scholarly journals Reintroduction of the archaic variant of NOVA1 in cortical organoids alters neurodevelopment

Science ◽  
2021 ◽  
Vol 371 (6530) ◽  
pp. eaax2537 ◽  
Author(s):  
Cleber A. Trujillo ◽  
Edward S. Rice ◽  
Nathan K. Schaefer ◽  
Isaac A. Chaim ◽  
Emily C. Wheeler ◽  
...  
Keyword(s):  
Neural Development ◽  
Modern Human ◽  
Protein Coding ◽  
Electrophysiological Properties ◽  
Induced Pluripotent Cells ◽  
Synaptic Markers ◽  
Functional Consequences ◽  
Induced Pluripotent ◽  
Human Induced Pluripotent Cells ◽  
And Function

The evolutionarily conserved splicing regulator neuro-oncological ventral antigen 1 (NOVA1) plays a key role in neural development and function. NOVA1 also includes a protein-coding difference between the modern human genome and Neanderthal and Denisovan genomes. To investigate the functional importance of an amino acid change in humans, we reintroduced the archaic allele into human induced pluripotent cells using genome editing and then followed their neural development through cortical organoids. This modification promoted slower development and higher surface complexity in cortical organoids with the archaic version of NOVA1. Moreover, levels of synaptic markers and synaptic protein coassociations correlated with altered electrophysiological properties in organoids expressing the archaic variant. Our results suggest that the human-specific substitution in NOVA1, which is exclusive to modern humans since divergence from Neanderthals, may have had functional consequences for our species’ evolution.

scholarly journals Histone Deacetylase 1 Deficiency Impairs Differentiation and Electrophysiological Properties of Cardiomyocytes Derived from Induced Pluripotent Cells

Stem Cells ◽  
2012 ◽  
Vol 30 (11) ◽  
pp. 2412-2422 ◽  
Author(s):  
Eneda Hoxha ◽  
Erin Lambers ◽  
Hehuang Xie ◽  
Alexandre De Andrade ◽  
Prasanna Krishnamurthy ◽  
...  
Keyword(s):  
Histone Deacetylase ◽  
Pluripotent Cells ◽  
Electrophysiological Properties ◽  
Histone Deacetylase 1 ◽  
Induced Pluripotent Cells ◽  
Induced Pluripotent

scholarly journals Non-coding RNAs Associated with Prader-Willi Syndrome Regulate Transcription of Neurodevelopmental Genes in Human Induced Pluripotent Stem Cells

2021 ◽  
Author(s):  
Monika Sledziowska ◽  
Matt Jones ◽  
Ruba Al Maghrabi ◽  
Daniel Hebenstreit ◽  
Paloma Garcia ◽  
...  
Keyword(s):  
Gene Expression ◽  
Specific Gene ◽  
Chromosome 15 ◽  
Prader Willi Syndrome ◽  
Transcriptional Signature ◽  
Induced Pluripotent Cells ◽  
Non Coding Rnas ◽  
Induced Pluripotent ◽  
Human Induced Pluripotent Cells ◽  
Aberrant Gene

Mutations and aberrant gene expression during cellular differentiation lead to neurodevelopmental disorders such as Prader-Willi syndrome (PWS) which results from the deletion of an imprinted locus on chromosome 15. We analysed chromatin-associated RNA in human induced pluripotent cells (iPSCs) upon depletion of hybrid small nucleolar long non-coding RNAs (sno-lncRNAs) and 5 snoRNA capped and polyadenylated long non-coding RNAs (SPA-lncRNAs) transcribed from the locus deleted in PWS. We found that rapid ablation of these lncRNAs affects transcription of specific gene classes. Downregulated genes contribute to neurodevelopment and neuronal maintenance while genes that are upregulated are predominantly involved in the negative regulation of cellular metabolism and apoptotic processes. Our data revealed the importance of SPA-lncRNAs and sno-lncRNAs in controlling gene expression in iPSCs and provided a platform for synthetic experimental approaches in PWS studies. We conclude that ncRNAs transcribed from the PWS locus are critical regulators of a transcriptional signature important for neuronal differentiation and development.

scholarly journals Engineered human induced pluripotent cells enable genetic code expansion in brain organoids

2021 ◽  
Author(s):  
Lea van Husen ◽  
Anna Maria Katsori ◽  
Birthe Meineke ◽  
Lars O Tjernberg ◽  
Sophia Schedin-Weiss ◽  
...  
Keyword(s):  
Genetic Code ◽  
Fluorescent Dyes ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Trna Synthetase ◽  
Induced Pluripotent Cells ◽  
Wide Range ◽  
Induced Pluripotent ◽  
Human Induced Pluripotent Cells ◽  
Tissue Models

Human induced pluripotent stem cell (hiPSC) technology has revolutionized human biology. A wide range of cell types and tissue models can be derived from hiPSCs to study complex human diseases. Here, we use PiggyBac mediated transgenesis to engineer hiPSCs with an expanded genetic code. We demonstrate that genomic integration of expression cassettes for a pyrrolysyl-tRNA synthetase (PylRS), pyrrolysyl-tRNA (PylT) and the target protein of interest enables site-specific incorporation of a non-canonical amino acid (ncAA) in response to amber stop codons. Neural stem cells, neurons and brain organoids derived from the engineered hiPSCs continue to express the amber suppression machinery and produce ncAA-bearing reporter. The incorporated ncAA can serve as a minimal bioorthogonal handle for further modifications by labeling with fluorescent dyes. Site-directed ncAA mutagenesis will open a wide range of applications to probe and manipulate proteins in brain organoids and other hiPSC-derived cell types and complex tissue models.

