Secretion of proteins and antibody fragments from transiently transfected endothelial progenitor cells.

Loree Heller,Richard Heller,Yu Jing,Reynald Thinard,Melanie Chevalier,Ruth Greferath,Sezgi Arpag,Claude Nicolau

doi:10.1111/jcmm.15511

Abstract

In neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, multiple sclerosis and amyotrophic lateral sclerosis, neuroinflammation can lead to blood‐brain barrier (BBB) breakdown. After intravenous or intra‐arterial injection into mice, endothelial progenitor cells (EPCs) home to the damaged BBB to promote neurovascular repair. Autologous EPCs transfected to express specific therapeutic proteins offer an innovative therapeutic option. Here, we demonstrate that EPC transfection by electroporation with plasmids encoding the reporter protein GFP or an anti‐β‐amyloid antibody fragment (Fab) leads to secretion of each protein. We also demonstrate the secreted anti‐β‐amyloid Fab protein functions in β‐amyloid aggregate solubilization.

Highlights

The blood-brain barrier (BBB) consists primarily of endothelial cells with pericytes, astrocytes and microglia on a basement membrane 1,2 and functions to regulate the transport of cells and proteins into the normal brain
We demonstrated that Endothelial progenitor cells (EPCs) can be transfected using electroporation at high efficiency with a minor loss of cell viability
human umbilical vein endothelial cells (HUVECs) viability was not assayed in the original PR0329 protocol

Summary

| INTRODUCTION

The blood-brain barrier (BBB) consists primarily of endothelial cells with pericytes, astrocytes and microglia on a basement membrane 1,2 and functions to regulate the transport of cells and proteins into the normal brain. We report here a novel targeted delivery system consisting of ex vivo transfected, autologous endothelial precursor cells (EPCs) capable of homing to the BBB and expressing therapeutic Fabs. These cells were obtained for the development of targeted cell-mediated gene therapy to a hypoxic site.[27] EPCs are able to repair the damaged BBB and blood-spinal cord barrier characteristic of neurodegenerative diseases such as AD,[28] ALS,[29,30] traumatic brain injury 31,32 or stroke 33,34 and will have a double function when combined with Fab production

| MATERIALS AND METHODS

Findings

| DISCUSSION