Triggering typical nemaline myopathy with compound heterozygous nebulin mutations reveals myofilament structural changes as pathomechanism

Lindqvist, Johan; Ma, Weikang; Li, Frank; Hernandez, Yaeren; Kolb, Justin; Kiss, Balazs; Tonino, Paola; van der Pijl, Robbert; Karimi, Esmat; Gong, Henry; Strom, Josh; Hourani, Zaynab; Smith III, John E.; Ottenheijm, Coen; Irving, Thomas; Granzier, Henk

doi:10.1038/s41467-020-16526-9

Title: Triggering typical nemaline myopathy with compound heterozygous nebulin mutations reveals myofilament structural changes as pathomechanism

Journal Article · Mon Jun 01 00:00:00 EDT 2020 · Nature Communications

DOI:https://doi.org/10.1038/s41467-020-16526-9· OSTI ID:1631850

Lindqvist, Johan ^[1];

^[2]; Li, Frank ^[1]; Hernandez, Yaeren ^[1]; Kolb, Justin ^[1]; Kiss, Balazs ^[1]; Tonino, Paola ^[1]; van der Pijl, Robbert ^[1]; Karimi, Esmat ^[1]; Gong, Henry ^[2]; Strom, Josh ^[1]; Hourani, Zaynab ^[1];

^[1]; Ottenheijm, Coen ^[1]; Irving, Thomas ^[2];

^[1]

Univ. of Arizona, Tucson, AZ (United States)
Illinois Inst. of Technology, Chicago, IL (United States)

Nebulin is a giant protein that winds around the actin filaments in the skeletal muscle sarcomere. Compound-heterozygous mutations in the nebulin gene (NEB) cause typical nemaline myopathy (NM), a muscle disorder characterized by muscle weakness with limited treatment options. We created a mouse model with a missense mutation p.Ser6366Ile and a deletion of NEB exon 55, the Compound-Het model that resembles typical NM. We show that Compound-Het mice are growth-retarded and have muscle weakness. Muscles have a reduced myofibrillar fractional-area and sarcomeres are disorganized, contain rod bodies, and have longer thin filaments. In contrast to nebulin-based severe NM where haplo-insufficiency is the disease driver, Compound-Het mice express normal amounts of nebulin. X-ray diffraction revealed that the actin filament is twisted with a larger radius, that tropomyosin and troponin behavior is altered, and that the myofilament spacing is increased. The unique disease mechanism of nebulin-based typical NM reveals novel therapeutic targets.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: A Foundation Building Strength; National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS); USDOE Office of Science (SC); National Institute of General Medical Sciences (NIGMS); National Institutes of Health (NIH)

Grant/Contract Number:: R01AR053897; AC02-06CH11357; 9 P41 GM103622; 1S10OD018090-01

OSTI ID:: 1631850

Journal Information:: Nature Communications, Vol. 11, Issue 1; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

Citation Metrics:

Cited by: 7 works

Citation information provided by
Web of Science

References (67)

