1. Scaffold-mediated CRISPR-Cas9 delivery system for acute myeloid leukemia therapy
- Author
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Michael W. Becker, Yamin Li, Yumei Chen, Qiaobing Xu, Tzu-Chieh Ho, Mark W. LaMere, Kam W. Leong, John M. Ashton, Hae-Won Kim, Haiyan Wang, Hye Sung Kim, Caroline Chen, Jing Gong, and Cal D. Palumbo
- Subjects
Chemokine ,02 engineering and technology ,03 medical and health sciences ,Genome editing ,hemic and lymphatic diseases ,Tumor Microenvironment ,Medicine ,Humans ,neoplasms ,Research Articles ,030304 developmental biology ,Cancer ,Gene Editing ,0303 health sciences ,Multidisciplinary ,biology ,business.industry ,Mesenchymal stem cell ,Myeloid leukemia ,SciAdv r-articles ,021001 nanoscience & nanotechnology ,medicine.disease ,Leukemia ,Leukemia, Myeloid, Acute ,medicine.anatomical_structure ,Ribonucleoproteins ,Applied Sciences and Engineering ,biology.protein ,Cancer research ,Bone marrow ,Stem cell ,CRISPR-Cas Systems ,0210 nano-technology ,business ,RNA, Guide, Kinetoplastida ,Research Article - Abstract
A scaffold-mediated system was developed to enhance the local delivery of Cas9 complexes for targeting leukemia stem cells in AML., 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.
- Published
- 2021