Oxidative stress pathogenically remodels the cardiac myocyte cytoskeleton via structural alterations to the microtubule lattice

Rebecca R. Goldblum; Mark R McClellan; Kyle White; Samuel Gonzalez; Brian R. Thompson; Hluechy X. Vang; Houda Cohen; Lee Ann Higgins; Todd W Markowski; Tzu-Yi Yang; Joseph M. Metzger; Melissa K. Gardner

doi:10.1016/j.devcel.2021.07.004

Oxidative stress pathogenically remodels the cardiac myocyte cytoskeleton via structural alterations to the microtubule lattice

Rebecca R. Goldblum, Mark R McClellan, Kyle White, Samuel Gonzalez, Brian R. Thompson, Hluechy X. Vang, Houda Cohen, Lee Ann Higgins, Todd W Markowski, Tzu-Yi Yang, Joseph M. Metzger, Melissa K. Gardner

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

Abstract

In the failing heart, the cardiac myocyte microtubule network is remodeled, which contributes to cellular contractile failure and patient death. However, the origins of this deleterious cytoskeletal reorganization are unknown. We now find that oxidative stress, a condition characteristic of heart failure, leads to cysteine oxidation of microtubules. Our electron and fluorescence microscopy experiments revealed regions of structural damage within the microtubule lattice that occurred at locations of oxidized tubulin. The incorporation of GTP-tubulin into these damaged, oxidized regions led to stabilized “hot spots” within the microtubule lattice, which suppressed the shortening of dynamic microtubules. Thus, oxidative stress may act inside of cardiac myocytes to facilitate a pathogenic shift from a sparse microtubule network into a dense, aligned network. Our results demonstrate how a disease condition characterized by oxidative stress can trigger a molecular oxidation event, which likely contributes to a toxic cellular-scale transformation of the cardiac myocyte microtubule network.

Original language	English (US)
Pages (from-to)	2252-2266.e6
Journal	Developmental Cell
Volume	56
Issue number	15
DOIs	https://doi.org/10.1016/j.devcel.2021.07.004
State	Published - Aug 9 2021

Bibliographical note

Funding Information:
The Gardner laboratory is supported by a National Institutes of Health grant NIGMS R35-GM126974 and R.R.G. was supported by the National Institute of Health Training Program in Muscle Research ( T32AR007612 ). J.M.M. is supported by National Institutes of Health grant NHLBI R01 HL123874 . Parts of this work were carried out in the Characterization Facility, University of Minnesota, a member of the National Science Foundation -funded Materials Research Facilities Network ( www.mrfn.org ) via the MRSEC program. The authors recognize the Center for Mass Spectrometry and Proteomics, a subunit of the Department of Biochemistry, Molecular Biology, and Biophysics at the University of Minnesota and various supporting agencies, which are listed here: https://cbs.umn.edu/cmsp/about . We thank members of the Gardner laboratory for helpful discussions, and Dr. Taylor Reid for software assistance and guidance. We thank Dr. Thomas Surrey for the generous gift of Mal3 constructs.

Funding Information:
The Gardner laboratory is supported by a National Institutes of Health grant NIGMS R35-GM126974 and R.R.G. was supported by the National Institute of Health Training Program in Muscle Research (T32AR007612). J.M.M. is supported by National Institutes of Health grant NHLBI R01 HL123874. Parts of this work were carried out in the Characterization Facility, University of Minnesota, a member of the National Science Foundation-funded Materials Research Facilities Network (www.mrfn.org) via the MRSEC program. The authors recognize the Center for Mass Spectrometry and Proteomics, a subunit of the Department of Biochemistry, Molecular Biology, and Biophysics at the University of Minnesota and various supporting agencies, which are listed here: https://cbs.umn.edu/cmsp/about. We thank members of the Gardner laboratory for helpful discussions, and Dr. Taylor Reid for software assistance and guidance. We thank Dr. Thomas Surrey for the generous gift of Mal3 constructs. Conceptualization, R.R.G. and M.K.G.; methodology, all authors; software, R.R.G. S.J.G. and M.K.G.; formal analysis, R.R.G. M.M. K.W. S.J.G. L.H. and M.K.G; investigation, R.R.G. M.M. L.H. T.M. T.-Y.Y. and K.W; resources, B.R.T, H.V. H.C, J.M.M. S.J.G. L.H. T.M. and T.-Y.Y.; data curation, M.K.G.; writing ? original draft, R.R.G; writing ? review and editing, all authors; supervision, M.K.G. and J.M.; funding acquisition, M.K.G. and J.M. The authors declare no competing interests.

Publisher Copyright:
© 2021 Elsevier Inc.

