Decomposition of Multiexponential Relaxation for Myelin Water Imaging and More

Eric Y. Zhao March 25, 2013

Abstract Magnetic resonance imaging provides high resolution radiological images with a variety of possible contrasts. Among these are modes of contrast calculated from the bulk of sampled information, such as from the time course of T1 and T2 decay processes. Mathematical methods have been developed for the approximation of multiple exponential relaxation schemes into weighted components, each of which represents nuclei of water molecules in dierent cellular environments. In particular, the signal fraction of short T2 relaxation has been found to relate to localization and quantication of water trapped between myelin membranes. This paper overlays key concepts important in the development of these methods: the theory of multiexponential analysis, the implementation and ndings in myelin water experiments, and the validation of these ndings in model systems.

Contents
1 A Conceptual Overview of Myelin Water Imaging 1

2 Methods of Multiexponential T2 Component Analysis and Myelin Water Quantitation 2.1 2.2 2.3 Non-negative Least Squares . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Quantication and Visualization of Myelin Water Fraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Filter-based Myelin Imaging by Linear Combination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

3 3 6 8

3 Histopathology Studies of Myelin Water Imaging

4 Research Applications of Myelin Water Imaging

5 Conclusion

11

A Conceptual Overview of Myelin Water Imaging

Magnetic resonance imaging (MRI) is a versatile and information rich imaging modality capable of delivering excellent diagnostic yield. Its means of signal detection is based on nuclear magnetic resonance (NMR), and relies largely on the magnetization of localized nuclear spins within the imaged object. One strength of MRI is its ability to provide a wide variety of dierent contrast modes for example proton density, T1 , and T2 weighted contrasts by varying simple imaging parameters (Wehrli et al., 1984). Contrast enhancing agents such as gadolinium and manganese can be used to achieve additional contrast between specic tissue or organ types. These techniques can reveal dierent properties of tissue and its aqueous and biochemical components. Image maps constructed from scan data can provide sources of contrast not attainable by conventional MRI. A map is matrix of calculated values corresponding to spatial regions of the imaged object. For example, in magnetic resonance spectroscopy (MRS), each voxel contains information about the NMR spectrum of the contents localized in its volume. Within this spectrum, dierent chemical species fall under peaks at dierent chemical shifts. Obtaining the area under these peaks allows for relative voxelwise quantication of these chemical species. For example, studies have used MRS to visualize and quantify myo-inositol in brain regions, which has been linked to gliosis (Ross et al., 1998). Like in MRS, inversion recovery (IR) and spin echo (SE) pulse sequences can also generate spectra, but in temporal rather than in frequency space. IR sequences involve an initial 180 inversion pulse followed by a 90 stimulating pulse occuring with varying latency and thereby allowing the magnetization vector to relax to dierent levels by T1 relaxation. IR signal amplitudes can therefore be used to measure the time course of spin-lattice (T1 ) relaxation (Rosen et al., 1984). SE sequences contain deliberately timed 180 pulses which induce local spin rephasing, producing a train of echoes. The time course of these echoes is largely unaected by the eects of magnetic eld inhomogeneity (T2 eects), and can thus be used to approximate T2 . T1 and T2 relaxation processes can be modeled as multiexponential functions with a spectrum of time constants (Figure 1). Methods of mathematical analysis have been developed which can resolve components of decay and compare their relative contributions to the aggregate function. The result is a set of signal fractions of selected T1 and T2 domains. These signal fractions relate to nuclei within captured voxels which exhibit dierent relaxation characteristics. The T2 times of nonaqueous cellular structures are on the order of microseconds. Aqueous components, however, have T2 components over 10 ms, and are therefore detectable via MRI. Vasilescu et al. (1978) proposed that dierent observable T2 components arise from water molecules in dierent microscopic environments. It has been suggested that the signal fraction of short T2 relaxation arises from water trapped between membranes of the myelin sheath (Vasilescu et al., 1978; Mackay et al., 1994). Supporting evidence by Whittall et al. (1997) found that the signal fraction of T2 relaxation between 10 ms and 50 ms was signicantly higher in white matter structures than in gray matter structures. Validation of these ndings was later reported by correlation of this signal fraction with the localization (Moore et al., 2000) and density (Laule et al., 2006, 2008) of myelin in histopathology studies. This paper will outline the theory and ndings involved in the decomposition of multiexponential relaxation processes and their application to myelin water imaging. It will also discuss the evidence supporting myelin water imaging techniques and their current and potential applications.

