Reza Rahmanzadeh1,2, Po-Jui Lu1,2, Muhamed Barakovic1,2, Riccardo Galbusera1,2, Matthias Weigel1,2,3, Pietro Maggi4, Thanh D. Nguyen 5, Simona Schiavi6, Francesco La Rosa 7,8, Daniel S. Reich9, Pascal Sati9, Yi Wang5, Meritxell Bach-Cuadra7,8, Ernst-Wilhelm Radue1, Jens Kuhle2, Ludwig Kappos2, and Cristina Granziera1,2
1Translational Imaging in Neurology (ThINk) Basel, Department of Medicine and Biomedical Engineering, University Hospital Basel and University of Basel, Basel, Switzerland, 2Neurologic Clinic and Policlinic, Departments of Medicine, Clinical Research and Biomedical Engineering, University Hospital Basel and University of Basel, Basel, Switzerland, 3Radiological Physics, Department of Radiology, University Hospital Basel, Basel, Switzerland, 4Department of Neurology, Lausanne University Hospital, Lausanne, Switzerland, 5Department of Radiology, Weill Cornell Medical College, New York, NY, United States, 6Department of Computer Science, University of Verona, Verona, Italy, 7Signal Processing Laboratory (LTSS), Ecole polytechnique fédérale de Lausanne (EPFL), Lausanne, Switzerland, 8Radiology Department, Center for Biomedical Imaging, Lausanne University and University Hospital, Lausanne, Switzerland, 9National Institute of Neurological Disorders and Stroke, Translational Neuroradiology Section, Division of Neuroimmunology and Neurovirology, Bethesda, MD, United States
Synopsis
The interplay between axonal and myelin damage in
multiple sclerosis (MS) is poorly understood. This study aimed to
evaluate the concomitant presence of axonal and myelin injury in
living MS patients by using myelin and multi-shell diffusion MRI.
Confirming neuropathological findings, our results show that (i)
axonal and myelin damage exists in MS lesions and spreads out from
the lesions in a centrifugal way, (ii) the extent of myelin
and axonal damage differs among lesion subtypes and according to
lesion anatomical locations and (iii) axonal (and not
myelin) damage differs between relapsing-remitting and progressive MS
patients.
Introduction
Multiple
sclerosis (MS) is an autoimmune disease of the central nervous system, which is
characterized by myelin and axonal damage1. Neuropathological
studies in postmortem brains showed axonal and myelin destruction in MS lesions
and in normal-appearing (NA) brain tissue as well as variations in the extent
of damage and repair among lesions at different locations2-6.
Nevertheless, only few studies have addressed the complex interplay between
myelin and axonal damage in living MS patients in small-sized
relapsing-remitting MS (RRMS) cohorts7. In the current work, we
have studied a large cohort of RRMS and progressive MS (PMS) patients and
performed a comprehensive assessment of axonal and myelin damage in focal
lesions and NA tissue. Specifically, we have evaluated axonal and myelin damage in:
(1) cortical/white
matter lesions (CLs, WMLs) and NA gray and white matter tissue (NAWM, NAGM);
(2) peri-plaque WM; (3)
lesions in different locations (periventricular vs juxtacortical) and lesions exhibiting ongoing chronic inflammation activity vs other lesions; (4) lesions in RRMS and PMS patients.METHODS
Sixty MS patients (41 RRMS and 19 PMS) and 35
healthy controls (HC) underwent multi-parametric magnetic resonance
imaging (MRI). MRI
was acquired in a 3T Prisma system (Siemens Healthcare, Germany)
using a 64-channel head and
neck coil.
The MRI protocol included:
(i) 3D FLAIR (TR/TE/TI=5000/386/1800
ms) and
MP2RAGE (TR/TI1/
TI2=5000/700/2500 ms)
with 1 mm3
isotropic
spatial resolution; (ii) Myelin Water Imaging (TR/TE/resolution= 7.5
ms/0.54 ms/0.93*0.93*5 mm3);
multi-shell diffusion (TR/TE/resolution= 4.5 s/75 ms/ 1.8 mm3
isotropic)
and 3D EPI
(TR/TE/resolution= 64 ms/ 35
ms/0.67*0.67*0.67 mm3).
