Adenylosuccinate lyase (ADSL) functions in de novo purine synthesis (DNPS) and the purine nucleotide cycle. ADSL deficiency (ADSLD) causes numerous neurodevelopmental pathologies, including microcephaly and autism spectrum disorder. ADSLD patients have normal serum purine nucleotide levels but exhibit accumulation of dephosphorylated ADSL substrates, S-Ado, and SAICAr, the latter being implicated in neurotoxic effects through unknown mechanisms. We examined the phenotypic effects of ADSL depletion in human cells and their relation to phenotypic outcomes. Using specific interventions to compensate for reduced purine levels or modulate SAICAr accumulation, we found that diminished AMP levels resulted in increased DNA damage signaling and cell cycle delays, while primary ciliogenesis was impaired specifically by loss of ADSL or administration of SAICAr. ADSL-deficient chicken and zebrafish embryos displayed impaired neurogenesis and microcephaly. Neuroprogenitor attrition in zebrafish embryos was rescued by pharmacological inhibition of DNPS, but not increased nucleotide concentration. Zebrafish also displayed phenotypes commonly linked to ciliopathies. Our results suggest that both reduced purine levels and impaired DNPS contribute to neurodevelopmental pathology in ADSLD and that defective ciliogenesis may influence the ADSLD phenotypic spectrum.

CIBER de Diabetes y Enfermedades Metabólicas Asociadas Instituto de Salud Carlos 3 Madrid Spain

Department of Cell Biology Instituto de Biología Molecular de Barcelona Barcelona Spain

Department of Experimental and Clinical Pharmacology and Pharmacogenomics University of Tübingen Tübingen Germany

Department of Paediatrics and Inherited Metabolic Disorders 1st Faculty of Medicine Charles University and General University Hospital Prague Prague Czech Republic

ICREA Barcelona Spain

Institute for Research in Biomedicine The Barcelona Institute of Science and Technology Barcelona Spain

Institute of Biochemistry and Molecular Biology Ulm University Ulm Germany

Institute of Complex Systems Universitat de Barcelona Barcelona Spain

National Cancer Institute Center for Cancer Research Radiation Oncology Branch Bethesda United States

Section of General Linguistics Universitat de Barcelona Barcelona Spain

Universitat Rovira i Virgili Department of Electronic Engineering IISPV Tarragona Spain

Zobrazit více v PubMed

Agarwal S, Chakravarthi B, Behring M, Kim HG, Chandrashekar DS, Gupta N, Bajpai P, Elkholy A, Balasubramanya SAH, Hardy C, Diffalha SA, Varambally S, Manne U. PAICS, a Purine Nucleotide Metabolic Enzyme, is Involved in Tumor Growth and the Metastasis of Colorectal Cancer. Cancers. 2020;12:E772. doi: 10.3390/cancers12040772. PubMed DOI PMC

Al-Dosari MS, Shaheen R, Colak D, Alkuraya FS. Novel CENPJ mutation causes Seckel syndrome. Journal of Medical Genetics. 2010;47:411–414. doi: 10.1136/jmg.2009.076646. PubMed DOI

Bangs F, Anderson KV. Primary Cilia and Mammalian Hedgehog Signaling. Cold Spring Harbor Perspectives in Biology. 2017;9:a028175. doi: 10.1101/cshperspect.a028175. PubMed DOI PMC

Baresova V, Krijt M, Skopova V, Souckova O, Kmoch S, Zikanova M. CRISPR-Cas9 induced mutations along de novo purine synthesis in HeLa cells result in accumulation of individual enzyme substrates and affect purinosome formation. Molecular Genetics and Metabolism. 2016;119:270–277. doi: 10.1016/j.ymgme.2016.08.004. PubMed DOI

Bazzi H, Anderson KV. Acentriolar mitosis activates a p53-dependent apoptosis pathway in the mouse embryo. PNAS. 2014;111:E1491–E1500. doi: 10.1073/pnas.1400568111. PubMed DOI PMC

Benedict B, van Harn T, Dekker M, Hermsen S, Kucukosmanoglu A, Pieters W, Delzenne-Goette E, Dorsman JC, Petermann E, Foijer F, Te Riele H. Loss of p53 suppresses replication-stress-induced DNA breakage in G1/S checkpoint deficient cells. eLife. 2018;7:e37868. doi: 10.7554/eLife.37868. PubMed DOI PMC

