{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T19:28:06Z","timestamp":1786562886752,"version":"build-2736575974"},"reference-count":48,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T00:00:00Z","timestamp":1773187200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T00:00:00Z","timestamp":1773187200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["00x0ma614"],"award-info":[{"award-number":["00x0ma614"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["88887.311601\/2018\u201000\u20102629\/2018"],"award-info":[{"award-number":["88887.311601\/2018\u201000\u20102629\/2018"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004586","name":"Funda\u00e7\u00e3o Carlos Chagas Filho de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro","doi-asserted-by":"publisher","award":["E26\/010.001706\/2019"],"award-info":[{"award-number":["E26\/010.001706\/2019"]}],"id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004586","name":"Funda\u00e7\u00e3o Carlos Chagas Filho de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro","doi-asserted-by":"publisher","award":["E26\/010.100623\/2018"],"award-info":[{"award-number":["E26\/010.100623\/2018"]}],"id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004586","name":"Funda\u00e7\u00e3o Carlos Chagas Filho de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro","doi-asserted-by":"publisher","award":["E26\/211.815\/2021"],"award-info":[{"award-number":["E26\/211.815\/2021"]}],"id":[{"id":"10.13039\/501100004586","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["402419\/2022\u20107"],"award-info":[{"award-number":["402419\/2022\u20107"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["faseb.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FASEB Journal"],"published-print":{"date-parts":[[2026,3,31]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Trypanosoma cruzi<\/jats:italic>\n                    , the causative agent of Chagas disease, possesses glycosomes\u2014unique organelles that house key metabolic enzymes, several of which are promising therapeutic targets. Among them, phosphoenolpyruvate carboxykinase (PEPCK) plays a central role in succinic fermentation, the main pathway for NAD\n                    <jats:sup>+<\/jats:sup>\n                    regeneration within the organelle. Using CRISPR\/Cas9 editing, the PEPCK gene was disrupted in\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>T. cruzi<\/jats:italic>\n                    <\/jats:styled-content>\n                    , producing single\u2010allele knockout epimastigotes (TcPEPCK\u2010sKO) with reduced PEPCK expression and enzyme activity. In a high glucose environment, PEPCK disruption impaired glucose consumption and mitochondrial respiration, particularly oxidative phosphorylation, reducing dependence on mitochondrial ATP production when glucose was supplied. To compensate, pyruvate phosphate dikinase was upregulated, increasing alanine production, possibly to maintain redox balance in glycosomes. Despite this metabolic adaptation, the growth of TcPEPCK\u2010sKO epimastigotes was partially reduced compared with non\u2010deleted parasites. In contrast, under low glucose conditions, PEPCK activity was not critical for mitochondrial bioenergetics, ATP production, or proliferation. Although TcPEPCK\u2010sKO epimastigotes exhibited a minor reduction in growth in high glucose medium, their differentiation (metacyclogenesis) and invasion were severely compromised. However, once inside the host cell, TcPEPCK\u2010sKO amastigotes increased their replication, leading to enhanced trypomastigote production. The same was observed in in\u00a0vivo infection, where TcPEPCK\u2010sKO infection in IFN\u03b3\u2010deficient mice caused uncontrolled parasitemia and severe pathology, highlighting the critical role of PEPCK in host\u2010pathogen interactions. However, an intact immune system effectively contained TcPEPCK\u2010sKO infection. Taken together, our findings demonstrate that glycosomal PEPCK is crucial for coupling glycolysis to mitochondrial bioenergetics, enabling the parasite differentiation within the insect vector and controlling the infection of mammalian host cells.\n                  <\/jats:p>","DOI":"10.1096\/fj.202503541r","type":"journal-article","created":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T14:58:07Z","timestamp":1773241087000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Glycosomal Phosphoenolpyruvate Carboxykinase\n                    <scp>CRISPR<\/scp>\n                    \/Cas9\u2010Deletion and Its Role in\n                    <i>Trypanosoma cruzi<\/i>\n                    Metacyclogenesis and Infectivity in Mammalian Host"],"prefix":"10.1096","volume":"40","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3312-5171","authenticated-orcid":false,"given":"Carolina S. D.","family":"Vieira","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Intera\u00e7\u00e3o de Tripanossomat\u00eddeos e Vetores, Departamento de Bioqu\u00edmica, IBRAG Universidade do Estado do Rio de Janeiro  Rio de Janeiro Brazil"},{"name":"Department of Cellular Biology, Center for Tropical and Emerging Global Diseases University of Georgia  Athens Georgia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7111-4687","authenticated-orcid":false,"given":"Wei","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Cellular Biology, Center for Tropical and Emerging Global Diseases University of Georgia  Athens Georgia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0045-0453","authenticated-orcid":false,"given":"Fernando","family":"Sanchez\u2010Valdez","sequence":"additional","affiliation":[{"name":"Department of Cellular Biology, Center for Tropical and Emerging Global Diseases University of Georgia  Athens Georgia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0583-371X","authenticated-orcid":false,"given":"Jihyun","family":"Lim","sequence":"additional","affiliation":[{"name":"Department of Cellular Biology, Center for Tropical and Emerging Global Diseases University of Georgia  Athens Georgia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0441-1355","authenticated-orcid":false,"given":"Brooke E.","family":"White","sequence":"additional","affiliation":[{"name":"Department of Cellular Biology, Center for Tropical and Emerging Global Diseases University of Georgia  Athens Georgia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-9090-6874","authenticated-orcid":false,"given":"Camilla G. S.","family":"Souza","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Intera\u00e7\u00e3o de Tripanossomat\u00eddeos e Vetores, Departamento de Bioqu\u00edmica, IBRAG Universidade do Estado do Rio de Janeiro  Rio de