Scaffold-mediated CRISPR-Cas9 delivery system for acute myeloid leukemia therapy

Abstract

Leukemia stem cells (LSCs) sustain the disease and contribute to relapse in acute myeloid leukemia (AML). Therapies that ablate LSCs may increase the chance of eliminating this cancer in patients. To this end, we used a bioreducible lipidoid-encapsulated Cas9/single guide RNA (sgRNA) ribonucleoprotein [lipidoid nanoparticle (LNP)–Cas9 RNP] to target the critical gene interleukin-1 receptor accessory protein (IL1RAP) in human LSCs. To enhance LSC targeting, we loaded LNP-Cas9 RNP and the chemokine CXCL12α onto mesenchymal stem cell membrane–coated nanofibril (MSCM-NF) scaffolds mimicking the bone marrow microenvironment. In vitro, CXCL12α release induced migration of LSCs to the scaffolds, and LNP-Cas9 RNP induced efficient gene editing. IL1RAP knockout reduced LSC colony-forming capacity and leukemic burden. Scaffold-based delivery increased the retention time of LNP-Cas9 in the bone marrow cavity. Overall, sustained local delivery of Cas9/IL1RAP sgRNA via CXCL12α-loaded LNP/MSCM-NF scaffolds provides an effective strategy for attenuating LSC growth to improve AML therapy.

Research Progress on Gene Editing Based on Nano-Drug Delivery Vectors for Tumor Therapy

“…Finally, materials used in clinical trials and even clinical treatment must have a high level of safety ( Kamali Shahri et al, 2021 ; Martín-Sabroso et al, 2021 ; Phillips and Mousa, 2022 ). In addition, the use of nanoparticles to deliver CRISPR/Cas9 to acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) genes in animal models has been shown to be a promising strategy for the treatment of leukemia ( Liu et al, 2018 ; Ho et al, 2021 ; Ren et al, 2022 ). However, there are few reports about non-solid tumors.…”

Cell Membrane-Cloaked Nanotherapeutics for Targeted Drug Delivery

“…For example, Chen et al [122] demonstrated that pancreatic beta-cell membrane-coated nanofiber scaffolds enhanced cell proliferation and insulin secretion function in vitro. Recently, we reported a scaffold-mediated CRISPR/Cas9 local delivery system for acute myeloid leukemia therapy [123]. Mesenchymal stem cell membrane-coated nanofiber scaffolds enabled leukemia stem cell targeting as they were injected in the bone marrow.…”

Utilization of CRISPR-Mediated Tools for Studying Functional Genomics in Hematological Malignancies: An Overview on the Current Perspectives, Challenges, and Clinical Implications

“…Authors such as Ho et al, applied the gene-editing process of CRISPR/Cas systems in knocking out interleukin-1 receptor accessory protein (IL1RAP) genes in human leukemia stem cells in an attempt to abolish relapses of leukemia in AML patients by designing a bone marrow scaffold-mediated delivery of the therapeutic CRISPR system (Ho et al, 2021). Certain studies have utilized CRISPR-mediated gene knockouts in identifying the significant role of KAT7 genes for the survival of AML cells (Au et al, 2021).…”

“…An attempt to alter the abnormal gene function in AMLrelated disease led researchers to explore therapies that possibly ablate leukemia stem cells (LSCs), thus increasing the chances of eliminating this cancer in patients. A latest study conducted by Tzu-Chieh Ho et al, showcased scaffold-mediated CRISPR-Cas9 delivery system for targeting the gene interleukin-1 receptor accessory protein (IL1RAP) in human LSCs (Ho et al, 2021). This approach was mediated using a bio-reducible lipidoidencapsulated Cas9/single guide RNA (sgRNA) ribonucleoprotein-induced efficient gene editing.…”