{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T05:57:48Z","timestamp":1790488668780,"version":"4.1.0"},"posted":{"date-parts":[[2026,9,23]]},"group-title":"Biochemistry","reference-count":60,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nd\/4.0\/"}],"funder":[{"award":["672623)"],"award-info":[{"award-number":["672623)"]}],"id":[{"id":"https:\/\/ror.org\/018mejw64","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,23]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Direct reprogramming (DR) converts one differentiated cell identity into another and is a promising strategy for regenerative approaches when cells are lost because of injury or disease. Transcription factors (TFs) that specify a given cell type can drive DR, but inhibitory mechanisms restrict this conversion in most cell types. To identify such barriers\n                  <jats:italic>in vivo<\/jats:italic>\n                  , we use\n                  <jats:italic>Caenorhabditis elegans<\/jats:italic>\n                  and the zinc finger TF CHE-1, which is required to specify the glutamatergic taste neuron fate of ASE neurons. Ectopic CHE-1 expression can induce DR of different cell types to ASE neurons upon RNAi-mediated depletion of barrier genes.\n                <\/jats:p>\n                <jats:p>\n                  Here we characterize the \u03b1 subunit of the mitochondrial isocitrate dehydrogenase 3 complex (IDH3), IDHA-1, as a barrier to DR of germ cells into neurons. RNAi against\n                  <jats:italic>idha-1<\/jats:italic>\n                  produced consistent ASE fate reporter expression, accompanied by morphological changes to neuron-like structures in the germline upon ectopic CHE-1 expression. Using different neuronal gene expression reporters, single-molecule FISH, and antibody staining, we confirm that this germ cell conversion (GeCo) produces neuron-like cells. We found that loss of IDH3 activity causes metabolic perturbations that result in a variety of direct and indirect effects on DR of germ cells. One arm implicates the hypoxia-inducible factor HIF-1, which promotes GeCo. Its loss causes a significant decrease, while the\n                  <jats:italic>vhl-1<\/jats:italic>\n                  mutant background, in which HIF-1 protein is stabilized, leads to enhanced GeCo. Another arm implicates epigenetic changes leading to a detectable loss of the repressive histone marks H3K27me3 and H3K9me3 upon\n                  <jats:italic>idha-1<\/jats:italic>\n                  depletion. Furthermore, stable-isotope-resolved metabolomics shows strong citrate accumulation without a noteworthy drop in \u03b1-Ketoglutarate (\u03b1KG) levels, indicating that compensatory pathways maintain \u03b1KG levels. Genetics and metabolomics analyses confirmed that Glutamate anaplerosis contributes to compensating for the loss of IDHA-1. Additionally, we found that depletion of glucose transporter FDGT-2 nearly doubles GeCo efficiency. Overall, our findings define the TCA cycle as an\n                  <jats:italic>in vivo<\/jats:italic>\n                  safeguard of germ cell identity and show that multiple metabolic inputs converge to keep germ cells refractory to TF-induced DR to neuronal cells.\n                <\/jats:p>","DOI":"10.64898\/2026.09.22.753437","type":"posted-content","created":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T17:15:12Z","timestamp":1790183712000},"source":"Crossref","is-referenced-by-count":0,"title":["Depletion of the isocitrate dehydrogenase 3 subunit IDHA-1 licenses germ cell-to-neuron direct reprogramming"],"prefix":"10.64898","author":[{"given":"Nida ul","family":"Fatima","sequence":"first","affiliation":[{"name":"Department of Biology, Institute of Cell and Systems Biology, Universit\u00e4t Hamburg, 20146 Hamburg, Germany"},{"name":"Max-Delbr\u00fcck Center for Molecular Medicine, Berlin Institute for Medical Systems Biology, 10115 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Reid","sequence":"additional","affiliation":[{"name":"Max-Delbr\u00fcck Center for Molecular Medicine, Berlin Institute for Medical Systems Biology, 10115 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amin","family":"Shadfar","sequence":"additional","affiliation":[{"name":"Department of Biology, Institute of Cell and Systems Biology, Universit\u00e4t Hamburg, 20146 Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Blume","sequence":"additional","affiliation":[{"name":"Department of Biology, Institute of Cell and Systems Biology, Universit\u00e4t Hamburg, 20146 Hamburg, Germany"},{"name":"Max-Delbr\u00fcck Center for Molecular Medicine, Berlin Institute for Medical Systems Biology, 10115 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tobias","family":"Opialla","sequence":"additional","affiliation":[{"name":"Max-Delbr\u00fcck Center for Molecular Medicine, Berlin Institute for Medical Systems Biology, 10115 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Kempa","sequence":"additional","affiliation":[{"name":"Max-Delbr\u00fcck Center for Molecular Medicine, Berlin Institute for Medical Systems Biology, 10115 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7293-8629","authenticated-orcid":false,"given":"Baris","family":"Tursun","sequence":"additional","affiliation":[{"name":"Department of Biology, Institute of Cell and Systems Biology, Universit\u00e4t Hamburg, 20146 Hamburg, 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