{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,7]],"date-time":"2025-11-07T09:43:58Z","timestamp":1762508638399,"version":"3.41.2"},"reference-count":58,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,10,28]],"date-time":"2021-10-28T00:00:00Z","timestamp":1635379200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006126","name":"Hospital for Sick Children","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100006126","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:p>Inherited defects that abrogate the function of the adenosine deaminase (ADA) enzyme and consequently lead to the accumulation of toxic purine metabolites cause profound lymphopenia and severe combined immune deficiency. Additionally, neutropenia and impaired neutrophil function have been reported among ADA-deficient patients. However, due to the rarity of the disorder, the neutrophil developmental abnormalities and the mechanisms contributing to them have not been characterized. Induced pluripotent stem cells (iPSC) generated from two unrelated ADA-deficient patients and from healthy controls were differentiated through embryoid bodies into neutrophils. ADA deficiency led to a significant reduction in the number of all early multipotent hematopoietic progenitors. At later stages of differentiation, ADA deficiency impeded the formation of granulocyte colonies in methylcellulose cultures, leading to a significant decrease in the number of neutrophils generated from ADA-deficient iPSCs. The viability and apoptosis of ADA-deficient neutrophils isolated from methylcellulose cultures were unaffected, suggesting that the abnormal purine homeostasis in this condition interferes with differentiation or proliferation. Additionally, there was a significant increase in the percentage of hyperlobular ADA-deficient neutrophils, and these neutrophils demonstrated significantly reduced ability to phagocytize fluorescent microspheres. Supplementing iPSCs and methylcellulose cultures with exogenous ADA, which can correct adenosine metabolism, reversed all abnormalities, cementing the critical role of ADA in neutrophil development. Moreover, chemical inhibition of the ribonucleotide reductase (RNR) enzyme, using hydroxyurea or a combination of nicotinamide and trichostatin A in iPSCs from healthy controls, led to abnormal neutrophil differentiation similar to that observed in ADA deficiency, implicating RNR inhibition as a potential mechanism for the neutrophil abnormalities. In conclusion, the findings presented here demonstrate the important role of ADA in the development and function of neutrophils while clarifying the mechanisms responsible for the neutrophil abnormalities in ADA-deficient patients. <\/jats:p>","DOI":"10.3389\/fimmu.2021.748519","type":"journal-article","created":{"date-parts":[[2021,10,28]],"date-time":"2021-10-28T04:44:10Z","timestamp":1635396250000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["The Use of Induced Pluripotent Stem Cells to Study the Effects of Adenosine Deaminase Deficiency on Human Neutrophil Development"],"prefix":"10.3389","volume":"12","author":[{"given":"Michael","family":"Tsui","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weixian","family":"Min","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephanie","family":"Ng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kerry","family":"Dobbs","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luigi D.","family":"Notarangelo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yigal","family":"Dror","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eyal","family":"Grunebaum","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2021,10,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-07-366781","article-title":"Alterations in the Adenosine Metabolism and CD39\/CD73 Adenosinergic Machinery Cause Loss of Treg Cell Function and Autoimmunity in ADA-Deficient SCID","volume":"119","author":"Sauer","year":"2012","journal-title":"Blood"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/j.clim.2019.108321","article-title":"Morbidity in an Adenosine Deaminase-Deficient Patient During 27 Years of Enzyme Replacement Therapy","volume":"211","author":"Grunebaum","year":"2020","journal-title":"Clin Immunol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.74.12.5677","article-title":"Lymphospecific Toxicity in Adenosine Deaminase Deficiency and Purine Nucleoside Phosphorylase Deficiency: Possible Role of Nucleoside Kinase(s)","volume":"74","author":"Carson","year":"1977","journal-title":"Proc Natl Acad Sci USA"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.75.1.472","article-title":"Deoxyadenosine Triphosphate as a Potentially Toxic Metabolite in Adenosine Deaminase Deficiency","volume":"75","author":"Cohen","year":"1978","journal-title":"Proc Natl Acad Sci USA"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1172\/JCI10360","article-title":"Adenosine Deaminase Deficiency Increases Thymic Apoptosis and Causes Defective T Cell Receptor Signaling","volume":"108","author":"Apasov","year":"2001","journal-title":"J Clin