Biological Systems Engineering University of Nebraska

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Engineering the Cell Microenvironment for
Nonviral Gene Delivery

Beth Duensing, Heidi Gengenbach
 

Inefficiency is a critical factor limiting the use of gene delivery in therapeutic applications including gene therapy to treat genetic deficiencies or tissue engineering matrices for treatment of organ loss and failure. The inefficiency of gene delivery is primarily a result of the inability of nonviral vectors to overcome extra- and intracellular barriers, whose cellular components are largely unknown.

ECM

The extracellular matrix (ECM) is a complex structural entity surrounding and supporting cells within mammalian tissues, comprised of a variety of structural and specialized proteins. These proteins, in part through integrin binding, mediate cell adhesion and play a key role in cellular processes including migration, organization of the cytoskeleton, focal contacts, and signaling, which in turn affect proliferation, gene expression and apoptosis. Recent insights suggest that ECM components and receptors controlling cell adhesion, as well as their corresponding intracellular components, may influence the ability of cells to internalize nonviral DNA. However, the specific characteristics of the ECM, including its architecture and physical properties that may regulate the cellular response to gene delivery have not been investigated.

We are using self-assembled monolayers as controllable surfaces to investigate the effect of ECM proteins and other environmental factors on the ability of cells to take up DNA and express the corresponding protein. In particular, we examine how cells respond to their environment (presence/ orientation of ECM molecules and/or DNA complexes) in the context of DNA delivery, either through a traditional bolus approach or through substrate-mediated gene delivery for application to tissue engineering scaffolds.


Engineering Intracellular Signaling Pathways for Nonviral Gene Delivery


Novel Biomaterials


Single Cell Imaging