Interactions between Nebulin-like Motifs and Thin Filament Regulatory Proteins Ogut, Ozgur; Hossain, M. Moazzem; Jin, Jian-Ping Journal of Biological Chemistry, Vol. 278, Issue 5 https://doi.org/10.1074/jbc.M205853200	journal	November 2002
Expression of Distinct Classes of Titin Isoforms in Striated and Smooth Muscles by Alternative Splicing, and Their Conserved Interaction with Filamins Labeit, Siegfried; Lahmers, Sunshine; Burkart, Christoph Journal of Molecular Biology, Vol. 362, Issue 4 https://doi.org/10.1016/j.jmb.2006.07.077	journal	September 2006
Tropomodulins and Leiomodins: Actin Pointed End Caps and Nucleators in Muscles Fowler, Velia M.; Dominguez, Roberto Biophysical Journal, Vol. 112, Issue 9 https://doi.org/10.1016/j.bpj.2017.03.034	journal	May 2017
Titin/connectin-based modulation of the Frank-Starling mechanism of the heart Fukuda, Norio; Granzier, Henk L. Journal of Muscle Research and Cell Motility, Vol. 26, Issue 6-8 https://doi.org/10.1007/s10974-005-9038-1	journal	February 2006
Extensibility and Symmetry of Actin Filaments in Contracting Muscles Bordas, J.; Svensson, A.; Rothery, M. Biophysical Journal, Vol. 77, Issue 6 https://doi.org/10.1016/S0006-3495(99)77150-X	journal	December 1999
The Filament Lattice of Striated Muscle Millman, Barry M. Physiological Reviews, Vol. 78, Issue 2 https://doi.org/10.1152/physrev.1998.78.2.359	journal	April 1998
Deleting nebulin's C-terminus reveals its importance to sarcomeric structure and function and is sufficient to invoke nemaline myopathy Li, Frank; Barton, Elisabeth R.; Granzier, Henk Human Molecular Genetics, Vol. 28, Issue 10 https://doi.org/10.1093/hmg/ddz016	journal	January 2019
Altered Contractility of Skeletal Muscle in Mice Deficient in Titin’s M-Band Region Ottenheijm, Coen A. C.; Hidalgo, Carlos; Rost, Katharina Journal of Molecular Biology, Vol. 393, Issue 1 https://doi.org/10.1016/j.jmb.2009.08.009	journal	October 2009
Nebulin-deficient mice exhibit shorter thin filament lengths and reduced contractile function in skeletal muscle Bang, Marie-Louise; Li, Xiaodong; Littlefield, Ryan Journal of Cell Biology, Vol. 173, Issue 6 https://doi.org/10.1083/jcb.200603119	journal	June 2006
Architecture of the sarcomere matrix of skeletal muscle: immunoelectron microscopic evidence that suggests a set of parallel inextensible nebulin filaments anchored at the Z line Wang, K. The Journal of Cell Biology, Vol. 107, Issue 6 https://doi.org/10.1083/jcb.107.6.2199	journal	December 1988
Dominantly inherited distal nemaline/cap myopathy caused by a large deletion in the nebulin gene Kiiski, Kirsi J.; Lehtokari, Vilma-Lotta; Vihola, Anna K. Neuromuscular Disorders, Vol. 29, Issue 2 https://doi.org/10.1016/j.nmd.2018.12.007	journal	February 2019
Altered myofilament function depresses force generation in patients with nebulin-based nemaline myopathy (NEM2) Ottenheijm, Coen A. C.; Hooijman, Pleuni; DeChene, Elizabeth T. Journal of Structural Biology, Vol. 170, Issue 2 https://doi.org/10.1016/j.jsb.2009.11.013	journal	May 2010
SMASH – semi-automatic muscle analysis using segmentation of histology: a MATLAB application Smith, Lucas R.; Barton, Elisabeth R. Skeletal Muscle, Vol. 4, Issue 1 https://doi.org/10.1186/2044-5040-4-21	journal	January 2014
Tissue Triage and Freezing for Models of Skeletal Muscle Disease Meng, Hui; Janssen, Paul M. L.; Grange, Robert W. Journal of Visualized Experiments, Issue 89 https://doi.org/10.3791/51586	journal	January 2014
Nemaline Myopathy: a new Congenital Myopathy Shy, G. Milton; Engel, W. King; Somers, J. E. Brain, Vol. 86, Issue 4 https://doi.org/10.1093/brain/86.4.793	journal	January 1963
Nemaline myopathy in the Ashkenazi Jewish population is caused by a deletion in the nebulin gene Anderson, SylviaL.; Ekstein, Josef; Donnelly, MaryC. Human Genetics, Vol. 115, Issue 3 https://doi.org/10.1007/s00439-004-1140-8	journal	June 2004
Myosin Head Configurations in Resting and Contracting Murine Skeletal Muscle Ma, Weikang; Gong, Henry; Irving, Thomas International Journal of Molecular Sciences, Vol. 19, Issue 9 https://doi.org/10.3390/ijms19092643	journal	September 2018
