TAZ/TEAD complex regulates TGF-β1-mediated fibrosis in iPSC-derived renal organoids

AbstractChronic kidney disease (CKD) progresses by replacement of functional tissue compartments with fibrosis, representing a maladaptive repair process. Shifting kidney repair towards a physiologically-intact architecture, rather than fibrosis, is key to blocking CKD progression. In this study, we developed a fibrosis model that uses human induced pluripotent stem cell (iPSC)-based three-dimensional renal organoids, in which exogenous TGF-β1 induces production of extracellular matrix. In these organoids, TGF- β1 increased transcription factor tafazzin (TAZ) expression. Further, in human kidney biopsies, nuclear TAZ expression was markedly increased in mild and moderate fibrosis. In cultured renal tubular cells expressing a fibrogenic program, TAZ formed a trimeric complex with phosphorylated mothers against decapentaplegic homolog 3 (p-SMAD3) and TEA domain protein (TEAD)-4. Overexpression of TEAD4 protein suppressed collagen-1α1 (COL1A1) promoter activity, and expression of TAZ attenuated this inhibition. INT-767, a dual bile acid receptor agonist binding farnesoid X receptor (FXR) and the Takeda G protein-coupled receptor 5 (TGR5), decreased the TGF-β1-induced increase in p-SMAD3 and TAZ, and preserved renal organoid architecture. These data demonstrate, in an iPSC-derived renal organoid fibrosis model, that INT767 prevents fibrosis programs early in the course of tubular injury through modulation of the TEAD4/TAZ pathway.

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Permissive effect of GSK3β on profibrogenic plasticity of renal tubular cells in progressive chronic kidney disease

Cell Death and Disease ◽

10.1038/s41419-021-03709-5 ◽

2021 ◽

Vol 12 (5) ◽

Author(s):

Bohan Chen ◽

Pei Wang ◽

Xianhui Liang ◽

Chunming Jiang ◽

Yan Ge ◽

...

Keyword(s):

Chronic Kidney Disease ◽

Kidney Disease ◽

Renal Fibrosis ◽

Binding Protein ◽

Renal Tubules ◽

Genetic Ablation ◽

Renal Tubular Cells ◽

Renal Tubular ◽

Renal Fibrogenesis ◽

Tgf Β1

AbstractRenal tubular epithelial cells (TECs) play a key role in renal fibrogenesis. After persistent injuries that are beyond self-healing capacity, TECs will dedifferentiate, undergo growth arrest, convert to profibrogenic phenotypes, and resort to maladaptive plasticity that ultimately results in renal fibrosis. Evidence suggests that glycogen synthase kinase (GSK) 3β is centrally implicated in kidney injury. However, its role in renal fibrogenesis is obscure. Analysis of publicly available kidney transcriptome database demonstrated that patients with progressive chronic kidney disease (CKD) exhibited GSK3β overexpression in renal tubulointerstitium, in which the predefined hallmark gene sets implicated in fibrogenesis were remarkably enriched. In vitro, TGF-β1 treatment augmented GSK3β expression in TECs, concomitant with dedifferentiation, cell cycle arrest at G2/M phase, excessive accumulation of extracellular matrix, and overproduction of profibrotic cytokines like PAI-1 and CTGF. All these profibrogenic phenotypes were largely abrogated by GSK3β inhibitors or by ectopic expression of a dominant-negative mutant of GSK3β but reinforced in cells expressing the constitutively active mutant of GSK3β. Mechanistically, GSK3β suppressed, whereas inhibiting GSK3β facilitated, the activity of cAMP response element-binding protein (CREB), which competes for CREB-binding protein, a transcriptional coactivator essential for TGF-β1/Smad signaling pathway to drive TECs profibrogenic plasticity. In vivo, in mice with folic acid-induced progressive CKD, targeting of GSK3β in renal tubules via genetic ablation or by microdose lithium mitigated the profibrogenic plasticity of TEC, concomitant with attenuated interstitial fibrosis and tubular atrophy. Collectively, GSK3β is likely a pragmatic therapeutic target for averting profibrogenic plasticity of TECs and improving renal fibrosis.

