Pharmacological Transdifferentiation of Human Nasal Olfactory Stem Cells into Dopaminergic Neurons

Abstract

The discovery of novel drugs for neurodegenerative diseases has been a real challenge over the last decades. The development of patient- and/or disease-specific in vitro models represents a powerful strategy for the development and validation of lead candidates in preclinical settings. The implementation of a reliable platform modeling dopaminergic neurons will be an asset in the study of dopamine-associated pathologies such as Parkinson’s disease. Disease models based on cell reprogramming strategies, using either human-induced pluripotent stem cells or transcription factor-mediated transdifferentiation, are among the most investigated strategies. However, multipotent adult stem cells remain of high interest to devise direct conversion protocols and establish in vitro models that could bypass certain limitations associated with reprogramming strategies. Here, we report the development of a six-step chemically defined protocol that drives the transdifferentiation of human nasal olfactory stem cells into dopaminergic neurons. Morphological changes were progressively accompanied by modifications matching transcript and protein dopaminergic signatures such as LIM homeobox transcription factor 1 alpha (LMX1A), LMX1B, and tyrosine hydroxylase (TH) expression, within 42 days of differentiation. Phenotypic changes were confirmed by the production of dopamine from differentiated neurons. This new strategy paves the way to develop more disease-relevant models by establishing reprogramming-free patient-specific dopaminergic cell models for drug screening and/or target validation for neurodegenerative diseases.

Dual control exerted by dopamine in blood-progenitor cell cycle regulation in Drosophila

“…We undertook an antibody based staining approach to investigate if blood cells of the Drosophila larval lymph gland showed any levels of dopamine. We performed an analysis of this monoamine in a late 3 rd instar (96-102h A fter E gg L aying, AEL) lymph gland hematopoietic tissue, using an antibody raised against dopamine that is routinely used to detect dopamine in neuronal cultures (Banks et al, 2017, Chabrat et al, 2019) and also been reported in invertebrate hemocytes (Wu SF et al, 2015). The staining was undertaken in a genetic background that allowed comparative analysis of dopamine in progenitor cells (using a progenitor marker domeMESO-Gal4, UAS-GFP ) which are Dome + GFP + as opposed to differentiating cells that are Dome - GFP - .…”

Direct Conversion of Human Fibroblasts into Adipocytes Using a Novel Small Molecular Compound: Implications for Regenerative Therapy for Adipose Tissue Defects

“…Even terminally differentiated somatic cells can be epigenetically reprogrammed, leading to phenotype conversion of the cells into a different lineage [ 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 ]. In this study, we developed a procedure to induce adipocytes from HDFs by stimulating the cells with a single small molecule, STK287794.…”

“…Fibroblasts can be obtained from patients with minimal invasiveness and expanded to a sufficient number in culture. Instead of transducing transcription factor genes, culturing of fibroblasts in the presence of certain chemical compounds may also induce direct reprogramming [ 15 , 16 , 17 , 18 , 19 , 20 ]. These transgene-free procedures may be more appropriate for cell processing for regenerative therapy compared with gene transduction because the cells induced may be less tumorigenic due to the absence of genetic aberrations associated with gene transduction.…”

Direct Reprogramming of Somatic Cells to Neurons: Pros and Cons of Chemical Approach

“…However, it was showed that a single TF such as ASCL1, using a novel protein intracellular delivery technology, in combination with the small molecules LDN193189, SB431542, DAPT and valproic acid can rapidly reprogram astrocytes into mature GABAergic and glutamatergic interneurons with high efficiency [ 47 ]. Moreover, Chabrat et al developed a novel in vitro model of dopaminergic-like neurons derived from human nasal olfactory stem cells through a six step transdifferentiation protocol based on a specific combination of signaling pathway modulators [ 60 ]. Indeed, chemical cocktails offer the possibility of fine-tuning their effects by altering their concentrations and combinations.…”