The nebulin SH3 domain is dispensable for normal skeletal muscle structure but is required for effective active load bearing in mouse Yamamoto, D. L.; Vitiello, C.; Zhang, J. Journal of Cell Science, Vol. 126, Issue 23 https://doi.org/10.1242/jcs.137026	journal	September 2013
Nebulin, a helical actin binding protein. Pfuhl, M.; Winder, S. J.; Pastore, A. The EMBO Journal, Vol. 13, Issue 8 https://doi.org/10.1002/j.1460-2075.1994.tb06446.x	journal	April 1994
Complete genomic structure of the human nebulin gene and identification of alternatively spliced transcripts Donner, Kati; Sandbacka, Maria; Lehtokari, Vilma-Lotta European Journal of Human Genetics, Vol. 12, Issue 9 https://doi.org/10.1038/sj.ejhg.5201242	journal	July 2004
Nebulin Alters Cross-bridge Cycling Kinetics and Increases Thin Filament Activation: A NOVEL MECHANISM FOR INCREASING TENSION AND REDUCING TENSION COST Chandra, Murali; Mamidi, Ranganath; Ford, Steven Journal of Biological Chemistry, Vol. 284, Issue 45 https://doi.org/10.1074/jbc.M109.049718	journal	September 2009
Deleting exon 55 from the nebulin gene induces severe muscle weakness in a mouse model for nemaline myopathy Ottenheijm, Coen A. C.; Buck, Danielle; de Winter, Josine M. Brain, Vol. 136, Issue 6 https://doi.org/10.1093/brain/awt113	journal	May 2013
Nebulin as a length regulator of thin filaments of vertebrate skeletal muscles: correlation of thin filament length, nebulin size, and epitope profile Kruger, M. The Journal of Cell Biology, Vol. 115, Issue 1 https://doi.org/10.1083/jcb.115.1.97	journal	October 1991
Nebulin deficiency in adult muscle causes sarcomere defects and muscle-type-dependent changes in trophicity: novel insights in nemaline myopathy Li, Frank; Buck, Danielle; De Winter, Josine Human Molecular Genetics, Vol. 24, Issue 18 https://doi.org/10.1093/hmg/ddv243	journal	June 2015
The giant protein titin regulates the length of the striated muscle thick filament Tonino, Paola; Kiss, Balazs; Strom, Josh Nature Communications, Vol. 8, Issue 1 https://doi.org/10.1038/s41467-017-01144-9	journal	October 2017
Rate of force generation in muscle: correlation with actomyosin ATPase activity in solution. Brenner, B.; Eisenberg, E. Proceedings of the National Academy of Sciences, Vol. 83, Issue 10 https://doi.org/10.1073/pnas.83.10.3542	journal	May 1986
Evidence that nebulin is a protein-ruler in muscle thin filaments Labeit, S.; Gibson, T.; Lakey, A. FEBS Letters, Vol. 282, Issue 2 https://doi.org/10.1016/0014-5793(91)80503-U	journal	May 1991
Ultrastructure of skeletal muscle fibers studied by a plunge quick freezing method: myofilament lengths Sosa, H.; Popp, D.; Ouyang, G. Biophysical Journal, Vol. 67, Issue 1 https://doi.org/10.1016/S0006-3495(94)80479-5	journal	July 1994
An Atomic Model of Actin Filaments Cross-Linked by Fimbrin and Its Implications for Bundle Assembly and Function Volkmann, Niels; DeRosier, David; Matsudaira, Paul The Journal of Cell Biology, Vol. 153, Issue 5 https://doi.org/10.1083/jcb.153.5.947	journal	May 2001
Nebulin interactions with actin and tropomyosin are altered by disease-causing mutations Marttila, Minttu; Hanif, Mubashir; Lemola, Elina Skeletal Muscle, Vol. 4, Issue 1 https://doi.org/10.1186/2044-5040-4-15	journal	January 2014
Nebulin and titin modulate cross-bridge cycling and length-dependent calcium sensitivity Mijailovich, Srboljub M.; Stojanovic, Boban; Nedic, Djordje The Journal of General Physiology, Vol. 151, Issue 5 https://doi.org/10.1085/jgp.201812165	journal	April 2019
Novel mutations in NEB cause abnormal nebulin expression and markedly impaired muscle force generation in severe nemaline myopathy Lawlor, Michael W.; Ottenheijm, Coen A.; Lehtokari, Vilma-Lotta Skeletal Muscle, Vol. 1, Issue 1 https://doi.org/10.1186/2044-5040-1-23	journal	January 2011
Myopathology in congenital myopathies Sewry, C. A.; Wallgren-Pettersson, C. Neuropathology and Applied Neurobiology, Vol. 43, Issue 1 https://doi.org/10.1111/nan.12369	journal	February 2017
Distal myopathy caused by homozygous missense mutations in the nebulin gene Wallgren-Pettersson, C.; Lehtokari, V. -L.; Kalimo, H. Brain, Vol. 130, Issue 6 https://doi.org/10.1093/brain/awm094	journal	April 2007
