Systemic delivery of CRISPR/Cas9 with PEG-PLGA nanoparticles for chronic myeloid leukemia targeted therapy

Abstract

Chronic myeloid leukemia (CML), which is characterized by the Philadelphia translocation, which fuses breakpoint cluster region (BCR) sequences from chromosome 22 upstream of the Abelson murine leukemia viral oncogene homolog (ABL) on chromosome 9, requires specific and efficient treatment. The CRISPR/Cas9 system, with its mechanism of specific DNA complementary recognition by engineered guide RNA (gRNA), allows the development of novel therapeutics for CML. To achieve targeted therapy of CML with the CRISPR/Cas9 system, we encapsulated a CRISPR/Cas9 plasmid (pCas9) expressing gRNA targeting the overhanging fusion region of the BCR-ABL gene (pCas9/gBCR-ABL) with poly(ethylene glycol)-b-poly(lactic acid-co-glycolic acid) (PEG-PLGA)-based cationic lipid-assisted polymeric nanoparticles (CLANs), which specifically disrupted the CML-related BCR-ABL gene while sparing the BCR and ABL genes in normal cells. After intravenous injection, CLANs carrying pCas9/gBCR-ABL (CLANpCas9/gBCR-ABL) efficiently knocked out the BCR-ABL fusion gene of CML cells and improved the survival of a CML mouse model, indicating that the combination of the CRISPR/Cas9 system with nanocarriers is a promising strategy for targeted treatment of CML.

PLGA-Based Nanoparticles in Cancer Treatment

“…Mice that were i.v. injected with the nanocarrier showed a reduced number of myelogenous leukemia cells (K562) in blood and bone marrow and a significant prolonged survival rate after the treatment compared to naive and vehicle controls ( Liu et al, 2018 ).…”

Non-viral delivery of CRISPR/Cas9 complex using CRISPR-GPS nanocomplexes

“…In order to address these challenges, transient delivery of sgRNA along with the Cas9 mRNA or Cas9 protein has been achieved using non-viral methods such as electroporation, 15,16 hydrodynamic injection, 17 microinjection, 18 lipids, [19][20][21][22] peptides, [23][24][25][26][27][28] polyethylenimine in combination with other agents such as DNA nanoclew, graphene oxide, cholesterol, [29][30][31] gold nanoparticles and other nanostructures, [32][33][34][35][36][37][38] extracellular vesicles, [39][40][41][42] virus-like particles, 43 and biolistic delivery in plants. 44,45 Apart from these methods, many hybrid frameworks have also been used to deliver CRISPR/Cas9.…”

Effective downregulation of BCR-ABL tumorigenicity by RNA targeted CRISPR-Cas13a

“…Earlier, in a study by García-Tuñón et al , it was observed that CRISPR Cas9 was able to effectively control the oncogenic capabilities of BCR-ABL chimeric gene in custom Boff-p210 cell line carrying BCR-ABL fusion transgene 9 . In a recent study, Liu et al demonstrated effective down regulation of BCR-ABL gene using CRISPR RNA guided Fokl Nucleases in both cell lines and murine xenograft model 30 . However, García-Tuñón et al , in their other research article on CRISPR, published recently, highlighted the limitation of CRISPR/Cas9 in effectively knocking out target DNA without a null variation inducing homology directed repair template in the construct.…”