GLORIA

GEOMAR Library Ocean Research Information Access

X

Your email was sent successfully. Check your inbox.

An error occurred while sending the email. Please try again.

Proceed reservation?

Export
  • 1
    Online Resource
    Online Resource
    Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy
    Wiley ; 2022
    In:  Journal of Cachexia, Sarcopenia and Muscle Vol. 13, No. 6 ( 2022-12), p. 3106-3121
    Details
    In: Journal of Cachexia, Sarcopenia and Muscle, Wiley, Vol. 13, No. 6 ( 2022-12), p. 3106-3121
    Abstract: Human pluripotent stem cell‐derived muscle models show great potential for translational research. Here, we describe developmentally inspired methods for the derivation of skeletal muscle cells and their utility in skeletal muscle tissue engineering with the aim to model skeletal muscle regeneration and dystrophy in vitro. Methods Key steps include the directed differentiation of human pluripotent stem cells to embryonic muscle progenitors followed by primary and secondary foetal myogenesis into three‐dimensional muscle. To simulate Duchenne muscular dystrophy (DMD), a patient‐specific induced pluripotent stem cell line was compared to a CRISPR/Cas9‐edited isogenic control line. Results The established skeletal muscle differentiation protocol robustly and faithfully recapitulates critical steps of embryonic myogenesis in two‐dimensional and three‐dimensional cultures, resulting in functional human skeletal muscle organoids (SMOs) and engineered skeletal muscles (ESMs) with a regeneration‐competent satellite‐like cell pool. Tissue‐engineered muscle exhibits organotypic maturation and function (up to 5.7 ± 0.5 mN tetanic twitch tension at 100 Hz in ESM). Contractile performance could be further enhanced by timed thyroid hormone treatment, increasing the speed of contraction (time to peak contraction) as well as relaxation (time to 50% relaxation) of single twitches from 107 ± 2 to 75 ± 4 ms ( P   〈  0.05) and from 146 ± 6 to 100 ± 6 ms ( P   〈  0.05), respectively. Satellite‐like cells could be documented as largely quiescent PAX7 + cells (75 ± 6% Ki67 − ) located adjacent to muscle fibres confined under a laminin‐containing basal membrane. Activation of the engineered satellite‐like cell niche was documented in a cardiotoxin injury model with marked recovery of contractility to 57 ± 8% of the pre‐injury force 21 days post‐injury ( P   〈  0.05 compared to Day 2 post‐injury), which was completely blocked by preceding irradiation. Absence of dystrophin in DMD ESM caused a marked reduction of contractile force (−35 ± 7%, P   〈  0.05) and impaired expression of fast myosin isoforms resulting in prolonged contraction (175 ± 14 ms, P   〈  0.05 vs. gene‐edited control) and relaxation (238 ± 22 ms, P   〈  0.05 vs. gene‐edited control) times. Restoration of dystrophin levels by gene editing rescued the DMD phenotype in ESM. Conclusions We introduce human muscle models with canonical properties of bona fide skeletal muscle in vivo to study muscle development, maturation, disease and repair.
