Tubulin acetylation increases cytoskeletal stiffness to regulate mechanotransduction in striated muscle

Coleman, Andrew; Joca, Humberto C.; Shi, Guoli; Lederer, W. Jonathan; Ward, Christopher W.

doi:10.1101/2020.06.10.144931

Cited by 7 publications

(7 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…51 Dystrophin functions as an intermediary between membrane and cytoskeleton biology, and it is known that the lack of dystrophin in DMD results in cytoskeletal disruptions, such as microtubule dysregulation and increased cytoskeletal stiffness, 71 as well as abnormalities in plasma membrane structure and function resulting in calcium overload. 72,73 A stiff cytoskeleton may result in increased cellular viscoelastic resistance leading to systolic and diastolic cardiac dysfunction, 74 possibly by directly impairing the contraction cycle. Similarly, elevated intracellular calcium due to sarcolemmal damage in DMD results in cardiomyocyte hypercontraction, and thus decreased sarcomere lengths as well as low contractility and slow relaxation.…”

Section: Discussionmentioning

confidence: 99%

Cardiac Myoediting Attenuates Cardiac Abnormalities in Human and Mouse Models of Duchenne Muscular Dystrophy

Atmanli

Chai

Cui

et al. 2021

Circulation Research

View full text Add to dashboard Cite

Rationale: Absence of dystrophin in Duchenne muscular dystrophy (DMD) results in the degeneration of skeletal and cardiac muscles. Owing to advances in respiratory management of DMD patients, cardiomyopathy has become a significant aspect of the disease. While CRISPR/Cas9 genome editing technology holds great potential as a novel therapeutic avenue for DMD, little is known about the potential of DMD correction using CRISPR/Cas9 technology to mitigate cardiac abnormalities in DMD. Objective: To define the effects of CRISPR/Cas9 genome editing on structural, functional and transcriptional abnormalities in DMD-associated cardiac disease. Methods and Results: We generated induced pluripotent stem cells (iPSCs) from a patient with a deletion of exon 44 of the DMD gene (ΔEx44) and his healthy brother. We targeted exon 45 of the DMD gene by CRISPR/Cas9 genome editing to generate corrected DMD (cDMD) iPSC lines, wherein the DMD open reading frame was restored via reframing (RF) or exon skipping (ES). While DMD cardiomyocytes (CMs) demonstrated morphologic, structural and functional deficits compared to control CMs, CMs from both cDMD lines were similar to control CMs. Bulk RNA-sequencing of DMD CMs showed transcriptional dysregulation consistent with dilated cardiomyopathy, which was mitigated in cDMD CMs. We then corrected dysfunctional DMD CMs by adenoviral delivery of Cas9/gRNA and showed that correction of DMD CMs post-differentiation reduces their arrhythmogenic potential. Single-nucleus RNA-sequencing of hearts of DMD mice showed transcriptional dysregulation in CMs and fibroblasts, which in corrected mice was reduced to similar levels as wildtype mice. Conclusions: We show that CRISPR/Cas9-mediated correction of DMD ΔEx44 mitigates structural, functional and transcriptional abnormalities consistent with dilated cardiomyopathy irrespective of how the protein reading frame is restored. We show that these effects extend to postnatal editing in iPSC-CMs and mice. These findings provide key insights into the utility of genome editing as a novel therapeutic for DMD-associated cardiomyopathy.

show abstract

Section: Discussionmentioning

confidence: 99%

Cardiac Myoediting Attenuates Cardiac Abnormalities in Human and Mouse Models of Duchenne Muscular Dystrophy

Atmanli

Chai

Cui

et al. 2021

Circulation Research

View full text Add to dashboard Cite

show abstract

“…The periodic, lateral reinforcement of microtubules increases their stability, leading to longer-lived microtubules and providing a dynamic cross-link with the sarcomere, increasing the viscoelastic resistance to myocyte motion and the ability of microtubules to transduce mechanical stress. Increased microtubule lifetimes also promote microtubule acetylation, which itself increases the ability of microtubules to withstand mechanical stress 35 and increases myocyte viscoelasticity 36 . In the setting of heart disease, the increased abundance of both desmin intermediate filaments and detyrosinated microtubules thus promotes a feed-forward substrate for enhanced mechanotransduction and myocardial stiffening.…”

Section: Discussionmentioning

confidence: 99%

Desmin intermediate filaments and tubulin detyrosination stabilize growing microtubules in the cardiomyocyte

