Leukocyte Nuclear Morphology Alterations in Dilated Cardiomyopathy Caused by a Lamin AC Truncating Mutation (LMNA/Ser431*) Are Modified by the Presence of a LAP2 Missense Polymorphism (TMPO/Arg690Cys)
Abstract
:1. Introduction
2. Results
2.1. Molecular Diagnosis
2.2. Protein Modeling Predicts the LAP2α Arg690Cys Polymorphism Destabilizes the LAP2α Homodimer
2.3. Effect of LAP2α Arg690Cys on LAP2α and Lamin AC Recognition in Silico
2.4. Immunostaining Analysis
3. Discussion
4. Materials and Methods
4.1. Subjects and Clinical Evaluation
4.2. DNA Analysis
4.3. Protein Modeling
4.4. Leukocyte Purification and Immunofluorescence (IF) Analysis
4.5. Protein Extraction and Immunoblot Analysis
4.6. Immunoprecipitation (IP) Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Tayal, U.; Prasad, S.; Cook, S.A. Genetics and genomics of dilated cardiomyopathy and systolic heart failure. Genome Med. 2017, 9, 20. [Google Scholar] [CrossRef] [Green Version]
- Reichart, D.; Magnussen, C.; Zeller, T.; Blankenberg, S. Dilated cardiomyopathy: From epidemiologic to genetic phenotypes: A translational review of current literature. J. Intern. Med. 2019, 286, 362–372. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- McNally, E.M.; Mestroni, L. Dilated Cardiomyopathy: Genetic Determinants and Mechanisms. Circ. Res. 2017, 121, 731–748. [Google Scholar] [CrossRef] [PubMed]
- Barp, A.; Bello, L.; Politano, L.; Melacini, P.; Calore, C.; Polo, A.; Vianello, S.; Sorarù, G.; Semplicini, C.; Pantic, B.; et al. Genetic Modifiers of Duchenne Muscular Dystrophy and Dilated Cardiomyopathy. PLoS ONE 2015, 10, e0141240. [Google Scholar] [CrossRef] [PubMed]
- Verdonschot, J.A.J.; Robinson, E.L.; James, K.N.; Mohamed, M.W.; Claes, G.R.F.; Casas, K.; Vanhoutte, E.K.; Hazebroek, M.R.; Kringlen, G.; Pasierb, M.M.; et al. Mutations in PDLIM5 are rare in dilated cardiomyopathy but are emerging as potential disease modifiers. Mol. Genet. Genom. Med. 2020, 8, e1049. [Google Scholar] [CrossRef] [Green Version]
- Gacita, A.M.; Fullenkamp, D.E.; Ohiri, J.; Pottinger, T.; Puckelwartz, M.J.; Nobrega, M.A.; McNally, E.M. Genetic Variation in Enhancers Modifies Cardiomyopathy Gene Expression and Progression. Circulation 2021, 143, 1302–1316. [Google Scholar] [CrossRef]
- Hershberger, R.E.; Siegfried, J.D. Update 2011: Clinical and Genetic Issues in Familial Dilated Cardiomyopathy. J. Am. Coll. Cardiol. 2011, 57, 1641–1649. [Google Scholar] [CrossRef] [Green Version]
- Arbustini, E.; Pilotto, A.; Repetto, A.; Grasso, M.; Negri, A.; Diegoli, M.; Campana, C.; Scelsi, L.; Baldini, E.; Gavazzi, A.; et al. Autosomal dominant dilated cardiomyopathy with atrioventricular block: A lamin A/C defect-related disease. J. Am. Coll. Cardiol. 2002, 39, 981–990. [Google Scholar] [CrossRef] [Green Version]
- Van Tienen, F.H.; Lindsey, P.J.; Kamps, M.A.; Krapels, I.P.; Ramaekers, F.; Brunner, H.G.; van den Wijngaard, A.; Broers, J.L. Assessment of fibroblast nuclear morphology aids interpretation of LMNA variants. Eur. J. Hum. Genet. 2019, 27, 389–399. [Google Scholar] [CrossRef] [Green Version]
- Wada, K.; Misaka, T.; Yokokawa, T.; Kimishima, Y.; Kaneshiro, T.; Oikawa, M.; Yoshihisa, A.; Takeishi, Y. Blood-Based Epigenetic Markers of FKBP5 Gene Methylation in Patients with Dilated Cardiomyopathy. J. Am. Heart Assoc. 2021, 10, e021101. [Google Scholar] [CrossRef]
