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      Semi-automatic detection of myocardial trabeculation using cardiovascular magnetic resonance: correlation with histology and reproducibility in a mouse model of non-compaction

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          Abstract

          Background

          The definition of left ventricular (LV) non-compaction is controversial, and discriminating between normal and excessive LV trabeculation remains challenging. Our goal was to quantify LV trabeculation on cardiovascular magnetic resonance (CMR) images in a genetic mouse model of non-compaction using a dedicated semi-automatic software package and to compare our results to the histology used as a gold standard.

          Methods

          Adult mice with ventricular non-compaction were generated by conditional trabecular deletion of Nkx2–5. Thirteen mice (5 controls, 8 Nkx2–5 mutants) were included in the study. Cine CMR series were acquired in the mid LV short axis plane (resolution 0.086 × 0.086x1mm 3) (11.75 T). In a sub set of 6 mice, 5 to 7 cine CMR were acquired in LV short axis to cover the whole LV with a lower resolution (0.172 × 0.172x1mm3). We used semi-automatic software to quantify the compacted mass (M c), the trabeculated mass (M t) and the percentage of trabeculation (M t/M c) on all cine acquisitions . After CMR all hearts were sliced along the short axis and stained with eosin, and histological LV contouring was performed manually, blinded from the CMR results, and M t, M c and M t/M c were quantified. Intra and interobserver reproducibility was evaluated by computing the intra class correlation coefficient (ICC).

          Results

          Whole heart acquisition showed no statistical significant difference between trabeculation measured at the basal, midventricular and apical parts of the LV. On the mid-LV cine CMR slice, the median M t was 0.92 mg (range 0.07–2.56 mg), M c was 12.24 mg (9.58–17.51 mg), M t/M c was 6.74% (0.66–17.33%). There was a strong correlation between CMR and the histology for M t, M c and M t/ M c with respectively: r 2 = 0.94 ( p < 0.001), r 2 = 0.91 ( p < 0.001), r 2 = 0.83 ( p < 0.001). Intra- and interobserver reproducibility was 0.97 and 0.8 for M t; 0.98 and 0.97 for M c; 0.96 and 0.72 for M t/M c, respectively and significantly more trabeculation was observed in the M c Mutant mice than the controls.

          Conclusion

          The proposed semi-automatic quantification software is accurate in comparison to the histology and reproducible in evaluating M c, M t and M t/ M c on cine CMR.

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          Most cited references17

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          Measurement of trabeculated left ventricular mass using cardiac magnetic resonance imaging in the diagnosis of left ventricular non-compaction.

          To describe a method for measuring trabeculated left ventricular (LV) mass using cardiac magnetic resonance imaging and to assess its value in the diagnosis of left ventricular non-compaction (LVNC). Between January 2003 and 2008, we prospectively included 16 patients with LVNC. During the mean period, we included 16 patients with dilated cardiomyopathy (DCM), 16 patients with hypertrophic cardiomyopathy (HCM), and 16 control subjects. Left ventricular volumes, LV ejection fraction, and trabeculated LV mass were measured in the four different populations. The percentage of trabeculated LV mass was almost three times higher in the patients with LVNC (32 +/- 10%), compared with those with DCM (11 +/- 4%, P < 0.0001), HCM (12 +/- 4%, P < 0.0001), and controls (12 +/- 5%, P < 0.0001). A value of trabeculated LV mass above 20% of the global mass of the LV predicted the diagnosis of LVNC with a sensitivity of 93.7% [95% confidence interval (CI), 71.6-98.8%] and a specificity of 93.7% (95% CI, 83.1-97.8%; kappa = 0.84). The method described is reproducible and provides an assessment of the global amount of LV trabeculation. A trabeculated LV mass above 20% of the global LV mass is highly sensitive and specific for the diagnosis of LVNC.
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            The cardiac homeobox gene Csx/Nkx2.5 lies genetically upstream of multiple genes essential for heart development.

            Csx/Nkx2.5 is a vertebrate homeobox gene with a sequence homology to the Drosophila tinman, which is required for the dorsal mesoderm specification. Recently, heterozygous mutations of this gene were found to cause human congenital heart disease (Schott, J.-J., Benson, D. W., Basson, C. T., Pease, W., Silberbach, G. M., Moak, J. P., Maron, B. J., Seidman, C. E. and Seidman, J. G. (1998) Science 281, 108-111). To investigate the functions of Csx/Nkx2.5 in cardiac and extracardiac development in the vertebrate, we have generated and analyzed mutant mice completely null for Csx/Nkx2.5. Homozygous null embryos showed arrest of cardiac development after looping and poor development of blood vessels. Moreover, there were severe defects in vascular formation and hematopoiesis in the mutant yolk sac. Interestingly, TUNEL staining and PCNA staining showed neither enhanced apoptosis nor reduced cell proliferation in the mutant myocardium. In situ hybridization studies demonstrated that, among 20 candidate genes examined, expression of ANF, BNP, MLC2V, N-myc, MEF2C, HAND1 and Msx2 was disturbed in the mutant heart. Moreover, in the heart of adult chimeric mice generated from Csx/Nkx2.5 null ES cells, there were almost no ES cell-derived cardiac myocytes, while there were substantial contributions of Csx /Nkx2.5-deficient cells in other organs. Whole-mount &bgr;-gal staining of chimeric embryos showed that more than 20% contribution of Csx/Nkx2. 5-deficient cells in the heart arrested cardiac development. These results indicate that (1) the complete null mutation of Csx/Nkx2.5 did not abolish initial heart looping, (2) there was no enhanced apoptosis or defective cell cycle entry in Csx/Nkx2.5 null cardiac myocytes, (3) Csx/Nkx2.5 regulates expression of several essential transcription factors in the developing heart, (4) Csx/Nkx2.5 is required for later differentiation of cardiac myocytes, (5) Csx/Nkx2. 5 null cells exert dominant interfering effects on cardiac development, and (6) there were severe defects in yolk sac angiogenesis and hematopoiesis in the Csx/Nkx2.5 null embryos.
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              Nkx2-5 pathways and congenital heart disease; loss of ventricular myocyte lineage specification leads to progressive cardiomyopathy and complete heart block.

