Correction for New perspectives of the cardiac cellular landscape: mapping cellular mediators of cardiac fibrosis using single-cell transcriptomics

Single-cell transcriptomics enables inference of context-dependent phenotypes of individual cells and determination of cellular diversity of complex tissues. Cardiac fibrosis is a leading factor in the development of heart failure and a major cause of morbidity and mortality worldwide with no effective treatment. Single-cell RNA-sequencing (scRNA-seq) offers a promising new platform to identify new cellular and molecular protagonists that may drive cardiac fibrosis and development of heart failure. This review will summarize the application scRNA-seq for understanding cardiac fibrosis and development of heart failure. We will also discuss some key considerations in interpreting scRNA-seq data and some of its limitations.

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Single-cell analysis of the long noncoding RNA transcriptome identifies novel therapeutic targets for cardiac fibrosis

Archives of Cardiovascular Diseases Supplements ◽

10.1016/j.acvdsp.2020.03.105 ◽

2020 ◽

Vol 12 (2-4) ◽

pp. 243

Author(s):

P. Aghagolzadeh ◽

R. Bernasconi ◽

M. Nemir ◽

H. Khalil ◽

C. Pulido ◽

...

Keyword(s):

Single Cell ◽

Long Noncoding Rna ◽

Noncoding Rna ◽

Cardiac Fibrosis ◽

Single Cell Analysis ◽

Therapeutic Targets ◽

Cell Analysis ◽

Novel Therapeutic

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Cardiac Fibroblast Diversity

Annual Review of Physiology ◽

10.1146/annurev-physiol-021119-034527 ◽

2020 ◽

Vol 82 (1) ◽

pp. 63-78 ◽

Cited By ~ 8

Author(s):

Michelle D. Tallquist

Keyword(s):

Extracellular Matrix ◽

Single Cell ◽

Future Development ◽

Cardiac Fibrosis ◽

Cardiac Fibroblasts ◽

Pathological Condition ◽

Cardiac Fibroblast ◽

Lineage Tracing ◽

Single Cell Sequencing ◽

Disease States

Cardiac fibrosis is a pathological condition that occurs after injury and during aging. Currently, there are limited means to effectively reduce or reverse fibrosis. Key to identifying methods for curbing excess deposition of extracellular matrix is a better understanding of the cardiac fibroblast, the cell responsible for collagen production. In recent years, the diversity and functions of these enigmatic cells have been gradually revealed. In this review, I outline current approaches for identifying and classifying cardiac fibroblasts. An emphasis is placed on new insights into the heterogeneity of these cells as determined by lineage tracing and single-cell sequencing in development, adult, and disease states. These recent advances in our understanding of the fibroblast provide a platform for future development of novel therapeutics to combat cardiac fibrosis.

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Cardiac Fibrosis and Fibroblasts

Cells ◽

10.3390/cells10071716 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1716

Author(s):

Hitoshi Kurose

Keyword(s):

Extracellular Matrix ◽

Single Cell ◽

Inflammatory Cytokines ◽

Cardiac Dysfunction ◽

Cardiac Fibrosis ◽

Structural Integrity ◽

New Technologies ◽

Time Dependent ◽

Cardiac Rupture ◽

Fibroblast Heterogeneity

Cardiac fibrosis is the excess deposition of extracellular matrix (ECM), such as collagen. Myofibroblasts are major players in the production of collagen, and are differentiated primarily from resident fibroblasts. Collagen can compensate for the dead cells produced by injury. The appropriate production of collagen is beneficial for preserving the structural integrity of the heart, and protects the heart from cardiac rupture. However, excessive deposition of collagen causes cardiac dysfunction. Recent studies have demonstrated that myofibroblasts can change their phenotypes. In addition, myofibroblasts are found to have functions other than ECM production. Myofibroblasts have macrophage-like functions, in which they engulf dead cells and secrete anti-inflammatory cytokines. Research into fibroblasts has been delayed due to the lack of selective markers for the identification of fibroblasts. In recent years, it has become possible to genetically label fibroblasts and perform sequencing at single-cell levels. Based on new technologies, the origins of fibroblasts and myofibroblasts, time-dependent changes in fibroblast states after injury, and fibroblast heterogeneity have been demonstrated. In this paper, recent advances in fibroblast and myofibroblast research are reviewed.

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Targeted Ablation of Periostin-Expressing Activated Fibroblasts Prevents Adverse Cardiac Remodeling in Mice

Circulation Research ◽

10.1161/circresaha.116.308643 ◽

2016 ◽

Vol 118 (12) ◽

pp. 1906-1917 ◽

Cited By ~ 94

Author(s):

Harmandeep Kaur ◽

Mikito Takefuji ◽

C.Y. Ngai ◽

Jorge Carvalho ◽

Julia Bayer ◽

...

