Autologous induced pluripotent stem cell-derived four-organ-chip

Microphysiological systems play a pivotal role in progressing toward a global paradigm shift in drug development. Here, we designed a four-organ-chip interconnecting miniaturized human intestine, liver, brain and kidney equivalents. All four organ models were predifferentiated from induced pluripotent stem cells from the same healthy donor and integrated into the microphysiological system. The coculture of the four autologous tissue models in one common medium deprived of tissue specific growth factors was successful over 14-days. Although there were no added growth factors present in the coculture medium, the intestine, liver and neuronal model maintained defined marker expression. Only the renal model was overgrown by coexisting cells and did not further differentiate. This model platform will pave the way for autologous coculture cross-talk assays, disease induction and subsequent drug testing.

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Towards an autologous iPSC-derived patient-on-a-chip

10.1101/376970 ◽

2018 ◽

Cited By ~ 3

Author(s):

Anja Patricia Ramme ◽

Leopold Koenig ◽

Tobias Hasenberg ◽

Christine Schwenk ◽

Corinna Magauer ◽

...

Keyword(s):

Drug Testing ◽

Induced Pluripotent Stem Cell ◽

Cell Types ◽

Organ Differentiation ◽

Stem Cell Source ◽

Organ Systems ◽

On Chip ◽

Induced Pluripotent ◽

Organ Models ◽

Model Platform

AbstractMicrophysiological systems are fundamental for progressing towards a global paradigm shift in drug development through the generation of patient-on-a-chip models. An increasing number of single- and multi-organ systems have been adopted by the pharmaceutical and cosmetic industries for predictive substance testing. These models run on heterogeneous tissues and cell types from different donors. However, a patient is an individual. Therefore, patient-on-a-chip systems need to be built from tissues from one autologous source. Individual on-chip organ differentiation from a single induced pluripotent stem cell source could provide a solution to this challenge.We designed a four-organ chip based on human physiology. It enables the interconnection of miniaturized human intestine, liver, brain and kidney equivalents. All four organ models were predifferentiated from induced pluripotent stem cells from the same healthy donor and integrated into the microphysiological system. The cross talk led to further differentiation over a 14-day cultivation period under pulsatile blood flow conditions in one common medium deprived of growth factors. This model platform will pave the way for disease induction and subsequent drug testing.

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Drug Testing in Human‐Induced Pluripotent Stem Cell–Derived Cardiomyocytes From a Patient With Short QT Syndrome Type 1

Clinical Pharmacology & Therapeutics ◽

10.1002/cpt.1449 ◽

2019 ◽

Vol 106 (3) ◽

pp. 642-651 ◽

Cited By ~ 4

Author(s):

Zhihan Zhao ◽

Xin Li ◽

Ibrahim El‐Battrawy ◽

Huan Lan ◽

Rujia Zhong ◽

...

Keyword(s):

Stem Cell ◽

Pluripotent Stem Cell ◽

Drug Testing ◽

Induced Pluripotent Stem Cell ◽

Syndrome Type ◽

Short Qt Syndrome ◽

Qt Syndrome ◽

Induced Pluripotent

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Genome-wide molecular effects of the neuropsychiatric 16p11 CNVs in an iPSC-to-iN neuronal model

10.1101/2020.02.09.940965 ◽

2020 ◽

Author(s):

Thomas R. Ward ◽

Xianglong Zhang ◽

Louis C. Leung ◽

Bo Zhou ◽

Kristin Muench ◽

...

Keyword(s):

Gene Expression ◽

Dna Methylation ◽

Cell Model ◽

Copy Number Variants ◽

Induced Pluripotent Stem Cell ◽

Neuronal Model ◽

Autism Spectrum ◽

Genome Wide ◽

Induced Pluripotent ◽

Methylation Patterns

AbstractCopy number variants (CNVs), either deletions or duplications, at the 16p11.2 locus in the human genome are known to increase the risk for autism spectrum disorders (ASD), schizophrenia, and for several other developmental conditions. Here, we investigate the global effects on gene expression and DNA methylation using a 16p11.2 CNV patient-derived induced pluripotent stem cell (iPSC) to induced neuron (iN) cell model system. This approach revealed genome-wide and cell-type specific alterations to both gene expression and DNA methylation patterns and also yielded specific leads on genes potentially contributing to some of the known 16p11.2 patient phenotypes. PCSK9 is identified as a possible contributing factor to the symptoms seen in carriers of the 16p11.2 CNVs. The protocadherin (PCDH) gene family is found to have altered DNA methylation patterns in the CNV patient samples. The iPSC lines used for this study are available through a repository as a resource for research into the molecular etiology of the clinical phenotypes of 16p11.2 CNVs and into that of neuropsychiatric and neurodevelopmental disorders in general.

