Objective: Here we study the effect of microRNAs (miRs) biogenesis on the dopamine (DA) neuronal maintenance and protection from different stressor-induced insults.

Background: Despite many years of extensive research, Parkinson’s disease (PD) remains one of the commonest neurodegenerative disorders with mostly unknown etiology. Our recent data show that miRs, small non-coding RNAs regulating translation and stability of mRNA targets, may be implicated in PD pathology. We have demonstrated that selective homo- and heterozygous deletions of miRs processing enzyme Dicer in vivo cause development of PD-like symptoms. In addition, pharmacological stimulation of miRs biogenesis with enoxacin results in improved survival of primary DA neurons and attenuates their vulnerability to endoplasmic reticulum stress (1).

Methods: To elucidate the role of miRs biogenesis pathway in survival and stress resistance of DA neurons, we used several approaches to either boost or impair miRs maturation process. To stimulate miRs biogenesis, we overexpress Dicer and its interaction proteins, such as Trbp and Pact, in primary DA cultures using corresponding lentiviral vectors (LV). For impairing miRs maturation, we utilize Cre-recombinase delivered by LV to delete “floxed” Dicer1 gene allele in primary DA cultures from corresponding genetically modified mice. Alternatively, we are also setting up the single vector LV delivery system utilizing CRISPR/Cas9 gene editing to target Dicer and other members of miRs biogenesis machinery. Cells transduced with LV are further treated with different stressors, such as alpha-synuclein preformed fibrils, and their survival is analyzed by immunohistochemistry against tyrosine hydroxylase.

Results: Currently we are assessing the effect of overexpression of Dicer and other proteins involved in miR biogenesis on survival and protection of DA neurons from various PD-associated stressors. Additionally, obtained DA neurons with homo- and heterozygous deletion of Dicer are tested for their ability to withstand various stress conditions.

Conclusions: In summary, our data will assess the role of miR biogenesis in supporting survival and stress-resistance of primary DA neurons and evaluate the possibility of targeting Dicer and its interaction proteins for PD therapy.

References: 1) Chmielarz P, Konovalova J, Najam SS, Alter H, Piepponen TP, Erfle H, et al. Dicer and microRNAs protect adult dopamine neurons. Cell Death Dis. 2017;8(5):e2813.

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MDS Abstracts - https://www.mdsabstracts.org/abstract/investigating-the-role-of-microrna-biogenesis-pathway-in-neuroprotection-in-primary-neuronal-culture-model-of-parkinsons-disease/