Keywords

H9c2
cardiomyocytes
cysteine oxidation
microtubule dynamics
microtubule repair
oxidative stress
rescue
tubulin

PubMed: MeSH publication types

Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

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10.1016/j.devcel.2021.07.004

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Goldblum, R. R., McClellan, M. R., White, K., Gonzalez, S., Thompson, B. R., Vang, H. X., Cohen, H., Higgins, L. A., Markowski, T. W., Yang, T.-Y., Metzger, J. M., & Gardner, M. K. (2021). Oxidative stress pathogenically remodels the cardiac myocyte cytoskeleton via structural alterations to the microtubule lattice. Developmental Cell, 56(15), 2252-2266.e6. https://doi.org/10.1016/j.devcel.2021.07.004

Goldblum, RR, McClellan, MR, White, K, Gonzalez, S, Thompson, BR , Vang, HX , Cohen, H , Higgins, LA , Markowski, TW, Yang, T-Y, Metzger, JM & Gardner, MK 2021, 'Oxidative stress pathogenically remodels the cardiac myocyte cytoskeleton via structural alterations to the microtubule lattice', Developmental Cell, vol. 56, no. 15, pp. 2252-2266.e6. https://doi.org/10.1016/j.devcel.2021.07.004

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title = "Oxidative stress pathogenically remodels the cardiac myocyte cytoskeleton via structural alterations to the microtubule lattice",

abstract = "In the failing heart, the cardiac myocyte microtubule network is remodeled, which contributes to cellular contractile failure and patient death. However, the origins of this deleterious cytoskeletal reorganization are unknown. We now find that oxidative stress, a condition characteristic of heart failure, leads to cysteine oxidation of microtubules. Our electron and fluorescence microscopy experiments revealed regions of structural damage within the microtubule lattice that occurred at locations of oxidized tubulin. The incorporation of GTP-tubulin into these damaged, oxidized regions led to stabilized “hot spots” within the microtubule lattice, which suppressed the shortening of dynamic microtubules. Thus, oxidative stress may act inside of cardiac myocytes to facilitate a pathogenic shift from a sparse microtubule network into a dense, aligned network. Our results demonstrate how a disease condition characterized by oxidative stress can trigger a molecular oxidation event, which likely contributes to a toxic cellular-scale transformation of the cardiac myocyte microtubule network.",

keywords = "H9c2, cardiomyocytes, cysteine oxidation, microtubule dynamics, microtubule repair, oxidative stress, rescue, tubulin",

author = "Goldblum, {Rebecca R.} and McClellan, {Mark R} and Kyle White and Samuel Gonzalez and Thompson, {Brian R.} and Vang, {Hluechy X.} and Houda Cohen and Higgins, {Lee Ann} and Markowski, {Todd W} and Tzu-Yi Yang and Metzger, {Joseph M.} and Gardner, {Melissa K.}",

note = "Funding Information: The Gardner laboratory is supported by a National Institutes of Health grant NIGMS R35-GM126974 and R.R.G. was supported by the National Institute of Health Training Program in Muscle Research ( T32AR007612 ). J.M.M. is supported by National Institutes of Health grant NHLBI R01 HL123874 . Parts of this work were carried out in the Characterization Facility, University of Minnesota, a member of the National Science Foundation -funded Materials Research Facilities Network ( www.mrfn.org ) via the MRSEC program. The authors recognize the Center for Mass Spectrometry and Proteomics, a subunit of the Department of Biochemistry, Molecular Biology, and Biophysics at the University of Minnesota and various supporting agencies, which are listed here: https://cbs.umn.edu/cmsp/about . We thank members of the Gardner laboratory for helpful discussions, and Dr. Taylor Reid for software assistance and guidance. We thank Dr. Thomas Surrey for the generous gift of Mal3 constructs. Funding Information: The Gardner laboratory is supported by a National Institutes of Health grant NIGMS R35-GM126974 and R.R.G. was supported by the National Institute of Health Training Program in Muscle Research (T32AR007612). J.M.M. is supported by National Institutes of Health grant NHLBI R01 HL123874. Parts of this work were carried out in the Characterization Facility, University of Minnesota, a member of the National Science Foundation-funded Materials Research Facilities Network (www.mrfn.org) via the MRSEC program. The authors recognize the Center for Mass Spectrometry and Proteomics, a subunit of the Department of Biochemistry, Molecular Biology, and Biophysics at the University of Minnesota and various supporting agencies, which are listed here: https://cbs.umn.edu/cmsp/about. We thank members of the Gardner laboratory for helpful discussions, and Dr. Taylor Reid for software assistance and guidance. We thank Dr. Thomas Surrey for the generous gift of Mal3 constructs. Conceptualization, R.R.G. and M.K.G.; methodology, all authors; software, R.R.G. S.J.G. and M.K.G.; formal analysis, R.R.G. M.M. K.W. S.J.G. L.H. and M.K.G; investigation, R.R.G. M.M. L.H. T.M. T.-Y.Y. and K.W; resources, B.R.T, H.V. H.C, J.M.M. S.J.G. L.H. T.M. and T.-Y.Y.; data curation, M.K.G.; writing ? original draft, R.R.G; writing ? review and editing, all authors; supervision, M.K.G. and J.M.; funding acquisition, M.K.G. and J.M. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",

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doi = "10.1016/j.devcel.2021.07.004",

language = "English (US)",

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T1 - Oxidative stress pathogenically remodels the cardiac myocyte cytoskeleton via structural alterations to the microtubule lattice

AU - Goldblum, Rebecca R.

