Therapeutic genome editing of triple-negative breast tumors using a noncationic and deformable nanolipogel

Abstract

Triple-negative breast cancer (TNBC), which has the highest mortality rate of all breast cancer, is in urgent need of a therapeutic that hinders the spread and growth of cancer cells. CRISPR genome editing holds the promise of a potential cure for many genetic diseases, including TNBC; however, its clinical translation is being challenged by the lack of safe and effective nonviral delivery systems for in vivo therapeutic genome editing. Here we report the synthesis and application of a noncationic, deformable, and tumor-targeted nanolipogel system (tNLG) for CRISPR genome editing in TNBC tumors. We have demonstrated that tNLGs mediate a potent CRISPR knockout of Lipocalin 2 (Lcn2), a known breast cancer oncogene, in human TNBC cells in vitro and in vivo. The loss of Lcn2 significantly inhibits the migration and the mesenchymal phenotype of human TNBC cells and subsequently attenuates TNBC aggressiveness. In an orthotopic TNBC model, we have shown that systemically administered tNLGs mediated >81% CRISPR knockout of Lcn2 in TNBC tumor tissues, resulting in significant tumor growth suppression (>77%). Our proof-of-principle results provide experimental evidence that tNLGs can be used as a safe, precise, and effective delivery approach for in vivo CRISPR genome editing in TNBC.

Biological Functions and Therapeutic Potential of Lipocalin 2 in Cancer

“…Effective carriers for delivering either the plasmids containing the guide RNA (gRNA) and Cas9 DNA sequences or the gRNA/Cas-9 ribonucleoprotein complex are needed. To knock out the LCN2 gene from triple-negative breast cancer cells, Guo et al [85] developed a DOPC (1,2-Dioleoyl-sn-Glycero-3-Phosphocholine)-based nanolipogel formulation and alginate hydrogel to co-deliver plasmids with gRNAs and Cas9 sequences. Three different CRISPR plasmids were encapsulated in DOPC and sequestered within alginate hydrogel conjugated to an intercellular adhesion molecule (ICAM-1) antibody for breast cancer cell specificity [85,86].…”

“…To knock out the LCN2 gene from triple-negative breast cancer cells, Guo et al [85] developed a DOPC (1,2-Dioleoyl-sn-Glycero-3-Phosphocholine)-based nanolipogel formulation and alginate hydrogel to co-deliver plasmids with gRNAs and Cas9 sequences. Three different CRISPR plasmids were encapsulated in DOPC and sequestered within alginate hydrogel conjugated to an intercellular adhesion molecule (ICAM-1) antibody for breast cancer cell specificity [85,86]. The combination of alginate as a noncationic, low-toxicity component in the nanoformulation decreases the possible cationic charge-induced toxicity of the nanoparticles [85,87].…”

“…Three different CRISPR plasmids were encapsulated in DOPC and sequestered within alginate hydrogel conjugated to an intercellular adhesion molecule (ICAM-1) antibody for breast cancer cell specificity [85,86]. The combination of alginate as a noncationic, low-toxicity component in the nanoformulation decreases the possible cationic charge-induced toxicity of the nanoparticles [85,87]. The delivery of this gene editing system allowed for a specific interaction between the nanoparticles and the triple-negative breast cancer cells, resulting in a knockout of the LCN2 gene.…”

Gene interfered-ferroptosis therapy of cancer

“…The detection of Lcn2 expression in 13 various mouse tissues in this study also revealed that the expression of Lcn2 is much higher in tumor than in other normal tissues ( Figures 6F and 7F). A recent study revealed that a CRISPR/Cas9-mediated knockdown of Lcn2 significantly inhibits migration of human triple-negative breast cancer (TNBC) cells of the mesenchymal phenotype, weakens TNBC aggressiveness, and suppress in vivo TNBC tumor growth [53]. Therefore, we deduced that inhibition of the two genes should be fetal for the IONPs-treated cancer cells, because their inhibition would prevent cell from exporting IONPs-produced iron ions, which can thus enhance IONPs-induced ferroptosis.…”

Effect of an alkyl spacer on the morphology and internalization of MUC1 aptamer‐naphthalimide amphiphiles for targeting and imaging triple negative breast cancer cells

“…2 Due to the lack of many primary therapeutic targets, there are few targeted therapy options available, leaving only surgery, chemotherapeutics, and radiation therapy as viable treatment options. [2][3][4][5] This route tends to be less effective with more pronounced side effects. As such, discovery of targeted delivery strategies is critical to improving breast cancer patient outcomes and experiences.…”