Invest"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2012.00265","article-title":"Autoimmune Dysregulation and Purine Metabolism in Adenosine Deaminase Deficiency","volume":"3","author":"Sauer","year":"2012","journal-title":"Front Immunol"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2007-05-092429","article-title":"Altered Intracellular and Extracellular Signaling Leads to Impaired T-Cell Functions in ADA-SCID Patients","volume":"111","author":"Cassani","year":"2008","journal-title":"Blood"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1172\/JCI4320","article-title":"Nucleotide Pool Imbalance and Adenosine Deaminase Deficiency Induce Alterations of N-Region Insertions During V(D)J Recombination","volume":"103","author":"Gangi-Peterson","year":"1999","journal-title":"J Clin Invest"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2012.02.003","article-title":"Pulmonary Alveolar Proteinosis in Patients With Adenosine Deaminase Deficiency","volume":"129","author":"Grunebaum","year":"2012","journal-title":"J Allergy Clin Immunol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1007\/BF02026688","article-title":"Chondroosseous Dysplasia in Severe Combined Immunodeficiency Due to Adenosine Deaminase Deficiency (Chondroosseous Dysplasia in ADA Deficiency SCID)","volume":"21","author":"Chakravarti","year":"1991","journal-title":"Pediatr Radiol"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1016\/j.pediatrneurol.2007.03.011","article-title":"Neurologic Abnormalities in Patients With Adenosine Deaminase Deficiency","volume":"37","author":"Nofech-Mozes","year":"2007","journal-title":"Pediatr Neurol"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpeds.2003.10.055","article-title":"Bilateral Sensorineural Deafness in Adenosine Deaminase-Deficient Severe Combined Immunodeficiency","volume":"144","author":"Albuquerque","year":"2004","journal-title":"J Pediatr"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00416","article-title":"Early Enzyme Replacement Therapy Improves Hearing and Immune Defects in Adenosine Deaminase Deficient-Mice","volume":"10","author":"Xu","year":"2019","journal-title":"Front Immunol"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2018.04.029","article-title":"Neutropenia Among Patients With Adenosine Deaminase Deficiency","volume":"143","author":"Kim","year":"2019","journal-title":"J Allergy Clin Immunol"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-01-329359","article-title":"Myeloid Dysplasia and Bone Marrow Hypocellularity in Adenosine Deaminase-Deficient Severe Combined Immune Deficiency","volume":"118","author":"Sokolic","year":"2011","journal-title":"Blood"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V88.3.1104.1104","article-title":"Granulocyte Colony-Stimulating Factor Recruitment of CD34+ Progenitors to Peripheral Blood: Impaired Mobilization in Chronic Granulomatous Disease and Adenosine Deaminase\u2013Deficient Severe Combined Immunodeficiency Disease Patients","volume":"88","author":"Sekhsaria","year":"1996","journal-title":"Blood"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2018.08.024","article-title":"Consensus Approach for the Management of Severe Combined Immune Deficiency Caused by Adenosine Deaminase Deficiency","volume":"143","author":"Kohn","year":"2019","journal-title":"J Allergy Clin Immunol"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2009-03-209221","article-title":"ADA-Deficient SCID Is Associated With a Specific Microenvironment and Bone Phenotype Characterized by RANKL\/OPG Imbalance and Osteoblast Insufficiency","volume":"114","author":"Sauer","year":"2009","journal-title":"Blood"},{"key":"B19","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1007\/BF01804395","article-title":"Effects of Deoxyadenosine on Ribonucleotide Reductase in Adenosine Deaminase-Deficient Lymphocytes","volume":"14","author":"Takeda","year":"1991","journal-title":"J Inherit Metab Dis"},{"key":"B20","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/b978-0-12-152819-5.50019-1","article-title":"Regulation of Ribonucleotide Reductase","volume":"19","author":"Holmgren","year":"1981","journal-title":"Curr Top Cell Regul"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1586\/14737140.2.4.437","article-title":"Evolving Role of Ribonucleoside Reductase Inhibitors in Hematologic Malignancies","volume":"2","author":"Tsimberidou","year":"2002","journal-title":"Expert Rev Anticancer Ther"},{"key":"B22","doi-asserted-by":"publisher","first-page":"3213","DOI":"10.1038\/s41467-019-11214-9","article-title":"Acetylation Regulates Ribonucleotide Reductase Activity and Cancer Cell Growth","volume":"10","author":"Chen","year":"2019","journal-title":"Nat Commun"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199505183322001","article-title":"Effect of Hydroxyurea on the Frequency of Painful Crises in Sickle Cell Anemia. Investigators of the Multicenter Study of Hydroxyurea in Sickle Cell Anemia","volume":"332","author":"Charache","year":"1995","journal-title":"N Engl J Med"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V94.5.1550","article-title":"Safety