A Novel Therapeutic siRNA Nanoparticle Designed for Dual-Targeting CD44 and Gli1 of Gastric Cancer Stem Cells.

Abstract

PURPOSE:

Gastric cancer stem cells (CSCs) are important for the initiation, growth, recurrence, and metastasis of gastric cancer, due to their chemo-resistance and indefinite proliferation. Herein, to eliminate gastric CSCs, we developed novel CSC-targeting glioma-associated oncogene homolog 1 (Gli1) small interfering RNA (siRNA) nanoparticles that are specifically guided by a di-stearoyl-phosphatidyl-ethanolamine- hyaluronic-acid (DSPE-HA) single-point conjugate, as an intrinsic ligand of the CD44 receptor. We refer to these as targeting Gli1 siRNA nanoparticles.

METHODS:

We used the reductive amination reaction method for attaching amine groups of DSPE to aldehydic group of hyaluronic acid (HA) at the reducing end, to synthesize the DSPE-HA single-point conjugate. Next, targeting Gli1 siRNA nanoparticles were prepared using the layer-by-layer assembly method. We characterized the stem cellular features of targeting Gli1 siRNA nanoparticles, including their targeting efficiency, self-renewal capacity, the migration and invasion capacity of gastric CSCs, and the penetration ability of 3D tumor spheroids. Next, we evaluated the therapeutic efficacy of the targeting Gli1 siRNA nanoparticles by using in vivo relapsed tumor models of gastric CSCs.

RESULTS:

Compared with the multipoint conjugates, DSPE-HA single-point conjugates on the surface of nanoparticles showed significantly higher binding affinities with CD44. The targeting Gli1 siRNA nanoparticles significantly decreased Gli1 protein expression, inhibited CSC tumor spheroid and colony formation, and suppressed cell migration and invasion. Furthermore, in vivo imaging demonstrated that targeting Gli1 siRNA nanoparticles accumulated in tumor tissues, showing significant antitumor recurrence efficacy in vivo.

CONCLUSION:

In summary, our targeting Gli1 siRNA nanoparticles significantly inhibited CSC malignancy features by specifically blocking Hedgehog (Hh) signaling both in vitro and in vivo, suggesting that this novel siRNA delivery system that specifically eliminates gastric CSCs provides a promising targeted therapeutic strategy for gastric cancer treatment.

• Authors: Hongjuan Yao, Lan Sun, Jingcao Li, Xiaofei Zhou, Rui Li, Rongguang Shao, Yingge Zhang, Liang Li
• Journal: International journal of nanomedicine (Int J Nanomedicine) Vol. 15 Pg. 7013-7034 ( 2020) ISSN: 1178-2013 [Electronic] New Zealand
• PMID: 33061365 (Publication Type: Journal Article)
• Copyright: © 2020 Yao et al.

Chemical References

• CD44 protein, human
• GLI1 protein, human
• Hedgehog Proteins
• Hyaluronan Receptors
• Nanoconjugates
• Phosphatidylethanolamines
• RNA, Small Interfering
• Zinc Finger Protein GLI1
• 1,2-distearoylphosphatidylethanolamine
• Hyaluronic Acid

Topics

• Animals
• Cell Line, Tumor
• Cell Movement (drug effects, genetics)
• Genetic Therapy (methods)
• Hedgehog Proteins (metabolism)
• Humans
• Hyaluronan Receptors (genetics)
• Hyaluronic Acid (chemistry)
• Male
• Mice, Inbred BALB C
• Molecular Targeted Therapy (methods)
• Nanoconjugates (administration & dosage, chemistry)
• Nanoparticles (administration & dosage, chemistry)
• Neoplastic Stem Cells (drug effects, metabolism, pathology)
• Phosphatidylethanolamines (chemistry)
• RNA, Small Interfering (administration & dosage, genetics)
• Signal Transduction (drug effects, genetics)
• Stomach Neoplasms (drug therapy, genetics, pathology)
• Xenograft Model Antitumor Assays
• Zinc Finger Protein GLI1 (genetics)