Salomon

Okami

Heffler

et al. 2021

Preprint

View full text Add to dashboard Cite

The microtubule network of the cardiomyocyte exhibits specialized architecture, stability and mechanical behavior that accommodate the demands of working muscle cells. Stable, post-translationally detyrosinated microtubules are physical coupled to the sarcomere, the contractile apparatus of muscle, and resist sarcomere motion to regulate muscle mechanics and mechanosignaling. Control of microtubule growth and shrinkage dynamics represents a potential intermediate in the formation of a stable, physically coupled microtubule network, yet the molecular determinants that govern dynamics are unknown. Here we test the hypothesis that desmin intermediate filaments may stabilize growing microtubules at the sarcomere Z-disk in a detyrosination-dependent manner. Using a combination of biochemical assays and direct observation of microtubule plus-end dynamics in primary adult cardiomyocytes, we determine that: 1) tyrosination increases the frequency of microtubule depolymerization and reduces the pausing of microtubules at the Z-disk, leading to a more dynamic microtubule; and 2) desmin intermediate filaments stabilize both growing and shrinking microtubules specifically at the Z-disk and protect them from depolymerization. This stabilizes iteratively growing, detyrosinated microtubules between adjacent sarcomeres, which promotes the formation of high-energy microtubules that buckle between sarcomeres and elevates myocyte viscoelasticity. Our findings inform on how the tubulin code and intermediate filaments regulate microtubule dynamics, and provide mechanism to the establishment of a spatially organized, physically coupled, and long-lived microtubule network in the cardiomyocyte.

show abstract

“…4i). Increased acetylation due to the loss of MeCP2 may play a role in muscle function as hyperacetylation has been shown to increase stiffness and resistance in striatal muscles in vitro 54 . We continued to evaluate the possible impact of tubulin acetylation by examining the colon, a biomechanically-active organ that is often distended in Rett patients and may play a major role in digestive tract disruption that is a major complaint of these patients clinically 53 .…”

Section: Oral Vorinostat Exhibits Broad Efficacy In Rett Mice Even Af...mentioning

confidence: 99%

“…In addition, we explored the potential link of tubulin acetylation with muscle function and found that vorinostat treatment can rescue the disrupted α-tubulin acetylation observed in femoral muscle sections ( 4 ). Increased acetylation due to the loss of MeCP2 may play a role in muscle function as hyperacetylation has been shown to increase stiffness and resistance in striatal muscles in vitro 54 .…”

Section: Oral Vorinostat Exhibits Broad Efficacy In Rett Mice Even Af...mentioning

confidence: 99%

Target-agnostic discovery of Rett Syndrome therapeutics by coupling computational network analysis and CRISPR-enabled in vivo disease modeling

Novak¹,

Lin²,

Kaushal³

et al. 2022

Preprint

View full text Add to dashboard Cite

It is difficult to develop effective treatments for neurodevelopmental genetic disorders, such as Rett syndrome, which are caused by a single gene mutation but trigger changes in numerous other genes, and thereby also severely impair functions of organs beyond the central nervous system (CNS). This challenge is further complicated by the lack of sufficiently broad and biologically relevant drug screens, and the inherent complexity in identifying clinically relevant targets responsible for diverse phenotypes. Here, we combined human gene regulatory network-based computational drug prediction with in vivo screening in a population-level diversity, CRISPR-edited, Xenopus laevis tadpole model of Rett syndrome to carry out target-agnostic drug discovery, which rapidly led to the identification of the FDA-approved drug vorinostat as a potential repurposing candidate. Vorinostat broadly improved both CNS and non-CNS (e.g., gastrointestinal, respiratory, inflammatory) abnormalities in a pre-clinical mouse model of Rett syndrome. This is the first Rett syndrome treatment to demonstrate pre-clinical efficacy across multiple organ systems when dosed after the onset of symptoms, and network analysis revealed a putative therapeutic mechanism for its cross-organ normalizing effects based on its impact on acetylation metabolism and post-translational modifications of microtubules.

show abstract

Tubulin acetylation increases cytoskeletal stiffness to regulate mechanotransduction in striated muscle

Cited by 7 publications

References 26 publications

Cardiac Myoediting Attenuates Cardiac Abnormalities in Human and Mouse Models of Duchenne Muscular Dystrophy

Cardiac Myoediting Attenuates Cardiac Abnormalities in Human and Mouse Models of Duchenne Muscular Dystrophy

Desmin intermediate filaments and tubulin detyrosination stabilize growing microtubules in the cardiomyocyte

Target-agnostic discovery of Rett Syndrome therapeutics by coupling computational network analysis and CRISPR-enabled in vivo disease modeling

Contact Info

Product

Resources

About