crutchley • E. M. Honey • M. M. Lippert • G. Matthijs • L. J. Mienie • H. C. Schuman • B. C. Vorster • J. Jaeken Received: 11 October 2011 /Revised: 30 April 2012 /Accepted: 07 May 2012 # SSIEM and Springer-Verlag Berlin Heidelberg 2012 Abstract ALG6-CDG (formerly named CDG-Ic) (pheno- for the known c.998C>T (p.A333V) mutation and the novel type OMIM 603147, genotype OMIM 604566), is caused by c.1338dupA (p.V447SfsX44) mutation. Four more patients, defective endoplasmic reticulum a-1,3-glucosyltransferase presenting with classical neurological involvement were (E.C 2.4.1.267) in the N-glycan assembly pathway identified and were compound heterozygous for the known (Grunewald€ et al. 2000). It is the second most frequent c.257 + 5G>A splice mutation and the c.680G>A(p.G227E) N-glycosylation disorder after PMM2-CDG; some 37 patients missense mutation. The patients belong to a semi-isolated have been reported with 21 different ALG6 gene mutations Caucasian community that may have originated from Euro- (Haeuptle & Hennet 2009; Al-Owain 2010). We report on the pean pioneers who colonized South Africa in the seventeenth/ clinical and biochemical findings of five novel Caucasian eighteenth centuries. South African patients. The first patient had a severe neuro- gastrointestinal presentation. He was compound heterozygous Introduction Communicated by: Eva Morava, MD PhD ALG6-CG (phenotype OMIM 603147, genotype OMIM Competing interests: none declared : : 604566) is a genetic disorder in the assembly of N-glycans M.
    [Show full text]
  • Viruses Like Sugars: How to Assess Glycan Involvement in Viral Attachment
    microorganisms Review Viruses Like Sugars: How to Assess Glycan Involvement in Viral Attachment Gregory Mathez and Valeria Cagno * Institute of Microbiology, Lausanne University Hospital, University of Lausanne, 1011 Lausanne, Switzerland; [email protected] * Correspondence: [email protected] Abstract: The first step of viral infection requires interaction with the host cell. Before finding the specific receptor that triggers entry, the majority of viruses interact with the glycocalyx. Identifying the carbohydrates that are specifically recognized by different viruses is important both for assessing the cellular tropism and for identifying new antiviral targets. Advances in the tools available for studying glycan–protein interactions have made it possible to identify them more rapidly; however, it is important to recognize the limitations of these methods in order to draw relevant conclusions. Here, we review different techniques: genetic screening, glycan arrays, enzymatic and pharmacological approaches, and surface plasmon resonance. We then detail the glycan interactions of enterovirus D68 and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), highlighting the aspects that need further clarification. Keywords: attachment receptor; viruses; glycan; sialic acid; heparan sulfate; HBGA; SARS-CoV-2; EV-D68 Citation: Mathez, G.; Cagno, V. Viruses Like Sugars: How to Assess 1. Introduction Glycan Involvement in Viral This review focuses on methods for assessing the involvement of carbohydrates in Attachment. Microorganisms 2021, 9, viral attachment and entry into the host cell. Viruses often bind to entry receptors that are 1238. https://doi.org/10.3390/ not abundant on the cell surface; to increase their chances of finding them, they initially microorganisms9061238 bind to attachment receptors comprising carbohydrates that are more widely expressed.
    [Show full text]
  • Prenatal Testing Requisition Form
    BAYLOR MIRACA GENETICS LABORATORIES SHIP TO: Baylor Miraca Genetics Laboratories 2450 Holcombe, Grand Blvd. -Receiving Dock PHONE: 800-411-GENE | FAX: 713-798-2787 | www.bmgl.com Houston, TX 77021-2024 Phone: 713-798-6555 PRENATAL COMPREHENSIVE REQUISITION FORM PATIENT INFORMATION NAME (LAST,FIRST, MI): DATE OF BIRTH (MM/DD/YY): HOSPITAL#: ACCESSION#: REPORTING INFORMATION ADDITIONAL PROFESSIONAL REPORT RECIPIENTS PHYSICIAN: NAME: INSTITUTION: PHONE: FAX: PHONE: FAX: NAME: EMAIL (INTERNATIONAL CLIENT REQUIREMENT): PHONE: FAX: SAMPLE INFORMATION CLINICAL INDICATION FETAL SPECIMEN TYPE Pregnancy at risk for specific genetic disorder DATE OF COLLECTION: (Complete FAMILIAL MUTATION information below) Amniotic Fluid: cc AMA PERFORMING PHYSICIAN: CVS: mg TA TC Abnormal Maternal Screen: Fetal Blood: cc GESTATIONAL AGE (GA) Calculation for AF-AFP* NTD TRI 21 TRI 18 Other: SELECT ONLY ONE: Abnormal NIPT (attach report): POC/Fetal Tissue, Type: TRI 21 TRI 13 TRI 18 Other: Cultured Amniocytes U/S DATE (MM/DD/YY): Abnormal U/S (SPECIFY): Cultured CVS GA ON U/S DATE: WKS DAYS PARENTAL BLOODS - REQUIRED FOR CMA -OR- Maternal Blood Date of Collection: Multiple Pregnancy Losses LMP DATE (MM/DD/YY): Parental Concern Paternal Blood Date of Collection: Other Indication (DETAIL AND ATTACH REPORT): *Important: U/S dating will be used if no selection is made. Name: Note: Results will differ depending on method checked. Last Name First Name U/S dating increases overall screening performance. Date of Birth: KNOWN FAMILIAL MUTATION/DISORDER SPECIFIC PRENATAL TESTING Notice: Prior to ordering testing for any of the disorders listed, you must call the lab and discuss the clinical history and sample requirements with a genetic counselor.
