# Micro RNA-based selection of hiPSC-derived dopaminergic neurons for genetic screening.

> **NIH NIH R03** · MASSACHUSETTS GENERAL HOSPITAL · 2020 · $168,000

## Abstract

Parkinson’s disease (PD) is a severe, second most common neurodegenerative disorder which is still poorly
understood and has few current treatment options. The clinical phenotype of PD is caused by the selective
degeneration of dopaminergic neurons in the substantia nigra pars compacta in the ventral midbrain. Though
several gene mutations associated with familial PD cases have been described, most PD cases are idiopathic,
where aging seems to be the major risk factor. For both familial and idiopathic PD, oxidative stress and
mitochondrial dysfunction are thought to underlie PD pathogenesis. How mitochondrial dysfunction effect
downstream neuronal targets and signaling pathways remains less well understood. Genetic screening is one of
the tools that are used to uncover new, potentially therapeutic targets for PD. Only few such genetic screens have
done to date, with none carried out in hiPSC-derived DA neurons, the neuronal population specifically affected
in PD. A major limitation with phenotypic screening in actual disease-affected neurons is that most
differentiation protocols result in heterogenous cell populations, confounding phenotypic readouts. Here we
propose to combine hiPSC-technology to generate 2 novel hiPSC mitophagy reporter lines with either expressing
a mitochondrial targeted, pH-sensitive fluophore (mt-Keima) to study effect PD-associated mutations on
mitophagy, or a hiPSC line expressing a combination of mt-Keima and an inducible, DNase inactive Cas9 fused
with the KRAB repressor domain for genome-wide screening purposes. In order to run future genetic and
phenotypic screens in homogenous, disease affected neuronal subtype populations, we aim to implement a cell
type-specific selection method based on cell type-specific microRNA expression. This miRNA-based molecular
switch will enable us to specifically select for dopaminergic neurons that we will use in phenotypic screens for
discovery of targets underlying disturbed mitophagy and mitochondrial dysfunction, associated with PD. The
outcome of this work will also provide a powerful new resource for genetic perturbation or phenotypic screens
for the broader research community.

## Key facts

- **NIH application ID:** 10042966
- **Project number:** 1R03NS118260-01
- **Recipient organization:** MASSACHUSETTS GENERAL HOSPITAL
- **Principal Investigator:** William Theodorus Hendriks
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $168,000
- **Award type:** 1
- **Project period:** 2020-09-15 → 2021-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10042966

## Citation

> US National Institutes of Health, RePORTER application 10042966, Micro RNA-based selection of hiPSC-derived dopaminergic neurons for genetic screening. (1R03NS118260-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10042966. Licensed CC0.

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