Multiscale Image Analysis of Calcium Dynamics in Cardiac Myocytes

Alexander Vallmitjana, Raúl Benítez


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Base Information

Volume

V55 - N2 / 2022 Ordinario

Reference

51076

DOI

http://dx.doi.org/10.7149/OPA.55.2.51076

Language

English

Keywords

Calcium Imaging, Fluorescence Microscopy, Image Processing

Abstract

Cardiac myocytes are the muscle cells that build up heart tissue and provide the mechanical action to pump blood by synchronously contracting at every heart beat. Heart muscle contraction is regulated by intracellular calcium concentration which exhibits a complex spatio-temporal dynamical behavior at the molecular, cellular and tissue levels. Details of such dynamical patterns are closely related to the mechanisms responsible for cardiovascular diseases, the single largest cause of death in the developed countries. The emerging field of translational cardiology focuses on the study of how such mechanisms connect and influence each other across spatial and temporal scales, eventually yielding to a certain clinical condition. To study such calcium dynamics in cardiac myocytes, we benefit from the recent advances in the field of experimental cell physiology. Fluorescence microscopy allows us to observe the distribution of calcium in the cell with a spatial resolution below one micron and a frame rate around one millisecond, thus providing a very accurate monitoring of calcium fluxes in the cell. The aim of the thesis summarized in this paper, was to develop image processing computational techniques for extracting quantitative data of physiological relevance from fluorescence confocal microscopy images at different scales. The two main subjects covered in the thesis were image segmentation and classification methods applied to fluorescence microscopy imaging of cardiac myocytes and calcium imaging. These methods were applied to a variety of problems involving different space and timescales, such as the localization of molecular receptors, the detection and characterization of spontaneous calcium-release events,and the propagation of calcium waves across a culture of cardiac cells. The following is a summary of the thesis as a consequence of having been awarded the 7th Justiniano Casas Award access it by the Sociedad Española de Óptica.

References

0

D. M. Bers, "Cardiac Excitation Contraction Coupling," Nature 415, 198-205 (2002).

1

A. Fabiato, "Calcium-Induced Release of Calcium from the Cardiac Sarcoplasmic-Reticulum," Am. J. Physiol. 245, C1-C14 (1983).

2

D. M. Jameson, Introduction to Fluorescence (Taylor & Francis, 2014).

3

A. Vallmitjana, "Multiscale Image Analysis of Calcium Dynamics in Cardiac Myocytes," PhD Thesis, Dep. d’Enginyeria Sist. Automàtica i Informàtica Ind. Univ. Politec. Catalunya (2017).

4

M. A. Walker, T. Kohl, S. E. Lehnart, J. L. Greenstein, W. J. Lederer, and R. L. Winslow, "On the Adjacency Matrix of RyR2 Cluster Structures," Plos Comput. Biol. 11, 1-21 (2015).

5

T. M. Hoang-Trong, A. Ullah, and M. S. Jafri, "Calcium Sparks in the Heart: Dynamics and Regulation.," Res. Rep. Biol. 6, 203-214(2015).

6

F. Hiess, A. Vallmitjana, R. Wang, H. Cheng, H. E. D. J. ter Keurs, J. Chen, L. Hove-Madsen, R. Benitez, S. R. W. Chen, and R. Benítez, "Distribution and function of cardiac ryanodine receptor clusters in live ventricular myocytes," J. Biol.Chem. 290, 20477-20487(2015).

7

C. Nolla-Colomer, "Anàlisi de patrons funcionals I estructurals en la regulació del calci en les cèl·lules cardíaques," PhD Thesis, Dep. d’Enginyeria Sist.Automàtica i Informàtica Ind. Univ. Politec. Catalunya (2021).

8

X. Zhong, B. Sun, A. Vallmitjana, T. Mi, W. Guo, M. Ni, R. Wang, A. Guo, H. J. Duff, A. M. Gillis, L.-S. Song, L. Hove-Madsen, R. Benitez, and S. R.W. W. Chen, "Suppression of Ryanodine Receptor Function Prolongs Ca2+ Release Refractoriness and Promotes Cardiac Alternans in Intact Hearts," Biochem. J. 473, 3951-3964 (2016).

