Deciphering the intricate crosstalk of the EFNA1-EPHA3 axis in orchestrating the proangiogenic niche of cervical adenocarcinoma: A promising therapy target.

e17508 Background: Cervical adenocarcinoma (ADC) shows more malignant phenotypes and poorer prognosis compared to squamous cell carcinoma (SCC). Anti-angiogenesis therapy is a promising therapeutic strategy for cervical cancer, yet it is not as efficient in ADC as that in SCC. Therefore, a better understanding of ADC angiogenesis is needed to develop precise anti-angiogenic therapies. Methods: A 120 patients clinical cohort of 24 ADC patients and 96 SCC patients were included. Single-cell RNA sequencing was performed on 42 samples (12 ADC and 21 SCC). Integrated 3D angiogenesis study system in vitro including vascularized ADC-derived tumoroids and tumoroids-on-a-chip and in vivo xenograft were established to explore the anti-angiogenic effect of targeting the EFNA1-EPHA3 axis by ifabotuzumab. Results: ADC patients displayed more aggressive clinical features (higher vaginal involvement, parametrial infiltration, uterine involvement, lymph node metastasis, adnexal involvement and recurrence incidence). Importantly, a higher incidence of lymphovascular space invasion (LVSI) in patients with ADC, suggesting ADC patients were more prone to angiogenesis compared with SCC patients. ADC epithelial cells and ADC-associated fibroblasts wrapped around tumour microvessels via the EFNA1-EPHA3 axis, forming a unique structure "proangiogenic niche". Based on the ADC-derived tumoroids we established, we constructed 3D vascular-like system integrated ADC-derived tumoroids and microfluidics-based tumoroids-on-a-chip, which successfully recapitulated the proangiogenic niche supported by the EFNA1-EPHA3 axis within ADC. For preclinical exploration, we validated the antiangiogenic effect of targeting the EFNA1-EPHA3 axis using ifabotuzumab based on in vitro vascular-like system integrated 3D ADC-derived tumoroid model, microfluidics-based tumoroids-on-a-chip and an in vivo xenograft model, highlighting the potential of ifabotuzumab in preclinical antiangiogenic therapy for ADCs. Conclusions: Our study inventively proposed "proangiogenic niche" connected by EFNA1-EPHA3 axis and highlighted ifabotuzumab as a promising anti-angiogenesis therapy for ADCs. Besides, we originally constructed integrated 3D angiogenesis study system.