Diabetes increases stiffness of live cardiomyocytes measured by atomic force microscopy nanoindentation

Benech, Juan Claudio; Benech, N.; Zambrana, Ana I.; Rauschert, Inés; Bervejillo, Verónica; Oddone, Natalia; Damián, Juan Pablo

doi:10.1152/ajpcell.00192.2013

Cited by 43 publications

(54 citation statements)

References 39 publications

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“…Elasticity measurements were obtained using the nano-indentation method with an atomic force microscope (BioScope Catalyst)(Bruker, Camarillo, CA USA), as described in [113]. The cells were cultured on Roboz slides (Cell Point Scientific, Gaithersburg, MD, USA) placed into an inverted microscope (Olympus IX81)(Olympus,Center Valley, PA, USA) coupled to the AFM.Before conducting the AFM experiments, the cell medium was changed from DMEM (1X) + GlutaMax with 10% serum and antibiotics to DMEM (1X) + GlutaMax.…”

Section: Methodsmentioning

confidence: 99%

Promoter hypermethylation as a mechanism for Lamin A/C silencing in a subset of neuroblastoma cells

et al. 2017

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Nuclear lamins support the nuclear envelope and provide anchorage sites for chromatin. They are involved in DNA synthesis, transcription, and replication. It has previously been reported that the lack of Lamin A/C expression in lymphoma and leukaemia is due to CpG island promoter hypermethylation. Here, we provide evidence that Lamin A/C is silenced via this mechanism in a subset of neuroblastoma cells. Moreover, Lamin A/C expression can be restored with a demethylating agent. Importantly, Lamin A/C reintroduction reduced cell growth kinetics and impaired migration, invasion, and anchorage-independent cell growth. Cytoskeletal restructuring was also induced. In addition, the introduction of lamin Δ50, known as Progerin, caused senescence in these neuroblastoma cells. These cells were stiffer and developed a cytoskeletal structure that differed from that observed upon Lamin A/C introduction. Of relevance, short hairpin RNA Lamin A/C depletion in unmethylated neuroblastoma cells enhanced the aforementioned tumour properties. A cytoskeletal structure similar to that observed in methylated cells was induced. Furthermore, atomic force microscopy revealed that Lamin A/C knockdown decreased cellular stiffness in the lamellar region. Finally, the bioinformatic analysis of a set of methylation arrays of neuroblastoma primary tumours showed that a group of patients (around 3%) gives a methylation signal in some of the CpG sites located within the Lamin A/C promoter region analysed by bisulphite sequencing PCR. These findings highlight the importance of Lamin A/C epigenetic inactivation for a subset of neuroblastomas, leading to enhanced tumour properties and cytoskeletal changes. Additionally, these findings may have treatment implications because tumour cells lacking Lamin A/C exhibit more aggressive behaviour.

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Section: Methodsmentioning

confidence: 99%

Promoter hypermethylation as a mechanism for Lamin A/C silencing in a subset of neuroblastoma cells

et al. 2017

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“…These data strongly suggest a "change (i.e. increase) in the number and/or the activation state of adhesion molecules present in the surface of the diabetic cardiomyocytes" [49] . However, at present if structures like costameres (including integrin linkage to the Z disk of the sarcomeres via the cytoskeleton) are affected by diabetes is still unknown.…”

Section: Mechanotransductionmentioning

confidence: 70%

“…As we shall see, pathologies like diabetes, deeply affects the nanomechanical properties of isolated cardiomyocytes, including adhesive force of myocyte sarcolemma [49] . These data strongly suggest a "change (i.e.…”