Breunig JJ, Sarkisian MR, Arellano JI, Morozov YM, Ayoub AE, Sojitra S, Wang B, Flavell RA, Rakic P, Town T. Primary cilia regulate hippocampal neurogenesis by mediating sonic hedgehog signaling. PNAS. 2008;105:13127–13132. doi: 10.1073/pnas.0804558105. PubMed DOI PMC

Burkhalter MD, Fralish GB, Premont RT, Caron MG, Philipp M. Grk5l controls heart development by limiting mTOR signaling during symmetry breaking. Cell Reports. 2013;4:625–632. doi: 10.1016/j.celrep.2013.07.036. PubMed DOI PMC

Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD. A versatile viral system for expression and depletion of proteins in mammalian cells. PLOS ONE. 2009;4:e6529. doi: 10.1371/journal.pone.0006529. PubMed DOI PMC

Chen P, Wang D, Chen H, Zhou Z, He X. The nonessentiality of essential genes in yeast provides therapeutic insights into a human disease. Genome Research. 2016;26:1355–1362. doi: 10.1101/gr.205955.116. PubMed DOI PMC

Cloëtta D, Thomanetz V, Baranek C, Lustenberger RM, Lin S, Oliveri F, Atanasoski S, Rüegg MA. Inactivation of mTORC1 in the developing brain causes microcephaly and affects gliogenesis. The Journal of Neuroscience. 2013;33:7799–7810. doi: 10.1523/JNEUROSCI.3294-12.2013. PubMed DOI PMC

Daignan-Fornier B, Pinson B. Yeast to Study Human Purine Metabolism Diseases. Cells. 2019;8:E67. doi: 10.3390/cells8010067. PubMed DOI PMC

Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, Naldini L. A third-generation lentivirus vector with a conditional packaging system. Journal of Virology. 1998;72:8463–8471. doi: 10.1128/JVI.72.11.8463-8471.1998. PubMed DOI PMC

Farooq M, Lindbæk L, Krogh N, Doganli C, Keller C, Mönnich M, Gonçalves AB, Sakthivel S, Mang Y, Fatima A, Andersen VS, Hussain MS, Eiberg H, Hansen L, Kjaer KW, Gopalakrishnan J, Pedersen LB, Møllgård K, Nielsen H, Baig SM, Tommerup N, Christensen ST, Larsen LA. RRP7A links primary microcephaly to dysfunction of ribosome biogenesis, resorption of primary cilia, and neurogenesis. Nature Communications. 2020;11:5816. doi: 10.1038/s41467-020-19658-0. PubMed DOI PMC

Fenton AR, Janowitz HN, McReynolds MR, Wang W, Hanna-Rose W. A Caenorhabditis elegans Model of Adenylosuccinate Lyase Deficiency Reveals Neuromuscular and Reproductive Phenotypes of Distinct Etiology. bioRxiv. 2017 doi: 10.1101/181719. PubMed DOI PMC

Fong CS, Mazo G, Das T, Goodman J, Kim M, O’Rourke BP, Izquierdo D, Tsou M-FB. 53BP1 and USP28 mediate p53-dependent cell cycle arrest in response to centrosome loss and prolonged mitosis. eLife. 2016;5:e16270. doi: 10.7554/eLife.16270. PubMed DOI PMC

Gabriel E, Wason A, Ramani A, Gooi LM, Keller P, Pozniakovsky A, Poser I, Noack F, Telugu NS, Calegari F, Šarić T, Hescheler J, Hyman AA, Gottardo M, Callaini G, Alkuraya FS, Gopalakrishnan J. CPAP promotes timely cilium disassembly to maintain neural progenitor pool. The EMBO Journal. 2016;35:803–819. doi: 10.15252/embj.201593679. PubMed DOI PMC