9

F. Hiess, P. Detampel, C. Nolla-Colomer, A. Vallmitjana, A. Ganguly, M. Amrein, H. E. D. J. ter Keurs, R. Benítez, L. Hove-Madsen, and S. R. W. W. Chen, "Dynamic and Irregular Distribution of RyR2 Clusters in the Periphery of Live Ventricular Myocytes," Biophys. J.114, 343-354 (2018).

10

E. Chudin, J. Goldhaber, A. Garfinkel, J. Weiss, and B. Kogan, "Intracellular Ca2+ dynamics and the stability of ventricular tachycardia,"Biophys. J. 77, 2930-2941 (1999).

11

H. Ishida, C. Genka, Y. Hirota, H. Nakazawa, and W. H. Barry, "Formation of Planar and Spiral Ca2+ Waves in Isolated Cardiac Myocytes," Biophys. J. 77, 2114-2122 (1999).

12

Y. Bai, P. P. Jones, J. Guo, X. Zhong, R. B. Clark, Q. Zhou, R. Wang, A. Vallmitjana, R. Benitez, L. Hove-madsen, L. Semeniuk, A. Guo, L. Song, H. J. Duff, and S. R. W. Chen, "Phospholamban knockout breaks arrhythmogenic Ca2+ waves and suppresses catecholaminergic polymorphic ventricular tachycardia in mice.," Circ. Res. 113, 517-526 (2013).

13

A. Herraiz-Martínez, J. Álvarez-García, A. Llach, C. E. Molina, J. Fernandes, A. Ferrero-Gregori, C. Rodríguez, A. Vallmitjana, R. Benítez, J. Padró, J. Martínez-González, J. Cinca, and L.Hove-Madsen, "Ageing is associated with deterioration of calcium homeostasis in isolated human right atrial myocytes," Cardiovasc. Res. 106, 76-86 (2015).

14

A. Herraiz-Martínez, A. Llach, C. Tarifa, J. Gandía, V. Jiménez-Sabado, E. Lozano-Velasco, S. A. S. A. Serra, A. Vallmitjana, E. Vázquez Ruiz De Castroviejo, R. Benítez, A. Aranega, C. Muñoz-Guijosa, D. Franco, J. Cinca, and L. Hove-Madsen, "The 4q25 variant rs13143308T links risk of atrial fibrillation to defective calcium homoeostasis," Cardiovasc. Res. 115, 578-589 (2019).

15

C. Nolla-Colomer, S. Casabella‐Ramon, V. Jimenez-Sabado, A. Vallmitjana, C. Tarifa, A. Herraiz-Martínez, A. Llach, M. Tauron, J. Montiel, J. Cinca, S. R. W. Chen, R. Benitez, and L. Hove-Madsen, "Β2-adrenergic Stimulation Potentiates Spontaneous Calcium Release by Increasing Signal Mass and Co-activation of Ryanodine Receptor Clusters," Acta Physiol. 234, e13736 (2021).

16

X. Zhong, A. Vallmitjana, B. Sun, Z. Xiao, W. Guo, J. Wei, M. Ni, Y. Chen, E. R. O’Brien, A. M. Gillis, M. Hoshijima, H. Takeshima, L. Hove-Madsen, R. Benitez, D. Belke, and S. R. W. Chen, "Reduced expression of cardiac ryanodine receptor protects against stress-induced ventricular tachyarrhythmia, but increases the susceptibility to cardiac alternans," Biochem. J. 475, 169-183 (2018).

17

A. Vallmitjana, A. Civera-Tregon, J. Hoenicka, F. Palau, and R. Benitez, "Motion estimation of subcellular structures from fluorescence microscopy images," Proc. Annu. Int. Conf. IEEE Eng. Med. Biol. Soc. EMBS 4419-4422 (2017).

18

A. Civera-Tregón, L. Domínguez, P. Martínez-Valero, C. Serrano, A. Vallmitjana, R. Benítez, J. Hoenicka, J. Satrústegui, and F. Palau, "Mitochondria and calcium defects correlate with axonal dysfunction in GDAP1-related Charcot-Marie-Tooth mouse model, "Neurobiol. Dis. 152, 105300 (2021).