Figure 1: Multiexponential decay curves are linear combinations of monoexponential functions. Spin-lattice and spin-spin relaxation follow monoexponential decay time courses in homogeneous samples. However, in non-homogeneous environments, such as those on cellular scales, relaxation progresses by a multi-exponential decay time course, seen as a non-linear trend on a logarithmic scale. This decay trend can be described by a linear combination of monoexponential functions, here shown as 1, 2, and 3. Figure adapted from Vasilescu et al. (1978)

Methods of Multiexponential T2 Component Analysis and Myelin Water Quantitation

The development of techniques to image myelin began with theoretical innovations in decomposing multiple exponential decay functions. Here, we describe the popular and robust non-negative least squares estimation approach, as described by Whittall and MacKay (1989). We will then outline how this technique was applied to the imaging of myelin water. We will also briey describe a method outlined by Vidarsson et al. (2005) which suppresses the signal from tissue exhibiting selected T2 relaxation time components, providing a direct and more rapid approach to myelin water imaging. 2.1 Non-negative Least Squares

Accurate determination of components in multiexponential decay is an area of active mathematical research. Central to this analysis is the transform between time space, t, and relaxation time space, T , which in MRI represents either T1 or T2 . This transforms between a multiexponential decay function, y (t), and its relaxation time spectrum, s(T ). A pulse sequence which samples amplitudes of y (t) discretely serves to obtain N measurements at times ti , where i = 1, 2, . . . , N . This gives rise to expected values of y (ti ) following the general equation
b

y (ti ) = yi =
a

s(T )eti /T dT, i = 1, 2, . . . , N,

which represents a summation of exponential functions possessing a continuous spectrum of relaxation times, T , each of which is weighed by its corresponding spectral value, s(T ). The typical shape of a multiple exponential decay curve is visualized in Figure 2A on a logarithmic scale. The general case can also be represented by the linear equation in Hilbert space y = As, (1)

where Aij is the exponential factor eti /Tj . Note that y arises from measurement, but s is unknown. This problem can thus be classied as an inverse problem. This distribution can be discretized by means of a spectrum of delta functions, each weighed by a corresponding sj such that the resulting spectrum preserves the total area under the curve. That is,
M

s(T ) =
j =1

sj (T Tj ),

which gives rise to a set of exponentials characterized by

b M

yi =
a M

j =1

sj (T Tj ) eti /T dT
b

=
j =1 M

sj
a

(T Tj )eti /T dT

=
j =1

sj eti /Tj . i = 1, 2, . . . , N.

Figure 2: Computational determination of multiple exponential decay components in known decay schemes with noise. A. Multiple exponential curves were generated from a nite number of selected monoexponential functions (left) and monoexponential functions with a continuous range of time constants (right). B. To each, random normally distributed zero-mean noise was introduced and a non-negative least squares method was used to deduce a discrete set of exponential functions which minimized the mist function. C. First order derivative constraints were added using a nite-dierence approximation to generate a smooth spectral distribution. Figure adapted from Whittall and MacKay (1989).

As expected, this simply gives us a discrete sum of single exponential terms, each with a dierent value of T . This is the equivalent of Equation 1 in the truncated space RN . Conventional approaches approximate decay curves using the smallest possible discrete set of exponentials (with sj > 0) which provide a satisfactory t. However, this approach is limited when applied to measured data with three or more nonzero components, and often fails to eectively resolve distinct exponential components with similar time constants. Indeed, the true spectrum of relaxation times is better represented by a set of exponential functions lying on a continuous domain of decay times, giving rise to a continuous spectrum, s(T ). In addition, it is necessary to utilize a method robust against random noise in the sampled data. Whittall and MacKay (1989) proposed a non-negative least squares (NNLS) approach to solving this inverse problem. This algorithm aims to minimize the least squares mist
N M 2

D=
i=1 j =1

Aij sj yi ,

or equivalently D = |As y|2 = |(As)1 y1 |2 + |(As)2 y2 |2 + . . . + |(As)N yN |2 . Here, s is selected to minimize the mist. As then gives a vector of theoretical values, yp , that best predicts the vector of measurements, y. In practice, the rows of A and the values of y are normalized by the standard deviation, i , of the corresponding measurement. This results in the expression D=
p N 2 i=1 (yy yi ) , 2 i

which is equivalent to the 2 statistic. Therefore, the proposed NNLS method aims to determine the spectrum vector s which, when transformed by A, provides a vector of predicted values, yp , which best minimizes the 2 statistic with the measured vector y. Figure 2B illustrates examples of best t linear combinations of monoexponential functions represented in the form of delta functions along the T -domain. 2 characterizes the noise of the measurement against the variance. For N measurements, 2 N represents a good t. If 2 >> N , then the determined t is not close enough and no adequate s(T ) can be determined. If 2 << N , then the t is too close, indicating that the prediction aligns with data produced as a consequence of noise and not signal alone, causing an artifact. This method, however, cannot yet generate a continuous spectrum for s(T ). To do this, additional constraints must be applied to the mist which must be minimized. The general form of additional constraints is D = |As y|2 + |Hs f |2 where A is an N M matrix and H is a K M matrix. Minimization requires that sj is found in a way that satises both constraints, whose relative weight is provided by the scaling factor . When = 0, the additional constraints disappear and we converge on the original solution. 5