Giaever G, Chu AM, Ni L, Connelly C, Riles L, Véronneau S, Dow S, Lucau-Danila A, Anderson K, André B, Arkin AP, Astromoff A, El-Bakkoury M, Bangham R, Benito R, Brachat S, Campanaro S, Curtiss M, Davis K, Deutschbauer A, Entian K-D, Flaherty P, Foury F, Garfinkel DJ, Gerstein M, Gotte D, Güldener U, Hegemann JH, Hempel S, Herman Z, Jaramillo DF, Kelly DE, Kelly SL, Kötter P, LaBonte D, Lamb DC, Lan N, Liang H, Liao H, Liu L, Luo C, Lussier M, Mao R, Menard P, Ooi SL, Revuelta JL, Roberts CJ, Rose M, Ross-Macdonald P, Scherens B, Schimmack G, Shafer B, Shoemaker DD, Sookhai-Mahadeo S, Storms RK, Strathern JN, Valle G, Voet M, Volckaert G, Wang C, Ward TR, Wilhelmy J, Winzeler EA, Yang Y, Yen G, Youngman E, Yu K, Bussey H, Boeke JD, Snyder M, Philippsen P, Davis RW, Johnston M. Functional profiling of the Saccharomyces cerevisiae genome. Nature. 2002;418:387–391. doi: 10.1038/nature00935. PubMed DOI

Gottifredi V, Shieh S, Taya Y, Prives C. p53 accumulates but is functionally impaired when DNA synthesis is blocked. PNAS. 2001;98:1036–1041. doi: 10.1073/pnas.021282898. PubMed DOI PMC

Gupta GD, Coyaud É, Gonçalves J, Mojarad BA, Liu Y, Wu Q, Gheiratmand L, Comartin D, Tkach JM, Cheung SWT, Bashkurov M, Hasegan M, Knight JD, Lin ZY, Schueler M, Hildebrandt F, Moffat J, Gingras AC, Raught B, Pelletier L. A Dynamic Protein Interaction Landscape of the Human Centrosome-Cilium Interface. Cell. 2015;163:1484–1499. doi: 10.1016/j.cell.2015.10.065. PubMed DOI PMC

Hamburger V, Hamilton HL. A series of normal stages in the development of the chick embryo. Journal of Morphology. 1951;88:49–92. doi: 10.1002/aja.1001950404. PubMed DOI

Han YG, Spassky N, Romaguera-Ros M, Garcia-Verdugo JM, Aguilar A, Schneider-Maunoury S, Alvarez-Buylla A. Hedgehog signaling and primary cilia are required for the formation of adult neural stem cells. Nature Neuroscience. 2008;11:277–284. doi: 10.1038/nn2059. PubMed DOI

Herrera A, Saade M, Menendez A, Marti E, Pons S. Sustained Wnt/β-catenin signalling causes neuroepithelial aberrations through the accumulation of aPKC at the apical pole. Nature Communications. 2014;5:4168. doi: 10.1038/ncomms5168. PubMed DOI

Holzer K, Drucker E, Roessler S, Dauch D, Heinzmann F, Waldburger N, Eiteneuer EM, Herpel E, Breuhahn K, Zender L, Schirmacher P, Ori A, Singer S. Proteomic Analysis Reveals GMP Synthetase as p53 Repression Target in Liver Cancer. The American Journal of Pathology. 2017;187:228–235. doi: 10.1016/j.ajpath.2016.09.022. PubMed DOI

Hoxhaj G, Hughes-Hallett J, Timson RC, Ilagan E, Yuan M, Asara JM, Ben-Sahra I, Manning BD. The mTORC1 Signaling Network Senses Changes in Cellular Purine Nucleotide Levels. Cell Reports. 2017;21:1331–1346. doi: 10.1016/j.celrep.2017.10.029. PubMed DOI PMC

Insolera R, Bazzi H, Shao W, Anderson KV, Shi SH. Cortical neurogenesis in the absence of centrioles. Nature Neuroscience. 2014;17:1528–1535. doi: 10.1038/nn.3831. PubMed DOI PMC

Jaeken J, Van den Berghe G. An infantile autistic syndrome characterised by the presence of succinylpurines in body fluids. Lancet (London, England) 1984;2:1058–1061. doi: 10.1016/S0140-6736(84)91505-8. PubMed DOI

Jaffe KM, Thiberge SY, Bisher ME, Burdine RD. Imaging cilia in zebrafish. Methods in Cell Biology. 2010;97:415–435. doi: 10.1016/S0091-679X(10)97022-2. PubMed DOI