    Type of Medium: Online Resource
    ISSN: 2190-5991 , 2190-6009
    URL: Issue
    URL: Article
    DOI: 10.1002/jcsm.v13.6
    DOI: 10.1002/jcsm.13094
    Language: English
    Publisher: Wiley
    Publication Date: 2022
    detail.hit.zdb_id: 2586864-0
    detail.hit.zdb_id: 2581863-6
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
  • 2
    Online Resource
    Online Resource
    Protocol to develop force-generating human skeletal muscle organoids
    Elsevier BV ; 2024
    In:  STAR Protocols Vol. 5, No. 1 ( 2024-03), p. 102794-
    Details
    In: STAR Protocols, Elsevier BV, Vol. 5, No. 1 ( 2024-03), p. 102794-
    Type of Medium: Online Resource
    ISSN: 2666-1667
    URL: Article
    DOI: 10.1016/j.xpro.2023.102794
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2024
    detail.hit.zdb_id: 3053335-1
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
  • 3
    Online Resource
    Online Resource
    Catecholamine-Dependent β-Adrenergic Signaling in a Pluripotent Stem Cell Model of Takotsubo Cardiomyopathy
    Elsevier BV ; 2017
    In:  Journal of the American College of Cardiology Vol. 70, No. 8 ( 2017-08), p. 975-991
    Details
    In: Journal of the American College of Cardiology, Elsevier BV, Vol. 70, No. 8 ( 2017-08), p. 975-991
    Type of Medium: Online Resource
    ISSN: 0735-1097
    URL: Article
    DOI: 10.1016/j.jacc.2017.06.061
    RVK:
    XA 17760
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2017
    detail.hit.zdb_id: 605507-2
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
  • 4
    Online Resource
    Online Resource
    p63RhoGEF regulates auto- and paracrine signaling in cardiac fibroblasts
    Elsevier BV ; 2015
    In:  Journal of Molecular and Cellular Cardiology Vol. 88 ( 2015-11), p. 39-54
    Details
    In: Journal of Molecular and Cellular Cardiology, Elsevier BV, Vol. 88 ( 2015-11), p. 39-54
    Type of Medium: Online Resource
    ISSN: 0022-2828
    URL: Article
    DOI: 10.1016/j.yjmcc.2015.09.009
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2015
    detail.hit.zdb_id: 80157-4
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
  • 5
    Online Resource
    Online Resource
    Comparative study of human-induced pluripotent stem cells derived from bone marrow cells, hair keratinocytes, and skin fibroblasts
    Oxford University Press (OUP) ; 2013
    In:  European Heart Journal Vol. 34, No. 33 ( 2013-9-1), p. 2618-2629
    Details
    In: European Heart Journal, Oxford University Press (OUP), Vol. 34, No. 33 ( 2013-9-1), p. 2618-2629
    Type of Medium: Online Resource
    ISSN: 1522-9645 , 0195-668X
    URL: Article
    DOI: 10.1093/eurheartj/ehs203
    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2013
    detail.hit.zdb_id: 2001908-7
    detail.hit.zdb_id: 603098-1
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
  • 6
    Online Resource
    Online Resource
    Extracellular Signal-Regulated Kinases 1 and 2 Regulate the Balance Between Eccentric and Concentric Cardiac Growth
    Ovid Technologies (Wolters Kluwer Health) ; 2011
    In:  Circulation Research Vol. 108, No. 2 ( 2011-01-21), p. 176-183
    Details
    In: Circulation Research, Ovid Technologies (Wolters Kluwer Health), Vol. 108, No. 2 ( 2011-01-21), p. 176-183
    Abstract: An increase in cardiac afterload typically produces concentric hypertrophy characterized by an increase in cardiomyocyte width, whereas volume overload or exercise results in eccentric growth characterized by cellular elongation and addition of sarcomeres in series. The signaling pathways that control eccentric versus concentric heart growth are not well understood. Objective: To determine the role of extracellular signal-regulated kinase 1 and 2 (ERK1/2) in regulating the cardiac hypertrophic response. Methods and Results: Here, we used mice lacking all ERK1/2 protein in the heart (Erk1 −/− Erk2 fl/fl-Cre ) and mice expressing activated mitogen-activated protein kinase kinase (Mek)1 in the heart to induce ERK1/2 signaling, as well as mechanistic experiments in cultured myocytes to assess cellular growth characteristics associated with this signaling pathway. Although genetic deletion of all ERK1/2 from the mouse heart did not block the cardiac hypertrophic response per se, meaning that the heart still increased in weight with both aging and pathological stress stimulation, it did dramatically alter how the heart grew. For example, adult myocytes from hearts of Erk1 −/− Erk2 fl/fl-Cre mice showed preferential eccentric growth (lengthening), whereas myocytes from Mek1 transgenic hearts showed concentric growth (width increase). Isolated adult myocytes acutely inhibited for ERK1/2 signaling by adenoviral gene transfer showed spontaneous lengthening, whereas infection with an activated Mek1 adenovirus promoted constitutive ERK1/2 signaling and increased myocyte thickness. A similar effect was observed in engineered heart tissue under cyclic stretching, where ERK1/2 inhibition led to preferential lengthening. Conclusions: Taken together, these data demonstrate that the ERK1/2 signaling pathway uniquely regulates the balance between eccentric and concentric growth of the heart.