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

Genes ◽

10.3390/genes12010112 ◽

2021 ◽

Vol 12 (1) ◽

pp. 112

Author(s):

Marta García-López ◽

Joaquín Arenas ◽

M. Esther Gallardo

Keyword(s):

Stem Cell ◽

Retinal Ganglion Cells ◽

Pluripotent Stem Cell ◽

Ganglion Cells ◽

Genetic Disorders ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Underlying Mechanism ◽

Retinal Ganglion ◽

Induced Pluripotent

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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Interleukin-22 attenuates renal tubular cells inflammation and fibrosis induced by TGF-β1 through Notch1 signaling pathway

Renal Failure ◽

10.1080/0886022x.2020.1753538 ◽

2020 ◽

Vol 42 (1) ◽

pp. 381-390 ◽

Cited By ~ 3

Author(s):

Rong Tang ◽

Xiangcheng Xiao ◽

Yang Lu ◽

Huihui Li ◽

Qiaoling Zhou ◽

...

Keyword(s):

Signaling Pathway ◽

Tubular Cells ◽

Interleukin 22 ◽

Renal Tubular Cells ◽

Notch1 Signaling ◽

Renal Tubular ◽

Notch1 Signaling Pathway ◽

Tgf Β1

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Single-cell RNA sequencing of human kidney

Scientific Data ◽

10.1038/s41597-019-0351-8 ◽

2020 ◽

Vol 7 (1) ◽

Cited By ~ 20

Author(s):

Jinling Liao ◽

Zhenyuan Yu ◽

Yang Chen ◽

Mengying Bao ◽

Chunlin Zou ◽

...

Keyword(s):

Single Cell ◽

Rna Sequencing ◽

Cell Biology ◽

Kidney Diseases ◽

Human Kidney ◽

High Quality ◽

Renal Tubular Cells ◽

Single Cell Rna Sequencing ◽

Normal Human ◽

Renal Tubular

AbstractA comprehensive cellular anatomy of normal human kidney is crucial to address the cellular origins of renal disease and renal cancer. Some kidney diseases may be cell type-specific, especially renal tubular cells. To investigate the classification and transcriptomic information of the human kidney, we rapidly obtained a single-cell suspension of the kidney and conducted single-cell RNA sequencing (scRNA-seq). Here, we present the scRNA-seq data of 23,366 high-quality cells from the kidneys of three human donors. In this dataset, we show 10 clusters of normal human renal cells. Due to the high quality of single-cell transcriptomic information, proximal tubule (PT) cells were classified into three subtypes and collecting ducts cells into two subtypes. Collectively, our data provide a reliable reference for studies on renal cell biology and kidney disease.

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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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Correction: Arg tyrosine kinase modulates TGF-β1 production in human renal tubular cells under high-glucose conditions (doi:10.1242/jcs.183640)

Journal of Cell Science ◽

10.1242/jcs.238832 ◽

2019 ◽

Vol 132 (19) ◽

pp. jcs238832

Author(s):

Barbara Torsello ◽

Cristina Bianchi ◽

Chiara Meregalli ◽

Vitalba Di Stefano ◽

Lara Invernizzi ◽

...

Keyword(s):

Tyrosine Kinase ◽

High Glucose ◽

Tubular Cells ◽

Renal Tubular Cells ◽

Arg Tyrosine Kinase ◽

Renal Tubular ◽

Tgf Β1

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Blocking core fucosylation of TGF-β1 receptors downregulates their functions and attenuates the epithelial-mesenchymal transition of renal tubular cells

AJP Renal Physiology ◽

10.1152/ajprenal.00426.2010 ◽

2011 ◽

Vol 300 (4) ◽

pp. F1017-F1025 ◽

Cited By ~ 37

Author(s):

Hongli Lin ◽

Dapeng Wang ◽

Taihua Wu ◽

Cui Dong ◽

Nan Shen ◽

...

