A therapeutic strategy for Cervical Gastric-type Adenocarcinoma by targeting CLU to relieve CLU-associated stress and sensitize chemotherapy

Objectives Gastric-type adenocarcinoma (GAS), an aggressive subtype of non-human papillomavirus (HPV)-associated (NHPVA) cervical adenocarcinomas (ADC), remains a treatment-refractory disease with poor prognosis. This study aims to explore the oncogenic mechanism and efficacious therapeutic target of GAS. Methods We included 19 NHPVA and 153 HPVA ADC patients from our center to investigate clinicopathological features. We collected 3 GAS and 2 usual-type endocervical adenocarcinoma (UEA) for scRNA-seq and TCR-seq. We conducted immunohistochemical staining (IHC) of 25 GAS and 25 UEA samples and multicolor IHC of 2 GAS samples for validation. We explored the efficacy of anti-CLU (OGX-011) and/or Cisplatin (DDP) for GAS based on GAS-derived tumoroids. Results (1) Based on clinical data, we verified clinicopathologically malignancy of GAS. (2) Through scRNA-seq, we delineated key cell subtypes including GAS epithelial cells, “GAS-enriched fibroblasts”, “GAS-associated γδT cells” and CD8 + Texs enduring heat stress and contributing to GAS aggressive phenotype. (3) Regarding validation, we verified CLU-associated heat stress, highlighted the potential role of CLU-associated stress in promoting immune escape, and established a 4-gene signature (CLU, PDGFB, TIGIT and C3) indicating poor prognosis of GAS induced by CLU-associated stress and immune escape. (4) Based on GAS-derived tumoroids retaining the histological features, CLU-associated stress and genetic profile of parental tumor, we validated the anti-tumor and sensitizing DDP efficacy of targeting CLU. Conclusion CLU-associated heat stress of key cell subtypes contributed to the malignant GAS microenvironment. Additionally, we pioneeringly constructed GAS-derived tumoroids and suggested that combining CLU-targeted treatment and DDP could improve the therapeutic efficacy for GAS.