    [Show full text]
  • Glycosylation of Immune Receptors in Cancer
    cells Review Glycosylation of Immune Receptors in Cancer Ruoxuan Sun, Alyssa Min Jung Kim and Seung-Oe Lim * Department of Medicinal Chemistry and Molecular Pharmacology, Purdue Institute of Drug Discovery, Purdue Center for Cancer Research, Purdue University, West Lafayette, IN 47907, USA; [email protected] (R.S.); [email protected] (A.M.J.K.) * Correspondence: [email protected]; Tel.: +1-765-494-3531 Abstract: Evading host immune surveillance is one of the hallmarks of cancer. Immune checkpoint therapy, which aims to eliminate cancer progression by reprogramming the antitumor immune response, currently occupies a solid position in the rapidly expanding arsenal of cancer therapy. As most immune checkpoints are membrane glycoproteins, mounting attention is drawn to asking how protein glycosylation affects immune function. The answers to this fundamental question will stimulate the rational development of future cancer diagnostics and therapeutic strategies. Keywords: glycosylation; N-glycan; immune receptor; immune checkpoint therapy; cancer 1. Introduction The immune system can eliminate malignancy at an early stage by recognizing the antigenic peptide epitope presented by neoplastically transformed cells [1,2]. However, a subset of tumor cells may evolve to coexist with antitumor immunity by a process defined as immunoediting [3]. To combat the immune attack and maintain the balance with the host, cancer cells develop a series of approaches such as (1) downregulation of antigen Citation: Sun, R.; Kim, A.M.J.; Lim, presentation to make them invisible to immunity/the immune system, (2) production S.-O. Glycosylation of Immune of immunomodulatory cytokines to establish an immune suppressive environment, and Receptors in Cancer. Cells 2021, 10, (3) expression of surface immune checkpoint proteins such as programmed cell death 1 1100.
    [Show full text]
  • Design of Glycosylation Sites by Rapid Synthesis and Analysis of Glycosyltransferases
    ARTICLES https://doi.org/10.1038/s41589-018-0051-2 Design of glycosylation sites by rapid synthesis and analysis of glycosyltransferases Weston Kightlinger 1,2,7, Liang Lin2,3,7, Madisen Rosztoczy3, Wenhao Li3, Matthew P. DeLisa 4,5, Milan Mrksich 1,2,3,6* and Michael C. Jewett 1,2* Glycosylation is an abundant post-translational modification that is important in disease and biotechnology. Current methods to understand and engineer glycosylation cannot sufficiently explore the vast experimental landscapes required to accurately predict and design glycosylation sites modified by glycosyltransferases. Here we describe a systematic platform for glycosyl- ation sequence characterization and optimization by rapid expression and screening (GlycoSCORES), which combines cell-free protein synthesis and mass spectrometry of self-assembled monolayers. We produced six N- and O-linked polypeptide-modify- ing glycosyltransferases from bacteria and humans in vitro and rigorously determined their substrate specificities using 3,480 unique peptides and 13,903 unique reaction conditions. We then used GlycoSCORES to optimize and design small glycosylation sequence motifs that directed efficient N-linked glycosylation in vitro and in the Escherichia coli cytoplasm for three heterolo- gous proteins, including the human immunoglobulin Fc domain. We find that GlycoSCORES is a broadly applicable method to facilitate fundamental understanding of glycosyltransferases and engineer synthetic glycoproteins. rotein glycosylation is the post-translational attachment of O-linked polypeptide N-acetylgalactosaminyltransferase (ppGalN- oligosaccharides (glycans), most commonly at asparagine AcT), O-linked N-acetylglucosamine transferase (OGT), and oligo- (N-linked) or serine and threonine (O-linked) amino acid side saccharyltransferase (OST) enzyme families. Such enzymes are of P 1,2 chains .
    [Show full text]
  • Genetic Predisposition to Fetal Alcohol Syndrome: Association with Congenital Disorders of N-Glycosylation
    nature publishing group Basic Science Investigation | Articles Genetic predisposition to fetal alcohol syndrome: association with congenital disorders of N-glycosylation María E. de la Morena-Barrio1, María J. Ballesta-Martínez2, Raquel López-Gálvez1, Ana I. Antón1, Vanessa López-González2, Laia Martínez-Ribot3,JoséPadilla1, Antonia Miñano1, Oscar García-Algar4, Miguel Del Campo5, Javier Corral1, Encarna Guillén-Navarro2 and Vicente Vicente1 BACKGROUND: Fetal alcohol syndrome (FAS) is caused by exposure, with the most serious consequences in those fetuses maternal alcohol consumption during pregnancy; although exposed to high concentrations during the initial weeks of additional factors might be involved, as development and pregnancy (6–9); however, additional factors may be involved severity are not directly related to alcohol intake. The as some exposed pregnancies develop FAS, whereas others do abnormal glycosylation caused by alcohol might play a role not, and the severity of patients is highly heterogeneous. The in FAS according to the clinical similarities shared with estimated prevalence of FAS is 1–3/1,000 live births and congenital disorders of glycosylation (CDG). Thus, mutations constitutes the principal cause of non-inherited mental underlying CDG, affecting genes involved in glycosylation, disability (6,7,10,11). could also be involved in FAS. Diverse pathophysiological mechanisms have been pro- METHODS: A panel of 74 genes involved in N-glycosylation posed to underlie FAS, but none of them completely clarifies was sequenced in 25 FAS patients and 20 controls with this disorder. Recently, a new pathogenetic model suggests prenatal alcohol exposure. Transferrin glycoforms were that glycosylation defects underlie FAS based on the evaluated by HPLC.
    [Show full text]