case_images: PMC10570525_01
case_images: Imaging results in the pelvic lesion tissue. The results of the MRI showed a mixed signal mass in the external cervical orifice and vaginal fornix (size about 7.4x7.9x8.5cm). (A, C: sagittal position; B, D: horizontal position). MRI, magnetic resonance imaging.
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case_images: The MRI findings strongly indicated a malignant process ( Figure 1 ).
case_images: The results of postoperative pathology and Sanger sequencing. (A) An ulcerated mass was seen at the external cervical orifice (size about 7cm x 8cm x 7cm) and presented as a tongue-shaped excrescence into the vagina. The cut surface was grayish solid with extensive hemorrhage and necrosis, invading the superficial layer of the cervical stroma. (B-D) Postoperative pathology of hematoxylin-eosin staining indicated that the tumor was a cERMS: Small, spindled, and round cells were located in a background of myxoid stroma suggestive of rhabdomyoblastic differentiation. The multinucleated giant cells could be seen in the lesion and nuclear fission is obvious (x400, Scale bar=20mm). (E-G) Immunochemical staining showed that the tissue was Desmin, MyoD1, and Myogenin positive (x400, Scale bar=20mm). (H) Sanger sequencing of tumor tissue did not detect DICER1 gene mutation in exons 24 and 25. The numerical annotations were the actual nucleotide order on the exons 24 and 25. cERMS, embryonal rhabdomyosarcoma of the cervix.
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case_images: The excised uterus and associated tumor specimens are presented in  Figure 2A . Pathological evaluation and subsequent immunohistochemical (IHC) staining confirmed the diagnosis of cERMS ( Figures 2B-G ). However, no DICER1 gene mutations were detected in these exons ( Figure 2H ).
case_images: The drug sensitivity test of PDOs. (A) Representative bright field images of normal tissue PDOs and cERMS PDOs prior to drug sensitivities (Scale bar=100mum). (B) Representative hematoxylin-eosin staining and immunochemical staining of Desmin images of cERMS PDOs prior to drug sensitivities (Scale bar=20mum). (C) Cell viability of PDOs treated with vincristine, actinomycin D, and cyclophosphamide. PDOs viability (%)=(OD Sample-OD Blank)/(OD Control-OD Blank)x100%. (D) The inhibition ratio of PDOs treated with the single or combination of vincristine, actinomycin D, or cyclophosphamide. PDOs Inhibition Ratio (%) = (1-(OD Sample-OD Blank)/(OD Control-OD Blank))x100%. (E) The cytotoxicity of the co-culture model of PBMCs and cERMS PDOs treated with the single PD-1 or combined with VAC. PDOs, patient-derived organoids; V, vincristine; A, actinomycin D; C, cyclophosphamide; PD-1, Programmed death protein 1; PBMCs, peripheral blood mononuclear cells; NS, no statistical significance; *P<0.05.
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case_images: Before drug sensitivity testing, the PDOs exhibited solid spheroid morphology with irregular edges ( Figure 3A ). HE and IHC staining revealed that the cultured PDOs preserved key phenotypic features of the parent cERMS, including nuclear pleomorphism, mitotic rate, and immunoreactive profiles ( Figure 3B ). To compare the drug sensitivities of the tested drugs, the half-inhibitory concentrations (IC50) of each drug were determined using "GraphPad Prism 8.0" ( Figure 3C ). The VAC combination therapy demonstrated a tumor inhibition rate of 95.87%, which was superior to monotherapy or the combination of any two drugs ( Figure 3D ). Thermo). Subsequently, the PBMCs were quantified and co-cultured with PDOs at a ratio of 50-100:1. In the co-culture model of PBMCs and cERMS PDOs, VAC and immune checkpoint inhibitors have synergistic anti-tumor effects. The results showed that the combined use of the Programmed death protein 1 (PD-1) inhibitor and VAC achieved a favorable therapeutic effect, with cytotoxicity reaching 34.49% ( Figure 3E ).
case_images: The diagnosis and treatment strategy timeline schematic.
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case_images: The diagnosis and treatment strategy timeline schematic are presented in  Figure 4 .