Jurecka A, Zikanova M, Tylki-Szymanska A, Krijt J, Bogdanska A, Gradowska W, Mullerova K, Sykut-Cegielska J, Kmoch S, Pronicka E. Clinical, biochemical and molecular findings in seven Polish patients with adenylosuccinate lyase deficiency. Molecular Genetics and Metabolism. 2008;94:435–442. doi: 10.1016/j.ymgme.2008.04.013. PubMed DOI

Jurecka A, Jurkiewicz E, Tylki-Szymanska A. Magnetic resonance imaging of the brain in adenylosuccinate lyase deficiency: a report of seven cases and a review of the literature. European Journal of Pediatrics. 2012;171:131–138. doi: 10.1007/s00431-011-1503-9. PubMed DOI

Jurecka A, Zikanova M, Kmoch S, Tylki-Szymańska A. Adenylosuccinate lyase deficiency. Journal of Inherited Metabolic Disease. 2015;38:231–242. doi: 10.1007/s10545-014-9755-y. PubMed DOI PMC

Keller KE, Tan IS, Lee YS. SAICAR stimulates pyruvate kinase isoform M2 and promotes cancer cell survival in glucose-limited conditions. Science (New York, N.Y.) 2012;338:1069–1072. doi: 10.1126/science.1224409. PubMed DOI PMC

Keller KE, Doctor ZM, Dwyer ZW, Lee YS. SAICAR induces protein kinase activity of PKM2 that is necessary for sustained proliferative signaling of cancer cells. Molecular Cell. 2014;53:700–709. doi: 10.1016/j.molcel.2014.02.015. PubMed DOI PMC

Kim MP, Zhang Y, Lozano G. Mutant p53: Multiple Mechanisms Define Biologic Activity in Cancer. Frontiers in Oncology. 2015;5:249. doi: 10.3389/fonc.2015.00249. PubMed DOI PMC

Lambrus BG, Daggubati V, Uetake Y, Scott PM, Clutario KM, Sluder G, Holland AJ. A USP28-53BP1-p53-p21 signaling axis arrests growth after centrosome loss or prolonged mitosis. The Journal of Cell Biology. 2016;214:143–153. doi: 10.1083/jcb.201604054. PubMed DOI PMC

Lin YN, Lee YS, Li SK, Tang TK. Loss of CPAP in developing mouse brain and its functional implication for human primary microcephaly. Journal of Cell Science. 2020;133:jcs243592. doi: 10.1242/jcs.243592. PubMed DOI

Mádrová L, Krijt M, Barešová V, Václavík J, Friedecký D, Dobešová D, Součková O, Škopová V, Adam T, Zikánová M. Mass spectrometric analysis of purine de novo biosynthesis intermediates. PLOS ONE. 2018;13:e0208947. doi: 10.1371/journal.pone.0208947. PubMed DOI PMC

Marjanović M, Sánchez-Huertas C, Terré B, Gómez R, Scheel JF, Pacheco S, Knobel PA, Martínez-Marchal A, Aivio S, Palenzuela L, Wolfrum U, McKinnon PJ, Suja JA, Roig I, Costanzo V, Lüders J, Stracker TH. CEP63 deficiency promotes p53-dependent microcephaly and reveals a role for the centrosome in meiotic recombination. Nature Communications. 2015;6:7676. doi: 10.1038/ncomms8676. PubMed DOI PMC

Marsac R, Pinson B, Saint-Marc C, Olmedo M, Artal-Sanz M, Daignan-Fornier B, Gomes JE. Purine Homeostasis Is Necessary for Developmental Timing, Germline Maintenance and Muscle Integrity in Caenorhabditis elegans. Genetics. 2019;211:1297–1313. doi: 10.1534/genetics.118.301062. PubMed DOI PMC

McIntyre RE, Lakshminarasimhan Chavali P, Ismail O, Carragher DM, Sanchez-Andrade G, Forment JV, Fu B, Del Castillo Velasco-Herrera M, Edwards A, van der Weyden L, Yang F, Ramirez-Solis R, Estabel J, Gallagher FA, Logan DW, Arends MJ, Tsang SH, Mahajan VB, Scudamore CL, White JK, Jackson SP, Gergely F, Adams DJ, Sanger Mouse Genetics Project Disruption of mouse Cenpj, a regulator of centriole biogenesis, phenocopies Seckel syndrome. PLOS Genetics. 2012;8:e1003022. doi: 10.1371/journal.pgen.1003022. PubMed DOI PMC

