Title

Novel siRNA Formulation to Effectively Knockdown Mutant p53 in Osteosarcoma.

Funding Source

National Institutes of Health, National Institute of General Medical Sciences, National Institute on Minority Health and Health Disparities, Louisiana Board of Regents

Grant Number

1G12RR026260-01,1R01CA127481,R21CA129776, 8UL1GM118967,R25GM060926, TL4GM118968,5G12MD007595, LEQSF (2007-12)-ENH-PKSFI-PRS-02,2007-11

Department

Department of Biology

Document Type

Article

Publication Date

6-2017

Abstract

Objectives: The tumor suppressor p53 plays a crucial role in the development of osteosarcoma. The primary objective of this study is to develop and optimize lipid based nanoparticle formulations that can carry siRNA and effectively silence mutant p53 in 318–1, a murine osteosarcoma cell line. Methods: The nanoparticles were composed of a mixture of two lipids (cholesterol and DOTAP) and either PLGA or PLGA-PEG and prepared by using an EmulsiFlex-B3 high pressure homogenizer. A series of studies that include using different nanoparticles, different amount of siRNAs, cell numbers, incubation time, transfection media volume, and storage temperature was performed to optimize the gene silencing efficiency. Key findings: Replacement of lipids by PLGA or PLGA-PEG decreased the particle size and overall cytotoxicity. Among all lipid-polymer nanoformulations, nanoparticles with 10% PLGA showed highest mutant p53 knockdown efficiency while maintaining higher cell viability when a nanoparticle to siRNA ratio equal to 6.8: 0.66 and 75 nM siRNA was used. With long term storage the mutant p53 knockdown efficiency decreased to a greater extent. Conclusions: This study warrants a future evaluation of this formulation for gene silencing efficiency of mutant p53 in tissue culture and animal models for the treatment of osteosarcoma.

Comments

DOI: 10.1371/journal.pone.0179168

PubMed ID: 28636657

This work is funded in part by the Louisiana Cancer Research Consortium, NIMHD grant number TL4GM118968 and 5G12MD007595, NIGMS grant number 8UL1GM118967 and R25GM060926, NIH grants 1R01CA127481, R21CA129776, 1G12RR026260-01, Louisiana Board of Regents RC/EEP (2007-11), and LEQSF (2007-12)-ENH-PKSFI-PRS-02, CUR from Xavier University of Louisiana and LBRN Pilot. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Link to Full Text

Share

COinS