A Duplex CRISPR-Cas9 Ribonucleoprotein Nanomedicine for Colorectal Cancer Gene Therapy

Abstract

Based on the high frequency of concurrent adenomatous polyposis coli (<i>APC</i>) and <i>KRAS</i> mutations and their strong cooperative interaction in human colorectal cancer (CRC) promotion, we herein develop a CRISPR-Cas9-based genome-editing nanomedicine to target both <i>APC</i> and <i>KRAS</i> mutations for the treatment of CRC. To this end, a hyaluronic acid (HA)-decorated phenylboronic dendrimer (HAPD) was designed for the targeted delivery of Cas9 ribonucleoprotein (RNP), by which both <i>APC</i> and <i>KRAS</i> genetic mutations harboring in CRC cells can be synergistically disrupted. Systemic administration of Cas9 RNP targeting <i>APC</i> and <i>KRAS</i> enabled by HAPD significantly inhibits tumor growth on xenografted and orthotopic CRC mouse models and also greatly prevents CRC-induced liver metastasis and lung metastasis. Thus, this duplex genome-editing system provides a promising gene therapy strategy for the treatment of human CRC and can be extended to other types of cancers with activated Wnt/β-catenin and RAS/extracellular signal-regulated kinase (ERK) pathways.