- Ferradini, V.; Cosma, J.; Romeo, F.; De Masi, C.; Murdocca, M.; Spitalieri, P.; Mannucci, S.; Parlapiano, G.; Di Lorenzo, F.; Martino, A.; et al. Clinical Features of LMNA-Related Cardiomyopathy in 18 Patients and Characterization of Two Novel Variants. J. Clin. Med. 2021, 10, 5075. [Google Scholar] [CrossRef]
- Sabatelli, P.; Lattanzi, G.; Ognibene, A.; Columbaro, M.; Capanni, C.; Merlini, L.; Maraldi, N.M.; Squarzoni, S. Nuclear alterations in autosomal-dominant Emery-Dreifuss muscular dystrophy. Muscle Nerve 2001, 24, 826–829. [Google Scholar] [CrossRef] [PubMed]
- Scaffidi, P.; Misteli, T. Lamin A-Dependent Nuclear Defects in Human Aging. Science 2006, 312, 1059–1063. [Google Scholar] [CrossRef] [Green Version]
- Shumaker, D.K.; Dechat, T.; Kohlmaier, A.; Adam, S.A.; Bozovsky, M.R.; Erdos, M.R.; Eriksson, M.; Goldman, A.E.; Khuon, S.; Collins, F.S.; et al. Mutant nuclear lamin A leads to progressive alterations of epigenetic control in premature aging. Proc. Natl. Acad. Sci. USA 2006, 103, 8703–8708. [Google Scholar] [CrossRef] [Green Version]
- Wang, Y.; Herron, A.J.; Worman, H.J. Pathology and nuclear abnormalities in hearts of transgenic mice expressing M371K lamin A encoded by an LMNA mutation causing Emery-Dreifuss muscular dystrophy. Hum. Mol. Genet. 2006, 15, 2479–2489. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Håkelien, A.M.; Delbarre, E.; Gaustad, K.G.; Buendia, B.; Collas, P. Expression of the myodystrophic R453W mutation of lamin A in C2C12 myoblasts causes promoter-specific and global epigenetic defects. Exp. Cell Res. 2008, 314, 1869–1880. [Google Scholar] [CrossRef]
- Park, Y.-E.; Hayashi, Y.K.; Goto, K.; Komaki, H.; Hayashi, Y.; Inuzuka, T.; Noguchi, S.; Nonaka, I.; Nishino, I. Nuclear changes in skeletal muscle extend to satellite cells in autosomal dominant Emery-Dreifuss muscular dystrophy/limb-girdle muscular dystrophy 1B. Neuromuscul. Disord. 2009, 19, 29–36. [Google Scholar] [CrossRef]
- Gupta, P.; Bilinska, Z.T.; Sylvius, N.; Boudreau, E.; Veinot, J.P.; Labib, S.; Bolongo, P.M.; Hamza, A.; Jackson, T.; Ploski, R.; et al. Genetic and ultrastructural studies in dilated cardiomyopathy patients: A large deletion in the lamin A/C gene is associated with cardiomyocyte nuclear envelope disruption. Basic Res. Cardiol. 2010, 105, 365–377. [Google Scholar] [CrossRef] [Green Version]
- Vahabikashi, A.; Adam, S.A.; Medalia, O.; Goldman, R.D. Nuclear lamins: Structure and function in mechanobiology. APL Bioeng. 2022, 6, 011503. [Google Scholar] [CrossRef] [PubMed]
- Perovanović, J.; Hoffman, E.P. Mechanisms of allelic and clinical heterogeneity of lamin A/C phenotypes. Physiol. Genom. 2018, 50, 694–704. [Google Scholar] [CrossRef]
- Gruenbaum, Y.; Margalit, A.; Goldman, R.D.; Shumaker, D.K.; Wilson, K.L. The nuclear lamina comes of age. Nat. Rev. Mol. Cell Biol. 2005, 6, 21–31. [Google Scholar] [CrossRef]
- Dechat, T.; Pfleghaar, K.; Sengupta, K.; Shimi, T.; Shumaker, D.K.; Solimando, L.; Goldman, R.D. Nuclear lamins: Major factors in the structural organization and function of the nucleus and chromatin. Genes Dev. 2008, 22, 832–853. [Google Scholar] [CrossRef] [Green Version]
