Profiling proliferative cells and their progeny in damaged murine hearts

Kretzschmar, Kai; Post, Yorick; Bannier-Hélaouët, Marie; Mattiotti, Andrea; Drost, Jarno; Basak, Onur; Li, Vivian; Born, Marise Ph.; Gunst, Quinn D.; Versteeg, Daniëlle; Kooijman, Lieneke; Elst, Stefan van der; Es, Johan H. van; Rooij, Eva van; Hoff, Maurice J.B. van den; Clevers, Hans

doi:10.1073/pnas.1805829115

Cited by 153 publications

(134 citation statements)

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“…Also, we found endothelial cells to express low amounts of mitochondrial associated proteins, which is surprising, given the central roles mitochondria play in the endothelial cell biology (Kluge et al, 2013). Consistent with past findings (Kretzschmar et al, 2018), IdU labeling was found to be most robust in endothelial cells and leukocytes.…”

Section: Integrated Analysis Reveals the Proportions And Relationshipsupporting

confidence: 86%

Assessing the Heterogeneity of Cardiac Non-myocytes and the Effect of Cell Culture with Integrative Single Cell Analysis

Davis²,

et al. 2020

Preprint

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Cardiac non-myocytes comprise a diverse and crucial cell population in the heart that plays dynamic roles in cardiac wound healing and growth. Non-myocytes broadly fall into four cell types: endothelium, fibroblasts, leukocytes, and pericytes. Here we characterize the diversity of the non-myocytes in vivo and in vitro using mass cytometry. By leveraging single-cell RNA sequencing we inform the design of a mass cytometry panel. To aid in annotation of the mass cytometry datasets, we utilize data integration with a neural network. We introduce approximately 460,000~ single cell proteomes of non-myocytes as well as 5,000~ CD31 negative single cell transcriptomes. Using our data, as well as previously reported datasets, we characterize cardiac non-myocytes with high depth in six mice, characterizing novel surface markers (CD9, CD200, Notch3, and FolR2). Further, we find that extended cell culture promotes the proliferation of CD45+CD11b+FolR2+IAIE-myeloid cells in addition to fibroblasts.

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Section: Integrated Analysis Reveals the Proportions And Relationshipsupporting

confidence: 86%

Assessing the Heterogeneity of Cardiac Non-myocytes and the Effect of Cell Culture with Integrative Single Cell Analysis

Davis²,

et al. 2020

Preprint

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“…Non-myocyte cardiac cell heterogeneity has been recently characterized based on FACS sorting different populations of immune, endothelial and cardiac fibroblast 14,28,29 or on the usage of lineage tracing models such as PDGFRα 30 or POSTN 9 reporter mice. Most of these studies, focus on the early phase after cardiac injury 30 and assessed other pathological conditions 9,28,29 .…”

Section: Discussionmentioning

confidence: 99%

“…In addition, our cell selection strategy was based on a different tracing model, potentially explaining some of the differences observed. For instance, Kretzschmar et al identified 11 clusters within 282 CF at 14dpi using Mki67-RFP reporter mice 28 . In this study, the authors described a subpopulation of activated CF located in the scar region with a similar function that IRCFs.…”

Section: Discussionmentioning

confidence: 99%

“…Future studies should addressed whether deletion of IRCF using Cre-inducible DTR transgenic mice models instead of deletion of the Cthrc1 hormone would result in a different outcome 32 . Based on their proliferation, a population of activated CF identified by their expression of Fstl1 and responsible for preventing cardiac rupture after MI has recently been characterized 28 which could also be part of our IRCF.…”

Section: Discussionmentioning

confidence: 99%

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Single-cell RNA-seq analysis reveals the crucial role of Collagen Triplex Helix Repeat Containing 1 (CTHRC1) cardiac fibroblasts for ventricular remodeling after myocardial infarction

Ruiz‐Villalba

Romero

Hernandez

et al. 2019

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Cardiac fibroblasts have a central role during the ventricular remodeling process associated with different types of cardiac injury. Recent studies have shown that fibroblasts do not respond homogeneously to heart damage, suggesting that the adult myocardium may contain specialized fibroblast subgroups with specific functions. Due to the limited set of bona fide fibroblast markers, a proper characterization of fibroblast population dynamics in response to cardiac damage is still missing. Using single-cell RNA-seq, we identified and characterized a fibroblast subpopulation that emerges in response to myocardial infarction (MI) in a murine model. These activated fibroblasts exhibit a clear pro-fibrotic signature, express high levels of the hormone CTHRC1 and of the immunomodulatory co-receptor CD200 and localize to the injured myocardium. Combining epigenomic profiling with functional assays, we show Sox9 and the non-canonical TGF-β signaling as important regulators mediating their response to cardiac damage. We show that the absence of CTHRC1, in this activated fibroblast subpopulation, results in pronounced lethality due to ventricular rupture in a mouse model of myocardial infarction. Finally, we find evidence for the existence of similar mechanisms in a pig preclinical model of MI and establish a correlation between CTHRC1 levels and cardiac function after MI.