AU - McClellan, Mark R

AU - White, Kyle

AU - Gonzalez, Samuel

AU - Thompson, Brian R.

AU - Vang, Hluechy X.

AU - Cohen, Houda

AU - Higgins, Lee Ann

AU - Markowski, Todd W

AU - Yang, Tzu-Yi

AU - Metzger, Joseph M.

AU - Gardner, Melissa K.

N1 - Funding Information: The Gardner laboratory is supported by a National Institutes of Health grant NIGMS R35-GM126974 and R.R.G. was supported by the National Institute of Health Training Program in Muscle Research ( T32AR007612 ). J.M.M. is supported by National Institutes of Health grant NHLBI R01 HL123874 . Parts of this work were carried out in the Characterization Facility, University of Minnesota, a member of the National Science Foundation -funded Materials Research Facilities Network ( www.mrfn.org ) via the MRSEC program. The authors recognize the Center for Mass Spectrometry and Proteomics, a subunit of the Department of Biochemistry, Molecular Biology, and Biophysics at the University of Minnesota and various supporting agencies, which are listed here: https://cbs.umn.edu/cmsp/about . We thank members of the Gardner laboratory for helpful discussions, and Dr. Taylor Reid for software assistance and guidance. We thank Dr. Thomas Surrey for the generous gift of Mal3 constructs. Funding Information: The Gardner laboratory is supported by a National Institutes of Health grant NIGMS R35-GM126974 and R.R.G. was supported by the National Institute of Health Training Program in Muscle Research (T32AR007612). J.M.M. is supported by National Institutes of Health grant NHLBI R01 HL123874. Parts of this work were carried out in the Characterization Facility, University of Minnesota, a member of the National Science Foundation-funded Materials Research Facilities Network (www.mrfn.org) via the MRSEC program. The authors recognize the Center for Mass Spectrometry and Proteomics, a subunit of the Department of Biochemistry, Molecular Biology, and Biophysics at the University of Minnesota and various supporting agencies, which are listed here: https://cbs.umn.edu/cmsp/about. We thank members of the Gardner laboratory for helpful discussions, and Dr. Taylor Reid for software assistance and guidance. We thank Dr. Thomas Surrey for the generous gift of Mal3 constructs. Conceptualization, R.R.G. and M.K.G.; methodology, all authors; software, R.R.G. S.J.G. and M.K.G.; formal analysis, R.R.G. M.M. K.W. S.J.G. L.H. and M.K.G; investigation, R.R.G. M.M. L.H. T.M. T.-Y.Y. and K.W; resources, B.R.T, H.V. H.C, J.M.M. S.J.G. L.H. T.M. and T.-Y.Y.; data curation, M.K.G.; writing ? original draft, R.R.G; writing ? review and editing, all authors; supervision, M.K.G. and J.M.; funding acquisition, M.K.G. and J.M. The authors declare no competing interests. Publisher Copyright: © 2021 Elsevier Inc.

PY - 2021/8/9

Y1 - 2021/8/9

N2 - In the failing heart, the cardiac myocyte microtubule network is remodeled, which contributes to cellular contractile failure and patient death. However, the origins of this deleterious cytoskeletal reorganization are unknown. We now find that oxidative stress, a condition characteristic of heart failure, leads to cysteine oxidation of microtubules. Our electron and fluorescence microscopy experiments revealed regions of structural damage within the microtubule lattice that occurred at locations of oxidized tubulin. The incorporation of GTP-tubulin into these damaged, oxidized regions led to stabilized “hot spots” within the microtubule lattice, which suppressed the shortening of dynamic microtubules. Thus, oxidative stress may act inside of cardiac myocytes to facilitate a pathogenic shift from a sparse microtubule network into a dense, aligned network. Our results demonstrate how a disease condition characterized by oxidative stress can trigger a molecular oxidation event, which likely contributes to a toxic cellular-scale transformation of the cardiac myocyte microtubule network.

AB - In the failing heart, the cardiac myocyte microtubule network is remodeled, which contributes to cellular contractile failure and patient death. However, the origins of this deleterious cytoskeletal reorganization are unknown. We now find that oxidative stress, a condition characteristic of heart failure, leads to cysteine oxidation of microtubules. Our electron and fluorescence microscopy experiments revealed regions of structural damage within the microtubule lattice that occurred at locations of oxidized tubulin. The incorporation of GTP-tubulin into these damaged, oxidized regions led to stabilized “hot spots” within the microtubule lattice, which suppressed the shortening of dynamic microtubules. Thus, oxidative stress may act inside of cardiac myocytes to facilitate a pathogenic shift from a sparse microtubule network into a dense, aligned network. Our results demonstrate how a disease condition characterized by oxidative stress can trigger a molecular oxidation event, which likely contributes to a toxic cellular-scale transformation of the cardiac myocyte microtubule network.

KW - H9c2

KW - cardiomyocytes

KW - cysteine oxidation

KW - microtubule dynamics

KW - microtubule repair

KW - oxidative stress

KW - rescue

KW - tubulin

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Oxidative stress pathogenically remodels the cardiac myocyte cytoskeleton via structural alterations to the microtubule lattice

Abstract

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Keywords

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