Notch Intracellular Domain Plasmid Delivery via Poly(Lactic-Co-Glycolic Acid) Nanoparticles to Upregulate Notch Pathway Molecules

Abstract

Notch signaling is a highly conserved signaling system that is required for embryonic development and regeneration of organs. When the signal is lost, maldevelopment occurs and leads to a lethal state. Delivering exogenous genetic materials encoding Notch into cells can reestablish downstream signaling and rescue cellular functions. In this study, we utilized the negatively charged and FDA approved polymer poly(lactic-co-glycolic acid) to encapsulate Notch Intracellular Domain-containing plasmid in nanoparticles. We show that primary human umbilical vein endothelial cells (HUVECs) readily uptake the nanoparticles with and without specific antibody targets. We demonstrated that our nanoparticles are non-toxic, stable over time, and compatible with blood. We further demonstrated that HUVECs could be successfully transfected with these nanoparticles in static and dynamic environments. Lastly, we elucidated that these nanoparticles could upregulate the downstream genes of Notch signaling, indicating that the payload was viable and successfully altered the genetic downstream effects.

Supramolecular Peptide Nanofiber/PLGA Nanocomposites for Enhancing Pulmonary Drug Delivery

“…A double emulsion method as described by Messerschmidt et al was employed for the synthesis of PLGA NPs . First, 100 mg of PLGA polymer (Polyscitech, West Lafayette, USA) was dissolved in dichloromethane at 100 mg/mL.…”

“…A double emulsion method as described by Messerschmidt et al was employed for the synthesis of PLGA NPs. 58 First, 100 mg of PLGA polymer (Polyscitech, West Lafayette, USA) was dissolved in dichloromethane at 100 mg/mL. 1% (w/w) Rhodamine B (Rho B) (Sigma-Aldrich, St. Louis, USA) was prepared as a water phase, which was later added dropwise into the oil-phase of the PLGA solution.…”

In vivo Evaluation of Non-viral NICD Plasmid-Loaded PLGA Nanoparticles in Developing Zebrafish to Improve Cardiac Functions

“…PLGA is popular as a drug carrier for proteins ( Golub et al, 2010 ; Allahyari and Mohit, 2016 ; Haji Mansor et al, 2018 ), hydrophobic drugs ( Shi et al, 2015 ; Allahyari and Mohit, 2016 ; Khan et al, 2016 ; Malinovskaya et al, 2017 ; Madani et al, 2018 ; Wilkosz et al, 2018 ; Lu et al, 2019 ; Choi et al, 2020 ) and hydrophilic drugs ( Dalpiaz et al, 2016 ). More recently, PLGA has been used to carry genetic information to cells ( Gvili et al, 2007 ; Cun et al, 2011 ; Amreddy et al, 2017 ; Guan and Rosenecker, 2017 ; Kalvanagh et al, 2019 ; Messerschmidt et al, 2021a ). The various payload types speak to the versatility of PLGA and its’ ability to protect and deliver the contents to the body.…”

“…The various payload types speak to the versatility of PLGA and its’ ability to protect and deliver the contents to the body. Additionally, PLGA can be functionalized to further target specific cell lines ( Kocbek et al, 2007 ; Fan et al, 2014 ; Shi et al, 2015 ; Khang et al, 2020 ; Messerschmidt et al, 2021a ).…”