    Type of Medium: Online Resource
    ISSN: 0009-7330 , 1524-4571
    URL: Article
    DOI: 10.1161/CIRCRESAHA.110.231514
    RVK:
    XA 10000
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2011
    detail.hit.zdb_id: 80100-8
    detail.hit.zdb_id: 1467838-X
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
  • 7
    Online Resource
    Online Resource
    Abstract 237: Antioxidant and Ryanodine Receptor Stabilizing Therapy Prevent Statin-Induced Contractile Dysfunction in a Tissue-Engineered Skeletal Muscle Model
    Ovid Technologies (Wolters Kluwer Health) ; 2012
    In:  Circulation Research Vol. 111, No. suppl_1 ( 2012-08-03)
    Details
    In: Circulation Research, Ovid Technologies (Wolters Kluwer Health), Vol. 111, No. suppl_1 ( 2012-08-03)
    Abstract: Rationale: Skeletal muscle toxicity of HMG-CoA-reductase inhibitors (statins) ranges from reversible myalgia to irreversible rhabdomyolysis. The underlying molecular mechanisms are not well defined. Tissue engineering models may help to gain insight into these clinically limiting pathologies. Objective: Here we aimed to model a phenotype of reversible myalgia in vitro to decipher mechanisms that contribute to the early stage of statin toxicity. Methods and Results: Engineered skeletal muscle (ESM) was generated from rat myoblasts, matrigel, and collagen. Isometrically suspended ESM developed 1.2±0.1 mN force under tetanic field stimulation (80 Hz; 200 mA; n=25). Exposure of ESM to statins for 5 days resulted in a loss of force and increased fatiguability in a concentration dependent manner. Cerivastatin was identified as the most potent statin with respect to muscle toxicity with a TC50 (=50% force reduction) of 0.02 µmol/L (n=25/group). Interestingly, at low cerivastatin concentration (0.01 µmol/L) contractile force of ESM was impaired without obvious signs for structural muscle damage (sarcomeric actin content, CK activity unchanged, n=4). Importantly, ESM dysfunction was fully reversible if challenged with TC50 statin concentrations (n=12-14/group). We reasoned that contractile dysfunction with increased fatiguability resulted from calcium leak via the ryanodine receptor. To test this hypothesis we co-administered S107 and observed a concentration-dependent inhibition of statin-induced force reduction (n=12) and calpain activitiy (a calcium-dependent protease, n=4-6). We further argued that RYR destabilization may have been caused by reactive nitrogen (RNS) and/or reactive oxygen species (ROS). Interestingly, the antioxidant N-acetyl cysteine (1 mmol/L, n=3/group), but not L-NAME (10 mmol/L; NO-synthase inhibition, n=6-12/group) prevented contractile dysfunction. Conclusion: We utilized a novel tissue engineered skeletal muscle model to decipher mechanisms of statin-induced muscle toxicity and provide evidence for a central role of ryanodine receptor leak, possibly caused by oxidative damage. Our data suggest that antioxidant and RYR-stabilizing approaches may be useful in counteracting statin myopathy.
