Development and preclinical evaluation of KDTV001, an adenovirus 5-vectored trivalent HPV therapeutic vaccine targeting HPV16/18/52.

BACKGROUND: Persistent high-risk HPV infections (e.g., HPV16/18/52) are directly linked to cervical cancer development, thus necessitating the development of effective multi-target therapeutic vaccines. METHODS: We developed KDTV001, a non-replicating adenovirus 5-vectored trivalent vaccine encoding engineered E6/E7 oncoproteins from HPV16/18/52. Its immunogenicity was evaluated in C57BL/6, CD1, HLA-transgenic mice, SD rats, and a human DC-T cell co-culture system. Assessments included IFN-γ ELISpot, flow cytometry, tumour challenge/rechallenge, single-cell RNA/TCR sequencing (scRNA/TCR-seq) of lymphocytes, and immune cell depletion. FINDINGS: KDTV001 demonstrated robust preclinical efficacy across multiple models, eliciting potent immune and cross-reactive responses in immunocompetent and HLA-transgenic mice. scRNA/TCR-seq revealed KDTV001 remodelled the tumour microenvironment, increasing cytotoxic CD8(+) T cells and promoting T cell clonal expansion and differentiation. A single dose induced complete TC-1 tumour regression in 60-80% of mice with durable protection (>200 days). Optimising the booster immunisation schedule mitigated the impact of pre-existing vector immunity on the immunogenicity of KDTV001. Human relevance was confirmed through antigen-specific T-cell activation and expansion in HLA-A transgenic mice and PBMC co-cultures. INTERPRETATION: KDTV001, the adenovirus 5-vectored trivalent therapeutic vaccine targeting HPV16/18/52, demonstrates potent immunogenicity, significant antitumour efficacy, and long-term protective immunity across diverse preclinical models, paving the way for clinical translation in treating HPV-associated malignancies. FUNDING: This study was funded by the Noncommunicable Chronic Diseases-National Science and Technology Major Project of China (Nos.2025ZD0544101), the Hubei Provincial Science and Technology Major Project of China (Nos.2023BCA004), and the National Clinical Research Center for Gynaecological Diseases Special Fund of China (Nos.2025LCPT01).