A Combinatorial Cell and Drug Delivery Strategy for Huntington's Disease Using Pharmacologically Active Microcarriers and RNAi Neuronally-Committed Mesenchymal Stromal Cells.

Emilie M André,Saikrishna Kandalam,Claudia N Montero-Menei,Gaëtan J Delcroix,Laurence Sindji

doi:10.3390/pharmaceutics11100526

Abstract

For Huntington’s disease (HD) cell-based therapy, the transplanted cells are required to be committed to a neuronal cell lineage, survive and maintain this phenotype to ensure their safe transplantation in the brain. We first investigated the role of RE-1 silencing transcription factor (REST) inhibition using siRNA in the GABAergic differentiation of marrow-isolated adult multilineage inducible (MIAMI) cells, a subpopulation of MSCs. We further combined these cells to laminin-coated poly(lactic-co-glycolic acid) PLGA pharmacologically active microcarriers (PAMs) delivering BDNF in a controlled fashion to stimulate the survival and maintain the differentiation of the cells. The PAMs/cells complexes were then transplanted in an ex vivo model of HD. Using Sonic Hedgehog (SHH) and siREST, we obtained GABAergic progenitors/neuronal-like cells, which were able to secrete HGF, SDF1 VEGFa and BDNF, of importance for HD. GABA-like progenitors adhered to PAMs increased their mRNA expression of NGF/VEGFa as well as their secretion of PIGF-1, which can enhance reparative angiogenesis. In our ex vivo model of HD, they were successfully transplanted while attached to PAMs and were able to survive and maintain this GABAergic neuronal phenotype. Together, our results may pave the way for future research that could improve the success of cell-based therapy for HDs.

Highlights

Huntington’s disease (HD) is a genetic neurodegenerative disorder caused by the abnormal repetition of CAG nucleotides in the Huntingtin (HTT) gene
RE-1 silencing transcription factor (REST) mRNA was quantified by room temperature (RT)-qPCR (Figure 2C) and down-regulated to 70.7% with the siREST at Day 5 when compared to the control cells, receiving a scrambled small interfering RNA (siRNA)
We here demonstrated the therapeutic potential of this novel combinatorial strategy using lipid nanocapsules (LNC)-delivered siREST and neuronally-committed Mesenchymal stromal cells (MSCs) transported by laminin-covered microcarriers delivering brain-derived neurotrophic factor (BDNF)

Summary

Introduction

Huntington’s disease (HD) is a genetic neurodegenerative disorder caused by the abnormal repetition of CAG nucleotides in the Huntingtin (HTT) gene. HD is characterized by a progressive degeneration of striatal GABAergic medium spiny projection neurons, followed by a progressive degeneration extending throughout the brain [3] This results in involuntary movements, cognitive impairment and psychiatric manifestations, culminating in death around 15–20 years after the onset of motor symptoms [4,5]. The trans-differentiation of MSCs into a neural/neuronal lineage, even if possible in vitro, is far less efficient in vivo and their functional maturity remains too scarce [12] The rationale for their transplantation does not lie on their capacity to replace the damaged neuronal cells, but on their ability to neuroprotect and repair by their paracrine effects on the surrounding environment [8]. It was recently shown that the silencing of REST, obtained by a recombinant lentivirus carrying a small interfering RNA (siRNA) for REST, induced a neural/neuronal differentiation of MSCs [15,16]

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