Keyword(s):

Transforming Growth Factor ◽

Epithelial Mesenchymal Transition ◽

Interstitial Fibrosis ◽

Cell Model ◽

Mesenchymal Transition ◽

Renal Tubular Cells ◽

Renal Tubular ◽

Core Fucosylation ◽

Tgf Β1

Posttranslational modification of proteins could regulate their multiple biological functions. Transforming growth factor-β receptor I and II (ALK5 and TGF-βRII), which are glycoproteins, play important roles in the renal tubular epithelial-mesenchymal transition (EMT). In the present study, we examined the role of core fucosylation of TGF-βRII and ALK5, which is regulated by α-1,6 fucosyltransferase (Fut8), in the process of EMT of cultured human renal proximal tubular epithelial (HK-2) cells. The typical cell model of EMT induced by TGF-β1 was constructed to address the role of core fucosylation in EMT. Core fucosylation was found to be essential for both TGF-βRII and ALK5 to fulfill their functions, and blocking it with Fut8 small interfering RNA greatly reduced the phosphorylation of Smad2/3 protein, caused the inactivation of TGF-β/Smad2/3 signaling, and resulted in remission of EMT. More importantly, even with high levels of expressions of TGF-β1, TGF-βRII, and ALK5, blocking core fucosylation also could attenuate the EMT of HK-2 cells. Thus blocking core fucosylation of TGF-βRII and ALK5 may attenuate EMT independently of the expression of these proteins. This study may provide new insight into the role of glycosylation in renal interstitial fibrosis. Furthermore, core fucosylation may be a novel potential therapeutic target for treatment of renal tubular EMT.

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Dynamic loading of human engineered heart tissue enhances contractile function and drives a desmosome-linked disease phenotype

Science Translational Medicine ◽

10.1126/scitranslmed.abd1817 ◽

2021 ◽

Vol 13 (603) ◽

pp. eabd1817

Author(s):

Jacqueline M. Bliley ◽

Mathilde C. S. C. Vermeer ◽

Rebecca M. Duffy ◽

Ivan Batalov ◽

Duco Kramer ◽

...

Keyword(s):

Dynamic Loading ◽

Disease Modeling ◽

Contractile Function ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Heart Tissue ◽

Disease Phenotype ◽

Heart Formation ◽

Induced Pluripotent ◽

And Function

The role that mechanical forces play in shaping the structure and function of the heart is critical to understanding heart formation and the etiology of disease but is challenging to study in patients. Engineered heart tissues (EHTs) incorporating human induced pluripotent stem cell (hiPSC)–derived cardiomyocytes have the potential to provide insight into these adaptive and maladaptive changes. However, most EHT systems cannot model both preload (stretch during chamber filling) and afterload (pressure the heart must work against to eject blood). Here, we have developed a new dynamic EHT (dyn-EHT) model that enables us to tune preload and have unconstrained contractile shortening of >10%. To do this, three-dimensional (3D) EHTs were integrated with an elastic polydimethylsiloxane strip providing mechanical preload and afterload in addition to enabling contractile force measurements based on strip bending. Our results demonstrated that dynamic loading improves the function of wild-type EHTs on the basis of the magnitude of the applied force, leading to improved alignment, conduction velocity, and contractility. For disease modeling, we used hiPSC-derived cardiomyocytes from a patient with arrhythmogenic cardiomyopathy due to mutations in the desmoplakin gene. We demonstrated that manifestation of this desmosome-linked disease state required dyn-EHT conditioning and that it could not be induced using 2D or standard 3D EHT approaches. Thus, a dynamic loading strategy is necessary to provoke the disease phenotype of diastolic lengthening, reduction of desmosome counts, and reduced contractility, which are related to primary end points of clinical disease, such as chamber thinning and reduced cardiac output.