19

L. Breiman, "Random forests," Mach. Learn. 45, 5-32 (2001).

20

M. Barriga, R. Cal, N. Cabello, A. Llach, A. Vallmitjana, R. Benitez, L. Badimon, J. Cinca, V. Llorente-Cortes, L. Hove-Madsen, and R. Benítez, "Low density lipoproteins promote unstable calcium handling linked to reduced SERCA2 and connexin 40 expression in cardiomyocytes," PLoS One 8, e58128-e58128 (2013).

21

C. E. Molina, A. Llach, A. Herraiz-Martínez, C. Tarifa, M. Barriga, R. F. Wiegerinck, J. Fernandes, N. Cabello, A. Vallmitjana, R. Benitez, J. Montiel, J. Cinca, and L. Hove-Madsen, "Prevention of adenosine A2A receptor activation diminishes beat-to-beat alternation in human atrial myocytes," Basic Res. Cardiol. 111, 5 (2016).

22

J. Rivera-Torres, C. J. C. J. Calvo, A. Llach, G. Guzmán-Martínez, R. Caballero, C. González-Gómez, L. J. L. J. Jiménez-Borreguero, J. A. Guadix, F. G. F. G. F. G. Osorio, C. López-Otín, A. Herraiz-Martínez, N. Cabello, A. Vallmitjana, R. Benítez, L. B. L. B. L. B. Gordon, J. Jalife, J. M. J. M. J. M. Pérez-Pomares, J. Tamargo, E. Delpón, L. Hove-Madsen, D. Filgueiras-Rama, and V. Andrés, "Cardiac electrical defects in progeroid mice and Hutchinson-Gilford progeria syndrome patients with nuclear lamina alterations," Proc. Natl. Acad. Sci. 113, E7250-E7259 (2016).

23

M. A. Colman, "The Multiple Mechanisms of Spatially Discordant Alternans in the Heart," Biophys. J. 118, 2336-2338 (2020).

24

A. Vallmitjana, M. Barriga, Z. Nenadic, A. Llach, E. Alvarez-Lacalle, L. Hove-Madsen, and R. Benitez, "Identification of intracellular calcium dynamics in stimulated cardiomyocytes," Annu Int Conf IEEE Eng Med Biol Soc. 2010, 68-71 (2010).

25

B. Sun, J. Yao, M. Ni, J. Wei, X. Zhong, W. Guo, L. Zhang, R. Wang, D. Belke, Y.-X. Chen, K. V. V. Lieve, A. K. Broendberg, T. M. Roston, I. Blankoff, J. A. Kammeraad, J. C. von Alvensleben, J. Lazarte, A. Vallmitjana, L. J. Bohne, R. A. Rose, R. Benitez, L. Hove-Madsen, C. Napolitano, R. A. Hegele, M. Fill, S. Sanatani, A. A. M. Wilde, J. D. Roberts, S. G. Priori, H. K. Jensen, and S. R. W. Chen,"Cardiac ryanodine receptor calcium release deficiency syndrome," Sci. Transl. Med. 13,eaba7287 (2021).

26

B. Sun, J. Wei, X. Zhong, W. Guo, J. Yao, R. Wang, A. Vallmitjana,R. Benitez, L. Hove-Madsen, and S. R. W. Chen, "The cardiac ryanodine receptor, but not sarcoplasmic reticulum Ca2+-ATPase, is a major determinant of Ca2+ alternans in intact mouse hearts," J. Biol. Chem. 293, jbc.RA118.003760 (2018).

27

J. Wei, J. Yao, D. Belke, W. Guo, X. Zhong, B. Sun, R. Wang, J. P. Estillore, A. Vallmitjana, R.Benitez, L. Hove-Madsen, E. Alvarez-Lacalle, B. Echebarria, and S. W. Chen, "Ca 2+-CaM Dependent Inactivation of RyR2 Underlies Ca 2+ Alternans in Intact Heart," Circ. Res. 128, 63-83 (2021).

28

A. Vallmitjana, M. Barriga, L. Hove-Madsen, and R. Benitez, "Multilevel analysis of calcium dynamics in stimulated cultures of cardiomyocytes,"Annu Int Conf IEEE Eng Med Biol Soc. 2013, 6514-6517 (2013).

29

L. Hove-Madsen, A. Vallmitjana, and R. Benitez, "Método implementado por ordenador para caracterización dinámica de células en cultivos celulares y programas informáticos para llevar a cabo el método," patent ES2532746B1 (2016).