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

Victoria L Messerschmidt; Uday Chintapula; Aneetta E Kuriakose; Samantha Laboy; Thuy Thi Dang Truong; LeNaiya A Kydd; Justyn Jaworski; Zui Pan; Hashem Sadek; Kytai T Nguyen; Juhyun Lee

doi:10.3389/fcvm.2021.707897

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

Front Cardiovasc Med. 2021 Sep 28:8:707897. doi: 10.3389/fcvm.2021.707897. eCollection 2021.

Authors

Victoria L Messerschmidt^{1

2}, Uday Chintapula^{1

2}, Aneetta E Kuriakose^{1

2}, Samantha Laboy¹, Thuy Thi Dang Truong¹, LeNaiya A Kydd¹, Justyn Jaworski¹, Zui Pan³, Hashem Sadek², Kytai T Nguyen^{1

2}, Juhyun Lee^{1

2}

Affiliations

¹ Department of Bioengineering, University of Texas at Arlington, Arlington, TX, United States.
² Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX, United States.
³ College of Nursing and Health Innovation, University of Texas at Arlington, Arlington, TX, United States.

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.

Keywords: Notch signaling; PLGA; gene delivery; nanoparticles; non-viral transfection.