Janeiro Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9589-5243","authenticated-orcid":false,"given":"Rick L.","family":"Tarleton","sequence":"additional","affiliation":[{"name":"Department of Cellular Biology, Center for Tropical and Emerging Global Diseases University of Georgia  Athens Georgia USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2247-8251","authenticated-orcid":false,"given":"Marcia C.","family":"Paes","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Intera\u00e7\u00e3o de Tripanossomat\u00eddeos e Vetores, Departamento de Bioqu\u00edmica, IBRAG Universidade do Estado do Rio de Janeiro  Rio de Janeiro Brazil"},{"name":"Instituto Nacional de Ci\u00eancia e Tecnologia\u2014Entomologia Molecular (INCT\u2010EM)  Rio de Janeiro Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3258-607X","authenticated-orcid":false,"given":"Natalia P. A.","family":"Nogueira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Intera\u00e7\u00e3o de Tripanossomat\u00eddeos e Vetores, Departamento de Bioqu\u00edmica, IBRAG Universidade do Estado do Rio de Janeiro  Rio de Janeiro Brazil"},{"name":"Department of Cellular Biology, Center for Tropical and Emerging Global Diseases University of Georgia  Athens Georgia USA"},{"name":"Instituto Nacional de Ci\u00eancia e Tecnologia\u2014Entomologia Molecular (INCT\u2010EM)  Rio de Janeiro Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,3,11]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1590\/S0074-02761909000200008"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.13-0726"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature09221"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.gheart.2015.06.001"},{"key":"e_1_2_10_6_1","volume-title":"Chagas Disease (American Trypanosomiasis)","author":"WHO","year":"2022"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/jeu.12338"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1258096"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02097-14"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.01788-17"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-018-8927-3"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/0305-0491(84)90380-8"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1042\/bse0510015"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1009495"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.2174\/1568005053174636"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens7020036"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-9861(90)90091-C"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1096\/fasebj.6.13.1397837"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0166-6851(95)00099-M"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molbiopara.2019.02.008"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00763064"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.exppara.2021.108102"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0000740"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.freeradbiomed.2017.03.027"},{"key":"e_1_2_10_25_1","first-page":"12","volume-title":"Mitochondrial Physiology Network","author":"Gnaiger E.","year":"2014"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molbiopara.2008.03.004"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1006\/meth.2001.1262"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1590\/S0074-02761988000100016"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1126\/scitranslmed.abb7656"},{"key":"e_1_2_10_30_1","article-title":"Quantitative 3D Imaging of Trypanosoma cruzi\u2014Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs","volume":"184","author":"Sanchez\u2010Valdez F.","year":"2022","journal-title":"Journal of Visualized Experiments"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00241-16"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.2741\/3931"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.02215-24"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0008728"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.124917"},{"issue":"3","key":"e_1_2_10_36_1","first-page":"443","article-title":"Proline Metabolism in Trypanosoma cruzi Epimastigotes","volume":"55","author":"Sylvester D.","year":"1976","journal-title":"Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.1080\/21505594.2020.1797274"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens11020268"},{"key":"e_1_2_10_39_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1550-7408.1989.tb01063.x"},{"key":"e_1_2_10_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molbiopara.2018.07.007"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.100548"},{"key":"e_1_2_10_42_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1110289"},{"key":"e_1_2_10_43_1","doi-asserted-by":"publisher","DOI":"10.1128\/msphere.00601-22"},{"key":"e_1_2_10_44_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.2000517"},{"key":"e_1_2_10_45_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M114.569434"},{"key":"e_1_2_10_46_1","doi-asserted-by":"publisher","DOI":"10.12688\/f1000research.6724.1"},{"key":"e_1_2_10_47_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mib.2021.07.017"},{"key":"e_1_2_10_48_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006747"},{"key":"e_1_2_10_49_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005511"}],"container-title":["The FASEB Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/faseb.onlinelibrary.wiley.com\/doi\/pdf\/10.1096\/fj.202503541R","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/faseb.onlinelibrary.wiley.com\/doi\/full-xml\/10.1096\/fj.202503541R","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/faseb.onlinelibrary.wiley.com\/doi\/pdf\/10.1096\/fj.202503541R","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T18:38:03Z","timestamp":1786559883000},"score":1,"resource":{"primary":{"URL":"https:\/\/faseb.onlinelibrary.wiley.com\/doi\/10.1096\/fj.202503541R"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,11]]},"references-count":48,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2026,3,31]]}},"alternative-id":["10.1096\/fj.202503541R"],"URL":"https:\/\/doi.org\/10.1096\/fj.202503541r","archive":["Portico"],"relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2024.12.20.629751","asserted-by":"object"}]},"ISSN":["0892-6638","1530-6860"],"issn-type":[{"value":"0892-6638","type":"print"},{"value":"1530-6860","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,11]]},"assertion":[{"value":"2025-10-02","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-02-27","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-03-11","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e71672"}}