McKeown CR, Cline HT. Nutrient restriction causes reversible G2 arrest in Xenopus neural progenitors. Development (Cambridge, England) 2019;146:dev178871. doi: 10.1242/dev.178871. PubMed DOI PMC

Mouchegh K, Zikánová M, Hoffmann GF, Kretzschmar B, Kühn T, Mildenberger E, Stoltenburg-Didinger G, Krijt J, Dvoráková L, Honzík T, Zeman J, Kmoch S, Rossi R. Lethal fetal and early neonatal presentation of adenylosuccinate lyase deficiency: observation of 6 patients in 4 families. The Journal of Pediatrics. 2007;150:57–61. doi: 10.1016/j.jpeds.2006.09.027. PubMed DOI

Murga M, Bunting S, Montaña MF, Soria R, Mulero F, Cañamero M, Lee Y, McKinnon PJ, Nussenzweig A, Fernandez-Capetillo O. A mouse model of ATR-Seckel shows embryonic replicative stress and accelerated aging. Nature Genetics. 2009;41:891–898. doi: 10.1038/ng.420. PubMed DOI PMC

O’Driscoll M, Ruiz-Perez VL, Woods CG, Jeggo PA, Goodship JA. A splicing mutation affecting expression of ataxia-telangiectasia and Rad3-related protein (ATR) results in seckel syndrome. Nature Genetics. 2003;33:497–501. doi: 10.1038/ng1129. PubMed DOI

Ogungbenro YA, Tena TC, Gaboriau D, Lalor P, Dockery P, Philipp M, Morrison CG. Centrobin controls primary ciliogenesis in vertebrates. The Journal of Cell Biology. 2018;217:1205–1215. doi: 10.1083/jcb.201706095. PubMed DOI PMC

Pantoja C, Serrano M. Murine fibroblasts lacking p21 undergo senescence and are resistant to transformation by oncogenic Ras. Oncogene. 1999;18:4974–4982. doi: 10.1038/sj.onc.1202880. PubMed DOI

Pinson B, Vaur S, Sagot I, Coulpier F, Lemoine S, Daignan-Fornier B. Metabolic intermediates selectively stimulate transcription factor interaction and modulate phosphate and purine pathways. Genes & Development. 2009;23:1399–1407. doi: 10.1101/gad.521809. PubMed DOI PMC

Reddy BA, van der Knaap JA, Bot AGM, Mohd-Sarip A, Dekkers DHW, Timmermans MA, Martens JWM, Demmers JAA, Verrijzer CP. Nucleotide biosynthetic enzyme GMP synthase is a TRIM21-controlled relay of p53 stabilization. Molecular Cell. 2014;53:458–470. doi: 10.1016/j.molcel.2013.12.017. PubMed DOI

Sir J-H, Barr AR, Nicholas AK, Carvalho OP, Khurshid M, Sossick A, Reichelt S, D’Santos C, Woods CG, Gergely F. A primary microcephaly protein complex forms a ring around parental centrioles. Nature Genetics. 2011;43:1147–1153. doi: 10.1038/ng.971. PubMed DOI PMC

Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS, Sabatini DM, Chen ISY, Hahn WC, Sharp PA, Weinberg RA, Novina CD. Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA (New York, N.Y.) 2003;9:493–501. doi: 10.1261/rna.2192803. PubMed DOI PMC

Stone TW, Roberts LA, Morris BJ, Jones PA, Ogilvy HA, Behan WM, Duley JA, Simmonds HA, Vincent MF, van den Berghe G. Succinylpurines induce neuronal damage in the rat brain. Advances in Experimental Medicine and Biology. 1998;431:185–189. doi: 10.1007/978-1-4615-5381-6_36. PubMed DOI

Stracker TH, Morrison CG, Gergely F. Molecular causes of primary microcephaly and related diseases: a report from the UNIA Workshop. Chromosoma. 2020;129:115–120. doi: 10.1007/s00412-020-00737-6. PubMed DOI