of Hydroxyurea in Children With Sickle Cell Anemia: Results of the HUG-KIDS Study, a Phase I\/II Trial","volume":"94","author":"Kinney","year":"1999","journal-title":"Pediatr Hydroxyurea Group Blood"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1136\/adc.81.5.437","article-title":"Preliminary Report of a Toxicity Study of Hydroxyurea in Sickle Cell Disease. French Study Group on Sickle Cell Disease","volume":"81","author":"de Montalembert","year":"1999","journal-title":"Arch Dis Child"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1007\/s00280-010-1481-z","article-title":"Phase II Trial of the Ribonucleotide Reductase Inhibitor 3-Aminopyridine-2-Carboxaldehydethiosemicarbazone Plus Gemcitabine in Patients With Advanced Biliary Tract Cancer","volume":"68","author":"Ocean","year":"2011","journal-title":"Cancer Chemother Pharmacol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1067\/mpd.2001.119590","article-title":"A Two-Year Pilot Trial of Hydroxyurea in Very Young Children With Sickle-Cell Anemia","volume":"139","author":"Wang","year":"2001","journal-title":"J Pediatr"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2003-07-2475","article-title":"Sustained Long-Term Hematologic Efficacy of Hydroxyurea at Maximum Tolerated Dose in Children With Sickle Cell Disease","volume":"103","author":"Zimmerman","year":"2004","journal-title":"Blood"},{"key":"B29","doi-asserted-by":"publisher","first-page":"1392","DOI":"10.1182\/blood-2014-06-583807","article-title":"Nuclear Hypersegmentation of Neutrophils, Eosinophils, and Basophils Due to Hydroxycarbamide (Hydroxyurea)","volume":"124","author":"Xu","year":"2014","journal-title":"Blood"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1111\/bcpt.12487","article-title":"The Effect of a Selective Inhibitor of Phosphodiesterase-9 on Oxidative Stress, Inflammation and Cytotoxicity in Neutrophils From Patients With Sickle Cell Anaemia","volume":"118","author":"Barbosa","year":"2016","journal-title":"Basic Clin Pharmacol Toxicol"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20141286","article-title":"Reticular Dysgenesis-Associated AK2 Protects Hematopoietic Stem and Progenitor Cell Development From Oxidative Stress","volume":"212","author":"Rissone","year":"2015","journal-title":"J Exp Med"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1172\/JCI80924","article-title":"Pathogenesis of ELANE-Mutant Severe Neutropenia Revealed by Induced Pluripotent Stem Cells","volume":"125","author":"Nayak","year":"2015","journal-title":"J Clin Invest"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1002\/humu.1380020415","article-title":"Two New Mutations at the Adenosine Deaminase (ADA) Locus (Q254X and Del Nt1050-54) Unusual for Not Being Missense Mutations","volume":"2","author":"Hirschhorn","year":"1993","journal-title":"Hum Mutat"},{"key":"B34","doi-asserted-by":"publisher","first-page":"1417","DOI":"10.1016\/j.jaci.2011.04.014","article-title":"Purine Metabolism, Immune Reconstitution, and Abdominal Adipose Tumor After Gene Therapy for Adenosine Deaminase Deficiency","volume":"127","author":"Grunebaum","year":"2011","journal-title":"J Allergy Clin Immunol"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1086\/302054","article-title":"Adenosine Deaminase Deficiency: Genotype-Phenotype Correlations Based on Expressed Activity of 29 Mutant Alleles","volume":"63","author":"Arredondo-Vega","year":"1998","journal-title":"Am J Hum Genet"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2008.07.041","article-title":"Disease-Specific Induced Pluripotent Stem Cells","volume":"134","author":"Park","year":"2008","journal-title":"Cell"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1111\/cei.13420","article-title":"Comparison of Elapegademase and Pegademase in ADA-Deficient Patients and Mice","volume":"200","author":"Murguia-Favela","year":"2020","journal-title":"Clin Exp Immunol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1002\/stem.2332","article-title":"Molecular Analysis of Neutrophil Differentiation From Human Induced Pluripotent Stem Cells Delineates the Kinetics of Key Regulators of Hematopoiesis","volume":"34","author":"Sweeney","year":"2016","journal-title":"Stem Cells"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0801812","article-title":"Chemotaxis and Phagocytosis in Neutrophils Is Independent of Coronin 1","volume":"182","author":"Combaluzier","year":"2009","journal-title":"J Immunol"},{"key":"B40","doi-asserted-by":"publisher","first-page":"189","DOI":"10.1007\/978-1-62703-845-4_12","article-title":"Generation of Functionally Mature Neutrophils From Induced Pluripotent Stem Cells","volume":"1124","author":"Sweeney","year":"2014","journal-title":"Methods Mol Biol"},{"key":"B41","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1002\/0471142956.cy0931s51","article-title":"Measurement of Phagocytosis and of the Phagosomal Environment in Polymorphonuclear Phagocytes by Flow Cytometry","volume":"9","author":"Simons","year":"2010","journal-title":"Curr Protoc