Effective drug and shRNA delivery for synergistic treatment of triple-negative breast cancer by sequentially targeting tumor hypoxia

Abstract

Clinical treatment of TNBC remains challenging, due to the lack of targeted therapies. As TNBC is highly hypoxic with higher HIF-1α expression than other subtypes, we fabricated hypoxia-responsive polymeric micelles co-loading drug and shRNA to treat TNBC by targeting hypoxic tumor microenvironment and subsequently targeting overexpressed HIF-1α under hypoxia. The micelles were assembled from methoxy-polyethylene glycol (mPEG) and poly-L-lysine (PLL) copolymer with AZO as a hypoxia-responsive bridge of mPEG and PLL. Once exposed to hypoxia, AZO bridge was cleaved, resulting in the disassembly of the micelles for rapid release. In vitro and in vivo results showed that the micelles enabled simultaneous delivery of drug and shRNA to hypoxic sites for site-specific rapid release, facilitated by sensitive response to hypoxia; hypoxia-responsive shRNA delivery effectively silenced HIF-1α and its downstream genes, which not only ameliorated the response of hypoxic tumor to drug, but also modulated tumor microenvironment for further improved drug and shRNA delivery; as a result, synergistic treatment of chemotherapy and HIF-1α targeted gene therapy inhibited the growth of primary TNBC tumor and its distant metastasis in a murine model of orthotopic TNBC. Together with their good biocompatibility, hypoxia-responsive polymeric micelles thus emerged as a safe, effective, and universally applicable drug and gene carrier for treatment of TNBC as well as other hypoxic tumors.