Time-programmed activation of CD47 disruption and immunogenic cell death with Cas9 ribonucleoprotein nanocapsule for improved cancer immunotherapy

Cancer immunotherapy refers to activating the body's antitumor immunity to fight cancer cells, showing great potential in long-term inhibition of tumor growth and recurrence. However, adequate activation of antitumor immunity remains challenging due to overactivated immune checkpoints and poor tumor immunogenicity. To address these challenges, herein, a Cas9 ribonucleoprotein (RNP) nanocapsule (Cas9NC) is described to synergistically activate antitumor immunity by time-programmed activation of CD47 disruption and immunogenic cell death (ICD) of tumor cells. Such Cas9NC is formed by coating RNP with a thin polymer shell made of multiple functional monomers, which not only endows Cas9NC with the capability to deliver payloads into tumor tissues, but also activates CRISPR-Cas9 system and ICD in a time-programmed manner. This time-programmed activation strategy significantly promotes the synergy of two payloads with different onset times. Additionally, CD47 disruption achieve superior therapeutic effects than antibody that achieve temporary conformational blockade. As a result, Cas9NC initiates robust and durable antitumor immune responses, and remarkably inhibits the growth of both primary and abscopal tumors, prevents the malignant tumor recurrence and metastasis. Collectively, this time-programmed activation strategy for synergistic CD47 disruption and ICD provides a promising approach for improved cancer immunotherapy.