{
  "abstract": "Background: TP53-mutated acute myeloid leukemia (AML) carries a dismal prognosis. Eprenetapopt (APR-246), a p53 reactivator, is under investigation as a targeted therapeutic strategy. Among mutations identified in our cohort, R248Q was the most frequent, a dominant-negative hotspot mutation considered pathogenic; structural analyses revealed no significant alterations, though data indicate impaired p53 function. Aim: This study aimed to evaluate the ability of APR-246 to reactivate the functional conformation of mutant p53 (R248Q) in Kasumi-1 cells. Materials and Methods: Kasumi-1 cells were cultured in RPMI with 20% FBS and treated with eprenetapopt (0, 2.5, 5, 10 µM) for 48–72h. Protein extracts were analyzed by Western blotting. Gene expression was assessed by RT-qPCR. Cell cycle and apoptosis were evaluated by flow cytometry using Annexin V/propidium iodide staining. Results: Increasing eprenetapopt doses reduced p53, p62, and LC3B-I/II levels, with concomitant upregulation of BAX, p21, and cleaved PARP1, indicating apoptosis and enhanced autophagic flux. RT-qPCR showed strong transcriptional reactivation of p53 targets after 10 µM eprenetapopt: CDKN1A mRNA increased ∼90-fold (p<0.001), correlating with cell cycle arrest, while BH3-only mediators BBC3/PUMA (∼5.5-fold) and PMAIP1/NOXA (∼6.3-fold) were upregulated (p<0.001). Conversely, BAX and MDM2 transcripts showed slight downregulation (p<0.001, p<0.05), suggesting cytotoxicity is driven mainly by the p21/PUMA/NOXA axis. Cell cycle analysis showed dose-dependent changes: untreated cells displayed normal distribution across G0/G1 and S phases with minimal Sub-G1 fragmentation, whereas increasing concentrations progressively reduced proliferative populations and induced G2/M arrest at intermediate doses (p<0.01, 5 µM), followed by a marked increase in Sub-G1 at the highest dose (p<0.001), reflecting genomic collapse. Apoptosis assays confirmed these findings: baseline apoptosis was below 15%, while treatment induced a dose-dependent shift toward Annexin V-positive populations, reaching ∼70% and ∼80% apoptosis at 5 and 10 µM, respectively (p<0.001 vs. control). Discussion: These findings indicate that eprenetapopt restores transcriptional and functional activity of mutant p53, driving G2/M arrest and robust apoptosis in Kasumi-1 cells. Marked CDKN1A induction confirms recovery of p53 transactivation capacity, consistent with the observed G2/M retention, while PUMA/NOXA upregulation indicates transcriptional reactivation of the intrinsic apoptotic program. Notably, BAX protein increased despite reduced mRNA, suggesting post-transcriptional regulation, possibly via PUMA/NOXA-mediated displacement of BAX from anti-apoptotic complexes (BCL-2/BCL-XL), reinforcing the central role of the p21/PUMA/NOXA axis over BAX transcriptional induction. Decreased MDM2 mRNA further suggests partial restoration of the p53-MDM2 feedback loop. Concurrent reduction of p62 and LC3B-I/II alongside decreased total p53 suggests autophagy acts as an auxiliary pathway favoring degradation of destabilized mutant p53 and amplifying apoptotic signaling. Conclusion: Collectively, these findings demonstrate that eprenetapopt effectively reactivates mutant p53 transcriptional function, translating into significant cytotoxicity in TP53-mutated AML cells, supporting its therapeutic potential in this poor-prognosis subgroup.",
  "abstract_shown": true,
  "affiliations": [
    "Universidade de São Paulo, São Paulo, SP, Brazil"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 5,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Letícia Machado Favery BERTOLINE"
    },
    {
      "affs": [
        0
      ],
      "name": "Maria Fernanda Lopes CARVALHO"
    },
    {
      "affs": [
        0
      ],
      "name": "Natália Sudan PARDUCCI"
    },
    {
      "affs": [
        0
      ],
      "name": "Keli LIMA"
    },
    {
      "affs": [
        0
      ],
      "name": "João Agostinho MACHADO NETO"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/dz5tanvvd4nky.bib",
  "cited_by": 0,
  "doi": "10.1016/j.htct.2026.107597",
  "fulltext_url": "http://www.sciencedirect.com/science/article/pii/S2531137926013441",
  "id": "dz5tanvvd4nky",
  "issns": [
    "2531-1379",
    "2531-1387"
  ],
  "landing_url": "https://doi.org/10.1016/j.htct.2026.107597",
  "language": "en",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "lcc",
      "RC633-647.5"
    ],
    [
      "openalex",
      "2730|Cancer-related Molecular Pathways"
    ]
  ],
  "openalex": "W7223106709",
  "pdf_url": null,
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-23",
  "publisher": "Elsevier",
  "ris": "https://research.draconitic.com/paper/dz5tanvvd4nky.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T13:43:25Z",
      "native_id": "530fc7ac1c744e8bac647f212239c76d",
      "source": "doaj",
      "url": "https://doi.org/10.1016/j.htct.2026.107597"
    }
  ],
  "subjects": [
    {
      "field": "medicine",
      "origin": "openalex:2730|Cancer-related Molecular Pathways",
      "subfield": "oncology"
    }
  ],
  "title": "FUNCTIONAL REACTIVATION OF MUTANT P53 R248Q BY EPRENETAPOPT TRIGGERS CELL CYCLE ARREST AND APOPTOSIS IN ACUTE MYELOID LEUKEMIA",
  "title_text": "FUNCTIONAL REACTIVATION OF MUTANT P53 R248Q BY EPRENETAPOPT TRIGGERS CELL CYCLE ARREST AND APOPTOSIS IN ACUTE MYELOID LEUKEMIA",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T13:43:25Z",
  "url": "https://research.draconitic.com/paper/dz5tanvvd4nky",
  "venue": "Hematology, Transfusion and Cell Therapy",
  "versions": []
}