              Human mutations in Nkx2-5 lead to progressive cardiomyopathy and conduction defects via unknown mechanisms. To define these pathways, we generated mice with a ventricular-restricted knockout of Nkx2-5, which display no structural defects but have progressive complete heart block, and massive trabecular muscle overgrowth found in some patients with Nkx2-5 mutations. At birth, mutant mice display a hypoplastic atrioventricular (AV) node and then develop selective dropout of these conduction cells. Transcriptional profiling uncovered the aberrant expression of a unique panel of atrial and conduction system-restricted target genes, as well as the ectopic, high level BMP-10 expression in the adult ventricular myocardium. Further, BMP-10 is shown to be necessary and sufficient for a major component of the ventricular muscle defects. Accordingly, loss of ventricular muscle cell lineage specification into trabecular and conduction system myocytes is a new mechanistic pathway for progressive cardiomyopathy and conduction defects in congenital heart disease.
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                Author and article information

                Contributors
                +336 42 54 79 51 , jfrandon38@gmail.com , julien.frandon@chu-nimes.fr
                stephanie.bricq@u-bourgogne.fr
                bentatouzakaria@gmail.com
                laetitia.marcadet@gmail.com
                pierreantoine.barral@ap-hm.fr
                MFinas@chu-grenoble.fr
                DFagret@chu-grenoble.fr
                frank.kober@univ-amu.fr
                gilbert.habib@ap-hm.fr
                monique.bernard@univ-amu.fr
                alain.Lalande@u-bourgogne.fr
                lucile.miquerol@univ-amu.fr
                alexis.jacquier@ap-hm.fr
                Journal
                J Cardiovasc Magn Reson
                J Cardiovasc Magn Reson
                Journal of Cardiovascular Magnetic Resonance
                BioMed Central (London )
                1097-6647
                1532-429X
                25 October 2018
                25 October 2018
                2018
                : 20
                : 70
                Affiliations
                [1 ]ISNI 0000 0001 2176 4817, GRID grid.5399.6, Aix-Marseille University, CNRS, CRMBM, ; Marseille, France
                [2 ]ISNI 0000 0001 0404 1115, GRID grid.411266.6, Department of Radiology, , Timone University Hospital, ; Marseille, France
                [3 ]ISNI 0000 0004 0593 8241, GRID grid.411165.6, Department of Radiology, , Nîmes University Hospital, ; Nîmes, France
                [4 ]ISNI 0000 0001 2298 9313, GRID grid.5613.1, Le2i, , Université de Bourgogne Franche-Comté, ; Dijon, France
                [5 ]ISNI 0000 0001 2176 4817, GRID grid.5399.6, CNRS UMR 7288, Developmental Biology Institute of Marseille, , Aix-Marseille University, ; Marseille, France
                [6 ]ISNI 0000 0004 0369 268X, GRID grid.450308.a, INSERM, U1039, Radiopharmaceutiques Biocliniques, , Université Grenoble Alpes, ; Grenoble, France
                [7 ]ISNI 0000 0001 0404 1115, GRID grid.411266.6, Department of Cardiology, APHM, , la Timone Hospital, ; Marseille, France
                [8 ]GRID grid.31151.37, Department of MRI, , University Hospital Francois Mitterrand, ; Dijon, France
                Author information
                http://orcid.org/0000-0001-7816-8178
                Article
                489
                10.1186/s12968-018-0489-0
                6201553
                30355287
                872540fd-8b30-4ec4-ae5e-4abe40fa8316
                © The Author(s). 2018

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 18 January 2018
                : 5 September 2018
                Funding
                Funded by: grant provided by the Société Française de Radiologie – French Society of Radiology – (SFR) together with the Collège des Enseignants en Radiologie de France – French Academic College of Radiology – (CERF)
                Funded by: france life imaging
                Award ID: ANR-11-INBS-0006
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2018

                Cardiovascular Medicine
                left ventricular non-compaction,semi-automatic segmentation,cmr,genetic mouse model

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