Julie Magat1,2,3, Maxime Yon1,2,3, Yann Bihan-Poudec4, and Valéry Ozenne1,2,3,5
1IHU Liryc, Electrophysiology and Heart Modeling Institute, Foundation Bordeaux Université, Bordeaux, France, 2Univ. Bordeaux, Centre de recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France, 3INSERM, Centre de recherche Cardio-Thoracique de Bordeaux, U1045, Bordeaux, France, 4Institut des Sciences Cognitives Marc Jeannerod, CNRS UMR 5229, Université Claude Bernard Lyon I, Bron, France, 5Centre de Résonance Magnétique des Systèmes Biologiques, UMR 5536, CNRS/Université de Bordeaux, Bordeaux, France
Synopsis
In this study, we developed
a groupwise
registration and tractography framework to investigate the global myofiber
arrangement of large mammalian sheep hearts. To
demonstrate the potential application of the proposed method, a novel
description of sub-regions in the intraventricular septum (IVS) is presented. The study focuses on one fiber-bundle in the posterior junction and
three fiber-bundles in the anterior junction.
Introduction
Knowledge
of the normal myocardial–myocyte
orientation could theoretically allow the definition of relevant quantitative biomarkers
in clinical routine to diagnose heart pathologies [1,2]. A whole heart
diffusion tensor template representative of the global myofiber organization
over species is therefore crucial for comparisons across populations. In this study,
we develop a groupwise registration
and tractography framework to investigate the global myofiber arrangement of large
mammalian sheep hearts. To demonstrate
the potential application of the proposed method, a novel description of
sub-regions in the intraventricular septum (IVS) is presented.Methods
This
protocol was approved by the Animal Research Ethics Committee in accordance
with the European rules for animal experimentation. The
hearts of 4 female sheeps (ovine) were explanted via sternal thoracotomy under general anesthesia. Three hearts were dedicated
to MR experiments and one heart was dissected for macroscopic
visualization. The three hearts (~12×8×6 cm3, heart
weight=150±10 g) were perfusion-fixed in 10% formaldehyde containing
2 mL of gadoterate meglumin. Before imaging, the heart were
removed from the solution and immersed in Fomblin. The experiments
were performed at 9.4T (Bruker BioSpin MRI system, Ettlingen Germany) with an
open bore access of 30 cm and a 200-mm inner diameter gradient (300 mT/m).
A 3D
FLASH sequence (TE/TR/FA = 9ms, 30ms, 30°) at 150
μm3 voxel resolution was applied to get anatomical images. Diffusion-weighted
(DW) images were acquired using a 3D diffusion-weighted spin-echo sequence (TE/TR
= 22 ms, 500 ms) at 600 μm3 voxel resolution with 6 diffusion encoded directions, a single
b-value of 1000 s/mm2 and three b0 image. This sample
preparation and acquisition is described with additional details in Magat et
al. [3].
A group-wise registration [4] of anatomical
and diffusion-weighted images was performed to generate anatomical and diffusion tensor templates (Fig. 1). Diffusion tensor metrics (eigenvalues,
eigenvectors, fractional anisotropy …) were computed [5] to provide a quantitative
and spatially-resolved analysis of cardiac microstructure. Then deterministic and probabilistic tractography [5]
was performed in order to : i) visualize the myofiber architecture, ii) segment sub-areas depicting the same fiber organization, iii) dilineate seed and tract
editing. Finally, dissection was performed to confirm the existence of
macroscopic structures identified in the diffusion tensor template (see [6]).Results
Fig. 2 shows the anatomical templates made with
different transformation models during the template creation.Misregistration
areas were more visible with rigid or affine transformations than with the symmetric
normalization (SyN) transformation model. Several
differences are evident through visual inspection: (i) a sharper definition of
the myocardial wall (Fig. 3A-F); (ii) an enhanced definition of the division of
the His bundle in left and right bundles (Fig. 3B); (iii) a more accurate
delineation of the leaflets of the tricuspid valve (Fig. 3B, 3E, 3F).
Fig.
3 shows color-coded FA (cFA) maps also known as Red-Green-Blue colormap maps
overlaid on anatomical data for the average diffusion tensor images. While a
smooth gradient of color from green to red (or vice versa) indicates a normal
arrangement of the cardiac myofibers, the existence of small sub-regions in the
LV indicates either a rapid change in orientation or apparent boundaries. In
particular: fiber arrangements in Inferior-Superior (IS) orientation are
visible: i) from the basal area to the mid-ventricular level close to the
papillary muscle, ii) at the intersection of the LV and RV from the basal area
to mid-ventricular level, iii) in the basal area of the IVS, divided into
two fiber-bundles that go on the
one hand to the pulmonary artery and on the other hand to the aorta.
To
further investigate the existence of abrupt changes in orientation, Fig.4 focuses
the analysis on the posterior wall. Fiber orientation can be visually
identified on the anatomical template images due to 150 µm resolution and the
T2* contrast (Fig 4A,B). Fiber orientation (Fig 4D,E) changes abruptly between
adjacent voxels in the IVS and in the middle of the LV/RV junction (blue)
depicting a triangular shape in the coronal view, denoted as the posterior
singularity.
Fig.
5 focuses the analysis on the anterior wall. We observe a division of the fibers
inside the IVS into the fiber-bundles going through
the aorta (yellow arrow) and the pulmonary artery (purple arrow). The third
fascicle of fibers (in pink), located close to the endocardium in the LV, is also
noticeable with an orientation differing from the surrounding structures.Discussion and conclusion
One
novelty of the proposed template were to cover both ventricles, the atria, and
a fraction of the arteries at an unprecedented spatial resolution of 600 µm
isotropic for the DTI acquisition. To assess the accuracy of the model, image-based
quantitative measurements and quality check on quantitative DTI metrics (see [6]) were done. To demonstrate the benefits of
the tractography to depict the cardiac fiber organization, the structural
organization of the anterior and posterior junction was characterized. The
study focuses on one fiber-bundle in the posterior
junction and three fiber-bundles in the
anterior junction. We found no reference and possible comparison in the
literature regarding the global myofiber organization in this region.
Nevertheless, the findings must be balanced for two differents reasons:
i) the fix state of the ex-vivo samples and ii) the low number of datasets.Acknowledgements
The authors are grateful for the help provided via the Github or Discourse
platform by Philip Cook and Nick Tustison regarding the use of ANTs and Max
Pietsh, Robert Smith and Donald Tournier regarding the use of MRtrix3 software.
The authors thank Girish Ramlugun for
proofreading
A large fraction of the datasets analyzed during the
current study has already been released and more are available upon reasonable
request. Data are available at this link (https://zenodo.org/record/5156088).
All the processing were fully automatic, reproducible and executed using shell scripts
without any input from the user with the exception of long axis alignement and ROIs
and seeds definition. The corresponding transformations, ROIs and seed have
been shared for reproducibility purposed.
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6 Julie Magat,
Maxime Yon, Yann Bihan-Poudec, Valéry Ozenne. A groupwise registration and tractography
framework for cardiac myofiber architecture description by diffusion MRI: an
application to the ventricular junctions. bioRxiv 2021.10.05.463112