Nemaline myopathy: A clinical study of 143 cases Ryan, Monique M.; Schnell, Christina; Strickland, Corinne D. Annals of Neurology, Vol. 50, Issue 3 https://doi.org/10.1002/ana.1080	journal	January 2001
Identification of 45 novel mutations in the nebulin gene associated with autosomal recessive nemaline myopathy Lehtokari, Vilma-Lotta; Pelin, Katarina; Sandbacka, Maria Human Mutation, Vol. 27, Issue 9 https://doi.org/10.1002/humu.20370	journal	January 2006
Thick-Filament Extensibility in Intact Skeletal Muscle Ma, Weikang; Gong, Henry; Kiss, Balázs Biophysical Journal, Vol. 115, Issue 8 https://doi.org/10.1016/j.bpj.2018.08.038	journal	October 2018
The Complete Mouse Nebulin Gene Sequence and the Identification of Cardiac Nebulin Kazmierski, Steven T.; Antin, Parker B.; Witt, Christian C. Journal of Molecular Biology, Vol. 328, Issue 4 https://doi.org/10.1016/S0022-2836(03)00348-6	journal	May 2003
Muscle histopathology in nebulin-related nemaline myopathy: ultrastrastructural findings correlated to disease severity and genotype Malfatti, Edoardo; Lehtokari, Vilma-Lotta; Böhm, Johann Acta Neuropathologica Communications, Vol. 2, Issue 1 https://doi.org/10.1186/2051-5960-2-44	journal	April 2014
Genetics of the nemaline myopathies and the myotubular myopathies Wallgren-Pettersson, Carina Neuromuscular Disorders, Vol. 8, Issue 6 https://doi.org/10.1016/S0960-8966(98)00045-5	journal	August 1998
Nebulin stiffens the thin filament and augments cross-bridge interaction in skeletal muscle Kiss, Balázs; Lee, Eun-Jeong; Ma, Weikang Proceedings of the National Academy of Sciences, Vol. 115, Issue 41 https://doi.org/10.1073/pnas.1804726115	journal	September 2018
Nebulin increases thin filament stiffness and force per cross-bridge in slow-twitch soleus muscle fibers Kawai, Masataka; Karam, Tarek S.; Kolb, Justin The Journal of General Physiology, Vol. 150, Issue 11 https://doi.org/10.1085/jgp.201812104	journal	October 2018
Sarcomere Dysfunction in Nemaline Myopathy de Winter, Josine M.; Ottenheijm, Coen A. C. Journal of Neuromuscular Diseases, Vol. 4, Issue 2 https://doi.org/10.3233/JND-160200	journal	May 2017
Nebulin regulates thin filament length, contractility, and Z-disk structure in vivo Witt, Christian C.; Burkart, Christoph; Labeit, Dietmar The EMBO Journal, Vol. 25, Issue 16 https://doi.org/10.1038/sj.emboj.7601242	journal	August 2006
Sarcoplasmic reticulum calcium uptake and speed of relaxation are depressed in nebulin-free skeletal muscle Ottenheijm, Coen A. C.; Fong, Chi; Vangheluwe, Peter The FASEB Journal, Vol. 22, Issue 8 https://doi.org/10.1096/fj.07-104372	journal	August 2008
A nebulin super-repeat panel reveals stronger actin binding toward the ends of the super-repeat region: Nebulin Binding of Actin Laitila, Jenni; Lehtonen, Johanna; Lehtokari, Vilma-Lotta Muscle & Nerve, Vol. 59, Issue 1 https://doi.org/10.1002/mus.26350	journal	November 2018
Reduced myofibrillar connectivity and increased Z-disk width in nebulin-deficient skeletal muscle Tonino, P.; Pappas, C. T.; Hudson, B. D. Journal of Cell Science, Vol. 123, Issue 3 https://doi.org/10.1242/jcs.042234	journal	January 2010
Nebulin mutations in autosomal recessive nemaline myopathy: an update Pelin, Katarina; Donner, Kati; Holmberg, Maria Neuromuscular Disorders, Vol. 12, Issue 7-8 https://doi.org/10.1016/S0960-8966(02)00066-4	journal	October 2002
The exon 55 deletion in the nebulin gene – One single founder mutation with world-wide occurrence Lehtokari, Vilma-Lotta; Greenleaf, Rebecca S.; DeChene, Elizabeth T. Neuromuscular Disorders, Vol. 19, Issue 3 https://doi.org/10.1016/j.nmd.2008.12.001	journal	March 2009
Modulation of Muscle Atrophy, Fatigue and MLC Phosphorylation by MuRF1 as Indicated by Hindlimb Suspension Studies on MuRF1-KO Mice Labeit, Siegfried; Kohl, Christine H.; Witt, Christian C. Journal of Biomedicine and Biotechnology, Vol. 2010 https://doi.org/10.1155/2010/693741	journal	January 2010
