Hereditary Optic Neuropathies: Induced Pluripotent Stem Cell-Based 2D/3D Approaches

Inherited optic neuropathies share visual impairment due to the degeneration of retinal ganglion cells (RGCs) as the hallmark of the disease. This group of genetic disorders are caused by mutations in nuclear genes or in the mitochondrial DNA (mtDNA). An impaired mitochondrial function is the underlying mechanism of these diseases. Currently, optic neuropathies lack an effective treatment, and the implementation of induced pluripotent stem cell (iPSC) technology would entail a huge step forward. The generation of iPSC-derived RGCs would allow faithfully modeling these disorders, and these RGCs would represent an appealing platform for drug screening as well, paving the way for a proper therapy. Here, we review the ongoing two-dimensional (2D) and three-dimensional (3D) approaches based on iPSCs and their applications, taking into account the more innovative technologies, which include tissue engineering or microfluidics.

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Single Cell Sequencing of Induced Pluripotent Stem Cell Derived Retinal Ganglion Cells (iPSC-RGC) Reveals Distinct Molecular Signatures and RGC Subtypes

Genes ◽

10.3390/genes12122015 ◽

2021 ◽

Vol 12 (12) ◽

pp. 2015

Author(s):

Harini V. Gudiseva ◽

Vrathasha Vrathasha ◽

Jie He ◽

Devesh Bungatavula ◽

Joan M. O’Brien ◽

...

Keyword(s):

Gene Expression ◽

Stem Cell ◽

Single Cell ◽

Retinal Ganglion Cells ◽

Pluripotent Stem Cell ◽

Ganglion Cells ◽

Induced Pluripotent Stem Cell ◽

Retinal Ganglion ◽

Single Cell Sequencing ◽

Induced Pluripotent

We intend to identify marker genes with differential gene expression (DEG) and RGC subtypes in cultures of human-induced pluripotent stem cell (iPSC)-derived retinal ganglion cells. Single-cell sequencing was performed on mature and functional iPSC-RGCs at day 40 using Chromium Single Cell 3’ V3 protocols (10X Genomics). Sequencing libraries were run on Illumina Novaseq to generate 150 PE reads. Demultiplexed FASTQ files were mapped to the hg38 reference genome using the STAR package, and cluster analyses were performed using a cell ranger and BBrowser2 software. QC analysis was performed by removing the reads corresponding to ribosomal and mitochondrial genes, as well as cells that had less than 1X mean absolute deviation (MAD), resulting in 4705 cells that were used for further analyses. Cells were separated into clusters based on the gene expression normalization via PCA and TSNE analyses using the Seurat tool and/or Louvain clustering when using BBrowser2 software. DEG analysis identified subsets of RGCs with markers like MAP2, RBPMS, TUJ1, BRN3A, SOX4, TUBB3, SNCG, PAX6 and NRN1 in iPSC-RGCs. Differential expression analysis between separate clusters identified significant DEG transcripts associated with cell cycle, neuron regulatory networks, protein kinases, calcium signaling, growth factor hormones, and homeobox transcription factors. Further cluster refinement identified RGC diversity and subtype specification within iPSC-RGCs. DEGs can be used as biomarkers for RGC subtype classification, which will allow screening model systems that represent a spectrum of diseases with RGC pathology.

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Three-dimensional induced pluripotent stem-cell models of human brain angiogenesis

Microvascular Research ◽

10.1016/j.mvr.2020.104042 ◽

2020 ◽

Vol 132 ◽

pp. 104042 ◽

Cited By ~ 1

Author(s):

Raleigh M. Linville ◽

Diego Arevalo ◽

Joanna C. Maressa ◽

Nan Zhao ◽

Peter C. Searson

Keyword(s):

Stem Cell ◽

Human Brain ◽

Pluripotent Stem Cell ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Cell Models ◽

Induced Pluripotent ◽

Stem Cell Models ◽

Brain Angiogenesis

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Three-dimensional modeling of human airway development using induced pluripotent stem cell-derived teratomas

Journal of Translational Science ◽

10.15761/jts.1000242 ◽

2018 ◽

Vol 5 (1) ◽

Author(s):

Sansac C ◽

Belbachir A ◽

Ahmed E ◽

Ramirez JM ◽

Fort A ◽

...

Keyword(s):

Stem Cell ◽

Pluripotent Stem Cell ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Human Airway ◽

Three Dimensional Modeling ◽

Dimensional Modeling ◽

Induced Pluripotent ◽

Airway Development

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Phenotypic and Functional Characterization of Müller Glia Isolated from Induced Pluripotent Stem Cell‐Derived Retinal Organoids: Improvement of Retinal Ganglion Cell Function upon Transplantation

Stem Cells Translational Medicine ◽

10.1002/sctm.18-0263 ◽

2019 ◽

Cited By ~ 5

Author(s):

Karen Eastlake ◽

Weixin Wang ◽

Hari Jayaram ◽

Celia Murray‐Dunning ◽

Amanda J. F. Carr ◽

...

Keyword(s):

Stem Cell ◽

Ganglion Cell ◽

Pluripotent Stem Cell ◽

Cell Function ◽

Functional Characterization ◽

Induced Pluripotent Stem Cell ◽

Retinal Ganglion ◽

Induced Pluripotent ◽

Retinal Ganglion Cell Function

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Progress of Induced Pluripotent Stem Cell Technologies to Understand Genetic Epilepsy

International Journal of Molecular Sciences ◽

10.3390/ijms21020482 ◽

2020 ◽

Vol 21 (2) ◽

pp. 482 ◽

Cited By ~ 1

Author(s):

Bruno Sterlini ◽

Floriana Fruscione ◽

Simona Baldassari ◽

Fabio Benfenati ◽

Federico Zara ◽

...

Keyword(s):

Stem Cell ◽

Pluripotent Stem Cell ◽

Neurological Diseases ◽

Functional Characterization ◽

3D Structure ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Genetic Epilepsy ◽

Human Neurons ◽

Induced Pluripotent

The study of the pathomechanisms by which gene mutations lead to neurological diseases has benefit from several cellular and animal models. Recently, induced Pluripotent Stem Cell (iPSC) technologies have made possible the access to human neurons to study nervous system disease-related mechanisms, and are at the forefront of the research into neurological diseases. In this review, we will focalize upon genetic epilepsy, and summarize the most recent studies in which iPSC-based technologies were used to gain insight on the molecular bases of epilepsies. Moreover, we discuss the latest advancements in epilepsy cell modeling. At the two dimensional (2D) level, single-cell models of iPSC-derived neurons lead to a mature neuronal phenotype, and now allow a reliable investigation of synaptic transmission and plasticity. In addition, functional characterization of cerebral organoids enlightens neuronal network dynamics in a three-dimensional (3D) structure. Finally, we discuss the use of iPSCs as the cutting-edge technology for cell therapy in epilepsy.

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