- Perovanovic, J.; Dell’Orso, S.; Gnochi, V.F.; Jaiswal, J.K.; Sartorelli, V.; Vigouroux, C.; Mamchaoui, K.; Mouly, V.; Bonne, G.; Hoffman, E.P. Laminopathies disrupt epigenomic developmental programs and cell fate. Sci. Transl. Med. 2016, 8, 335ra58. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gerbino, A.; Procino, G.; Svelto, M.; Carmosino, M. Role of Lamin A/C Gene Mutations in the Signaling Defects Leading to Cardiomyopathies. Front. Physiol. 2018, 9, 1356. [Google Scholar] [CrossRef] [Green Version]
- Dauer, W.T.; Worman, H.J. The Nuclear Envelope as a Signaling Node in Development and Disease. Dev. Cell 2009, 17, 626–638. [Google Scholar] [CrossRef] [Green Version]
- Peric-Hupkes, D.; van Steensel, B. Role of the Nuclear Lamina in Genome Organization and Gene Expression. Cold Spring Harb. Symp. Quant. Biol. 2010, 75, 517–524. [Google Scholar] [CrossRef] [Green Version]
- Dechat, T.; Korbei, B.; Vaughan, O.A.; Vlcek, S.; Hutchison, C.J.; Foisner, R. Lamina-associated polypeptide 2alpha binds intranuclear A-type lamins. J. Cell Sci. 2000, 113, 3473–3484. [Google Scholar] [CrossRef] [PubMed]
- Crasto, S.; My, I.; Di Pasquale, E. The Broad Spectrum of LMNA Cardiac Diseases: From Molecular Mechanisms to Clinical Phenotype. Front. Physiol. 2020, 11, 761. [Google Scholar] [CrossRef] [PubMed]
- Wilson, K.L.; Foisner, R. Lamin-binding Proteins. Cold Spring Harb. Perspect. Biol. 2010, 2, a000554. [Google Scholar] [CrossRef] [Green Version]
- Naetar, N.; Ferraioli, S.; Foisner, R. Lamins in the nuclear interior—Life outside the lamina. J. Cell Sci. 2017, 130, 2087–2096. [Google Scholar] [CrossRef]
- Taylor, M.R.; Slavov, D.; Gajewski, A.; Vlcek, S.; Ku, L.; Fain, P.R.; Carniel, E.; Di Lenarda, A.; Sinagra, G.; Boucek, M.M.; et al. Thymopoietin (lamina-associated polypeptide 2) gene mutation associated with dilated cardiomyopathy. Hum. Mutat. 2005, 26, 566–574. [Google Scholar] [CrossRef] [PubMed]
- Rosas-Madrigal, S.; Villarreal-Molina, M.T.; Flores-Rivera, J.; Rivas-Alonso, V.; Macias-Kauffer, L.R.; Ordoñez, G.; Chima-Galán, M.D.C.; Acuña-Alonzo, V.; Macín-Pérez, G.; Barquera, R.; et al. Interaction of HLA Class II rs9272219 and TMPO rs17028450 (Arg690Cys) Variants Affects Neuromyelitis Optica Spectrum Disorder Susceptibility in an Admixed Mexican Population. Front. Genet. 2021, 12, 647343. [Google Scholar] [CrossRef] [PubMed]
- Saj, M.; Bilinska, Z.T.; Tarnowska, A.; Sioma, A.; Bolongo, P.; Sobieszczanska-Malek, M.; Michalak, E.; Golen, D.; Mazurkiewicz, L.; Malek, L.; et al. LMNA mutations in Polish patients with dilated cardiomyopathy: Prevalence, clinical characteristics, and in vitro studies. BMC Med. Genet. 2013, 14, 55. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Carmosino, M.; Torretta, S.; Procino, G.; Gerbino, A.; Forleo, C.; Favale, S.; Svelto, M. Role of nuclear Lamin A/C in cardiomyocyte functions. Biol. Cell 2014, 106, 346–358. [Google Scholar] [CrossRef]
- Wolf, C.M.; Wang, L.; Alcalai, R.; Pizard, A.; Burgon, P.G.; Ahmad, F.; Sherwood, M.; Branco, D.M.; Wakimoto, H.; Fishman, G.; et al. Lamin A/C haploinsufficiency causes dilated cardiomyopathy and apoptosis-triggered cardiac conduction system disease. J. Mol. Cell. Cardiol. 2008, 44, 293–303. [Google Scholar] [CrossRef] [Green Version]