Each Actin Subunit Has Three Nebulin Binding Sites Lukoyanova, Natalya; VanLoock, Margaret S.; Orlova, Albina Current Biology, Vol. 12, Issue 5 https://doi.org/10.1016/S0960-9822(02)00678-4	journal	March 2002
Congenital myopathies: disorders of excitation–contraction coupling and muscle contraction Jungbluth, Heinz; Treves, Susan; Zorzato, Francesco Nature Reviews Neurology, Vol. 14, Issue 3 https://doi.org/10.1038/nrneurol.2017.191	journal	February 2018
Nemaline myopathies: a current view Sewry, Caroline A.; Laitila, Jenni M.; Wallgren-Pettersson, Carina Journal of Muscle Research and Cell Motility, Vol. 40, Issue 2 https://doi.org/10.1007/s10974-019-09519-9	journal	June 2019
Thin filament length dysregulation contributes to muscle weakness in nemaline myopathy patients with nebulin deficiency Ottenheijm, C. A. C.; Witt, C. C.; Stienen, G. J. Human Molecular Genetics, Vol. 18, Issue 13 https://doi.org/10.1093/hmg/ddp168	journal	April 2009
Relationship between muscle fiber types and sizes and muscle architectural properties in the mouse hindlimb Burkholder, Thomas J.; Fingado, Brian; Baron, Stephanie Journal of Morphology, Vol. 221, Issue 2 https://doi.org/10.1002/jmor.1052210207	journal	August 1994
Nebulin plays a direct role in promoting strong actin-myosin interactions Bang, Marie-Louise; Caremani, Marco; Brunello, Elisabetta The FASEB Journal, Vol. 23, Issue 12 https://doi.org/10.1096/fj.09-137729	journal	December 2009
Mutation-specific effects on thin filament length in thin filament myopathy: Thin Filament Myopathy Winter, Josine M. de; Joureau, Barbara; Lee, Eun-Jeong Annals of Neurology, Vol. 79, Issue 6 https://doi.org/10.1002/ana.24654	journal	April 2016
New Insights into the Structural Roles of Nebulin in Skeletal Muscle Ottenheijm, Coen A. C.; Granzier, Henk Journal of Biomedicine and Biotechnology, Vol. 2010 https://doi.org/10.1155/2010/968139	journal	January 2010
Nemaline myopathy: current concepts. The ENMC International Consortium and Nemaline Myopathy. North, K. N.; Laing, N. G.; Wallgren-Pettersson, C. Journal of Medical Genetics, Vol. 34, Issue 9 https://doi.org/10.1136/jmg.34.9.705	journal	September 1997
Mutations in the nebulin gene associated with autosomal recessive nemaline myopathy Pelin, K.; Hilpela, P.; Donner, K. Proceedings of the National Academy of Sciences, Vol. 96, Issue 5 https://doi.org/10.1073/pnas.96.5.2305	journal	March 1999
Nebulin as a giant actin-binding template protein in skeletal muscle sarcomere Interaction of actin and cloned human nebulin fragments Jin, Jian-Ping; Wang, Kuan FEBS Letters, Vol. 281, Issue 1-2 https://doi.org/10.1016/0014-5793(91)80366-B	journal	April 1991
The BioCAT undulator beamline 18ID: a facility for biological non-crystalline diffraction and X-ray absorption spectroscopy at the Advanced Photon Source Fischetti, R.; Stepanov, S.; Rosenbaum, G. Journal of Synchrotron Radiation, Vol. 11, Issue 5 https://doi.org/10.1107/S0909049504016760	journal	August 2004
X-ray diffraction evidence for the extensibility of actin and myosin filaments during muscle contraction Wakabayashi, K.; Sugimoto, Y.; Tanaka, H. Biophysical Journal, Vol. 67, Issue 6 https://doi.org/10.1016/S0006-3495(94)80729-5	journal	December 1994
Mutation Update: The Spectra of Nebulin Variants and Associated Myopathies Lehtokari, Vilma-Lotta; Kiiski, Kirsi; Sandaradura, Sarah A. Human Mutation, Vol. 35, Issue 12 https://doi.org/10.1002/humu.22693	journal	November 2014
The velocity of unloaded shortening and its relation to sarcomere length and isometric force in vertebrate muscle fibres. Edman, K. A. The Journal of Physiology, Vol. 291, Issue 1 https://doi.org/10.1113/jphysiol.1979.sp012804	journal	June 1979
Vertical agarose gel electrophoresis and electroblotting of high-molecular-weight proteins Warren, Chad M.; Krzesinski, Paul R.; Greaser, Marion L. ELECTROPHORESIS, Vol. 24, Issue 11 https://doi.org/10.1002/elps.200305392	journal	June 2003
Developmental Control of Titin Isoform Expression and Passive Stiffness in Fetal and Neonatal Myocardium Lahmers, Sunshine; Wu, Yiming; Call, Douglas R. Circulation Research, Vol. 94, Issue 4 https://doi.org/10.1161/01.RES.0000115522.52554.86	journal	March 2004