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Metabolically-Driven Maturation of hiPSC-Cell Derived Cardiac Chip

10.1101/485169 ◽

2018 ◽

Cited By ~ 2

Author(s):

Nathaniel Huebsch ◽

Berenice Charrez ◽

Brian Siemons ◽

Steven C. Boggess ◽

Samuel Wall ◽

...

Keyword(s):

Pluripotent Stem Cell ◽

Disease Modeling ◽

Induced Pluripotent Stem Cell ◽

Left Ventricular ◽

Calcium Handling ◽

Acid Oxidation ◽

Microphysiological Systems ◽

Induced Pluripotent ◽

Drug Responsiveness

AbstractHuman induced pluripotent stem cell derived cardiomyocytes (hiPSC-CM) are a promising in vitro tool for drug development and disease modeling, but their immature electrophysiology limits diagnostic utility. Tissue engineering approaches involving aligned 3D cultures enhance hiPSC-CM structural maturation but are insufficient to induce mature electrophysiology. We hypothesized that mimicking post-natal switching of the heart’s primary ATP source from glycolysis to fatty acid oxidation could enhance electrophysiological maturation of hiPSC-CM. We combined hiPSC-CM with microfabricated culture chambers to form 3D cardiac microphysiological systems (MPS) that enhanced immediate microtissue alignment and tissue specific extracellular matrix (ECM) production. Using Robust Experimental design, we identified a maturation media that improved calcium handling in MPS derived from two genetically distinct hiPSC sources. Although calcium handling and metabolic maturation were improved in both genotypes, there was a divergent effect on action potential duration (APD): MPS that started with abnormally prolonged APD exhibited shorter APD in response to maturation media, whereas the same media prolonged the APD in MPS that started with aberrantly short APD. Importantly, the APD of both genotypes was brought near the range of 270-300ms observed in human left ventricular cardiomyocytes. Mathematical modeling explained these divergent phenotypes, and further predicted the response of matured MPS to drugs with known pro-arrhythmic effects. These results suggest that systematic combination of biophysical stimuli and metabolic cues can enhance the electrophysiological maturation of hiPSC-derived cardiomyocytes. However, they also reveal that maturation-inducing cues can have differential effects on electrophysiology depending on the baseline phenotype of hiPSC-CM. In silico models provide a valuable tool for predicting how changes in cellular maturation will manifest in drug responsiveness.

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

10.1101/2021.06.24.449576 ◽

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.

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Deficiency of the Lysosomal Protein CLN5 Alters Lysosomal Function and Movement

Biomolecules ◽

10.3390/biom11101412 ◽

2021 ◽

Vol 11 (10) ◽

pp. 1412

Author(s):

Indranil Basak ◽

Rachel A. Hansen ◽

Michael E. Ward ◽

Stephanie M. Hughes

Keyword(s):

