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Cervical cancer, the second most common cause of cancer death in women worldwide, is significantly associated with infection of high-risk human papillomaviruses (HPVs), especially the most common genotype, HPV 16. To date, there is no established noninvasive therapy to treat cervical cancer. Methods: Here, we report a novel affitoxin that targets HPV16 E7 protein, one of the primary target proteins in molecular targeted therapy for HPV-induced cervical cancer. The affitoxin, Z HPV16E7 affitoxin384 was generated by fusing the modified Pseudomonas Exotoxin A (PE38KDEL) to the HPV16 E7-specific affibody. The expressed and purified Z HPV16E7 affitoxin384 was characterized using numerous methods. SPR assay, indirect immunofluorescence assay, and near-infrared (NIR) optical imaging were respectively performed to assess the targeting ability of Z HPV16E7 affitoxin384 to HPV16 E7 protein both in vitro and in vivo . Cell viability assays and SiHa tumor-bearing nude mice were used to evaluate the efficacy of Z HPV16 E7 affitoxin384 in vitro and in vivo , respectively. Results: Using in vitro methods the SPR assay and indirect immunofluorescence assay showed that Z HPV16E7 affitoxin384 targeted HPV16 E7 with high binding affinity and specificity. Significant reduction of cell viability in HPV16 positive cells was observed in the presence of Z HPV16 E7 affitoxin384. By NIR optical imaging, Z HPV16 E7 affitoxin384 specifically targeted HPV16 positive tumors in vivo . Z HPV16E7 affitoxin384 showed significant in vivo antitumor efficacy in two kinds of tumor-bearing nude mouse models. Conclusions: Z HPV16E7 affitoxin384 is a potent anti-cervical cancer therapeutic agent that could be effective against HPV16 positive tumors in humans.

A novel HPV16 E7-affitoxin for targeted therapy of HPV16-induced human cervical cancer