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“…These observations may also explain why direct injection into the parenchyma of the heart, as done in the majority of animal studies 7-9 , is universally efficacious while pre-clinical studies using systemic vascular infusion of progenitor cells as performed in the majority of human clinical trials have shown mixed results 4,6,58,59 . Indeed, it is uncertain how systemic vascular delivery of progenitor cells could provide benefit to the heart given the mechanism of action we proposed here, as the vast majority of infused cells do not take up residence in the heart nor persist in the circulatory system 60,61 .Our study was designed to examine the mechanistic basis of adult cardiac stem cell therapy given the new consensus that direct cardiomyocyte regeneration is no longer a tenable mechanism 17,62 and that the adult mammalian heart is largely non-regenerative and does not contain a resident stem cell population [63][64][65] . As suggested 10 years ago 66 , we observed that the acute inflammatory response is the overwhelming mechanism of benefit behind cell therapy to the post-MI injured heart.…”

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confidence: 99%

An acute immune response underlies the benefit of cardiac adult stem cell therapy

Vagnozzi

Maillet

Sargent

et al. 2018

Preprint

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Clinical trials using adult stem cells to regenerate damaged heart tissue continue to this day 1-3 despite ongoing questions of efficacy and a lack of mechanistic understanding of the underlying biologic effect 4-6 . The rationale for these cell therapy trials is derived from animal studies that show a modest but reproducible improvement in cardiac function in models of cardiac ischemic injury 7-9 . Here we examined the mechanistic basis for cell therapy in mice after ischemia/reperfusion (I/R) injury, and while heart function was enhanced, it was not associated with new cardiomyocyte production. Cell therapy improved heart function through an acute sterile immune response characterized by the temporal and regional induction of CCR2 + and CX3CR1 + macrophages. Here we observed that intra-cardiac injection of 2 distinct types of progenitor cells, freeze/thaw-killed cells or a chemical inducer of the innate immune response similarly induced regional CCR2 + and CX3CR1 + macrophage accumulation and provided functional rejuvenation to the I/R-injured heart. Mechanistically, this selective macrophage response altered cardiac fibroblast activity and reduced border zone extracellular matrix (ECM) content and enhanced the mechanical properties of the injured area. The functional benefit of cardiac cell therapy is thus due to an acute inflammatory-based wound healing response that rejuvenates the mechanical properties of the infarcted area of the heart. Such results suggest a re-evaluation of strategies underlying cardiac cell therapy in current and planned human clinical trials.Initial animal studies of adult stem or progenitor cell therapy for cardiac regeneration reported improved heart function with robust de novo myogenesis derived directly from the injected cells [10][11][12][13] . Many independent groups have since repeated these studies with an ever-increasing variety of adult stem cells and while functional improvement is reproducibly observed, nearly all have failed to observe new cardiomyocyte formation from these injected cells [14][15][16][17] . At the same time, clinical trials with adult stem cells in patients with acute myocardial infarction (MI) injury or decompensated heart failure have expanded worldwide over the past 17 years, with thousands of patients enrolled and over $1 billion USD in funding utilized 4,6 . However, while results of these trials have been disappointing, this might simply reflect ineffective trial design because the true mechanistic basis of cell therapy remains unknown 6 . For example, more recent literature has shown that the vast majority of transplanted adult stem or progenitor cells simply die within days of delivery into the hearts of ischemia-injured animal models 18,19 , and as such it remains unclear how they might function in a therapeutic manner, although a paracrine hypothesis has been proposed whereby injected cells temporarily release protective growth factors or RNA species [20][21][22] .Over 15 types of adult stem cells show some level of efficacy in cardiac regenerat...

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Profiling proliferative cells and their progeny in damaged murine hearts

Cited by 153 publications

References 54 publications

Assessing the Heterogeneity of Cardiac Non-myocytes and the Effect of Cell Culture with Integrative Single Cell Analysis

Assessing the Heterogeneity of Cardiac Non-myocytes and the Effect of Cell Culture with Integrative Single Cell Analysis

Single-cell RNA-seq analysis reveals the crucial role of Collagen Triplex Helix Repeat Containing 1 (CTHRC1) cardiac fibroblasts for ventricular remodeling after myocardial infarction

An acute immune response underlies the benefit of cardiac adult stem cell therapy

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