Section: Mechanotransductionmentioning

confidence: 97%

Intrinsic nanomechanical changes in live diabetic cardiomyocytes

Benech

Benech²,

Zambrana³

et al. 2015

Cardiovasc Regen Med

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Patients with diabetes develop a cardiomyopathy that is independent of both coronary artery disease and hypertension and contributes to mortality and morbidity caused by diabetes. The mechanisms underlying the development of diabetic cardiomyopathy are poorly understood. Several reports showed that an early manifestation of diabetic myocardial dysfunction is an increased diastolic left ventricular (LV) stiffness. This increase is usually attributed to myocardial fibrosis or to myocardial deposition of advanced glycation end products. Alteration of the stiffness (resting tension) of the diabetic cardiomyocyte was also proposed to be an important factor contributing to increased LV stiffness. Some of these data were obtained from isolated cardiomyocytes from human frozen biopsy samples that had been thawed, mechanically disrupted, and incubated with Triton X-100, disrupting sarcolemmal and sarcoplasmic membranes. We therefore determined the stiffness of live isolated cardiomyocytes from control and streptozotocin-treated mice using atomic force microscopy (AFM) nanoindentation. We show that 3 months of type 1 diabetes provoked fragmentation and disorder of myocardial fibers, interstitial collagen deposition, reduction in SERCA2a calcium pump expression and changes in F-actin organization. Moreover, we show that live isolated diabetic cardiomyocytes are stiffer than control cardiomyocytes when tested in Tyrode buffer with different ionic compositions. Hence, it is very likely that intrinsic mechanical changes of cardiomyocytes are an important factor in increasing myocardial stiffness in vivo.

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“…Our data further illustrated the binding specificity between other ECM proteins and CM integrins, where CN binds to α 10 ‐ and α 11 ‐integrin subunits and LN binds to α 3 ‐ and α 7 ‐integrin subunits (Figure 3). These results may be helpful for studying cardiac disease with accumulation of ECM, such as diabetes, by studying particular interactions between ECM proteins and their integrins on CMs …”

Section: Discussionmentioning

confidence: 98%

“…These results may be helpful for studying cardiac disease with accumulation of ECM, such as diabetes, by studying particular interactions between ECM proteins and their integrins on CMs. 49,50 It has been reported that the conformation or function of integrins may be influenced by cations (such as Ca 2+ or Mg 2+ ). 23 Recent study showed that the increment in concentration of extracellular magnesium (from 0 to 5 mM) would enhance the integrins binding ability in human thyroid follicular carcinoma cell line, characterized by AFM and traditional bulk technique.…”

Section: Discussionmentioning

confidence: 99%

Characterization of adhesion properties of the cardiomyocyte integrins and extracellular matrix proteins using atomic force microscopy

Liu

Yang

et al. 2019

J of Molecular Recognition

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Integrins are transmembrane adhesion receptors that play important roles in the cardiovascular system by interacting with the extracellular matrix (ECM). However, direct quantitative measurements of the adhesion properties of the integrins on cardiomyocyte (CM) and their ECM ligands are lacking. In this study, we used atomic force microscopy (AFM) to quantify the adhesion force (peak force and mean force) and binding probability between CM integrins and three main heart tissue ECM proteins, ie, collagen (CN), fibronectin (FN), and laminin (LN). Functionalizing the AFM probes with ECM proteins, we found that the peak force (mean force) was 61.69 ± 5.5 pN (76.54 ± 4.0 pN), 39.26 ± 4.4 pN (59.84 ± 3.6 pN), and 108.31 ± 4.2 pN (129.63 ± 6.0 pN), respectively, for the bond of CN‐integrin, FN‐integrin, and LN‐integrin. The binding specificity between CM integrins and ECM proteins was verified by using monoclonal antibodies, where α10‐ and α11‐integrin bind to CN, α3‐ and α5‐integrin bind to FN, and α3‐ and α7‐integrin bind to LN. Furthermore, adhesion properties of CM integrins under physiologically high concentrations of extracellular Ca2+ and Mg2+ were tested. Additional Ca2+ reduced the adhesion mean force to 68.81 ± 4.0 pN, 49.84 ± 3.3 pN, and 119.21 ± 5.8 pN and binding probability to 0.31, 0.34, 0.40 for CN, FN, and LN, respectively, whereas Mg2+ caused very minor changes to adhesion properties of CM integrins. Thus, adhesion properties between adult murine CM integrins and its main ECM proteins were characterized, paving the way for an improved understanding of CM mechanobiology.

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Diabetes increases stiffness of live cardiomyocytes measured by atomic force microscopy nanoindentation

Abstract: Benech JC, Benech N, Zambrana AI, Rauschert I, Bervejillo V, Oddone N, Damián JP. Diabetes increases stiffness of live cardiomyocytes measured by atomic force microscopy nanoindentation. Am

Cited by 43 publications

References 39 publications

Promoter hypermethylation as a mechanism for Lamin A/C silencing in a subset of neuroblastoma cells

Promoter hypermethylation as a mechanism for Lamin A/C silencing in a subset of neuroblastoma cells

Intrinsic nanomechanical changes in live diabetic cardiomyocytes

Characterization of adhesion properties of the cardiomyocyte integrins and extracellular matrix proteins using atomic force microscopy

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