AUTHOR=Guzmán-Sastoque Paula , Sotelo Sebastián , Esmeral Natalia P. , Albarracín Sonia Luz , Sutachan Jhon-Jairo , Reyes Luis H. , Muñoz-Camargo Carolina , Cruz Juan C. , Bloch Natasha I. 

TITLE=Assessment of CRISPRa-mediated gdnf overexpression in an In vitro Parkinson’s disease model

JOURNAL=Frontiers in Bioengineering and Biotechnology

VOLUME=12

YEAR=2024

URL=https://www.frontiersin.org/journals/bioengineering-and-biotechnology/articles/10.3389/fbioe.2024.1420183

DOI=10.3389/fbioe.2024.1420183

ISSN=2296-4185

ABSTRACT=<sec><title>Introduction</title><p>Parkinson’s disease (PD) presents a significant challenge in medical science, as current treatments are limited to symptom management and often carry significant side effects. Our study introduces an innovative approach to evaluate the effects of <italic>gdnf</italic> overexpression mediated by CRISPRa in an <italic>in vitro</italic> model of Parkinson’s disease. The expression of <italic>gdnf</italic> can have neuroprotective effects, being related to the modulation of neuroinflammation and pathways associated with cell survival, differentiation, and growth.</p></sec><sec><title>Methods</title><p>We have developed a targeted delivery system using a magnetite nanostructured vehicle for the efficient transport of genetic material. This system has resulted in a substantial increase, up to 200-fold) in <italic>gdnf</italic> expression in an <italic>In vitro</italic> model of Parkinson’s disease using a mixed primary culture of astrocytes, neurons, and microglia.</p></sec><sec><title>Results and Discussion</title><p>The delivery system exhibits significant endosomal escape of more than 56%, crucial for the effective delivery and activation of the genetic material within cells. The increased <italic>gdnf</italic> expression correlates with a notable reduction in MAO-B complex activity, reaching basal values of 14.8 μU/μg of protein, and a reduction in reactive oxygen species. Additionally, there is up to a 34.6% increase in cell viability in an <italic>In vitro</italic> Parkinson’s disease model treated with the neurotoxin MPTP. Our study shows that increasing <italic>gdnf</italic> expression can remediate some of the cellular symptoms associated with Parkinson’s disease in an <italic>in vitro</italic> model of the disease using a novel nanostructured delivery system.</p></sec>