Batten Disease ◽

Induced Pluripotent Stem Cell ◽

Epileptic Seizures ◽

Neuronal Model ◽

Cell System ◽

Motor Deficits ◽

Glutamatergic Neurons ◽

Human Neurons ◽

Induced Pluripotent ◽

Acidic Organelles

Batten disease is a devastating, childhood, rare neurodegenerative disease characterised by the rapid deterioration of cognition and movement, leading to death within ten to thirty years of age. One of the thirteen Batten disease forms, CLN5 Batten disease, is caused by mutations in the CLN5 gene, leading to motor deficits, mental deterioration, cognitive impairment, visual impairment, and epileptic seizures in children. A characteristic pathology in CLN5 Batten disease is the defects in lysosomes, leading to neuronal dysfunction. In this study, we aimed to investigate the lysosomal changes in CLN5-deficient human neurons. We used an induced pluripotent stem cell system, which generates pure human cortical-like glutamatergic neurons. Using CRISPRi, we inhibited the expression of CLN5 in human neurons. The CLN5-deficient human neurons showed reduced acidic organelles and reduced lysosomal enzyme activity measured by microscopy and flow cytometry. Furthermore, the CLN5-deficient human neurons also showed impaired lysosomal movement—a phenotype that has never been reported in CLN5 Batten disease. Lysosomal trafficking is key to maintain local degradation of cellular wastes, especially in long neuronal projections, and our results from the human neuronal model present a key finding to understand the underlying lysosomal pathology in neurodegenerative diseases.

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Application of Patient-Specific iPSCs for Modelling and Treatment of X-Linked Cardiomyopathies

International Journal of Molecular Sciences ◽

10.3390/ijms22158132 ◽

2021 ◽

Vol 22 (15) ◽

pp. 8132

Author(s):

Jennifer Zhang ◽

Oscar Hou-In Chou ◽

Yiu-Lam Tse ◽

Kwong-Man Ng ◽

Hung-Fat Tse

Keyword(s):

Drug Testing ◽

Control Cell ◽

Induced Pluripotent Stem Cell ◽

Patient Specific ◽

Mortality And Morbidity ◽

Danon Disease ◽

Disease Phenotypes ◽

Induced Pluripotent ◽

And Control

Inherited cardiomyopathies are among the major causes of heart failure and associated with significant mortality and morbidity. Currently, over 70 genes have been linked to the etiology of various forms of cardiomyopathy, some of which are X-linked. Due to the lack of appropriate cell and animal models, it has been difficult to model these X-linked cardiomyopathies. With the advancement of induced pluripotent stem cell (iPSC) technology, the ability to generate iPSC lines from patients with X-linked cardiomyopathy has facilitated in vitro modelling and drug testing for the condition. Nonetheless, due to the mosaicism of the X-chromosome inactivation, disease phenotypes of X-linked cardiomyopathy in heterozygous females are also usually more heterogeneous, with a broad spectrum of presentation. Recent advancements in iPSC procedures have enabled the isolation of cells with different lyonisation to generate isogenic disease and control cell lines. In this review, we will summarise the current strategies and examples of using an iPSC-based model to study different types of X-linked cardiomyopathy. The potential application of isogenic iPSC lines derived from a female patient with heterozygous Danon disease and drug screening will be demonstrated by our preliminary data. The limitations of an iPSC-derived cardiomyocyte-based platform will also be addressed.

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Induced Pluripotency: A Powerful Tool for In Vitro Modeling

International Journal of Molecular Sciences ◽

10.3390/ijms21238910 ◽

2020 ◽

Vol 21 (23) ◽

pp. 8910 ◽

Cited By ~ 1

Author(s):

Romana Zahumenska ◽

Vladimir Nosal ◽

Marek Smolar ◽

Terezia Okajcekova ◽

Henrieta Skovierova ◽

...

Keyword(s):

Drug Testing ◽

Disease Modeling ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Cellular Mechanisms ◽

Major Health ◽

Induced Pluripotency ◽

Developmental Capacity ◽

Induced Pluripotent

One of the greatest breakthroughs of regenerative medicine in this century was the discovery of induced pluripotent stem cell (iPSC) technology in 2006 by Shinya Yamanaka. iPSCs originate from terminally differentiated somatic cells that have newly acquired the developmental capacity of self-renewal and differentiation into any cells of three germ layers. Before iPSCs can be used routinely in clinical practice, their efficacy and safety need to be rigorously tested; however, iPSCs have already become effective and fully-fledged tools for application under in vitro conditions. They are currently routinely used for disease modeling, preparation of difficult-to-access cell lines, monitoring of cellular mechanisms in micro- or macroscopic scales, drug testing and screening, genetic engineering, and many other applications. This review is a brief summary of the reprogramming process and subsequent differentiation and culture of reprogrammed cells into neural precursor cells (NPCs) in two-dimensional (2D) and three-dimensional (3D) conditions. NPCs can be used as biomedical models for neurodegenerative diseases (NDs), which are currently considered to be one of the major health problems in the human population.

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