Rescue of p53 functions by in vitro‐transcribed mRNA impedes the growth of high‐grade serous ovarian cancer

Abstract

BackgroundThe cellular tumor protein p53 (TP53) is a tumor suppressor gene that is frequently mutated in human cancers. Among various cancer types, the very aggressive high‐grade serous ovarian carcinoma (HGSOC) exhibits the highest prevalence of TP53 mutations, present in >96% of cases. Despite intensive efforts to reactivate p53, no clinical drug has been approved to rescue p53 function. In this study, our primary objective was to administer in vitro‐transcribed (IVT) wild‐type (WT) p53‐mRNA to HGSOC cell lines, primary cells, and orthotopic mouse models, with the aim of exploring its impact on inhibiting tumor growth and dissemination, both in vitro and in vivo.MethodsTo restore the activity of p53, WT p53 was exogenously expressed in HGSOC cell lines using a mammalian vector system. Moreover, IVT WT p53 mRNA was delivered into different HGSOC model systems (primary cells and patient‐derived organoids) using liposomes and studied for proliferation, cell cycle progression, apoptosis, colony formation, and chromosomal instability. Transcriptomic alterations induced by p53 mRNA were analyzed using RNA sequencing in OVCAR‐8 and primary HGSOC cells, followed by ingenuity pathway analysis. In vivo effects on tumor growth and metastasis were studied using orthotopic xenografts and metastatic intraperitoneal mouse models.ResultsReactivation of the TP53 tumor suppressor gene was explored in different HGSOC model systems using newly designed IVT mRNA‐based methods. The introduction of WT p53 mRNA triggered dose‐dependent apoptosis, cell cycle arrest, and potent long‐lasting inhibition of HGSOC cell proliferation. Transcriptome analysis of OVCAR‐8 cells upon mRNA‐based p53 reactivation revealed significant alterations in gene expression related to p53 signaling, such as apoptosis, cell cycle regulation, and DNA damage. Restoring p53 function concurrently reduces chromosomal instability within the HGSOC cells, underscoring its crucial contribution in safeguarding genomic integrity by moderating the baseline occurrence of double‐strand breaks arising from replication stress. Furthermore, in various mouse models, treatment with p53 mRNA reduced tumor growth and inhibited tumor cell dissemination in the peritoneal cavity in a dose‐dependent manner.ConclusionsThe IVT mRNA‐based reactivation of p53 holds promise as a potential therapeutic strategy for HGSOC, providing valuable insights into the molecular mechanisms underlying p53 function and its relevance in ovarian cancer treatment.

Research progress on the application of organoids in gynecological tumors

“…In addition, in the extraction of OC organoids, establishing organoids derived from omental histology facilitates the exploration of relevant metabolic mechanisms ( Lõhmussaar et al, 2020b ). By constructing a model of organoid diseases and constructing OC models with different gene expressions, we aimed to explore the relationship between genes and the occurrence, development, and prognosis of OC ( Iwahashi et al, 2022 ; Raab et al, 2024 ); identify ligands and receptors associated with poor survival in OC ( Carvalho et al, 2022 ); study chemotherapy resistance mechanisms ( Gorski et al, 2021 ); explore platinum-based chemotherapy response and survival markers for OC ( Wang Z. et al, 2022 ; Compadre et al, 2023 ); and explore the role of signaling pathways in OC ( Hoffmann et al, 2020 ). In terms of drug detection and screening, by constructing an OC organoid model and summarizing the characteristics of the tumor, drug screening experiments ( Nanki et al, 2020 ; Vernon et al, 2020 ; Wang W. et al, 2022 ; Zhang et al, 2022a , 2022b ; Liu et al, 2022 ; Cesari et al, 2023 ; Wan et al, 2023 ) and drug sensitivity tests ( Kopper et al, 2019 ; Lõhmussaar et al, 2020b ; Maenhoudt et al, 2020 ) can be conducted to explore the mechanisms of drug resistance and drug action in OC ( Zhang X. et al, 2023 ; Cao et al, 2023 ).…”

“…(HGSOC) cells, reduce chromosomal instability, and cause cell death (Raab et al, 2024). Iwahashi et al extracted samples from 121 patients with wild-type p53 or mutant p53 high-grade serous OC and constructed OC organoids using the matrix gel method.…”

“…Raab et al extracted samples from patients with epithelial OC and cultured serous OC organoids using the matrix gel method. They found that restoring p53 function can inhibit the proliferation of advanced serous ovarian cancer (HGSOC) cells, reduce chromosomal instability, and cause cell death ( Raab et al, 2024 ). Iwahashi et al extracted samples from 121 patients with wild-type p53 or mutant p53 high-grade serous OC and constructed OC organoids using the matrix gel method.…”

mRNA vaccine development and applications: A special focus on tumors (Review)

“…After treatment with the above mRNA, patient-derived ovarian organoids began to shrink and die, and primary tumors and metastases in mouse models of ovarian cancer almost completely disappeared. These findings suggest that IVT-mRNA-based protein replacement therapy may reactivate oncogenic proteins in cancer cells and may thus serve as a valuable therapeutic option for the treatment of cancer ( 230 ).…”