Novel therapeutic strategies targeting fibroblasts and fibrosis in heart disease

Gourdie, Robert G.; Dimmeler, Stefanie; Kohl, Peter

doi:10.1038/nrd.2016.89

Cited by 269 publications

(213 citation statements)

References 307 publications

(320 reference statements)

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“…For the heart in particular, excitable-nonexcitable cell cross-talk has potentially far-reaching conceptual, physiological, pathological, and therapeutic implications (36), because-in the long runit may allow one to steer atrial scars to remain electrically insulating postablation (e.g., by preventing heterocellular coupling) or in contrast, to preserve uniform ventricular conduction across small postinfarct scars (by locally increasing nonmyocyte-mediated AP conduction in the heart).…”

Section: Discussionmentioning

confidence: 99%

Electrotonic coupling of excitable and nonexcitable cells in the heart revealed by optogenetics

Quinn

Camelliti

Rog-Zielinska

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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186

177

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Electrophysiological studies of excitable organs usually focus on action potential (AP)-generating cells, whereas nonexcitable cells are generally considered as barriers to electrical conduction. Whether nonexcitable cells may modulate excitable cell function or even contribute to AP conduction via direct electrotonic coupling to AP-generating cells is unresolved in the heart: such coupling is present in vitro, but conclusive evidence in situ is lacking. We used genetically encoded voltage-sensitive fluorescent protein 2.3 (VSFP2.3) to monitor transmembrane potential in either myocytes or nonmyocytes of murine hearts. We confirm that VSFP2.3 allows measurement of cell type-specific electrical activity. We show that VSFP2.3, expressed solely in nonmyocytes, can report cardiomyocyte AP-like signals at the border of healed cryoinjuries. Using EM-based tomographic reconstruction, we further discovered tunneling nanotube connections between myocytes and nonmyocytes in cardiac scar border tissue. Our results provide direct electrophysiological evidence of heterocellular electrotonic coupling in native myocardium and identify tunneling nanotubes as a possible substrate for electrical cell coupling that may be in addition to previously discovered connexins at sites of myocyte-nonmyocyte contact in the heart. These findings call for reevaluation of cardiac nonmyocyte roles in electrical connectivity of the heterocellular heart.

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Section: Discussionmentioning

confidence: 99%

Electrotonic coupling of excitable and nonexcitable cells in the heart revealed by optogenetics

Quinn

Camelliti

Rog-Zielinska

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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186

177

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“…In addition to producing and secreting abundant ECM material, myofibroblasts acquire contractile activity through induction of genes such as Acta2 (α-smooth muscle actin [αSMA]), which allows these cells to physically remodel the ECM or scar after MI (7,8). While the physiologic function of myofibroblasts is generally beneficial, such as during acute wound healing, sustained activation and continued generation of these cells likely underlie inappropriate developmental remodeling and a wide range of adult fibrotic diseases (9)(10)(11). Thus, inhibition of myofibroblast conversion or their activity would be an attractive therapeutic approach in adult fibrotic disease states, including heart failure (12)(13)(14).…”