scholarly journals Self-organized Kidney Rudiments: Prospects for Better in vitro Nephrotoxicity Assays

2015 ◽  
Vol 10s1 ◽  
pp. BMI.S20056 ◽  
Author(s):  
Jamie A. Davies
Keyword(s):  
Stem Cells ◽  
In Vitro Assays ◽  
Limiting Factor ◽  
Induced Pluripotent Cells ◽  
Therapeutic Drugs ◽  
Vascular Regulation ◽  
Single Cell Type ◽  
Induced Pluripotent ◽  
Human Induced Pluripotent Cells

Kidneys are essential to life but vulnerable to a range of toxicants, including therapeutic drugs and their metabolites. Indeed, nephrotoxicity is often a limiting factor in both drug use and drug development. Most toxicants damage kidneys by one of four mechanisms: damage to the membrane and its junctions, oxidative stress and free radical generation, activation of inflammatory processes, and interference with vascular regulation. Traditionally, animal models were used in preclinical screening for nephrotoxicity, but these can be poorly predictive of human reactions. Animal screens have been joined by simple single-cell–type in vitro assays using primary or immortalized human cells, particularly proximal tubule cells as these are especially vulnerable to toxicants. Recent research, aimed mainly at engineering new kidneys for transplant purposes, has resulted in a method for constructing anatomically realistic mini-kidneys from renogenic stem cells. So far, this has been done only using renogenic stem cells obtained directly from mouse embryos but, in principle, it should be possible to make them from renogenically directed human-induced pluripotent cells. If this can be done, the resulting human-based mini-kidneys would be a promising system for detecting some types of nephrotoxicity and for developing nephroprotective drugs.

scholarly journals Engineered Human Induced Pluripotent Cells Enable Genetic Code Expansion in Brain Organoids

ChemBioChem ◽  
2021 ◽  
Author(s):  
Lea S. Husen ◽  
Anna‐Maria Katsori ◽  
Birthe Meineke ◽  
Lars O. Tjernberg ◽  
Sophia Schedin‐Weiss ◽  
...  
Keyword(s):  
Genetic Code ◽  
Pluripotent Cells ◽  
Induced Pluripotent Cells ◽  
Genetic Code Expansion ◽  
Induced Pluripotent ◽  
Human Induced Pluripotent Cells

scholarly journals Generation of human induced pluripotent cells on a human feeder layer

2009 ◽  
Vol 18 ◽  
pp. S-7-S-8
Author(s):  
M Dijon-Grinand ◽  
S Assou ◽  
L Nadal ◽  
C Monzo ◽  
S Hamamah ◽  
...  
Keyword(s):  
Feeder Layer ◽  
Pluripotent Cells ◽  
Induced Pluripotent Cells ◽  
Induced Pluripotent ◽  
Human Induced Pluripotent Cells

scholarly journals Engineered human induced pluripotent cells enable genetic code expansion in brain organoids

ChemBioChem ◽  
2021 ◽  
Author(s):  
Lea S. van Husen ◽  
Anna-Maria Katasori ◽  
Birthe Meineke ◽  
Lars O. Tjernberg ◽  
Sophia Schedin-Weiss ◽  
...  
Keyword(s):  
Genetic Code ◽  
Pluripotent Cells ◽  
Induced Pluripotent Cells ◽  
Genetic Code Expansion ◽  
Induced Pluripotent ◽  
Human Induced Pluripotent Cells

scholarly journals Immunoreactivity of Pluripotent Markers SSEA-5 and L1CAM in Human Tumors, Teratomas, and Induced Pluripotent Stem Cells

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Linda Cassidy ◽  
Meerim Choi ◽  
Jason Meyer ◽  
Rui Chang ◽  
Gail M. Seigel
Keyword(s):  
Cell Lines ◽  
Human Tumors ◽  
Histological Evaluation ◽  
Stem Cell Markers ◽  
Induced Pluripotent Cells ◽  
Undifferentiated Cells ◽  
Embryonic Antigen ◽  
Induced Pluripotent ◽  
Human Induced Pluripotent Cells ◽  
Pluripotent Marker

Pluripotent stem cell markers can be useful for diagnostic evaluation of human tumors. The novel pluripotent marker stage-specific embryonic antigen-5 (SSEA-5) is expressed in undifferentiated human induced pluripotent cells (iPSCs), but little is known about SSEA-5 expression in other primitive tissues (e.g., human tumors). We evaluated SSEA-5 immunoreactivity patterns in human tumors, cell lines, teratomas, and iPS cells together with another pluripotent cell surface marker L1 cell adhesion molecule (L1CAM). We tested two hypotheses: (1) SSEA-5 and L1CAM would be immunoreactive and colocalized in human tumors; (2) SSEA-5 and L1CAM immunoreactivity would persist in iPSCs following retinal differentiating treatment. SSEA-5 immunofluorescence was most pronounced in primitive tumors, such as embryonal carcinoma. In tumor cell lines, SSEA-5 was highly immunoreactive in Capan-1 cells, while L1CAM was highly immunoreactive in U87MG cells. SSEA-5 and L1CAM showed colocalization in undifferentiated iPSCs, with immunopositive iPSCs remaining after 20 days of retinal differentiating treatment. This is the first demonstration of SSEA-5 immunoreactivity in human tumors and the first indication of SSEA-5 and L1CAM colocalization. SSEA-5 and L1CAM warrant further investigation as potentially useful tumor markers for histological evaluation or as markers to monitor the presence of undifferentiated cells in iPSC populations prior to therapeutic use.

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