    Type of Medium: Online Resource
    ISSN: 0009-7330 , 1524-4571
    URL: Article
    DOI: 10.1161/res.111.suppl_1.A237
    RVK:
    XA 10000
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2012
    detail.hit.zdb_id: 80100-8
    detail.hit.zdb_id: 1467838-X
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
  • 8
    Online Resource
    Online Resource
    Abstract 10240: Key Pathomechanisms of Cardiomyopathy Due to TTN Truncation
    Ovid Technologies (Wolters Kluwer Health) ; 2021
    In:  Circulation Vol. 144, No. Suppl_1 ( 2021-11-16)
    Details
    In: Circulation, Ovid Technologies (Wolters Kluwer Health), Vol. 144, No. Suppl_1 ( 2021-11-16)
    Abstract: Introduction: Heterozygous truncating variants in TTN (TTNtv) coding for titin cause dilated cardiomyopathy (DCM) but the underlying pathomechanisms are unclear and disease management remains uncertain. Here, we aim to elucidate the key pathomechanisms of TTNtv-DCM. Hypothesis: Stable expression of truncated titin proteins and titin haploinsufficiency characterize TTNtv-DCM and represent potential targets for therapy. Methods and Results: We studied left ventricular tissues from 14 nonfailing donor hearts and 113 endstage failing DCM patients and identified a TTNtv in 22 DCM patients (19.5%) by next generation sequencing. Using titin protein gel electrophoresis and western blot we demonstrate, for the first time, titin haploinsufficiency in TTNtv-DCM hearts. Strikingly, all 21 adult TTNtv-DCM hearts of our cohort showed stable expression of truncated titin proteins. Expression was variable, up to one-half of the total titin protein pool, and negatively correlated to patient age at heart transplantation. Truncated titin proteins were not detected in sarcomeres but were present in intracellular aggregates. Deregulated ubiquitin-dependent protein quality control was apparent. Next, we produced human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), comparing wildtype controls to cells with a patient-derived, prototypical A-band TTNtv or a CRISPR/Cas9-edited M-band TTNtv. TTNtv hiPSC-CMs showed reduced wildtype titin expression and contained truncated titin proteins, whose proportion increased upon inhibition of proteasomal activity. In engineered heart muscle generated from hiPSC-CMs, depressed contractility caused by TTNtv could be reversed by correction of the mutation using CRISPR/Cas9, thereby eliminating truncated titin proteins and raising wildtype titin content. Functional improvement also occurred when titin protein content was increased by proteasome inhibition. Conclusions: Our study reveals the major pathomechanisms of TTNtv-DCM, which include titin haploinsufficiency as a life-long condition and truncated titin protein enrichment with aggregate formation, along with aberrant protein quality control. Results can be exploited for new therapies of TTNtv cardiomyopathies.
    Type of Medium: Online Resource
    ISSN: 0009-7322 , 1524-4539
    URL: Article
    DOI: 10.1161/circ.144.suppl_1.10240
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2021
    detail.hit.zdb_id: 1466401-X
    detail.hit.zdb_id: 80099-5
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
  • 9
    Online Resource
    Online Resource
    Abstract 5517: A Novel Tissue Engineered Skeletal Muscle Screening Platform to Assess the Myotoxic Potential of Statins
    Ovid Technologies (Wolters Kluwer Health) ; 2008
    In:  Circulation Vol. 118, No. suppl_18 ( 2008-10-28)
    Details
    In: Circulation, Ovid Technologies (Wolters Kluwer Health), Vol. 118, No. suppl_18 ( 2008-10-28)
    Abstract: Our study had two objectives: to develop a novel tissue engineered skeletal muscle drug screening platform and to assess myotoxicity of various statins and identify underlying mechanisms. Methods: Engineered skeletal muscle tissue (SMT) was generated from adult rat skeletal myoblasts, rat fibroblasts (1.25x106 cells/SMT), matrigel, and collagen. Reconstitution-mixtures (450 μl) were poured into circular moulds yielding ring-shaped constructs after 5 days. SMTs were cultured for additional 10 days on stretch devices and treated with increasing concentrations of statins or vehicle for the last 5 culture days. Force of contraction (FOC) was measured and SMTs subjected to either immune fluorescence (IF), HE-staining or Western blotting (WB). Results: SMTs displayed differentiated muscle bundles (actin-IF and HE) and elicited tetanic contractions with maximal force at 60 Hz (1.3±0.2 mN; n=5). Carbachol (1 μM) induced a reversible block of muscle contraction which was antagonized by pancuronium (10 μM) indicating the presence and functionality of nicotinergic acetylcholine receptors. Statin treatment caused a concentration-dependent decrease in FOC in the following order of potency (EC50): cerivastatin (10 nM), simvastatin (100 nM), atorvastatin (350 nM), pravastatin (1860 nM; n=6–8/group and concentration). Contractile failure was paralleled by sarcomere breakdown and apoptosis (IF and WB: reduced actin, increased caspase 3). To assess the mechanism of statin myotoxicity, we investigated the protective effects of mevalonic acid (MEV, 100 μM), farnesyl-pyrophosphate (F-PP, 10 μM), squalene (S, 10 μM), and geranyl-geranyl-pyrophophate (G-PP, 10 μM) in the presence of maximally toxic cerivastatin concentrations (1 μM; complete contractile failure). MEV prevented cerivastatin toxicity completely (FOC: 102.3±16% of vehicle; n=7). GG-PP partially prevented cerivastatin-induced contractile failure (FOC: 53.3±14% of vehicle; n=4). S and F-PP had no protective effect (n=4/group). Conclusion: SMT can be used to identify and compare statin myotoxicity in vitro and may be useful to dissect its underlying mechanisms. In addition, our data suggests that statin myotoxicity is at least in part independent of cholesterol depletion.