Cytom"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.1084\/jem.192.2.159","article-title":"Metabolic Consequences of Adenosine Deaminase Deficiency in Mice Are Associated With Defects in Alveogenesis, Pulmonary Inflammation, and Airway Obstruction","volume":"192","author":"Blackburn","year":"2000","journal-title":"J Exp Med"},{"key":"B43","doi-asserted-by":"publisher","first-page":"1467","DOI":"10.1016\/j.jaci.2013.11.029","article-title":"Pulmonary Alveolar Proteinosis in Adenosine Deaminase-Deficient Mice","volume":"133","author":"Dhanju","year":"2014","journal-title":"J\u00a0Allergy Clin Immunol"},{"key":"B44","article-title":"Role of Adenosine Deaminase in the Early Stages of Precursor T Cell Maturation","volume":"44","author":"Shore","year":"1981","journal-title":"Clin Exp Immunol"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20141528","article-title":"Adenosine Signaling Promotes Hematopoietic Stem and Progenitor Cell Emergence","volume":"212","author":"Jing","year":"2015","journal-title":"J\u00a0Exp Med"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2012-02-408591","article-title":"Gene Therapy\/Bone Marrow Transplantation in ADA-Deficient Mice: Roles of Enzyme-Replacement Therapy and Cytoreduction","volume":"120","author":"Carbonaro","year":"2012","journal-title":"Blood"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.18388\/abp.2003_3627","article-title":"Degradation of Extracellular Nucleotides and Their Analogs in HeLa and HUVEC Cell Cultures","volume":"50","author":"Gendaszewska-Darmach","year":"2003","journal-title":"Acta Biochim Pol"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.18.8373","article-title":"P53 Expression is Required for Thymocyte Apoptosis Induced by Adenosine Deaminase Deficiency","volume":"92","author":"Benveniste","year":"1995","journal-title":"Proc Natl Acad Sci USA"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.111.226845","article-title":"Regulation of Neutrophil Function by Adenosine","volume":"32","author":"Barletta","year":"2012","journal-title":"Arterioscler Thromb Vasc Biol"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0903647","article-title":"Adenosine Deaminase Modulation of Telomerase Activity and Replicative Senescence in Human CD8 T Lymphocytes","volume":"184","author":"Parish","year":"2010","journal-title":"J Immunol"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1098\/rsob.120134","article-title":"Neutrophil Heterogeneity in Health and Disease: A Revitalized Avenue in Inflammation and Immunity","volume":"2","author":"Beyrau","year":"2012","journal-title":"Open Biol"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.0505249","article-title":"Differential Expression of Adenosine Receptors in Human Neutrophils: Up-Regulation by Specific Th1 Cytokines and Lipopolysaccharide","volume":"79","author":"Fortin","year":"2006","journal-title":"J Leukoc Biol"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.116.021311","article-title":"Beneficial Role of Erythrocyte Adenosine A2B Receptor-Mediated AMP-Activated Protein Kinase Activation in High-Altitude Hypoxia","volume":"134","author":"Liu","year":"2016","journal-title":"Circulation"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)43528-4","article-title":"Adenosine Deaminase Impairment and Ribonucleotide Reductase Activity and Levels in HeLa Cells","volume":"255","author":"Lin","year":"1980","journal-title":"J Biol Chem"},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.1038\/nm.1913","article-title":"NAMPT Is Essential for the G-CSF-Induced Myeloid Differentiation via a NAD(+)-Sirtuin-1-Dependent Pathway","volume":"15","author":"Skokowa","year":"2009","journal-title":"Nat Med"},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.03.024","article-title":"Nicotinamide Riboside Promotes Sir2 Silencing and Extends Lifespan via Nrk and Urh1\/Pnp1\/Meu1 Pathways to NAD+","volume":"129","author":"Belenky","year":"2007","journal-title":"Cell"},{"key":"B57","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1046\/j.1365-2249.2003.02031.x","article-title":"Nicotinamide Is a Potent Inhibitor of Proinflammatory Cytokines","volume":"131","author":"Ungerstedt","year":"2003","journal-title":"Clin Exp Immunol"},{"key":"B58","doi-asserted-by":"publisher","DOI":"10.1002\/sctm.18-0255","article-title":"Neutrophils Derived From Genetically Modified Human Induced Pluripotent Stem Cells Circulate and Phagocytose Bacteria In Vivo","volume":"8","author":"Trump","year":"2019","journal-title":"Stem Cells Transl Med"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2021.748519\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,10,28]],"date-time":"2021-10-28T04:44:16Z","timestamp":1635396256000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2021.748519\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,28]]},"references-count":58,"alternative-id":["10.3389\/fimmu.2021.748519"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2021.748519","relation":{},"ISSN":["1664-3224"],"issn-type":[{"type":"electronic","value":"1664-3224"}],"subject":[],"published":{"date-parts":[[2021,10,28]]},"article-number":"748519"}}