Myelin
water fraction (MWF) and neurite density index (NDI) maps were
reconstructed as in 8,9. Paramagnetic rims were identified on
unwrapped phase images by 2 raters 10,11.
Lesions were automatically segmented 12 and manually corrected. CLs were manually
divided into intra-cortical and leuko-cortical lesions (ICLs and
LCLs). Peri-ventricular (PV) and juxta-cortical (JC) lesions were
defined as WM lesions located within 3mm
from the boundary between WM and GM and WM and ventricles,
respectively. We
also manually segmented the mirror NA contralateral region of 200
WMLs and evaluated the percentage of myelin (%MWF) and axonal density
(%NDI) reduction in WMLs.
MWF
and NDI were extracted in (i) WMLs, CLs; (ii) two consecutive 2-voxel
layers of peri-plaque WM (denoted as PP-1st and PP-2nd)
; (iii) NAWM and NAGM; (iv) WM and GM in HC (WM-HC and GM-HC) ; (v)
PV and JC lesions ; (vi) paramagnetic rims lesions and non-rim
lesions ; (vii) WMLs in RRMS and PMS patients.
Statistical
analysis was performed by using nonparametric Mann-Whitney test (for
two-group unpaired analyses) and Kruskal-Wallis test with Dunn’s
correction for multiple comparison (for more than two-group
analyses). RESULTS
We analyzed 1310 MS lesions (WMLs: 1106, CLs:
204).
In
WMLs, MWF and NDI were reduced compared to NAWM and WM-HC (p<
0.001) (Figure 1 A-D). The %MWF reduction in WMLs compared to the
mirror NAWM region was higher than the %NDI decrease (p< 0.0001).
As to cortical lesions, ICLs showed lower MWF and NDI compared to
NAGM (p< 0.01) whereas LCLs had lower NDI (p< 0.001) but equal
MWF to NAGM (Figure 1 E-G). In PP-1st and PP-2nd of WMLs, MWF and NDI were significantly
higher than in WMLs (p< 0.001) and lower than in WM-HC (p<0.001)
(Figure 2). In PV lesions, MWF and NDI were lower than in JC lesions
(p<0.0001) (Figure 3). In lesions with paramagnetic rims lesions,
both MWF and NDI were lower compared with other WM lesions without
paramagnetic rims (p<0.001) (Figure 4).
NDI
in lesions in PMS patients is lower than in RRMS (p<0.001) (Figure
5). However, the scatter plots (Figure 3&5) show that all WMLs
exhibit a proportional decrease in myelin and axonal content.DISCUSSION
Confirming neuropathological findings, our study
showed that WMLs and CLs exhibit significant myelin and axonal
injury compared to normal-appearing and healthy tissue. In WMLs,
myelin pathology outweighed axonal damage; in addition, diffuse
axonal pathology was present - though at a lower extent - in
peri-plaque NAWM. Adding to existing neuropathological knowledge, we
also found that axonal damage in ICLs was higher than LCLs, which may
indicate involvement of different pathogenic processes. Myelin
pathology also differed according to lesion location and lesion type:
PV lesions exhibited more axonal and myelin damage compared with JC
lesions, as expected from previous neuropathological2-5 and PET-studies6;
and paramagnetic rim lesions – which have been shown postmortem to
exhibit extensive axonal damage in the center and ongoing damage in
the surrounding tissue13 - exhibit as well more myelin and axonal reduction than lesions
without rim. Our data also provide new knowledge about myelin
and axonal damage in RRMS and PMS patients: axonal content in WMLs is
lower in PMS than in RRMS whereas myelin damage is similar. This may
well support the hypothesis that other mechanisms than demyelination
lead to axonal damage in PMS14,15.CONCLUSION
Confirming neuropathological findings, our results
in living patients show that (i) axonal and myelin damage exists in
cortical and WM MS lesions as well as in peri-plaque WM and that (ii)
the extent of myelin and axonal damage differs among lesion subtypes
and between PVLs and JCLs. We have also provided new evidence that
axonal (and not myelin) damage differs between RRMS and PMS.Acknowledgements
This
work was funded by the Swiss National Funds PZ00P3_154508,
PZ00P3_131914 and PP00P3_176984
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