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A Mechanism Leading to Changes in Copy Number Variations Affected by Transcriptional Level Might Be Involved in Evolution, Embryonic Development, Senescence, and Oncogenesis Mediated by Retrotransposons

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.618113 ◽

2021 ◽

Vol 9 ◽

Author(s):

Yunpeng Sui ◽

Shuanghong Peng

Keyword(s):

Embryonic Development ◽

Copy Number ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Copy Number Variations ◽

Mrna Splicing ◽

Transcriptional Level ◽

Underlying Mechanisms ◽

Induced Pluripotent ◽

Hiv 1

In recent years, more and more evidence has emerged showing that changes in copy number variations (CNVs) correlated with the transcriptional level can be found during evolution, embryonic development, and oncogenesis. However, the underlying mechanisms remain largely unknown. The success of the induced pluripotent stem cell suggests that genome changes could bring about transformations in protein expression and cell status; conversely, genome alterations generated during embryonic development and senescence might also be the result of genome changes. With rapid developments in science and technology, evidence of changes in the genome affected by transcriptional level has gradually been revealed, and a rational and concrete explanation is needed. Given the preference of the HIV-1 genome to insert into transposons of genes with high transcriptional levels, we propose a mechanism based on retrotransposons facilitated by specific pre-mRNA splicing style and homologous recombination (HR) to explain changes in CNVs in the genome. This mechanism is similar to that of the group II intron that originated much earlier. Under this proposed mechanism, CNVs on genome are dynamically and spontaneously extended in a manner that is positively correlated with transcriptional level or contract as the cell divides during evolution, embryonic development, senescence, and oncogenesis, propelling alterations in them. Besides, this mechanism explains several critical puzzles in these processes. From evidence collected to date, it can be deduced that the message contained in genome is not just three-dimensional but will become four-dimensional, carrying more genetic information.

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Direct SARS-CoV-2 infection of the human inner ear may underlie COVID-19-associated audiovestibular dysfunction

Communications Medicine ◽

10.1038/s43856-021-00044-w ◽

2021 ◽

Vol 1 (1) ◽

Author(s):

Minjin Jeong ◽

Karen E. Ocwieja ◽

Dongjun Han ◽

P. Ashley Wackym ◽

Yichen Zhang ◽

...

Keyword(s):

Inner Ear ◽

Hair Cells ◽

Molecular Mechanisms ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

In Vitro Models ◽

Cellular Models ◽

Induced Pluripotent ◽

Human Inner Ear

Abstract Background COVID-19 is a pandemic respiratory and vascular disease caused by SARS-CoV-2 virus. There is a growing number of sensory deficits associated with COVID-19 and molecular mechanisms underlying these deficits are incompletely understood. Methods We report a series of ten COVID-19 patients with audiovestibular symptoms such as hearing loss, vestibular dysfunction and tinnitus. To investigate the causal relationship between SARS-CoV-2 and audiovestibular dysfunction, we examine human inner ear tissue, human inner ear in vitro cellular models, and mouse inner ear tissue. Results We demonstrate that adult human inner ear tissue co-expresses the angiotensin-converting enzyme 2 (ACE2) receptor for SARS-CoV-2 virus, and the transmembrane protease serine 2 (TMPRSS2) and FURIN cofactors required for virus entry. Furthermore, hair cells and Schwann cells in explanted human vestibular tissue can be infected by SARS-CoV-2, as demonstrated by confocal microscopy. We establish three human induced pluripotent stem cell (hiPSC)-derived in vitro models of the inner ear for infection: two-dimensional otic prosensory cells (OPCs) and Schwann cell precursors (SCPs), and three-dimensional inner ear organoids. Both OPCs and SCPs express ACE2, TMPRSS2, and FURIN, with lower ACE2 and FURIN expression in SCPs. OPCs are permissive to SARS-CoV-2 infection; lower infection rates exist in isogenic SCPs. The inner ear organoids show that hair cells express ACE2 and are targets for SARS-CoV-2. Conclusions Our results provide mechanistic explanations of audiovestibular dysfunction in COVID-19 patients and introduce hiPSC-derived systems for studying infectious human otologic disease.

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