- Geiger, S.K.; Bär, H.; Ehlermann, P.; Wälde, S.; Rutschow, D.; Zeller, R.; Ivandic, B.T.; Zentgraf, H.; Katus, H.A.; Herrmann, H.; et al. Incomplete nonsense-mediated decay of mutant lamin A/C mRNA provokes dilated cardiomyopathy and ventricular tachycardia. J. Mol. Med. 2007, 86, 281–289. [Google Scholar] [CrossRef]
- Al-Saaidi, R.; Rasmussen, T.B.; Palmfeldt, J.; Nissen, P.H.; Beqqali, A.; Hansen, J.; Pinto, Y.M.; Boesen, T.; Mogensen, J.; Bross, P. The LMNA mutation p.Arg321Ter associated with dilated cardiomyopathy leads to reduced expression and a skewed ratio of lamin A and lamin C proteins. Exp. Cell Res. 2013, 319, 3010–3019. [Google Scholar] [CrossRef]
- Cai, Z.-J.; Lee, Y.-K.; Lau, Y.-M.; Ho, J.C.-Y.; Lai, W.-H.; Wong, N.L.-Y.; Huang, D.; Hai, J.-J.; Ng, K.-M.; Tse, H.-F.; et al. Expression of LMNA-R225X nonsense mutation results in dilated cardiomyopathy and conduction disorders (DCM-CD) in mice: Impact of exercise training. Int. J. Cardiol. 2020, 298, 85–92. [Google Scholar] [CrossRef] [Green Version]
- 1000 Genomes Project Consortium. A global reference for human genetic variation. Nature 2015, 526, 68–74. [Google Scholar] [CrossRef] [Green Version]
- Gesson, K.; Rescheneder, P.; Skoruppa, M.P.; von Haeseler, A.; Dechat, T.; Foisner, R. A-type lamins bind both hetero- and euchromatin, the latter being regulated by lamina-associated poly-peptide 2 alpha. Genome Res. 2016, 26, 462–473. [Google Scholar] [CrossRef]
- Lang, R.M.; Badano, L.P.; Mor-Avi, V.; Afilalo, J.; Armstrong, A.; Ernande, L.; Flachskampf, F.A.; Foster, E.; Goldstein, S.A.; Kuznetsova, T.; et al. Recommendations for Cardiac Chamber Quantification by Echocardiography in Adults: An Update from the American Society of Echocardiography and the European Association of Cardiovascular Imaging. Eur. Heart J. Cardiovasc. Imaging 2015, 16, 233–271. [Google Scholar] [CrossRef]
- Yang, H.; Wang, K. Genomic variant annotation and prioritization with ANNOVAR and wANNOVAR. Nat. Protoc. 2015, 10, 1556–1566. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Morales, A.; Kinnamon, D.D.; Jordan, E.; Platt, J.; Vatta, M.; Dorschner, M.O.; Starkey, C.A.; Mead, J.O.; Ai, T.; Burke, W.; et al. Variant Interpretation for Dilated Cardiomyopathy: Refinement of the American College of Medical Genetics and Genomics/ClinGen Guidelines for the DCM Precision Medicine Study. Circ. Genom. Precis. Med. 2020, 13, e002480. [Google Scholar] [CrossRef]
- Mackerell, A.D., Jr.; Feig, M.; Brooks, C.L., III. Extending the treatment of backbone energetics in protein force fields: Limitations of gas-phase quantum mechanics in reproducing protein conformational distributions in molecular dynamics simulations. J. Comput. Chem. 2004, 25, 1400–1415. [Google Scholar] [CrossRef]
- Krimm, I.; Östlund, C.; Gilquin, B.; Couprie, J.; Hossenlopp, P.; Mornon, J.-P.; Bonne, G.; Courvalin, J.-C.; Worman, H.J.; Zinn-Justin, S. The Ig-like Structure of the C-Terminal Domain of Lamin A/C, Mutated in Muscular Dystrophies, Cardiomyopathy, and Partial Lipodystrophy. Structure 2002, 10, 811–823. [Google Scholar] [CrossRef] [Green Version]
- MacIndoe, G.; Mavridis, L.; Venkatraman, V.; Devignes, M.-D.; Ritchie, D.W. HexServer: An FFT-based protein docking server powered by graphics processors. Nucleic Acids Res. 2010, 38, W445–W449. [Google Scholar] [CrossRef] [PubMed]