Figures / Tables (8)

Similar Records

Nebulin stiffens the thin filament and augments cross-bridge interaction in skeletal muscle

Journal Article · Mon Sep 24 00:00:00 EDT 2018 · Proceedings of the National Academy of Sciences of the United States of America · OSTI ID:1631850

Kiss, Balázs; Lee, Eun-Jeong; Ma, Weikang; +5 more

Comparative decline of the protein profiles of nebulin in response to denervation in skeletal muscle

Journal Article · Fri Oct 09 00:00:00 EDT 2015 · Biochemical and Biophysical Research Communications · OSTI ID:1631850

Wei, Jih-Hua; Chang, Nen-Chung; Chen, Sy-Ping; +3 more

Nebulin and titin modulate cross-bridge cycling and length-dependent calcium sensitivity

Journal Article · Thu Apr 04 00:00:00 EDT 2019 · Journal of General Physiology · OSTI ID:1631850

Mijailovich, Srboljub M.; Stojanovic, Boban; Nedic, Djordje; +4 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Cytoskeleton
Muscle
Neuromuscular disease

Title: Triggering typical nemaline myopathy with compound heterozygous nebulin mutations reveals myofilament structural changes as pathomechanism

Citation Formats

References (67)

Figures / Tables (8)

Similar Records

Related Subjects