Taha-Mehlitz S, Bianco G, Coto-Llerena M, Kancherla V, Bantug GR, Gallon J, Ercan C, Panebianco F, Eppenberger-Castori S, von Strauss M, Staubli S, Bolli M, Peterli R, Matter MS, Terracciano LM, von Flüe M, Ng CKY, Soysal SD, Kollmar O, Piscuoglio S. Adenylosuccinate lyase is oncogenic in colorectal cancer by causing mitochondrial dysfunction and independent activation of NRF2 and mTOR-MYC-axis. Theranostics. 2021;11:4011–4029. doi: 10.7150/thno.50051. PubMed DOI PMC

Tamiya S, Liu L, Kaplan HJ. Epithelial-mesenchymal transition and proliferation of retinal pigment epithelial cells initiated upon loss of cell-cell contact. Investigative Ophthalmology & Visual Science. 2010;51:2755–2763. doi: 10.1167/iovs.09-4725. PubMed DOI

Thisse B, Thisse C. In situ hybridization on whole-mount zebrafish embryos and young larvae. Methods in Molecular Biology (Clifton, N.J.) 2014;1211:53–67. doi: 10.1007/978-1-4939-1459-3_5. PubMed DOI

Vaz R, Hofmeister W, Lindstrand A. Zebrafish Models of Neurodevelopmental Disorders: Limitations and Benefits of Current Tools and Techniques. International Journal of Molecular Sciences. 2019;20:E1296. doi: 10.3390/ijms20061296. PubMed DOI PMC

Walia V, Cuenca A, Vetter M, Insinna C, Perera S, Lu Q, Ritt DA, Semler E, Specht S, Stauffer J, Morrison DK, Lorentzen E, Westlake CJ. Akt Regulates a Rab11-Effector Switch Required for Ciliogenesis. Developmental Cell. 2019;50:229–246. doi: 10.1016/j.devcel.2019.05.022. PubMed DOI PMC

Xavier GM, Seppala M, Barrell W, Birjandi AA, Geoghegan F, Cobourne MT. Hedgehog receptor function during craniofacial development. Developmental Biology. 2016;415:198–215. doi: 10.1016/j.ydbio.2016.02.009. PubMed DOI

Yamauchi T, Miyawaki K, Semba Y, Takahashi M, Izumi Y, Nogami J, Nakao F, Sugio T, Sasaki K, Pinello L, Bauer DE, Bamba T, Akashi K, Maeda T. Targeting leukemia-specific dependence on the de novo purine synthesis pathway. Leukemia. 2022;36:383–393. doi: 10.1038/s41375-021-01369-0. PubMed DOI

Yan M, Chakravarthy S, Tokuda JM, Pollack L, Bowman GD, Lee YS. Succinyl-5-aminoimidazole-4-carboxamide-1-ribose 5’-Phosphate (SAICAR) Activates Pyruvate Kinase Isoform M2 (PKM2) in Its Dimeric Form. Biochemistry. 2016;55:4731–4736. doi: 10.1021/acs.biochem.6b00658. PubMed DOI PMC

Zikánová M, Krijt J, Hartmannová H, Kmoch S. Preparation of 5-amino-4-imidazole-N-succinocarboxamide ribotide, 5-amino-4-imidazole-N-succinocarboxamide riboside and succinyladenosine, compounds usable in diagnosis and research of adenylosuccinate lyase deficiency. Journal of Inherited Metabolic Disease. 2005;28:493–499. doi: 10.1007/s10545-005-0493-z. PubMed DOI

Zurlo G, Liu X, Takada M, Fan C, Simon JM, Ptacek TS, Rodriguez J, von Kriegsheim A, Liu J, Locasale JW, Robinson A, Zhang J, Holler JM, Kim B, Zikánová M, Bierau J, Xie L, Chen X, Li M, Perou CM, Zhang Q. Prolyl hydroxylase substrate adenylosuccinate lyase is an oncogenic driver in triple negative breast cancer. Nature Communications. 2019;10:5177. doi: 10.1038/s41467-019-13168-4. PubMed DOI PMC

Zobrazit více v PubMed

figshare
10.25452/figshare.plus.c.5793614