- Pierce, B.G.; Wiehe, K.; Hwang, H.; Kim, B.-H.; Vreven, T.; Weng, Z. ZDOCK server: Interactive docking prediction of protein-protein complexes and symmetric multimers. Bioinformatics 2014, 30, 1771–1773. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Van Zundert, G.C.P.; Rodrigues, J.P.G.L.M.; Trellet, M.; Schmitz, C.; Kastritis, P.L.; Karaca, E.; Melquiond, A.S.J.; van Dijk, M.; De Vries, S.J.; Bonvin, A.M.J.J. The HADDOCK2.2 Web Server: User-Friendly Integrative Modeling of Biomolecular Complexes. J. Mol. Biol. 2016, 428, 720–725. [Google Scholar] [CrossRef] [Green Version]
- McCarthy, D.; Perry, J.; Holburn, C.; Kirk, A.; James, D.; Moore, S.; Holborow, E. Centrifugation of normal and rheumatoid arthritis blood on Ficoll-Hypaque and Ficoll-Nycodenz solutions. J. Immunol. Methods 1984, 73, 415–425. [Google Scholar] [CrossRef]
- Fotino, M.; Merson, E.J.; Allen, F.H., Jr. Micromethod for rapid separation of lymphocytes from peripheral blood. Ann. Clin. Lab. Sci. 1971, 1, 131–133. [Google Scholar]
- Ruitenberg, J.J.; Mulder, C.B.; Maino, V.C.; Landay, A.L.; Ghanekar, S.A. VACUTAINER® CPT™ and Ficoll density gradient separation perform equivalently in maintaining the quality and function of PBMC from HIV seropositive blood samples. BMC Immunol. 2006, 7, 11. [Google Scholar] [CrossRef] [PubMed]
- Arellanes-Robledo, J.; Reyes-Gordillo, K.; Ibrahim, J.; Leckey, L.; Shah, R.; Lakshman, M.R. Ethanol targets nucleoredoxin/dishevelled interactions and stimulates phosphatidylinositol 4-phosphate production in vivo and in vitro. Biochem. Pharmacol. 2018, 156, 135–146. [Google Scholar] [CrossRef] [PubMed]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
González-Garrido, A.; Rosas-Madrigal, S.; Rojo-Domínguez, A.; Arellanes-Robledo, J.; López-Mora, E.; Carnevale, A.; Arregui, L.; Rosendo-Gutiérrez, R.; Romero-Hidalgo, S.; Villarreal-Molina, M.T. Leukocyte Nuclear Morphology Alterations in Dilated Cardiomyopathy Caused by a Lamin AC Truncating Mutation (LMNA/Ser431*) Are Modified by the Presence of a LAP2 Missense Polymorphism (TMPO/Arg690Cys). Int. J. Mol. Sci. 2022, 23, 13626. https://doi.org/10.3390/ijms232113626
González-Garrido A, Rosas-Madrigal S, Rojo-Domínguez A, Arellanes-Robledo J, López-Mora E, Carnevale A, Arregui L, Rosendo-Gutiérrez R, Romero-Hidalgo S, Villarreal-Molina MT. Leukocyte Nuclear Morphology Alterations in Dilated Cardiomyopathy Caused by a Lamin AC Truncating Mutation (LMNA/Ser431*) Are Modified by the Presence of a LAP2 Missense Polymorphism (TMPO/Arg690Cys). International Journal of Molecular Sciences. 2022; 23(21):13626. https://doi.org/10.3390/ijms232113626
Chicago/Turabian StyleGonzález-Garrido, Antonia, Sandra Rosas-Madrigal, Arturo Rojo-Domínguez, Jaime Arellanes-Robledo, Enrique López-Mora, Alessandra Carnevale, Leticia Arregui, Rigoberto Rosendo-Gutiérrez, Sandra Romero-Hidalgo, and María Teresa Villarreal-Molina. 2022. "Leukocyte Nuclear Morphology Alterations in Dilated Cardiomyopathy Caused by a Lamin AC Truncating Mutation (LMNA/Ser431*) Are Modified by the Presence of a LAP2 Missense Polymorphism (TMPO/Arg690Cys)" International Journal of Molecular Sciences 23, no. 21: 13626. https://doi.org/10.3390/ijms232113626