Section: Introductionmentioning

confidence: 99%

Fibroblast-specific TGF-β–Smad2/3 signaling underlies cardiac fibrosis

Khalil¹,

Kanisicak²,

Prasad³

et al. 2017

Journal of Clinical Investigation

675

551

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reverse transcribed using random oligo-dT primers and a Verso cDNA Synthesis Kit (Thermo Fisher, AB1453) according to the manufacturer's instructions. Real-time PCR was performed using SsoAdvanced SYBR Green (Bio-Rad, 6090), and Rpl7 expression was used for normalization. The following primer sets were used to identify transcripts: collagen 1a1, 5′-AATGGCACGGCTGTGTGCGA and 5′-AACGGGTCCCCTTG-GGCCTT; collagen 3a1, 5′-TCCCCTGGAATCTGTGAATC and 5′-TGAGTCGAATTGGGGAGAAT; periostin, 5′-ACGGAGCTCAGG-GCTGAAGATG and 5′-GTTTGGGCCCTGATCCCGAC.Cell death analysis. At 90% confluence, primary skin fibroblasts were treated with 200 nM staurosporine for 36 hours or vehicle (DMSO). Cell death was determined by the Muse Count & Viability Assay (Millipore, MCH100102) as previously described (62). Briefly, the medium was collected with the trypsin-liberated cells, which were centrifuged and washed twice with PBS and then incubated with the Muse Count & Viability reagent. The cells were then quantified on a Muse cell analyzer (Millipore) at 5,000 counts per sample.Statistics. One-way ANOVA with post hoc Tukey's honest significant difference (HSD) or Student's t test was used to determine statistical significance, depending on the type of data analyzed and number of comparisons. P values of less than 0.05 were considered statistically significant. Averaged data are presented with SEM to indicate variability.Study approval. Mice were observed daily and cages changed weekly by certified veterinary technicians at Cincinnati Children's Hospital Medical Center. Mice were also closely assessed for their well-being, monitored by adequate physical activity and food intake on a daily basis. Housing conditions and husbandry conformed to AAALAC standards as well as the standard guidelines from the NIH Office of Laboratory Animal Welfare (http://grants.nih.gov/grants/olaw/animal_use. htm). The institution also retains ongoing certification by AAALAC.

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“…Meanwhile, chloride channels play essential roles in a variety of physiological functions including epithelial secretion, smooth muscle contraction, and sensory transduction [8]. Increasing evidence indicates that calcium-activated chloride channels (CaCCs) may participate in CF function [9][10][11] and be potential drug targets for the treatment of cardiovascular diseases [12].…”

Section: Introductionmentioning

confidence: 99%

Effects of the Calcium-Activated Chloride Channel Inhibitors T16Ainh-A01 and CaCCinh-A01 on Cardiac Fibroblast Function

Tian

Wang

et al. 2018

Cell Physiol Biochem

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Background/Aims: Calcium-activated chloride channels (CaCCs) regulate numerous physiological processes including cell proliferation, migration, and extracellular matrix secretion. T16Ainh-A01 and CaCCinh-A01 are selective inhibitors of CaCCs. But it is unknown whether these two compounds have functional effects on cardiac fibroblasts (CFs). Methods: Primary CFs were obtained by enzymatic dissociation of cardiomyocytes from neonatal rat hearts. Intracellular Ca²⁺ ([Ca²⁺]_i) and Cl^- ([Cl^-]_i) were measured using the fluorescent calcium indicators (Fluo-4 AM) and N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide respectively. The expression of anoctamin-1 (ANO1) and α-smooth muscle actin (α-SMA) was detected by quantitative RT-PCR, immunofluorescence, and western blotting. A hydroxyproline assay was used to examine collagen secretion. Cell proliferation, cell cycle distribution, and cell migration were assessed by Cell Counting Kit-8, flow cytometry, and Transwell assays, respectively. Results: ANO1 was preferentially expressed on the nuclear membrane and partially within intracellular compartments around the nucleus. T16Ainh-A01 and CaCCinh-A01 displayed different inhibitory effects on [Cl^-]_i in CFs. T16Ainh-A01 considerably decreased [Cl^-]_i in the nucleus, whereas CaCCinh-A01 reduced [Cl^-]_i in intracellular compartments around the nucleus, and both inhibitors exhibited a minimal effect on [Ca²⁺]_i in CFs. ANO1 and α-SMA expression levels were significantly repressed by CaCCinh-A01. T16Ainh-A01 showed a marked inhibitory effect on the mRNA levels of ANO1 and α-SMA, but had a negligible effect on ANO1 at the protein level. T16Ainh-A01 and CaCCinh-A01 led to the significant repression of cell proliferation, cell migration, and collagen secretion in CFs. Conclusion: Our findings indicate that T16Ainh-A01 and CaCCinh-A01 have the potential to inhibit the proliferation and collagen secretion of CFs and may serve as novel anti-fibrotic therapeutic drugs in the future.

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Novel therapeutic strategies targeting fibroblasts and fibrosis in heart disease

Cited by 269 publications

References 307 publications

Electrotonic coupling of excitable and nonexcitable cells in the heart revealed by optogenetics

Electrotonic coupling of excitable and nonexcitable cells in the heart revealed by optogenetics

Fibroblast-specific TGF-β–Smad2/3 signaling underlies cardiac fibrosis

Effects of the Calcium-Activated Chloride Channel Inhibitors T16Ainh-A01 and CaCCinh-A01 on Cardiac Fibroblast Function

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