    Type of Medium: Online Resource
    ISSN: 0009-7322 , 1524-4539
    URL: Article
    DOI: 10.1161/circ.118.suppl_18.S_565-d
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2008
    detail.hit.zdb_id: 1466401-X
    detail.hit.zdb_id: 80099-5
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
  • 10
    Online Resource
    Online Resource
    Intronic CRISPR Repair in a Preclinical Model of Noonan Syndrome–Associated Cardiomyopathy
    Ovid Technologies (Wolters Kluwer Health) ; 2020
    In:  Circulation Vol. 142, No. 11 ( 2020-09-15), p. 1059-1076
    Details
    In: Circulation, Ovid Technologies (Wolters Kluwer Health), Vol. 142, No. 11 ( 2020-09-15), p. 1059-1076
    Abstract: Noonan syndrome (NS) is a multisystemic developmental disorder characterized by common, clinically variable symptoms, such as typical facial dysmorphisms, short stature, developmental delay, intellectual disability as well as cardiac hypertrophy. The underlying mechanism is a gain-of-function of the RAS–mitogen-activated protein kinase signaling pathway. However, our understanding of the pathophysiological alterations and mechanisms, especially of the associated cardiomyopathy, remains limited and effective therapeutic options are lacking. Methods: Here, we present a family with two siblings displaying an autosomal recessive form of NS with massive hypertrophic cardiomyopathy as clinically the most prevalent symptom caused by biallelic mutations within the leucine zipper-like transcription regulator 1 ( LZTR1 ). We generated induced pluripotent stem cell–derived cardiomyocytes of the affected siblings and investigated the patient-specific cardiomyocytes on the molecular and functional level. Results: Patients’ induced pluripotent stem cell–derived cardiomyocytes recapitulated the hypertrophic phenotype and uncovered a so-far-not-described causal link between LZTR1 dysfunction, RAS–mitogen-activated protein kinase signaling hyperactivity, hypertrophic gene response and cellular hypertrophy. Calcium channel blockade and MEK inhibition could prevent some of the disease characteristics, providing a molecular underpinning for the clinical use of these drugs in patients with NS, but might not be a sustainable therapeutic option. In a proof-of-concept approach, we explored a clinically translatable intronic CRISPR (clustered regularly interspaced short palindromic repeats) repair and demonstrated a rescue of the hypertrophic phenotype. Conclusions: Our study revealed the human cardiac pathogenesis in patient-specific induced pluripotent stem cell–derived cardiomyocytes from NS patients carrying biallelic variants in LZTR1 and identified a unique disease-specific proteome signature. In addition, we identified the intronic CRISPR repair as a personalized and in our view clinically translatable therapeutic strategy to treat NS-associated hypertrophic cardiomyopathy.
    Type of Medium: Online Resource
    ISSN: 0009-7322 , 1524-4539
    URL: Article
    DOI: 10.1161/CIRCULATIONAHA.119.044794
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2020
    detail.hit.zdb_id: 1466401-X
    detail.hit.zdb_id: 80099-5
    Location Call Number Limitation Availability
    BibTip Others were also interested in ...
    Online Resource
Close ⊗
This website uses cookies and the analysis tool Matomo. More information can be found here...