Keyword(s):

Myocardial Infarction ◽

Angiotensin Ii ◽

Cardiac Function ◽

Single Cell ◽

Cardiac Remodeling ◽

Cardiac Fibrosis ◽

Cardiac Fibroblasts ◽

Transcriptional Analysis ◽

Cardiomyocyte Hypertrophy ◽

Cardiac Damage

Rationale: Activated cardiac fibroblasts (CF) are crucial players in the cardiac damage response; excess fibrosis, however, may result in myocardial stiffening and heart failure development. Inhibition of activated CF has been suggested as a therapeutic strategy in cardiac disease, but whether this truly improves cardiac function is unclear. Objective: To study the effect of CF ablation on cardiac remodeling. Methods and Results: We characterized subgroups of murine CF by single-cell expression analysis and identified periostin as the marker showing the highest correlation to an activated CF phenotype. We generated bacterial artificial chromosome–transgenic mice allowing tamoxifen-inducible Cre expression in periostin-positive cells as well as their diphtheria toxin-mediated ablation. In the healthy heart, periostin expression was restricted to valvular fibroblasts; ablation of this population did not affect cardiac function. After chronic angiotensin II exposure, ablation of activated CF resulted in significantly reduced cardiac fibrosis and improved cardiac function. After myocardial infarction, ablation of periostin-expressing CF resulted in reduced fibrosis without compromising scar stability, and cardiac function was significantly improved. Single-cell transcriptional analysis revealed reduced CF activation but increased expression of prohypertrophic factors in cardiac macrophages and cardiomyocytes, resulting in localized cardiomyocyte hypertrophy. Conclusions: Modulation of the activated CF population is a promising approach to prevent adverse cardiac remodeling in response to angiotensin II and after myocardial infarction.

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Primate heart regeneration via migration and fibroblast repulsion by human heart progenitors

10.1101/2020.07.03.183798 ◽

2020 ◽

Author(s):

Christine Schneider ◽

Kylie S. Foo ◽

Maria Teresa De Angelis ◽

Gianluca Santamaria ◽

Franziska Reiter ◽

...

Keyword(s):

Single Cell ◽

Human Heart ◽

Cardiac Fibrosis ◽

Lineage Tracing ◽

Heart Regeneration ◽

Cardiac Progenitors ◽

Cytoskeletal Dynamics ◽

Human Primate ◽

Muscular Regeneration

SUMMARYHuman heart regeneration is one of the most critical unmet clinical needs at a global level1. Muscular regeneration is hampered both by the limited renewing capacity of adult cardiomyocytes2-4 and the onset of cardiac fibrosis5,6, resulting in reduced compliance of the tissue. Primate have proven to be ideal models for pluripotent stem cell strategies for heart regeneration, but unravelling specific approaches to drive cell migration to the site of injury and inhibition of subsequent fibrosis have been elusive. Herein, by combining human cardiac progenitor lineage tracing and single-cell transcriptomics in injured non-human primate heart bio-mimics, we uncover the coordinated muscular regeneration of the primate heart via directed migration of human ventricular progenitors to sites of injury, subsequent fibroblast repulsion targeting fibrosis, and ultimate functional replacement of damaged cardiac muscle by differentiation and electromechanical integration. Single-cell RNAseq captured distinct modes of action, uncovering chemoattraction mediated by CXCL12/CXCR4 signalling and fibroblast repulsion regulated by SLIT2/ROBO1 guidance in organizing cytoskeletal dynamics. Moreover, transplantation of human cardiac progenitors into hypo-immunogenic CAG-LEA29Y transgenic porcine hearts following injury proved their chemotactic response and their ability to generate a remuscularized scar without the risk of arrhythmogenesis in vivo. Our study demonstrates that inherent developmental programs within cardiac progenitors are sequentially activated in the context of disease, allowing the cells to sense and counteract injury. As such, they may represent an ideal bio-therapeutic for functional heart rejuvenation.

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Electron microscopy of keratinocytes cultured on a collagen substrate used as an allograft for the treatment of chronic wounds

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100130158 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1104-1105

Author(s):

Debby A. Jennings ◽

Michael J. Morykwas ◽

Louis C. Argenta

Keyword(s):

Electron Microscopy ◽

Cell Suspension ◽

Single Cell ◽

Chronic Wounds ◽

Mechanical Stability ◽

Uv Light ◽

Type I ◽

Single Cell Suspension ◽

Collagen Substrate ◽

Dermal Collagen

Grafts of cultured allogenic or autogenic keratlnocytes have proven to be an effective treatment of chronic wounds and burns. This study utilized a collagen substrate for keratinocyte and fibroblast attachment. The substrate provided mechanical stability and augmented graft manipulation onto the wound bed. Graft integrity was confirmed by light and transmission electron microscopy.Bovine Type I dermal collagen sheets (100 μm thick) were crosslinked with 254 nm UV light (13.5 Joules/cm2) to improve mechanical properties and reduce degradation. A single cell suspension of third passage neonatal foreskin fibroblasts were plated onto the collagen. Five days later, a single cell suspension of first passage neonatal foreskin keratinocytes were plated on the opposite side of the collagen. The grafts were cultured for one month.The grafts were fixed in phosphate buffered 4% formaldehyde/1% glutaraldehyde for 24 hours. Graft pieces were then washed in 0.13 M phosphate buffer, post-fixed in 1% osmium tetroxide, dehydrated, and embedded in Polybed 812.

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