H can be set so as to minimize a desired quality of s. For example, if H = I (the unit matrix) and f = 0 (the zero vector), then the right side contraint minimizes |sj |2 , or the energy of the spectrum. Alternatively, H can be set to nite dierence approximations, which numerically approximate the derivatives of a given vector. Setting H to the nite dierence approximation of the second derivative,
M 2

|sj +2 2sj +1 + sj |2 ,
j =1

minimizes the energy of the curviture of sj . This constraint enforces a smoothing condition on s(T ). Increasing broadens the resulting peaks of spectrum while lowering the peak amplitudes. Figure 2C is an example of a best t continuous smoothed spectrum. (Whittall and MacKay, 1989) also described additional approaches utilizing linear programming. These techniques are beyond the scope of this paper, as the methods discussed here cover the fundamental concepts for disambiguating the components of dierent water compartments to T1 and T2 relaxation. This work was instrumental to the ability to quantify myelin water and water in other compartments as it proposed a robust method of analyzing multiexponential relaxation phenomena. 2.2 Quantication and Visualization of Myelin Water Fraction

Previous experiments employing NMR of excised animal nervous tissue have detected multiple components of spin-spin relaxation within myelinated bres and ascribed the components to water in dierent environments. Vasilescu et al. (1978) found that T2 decay could be approximated by three primary exponential components in frog (Rana temporaria ) sciatic nerve tissue. Of these, the longest T2 portion, comprising 21% of total water, was attributed to extracellular water as it most closely resembles T2 decay in dilute aqueous solution. The shortest T2 component was associated with characteristics of water bound to proteins and phospholipids and comprised 29% of total water. The remaining 50% was attributed to intracellular axoplasmic water. The association of water in the short T2 component with proteins and phospholipids suggests that this signal arises from water lying along cellular borders. It was thus hypothesized that this signal arises from water trapped tightly between myelin sheaths. Mackay et al. (1994) devised a method for visualizing this fraction of water in vivo. Their method employed a single-slice 32 echo pulse sequence in a 1.5 T clinical scanner. NNLS was used to isolate components of T2 decay as discrete delta functions. The use of smooth T2 distributions gave similar results. Three T2 components were identied, with the component representing water in myelin membranes between 10 ms and 55 ms. The authors suggested that the variability primarily arises from the diculty in accurately determining T2 component times, not from inherent variability of T2 in myelin water. Myelin water fraction (MWF) was calculated as the area under the curve in the myelin water T2 range divided by the total T2 distribution. Myelin water maps were generated where pixel intensity was determined by MWF on a logarithmic scale to provide better contrast in the appropriate range. MWF was found to be signicantly higher in white matter (15.6 8.1%) than in gray matter (3.8 6.4%). These values agreed with myelin water values determined in vitro. Additionally, 6

Figure 3: T2 components and myelin water fraction determined in various brain regions. T2 decay curves were generated from 32-echo MRI scans in vivo and decomposed into a spectrum of monoexponential components by a NNLS method. Regions of interest selected for various brain areas and the spectra was determined for each (C). Area under the T2 spectrum was used to determine water content (A) and the fraction of area under the short T2 component was used to determine myelin water fraction (B). Figure adapted from Whittall et al. (1997). MWF of lesions in four MS patients fell between 4.1% and 6.4%, signicantly lower than MWF in white matter. Additional work performed by Whittall et al. (1997) in characterizing myelin imaging in brain regions found higher MWF in all white matter regions than in gray matter regions. The highest MWF was found in the internal capsules and splenium of the corpus callosum, and lowest MWF in the cingulate gyrus, insular cortex, and cortical gray matter (Figure 3). This study employed an algorithm which determined a continuous spectrum of T2 times, and could thus assess the width of T2 peaks. Although much of the dierence in distribution width arises through the mathematical process of calculating the least squares mist, it may still be sensitive to inhomogeneity in the cellular environment. Such inhomogeneity was found in the long T2 component in the major forceps, posterior internal capsules, and splenium of the corpus callosum, regions with relatively low total water content. A strong positive linear relationship between long T2 fraction peak width and MWF suggested that myelin may play a dominant role in inhomogeneity at a cellular level. These studies demonstrate that the quantication and visualization of MWF is possible in vivo and that it can provide the potential to monitor the myelination state of white matter and MS lesions. In addition, the study of water components by T2 decomposition can aid in characterizing the microscopic anatomy of the central nervous system.

2.3

Filter-based Myelin Imaging by Linear Combination

Although NNLS analysis of spin-echo data oers the most robust measure of T1 and T2 signal fractions with respect to B0 inhomogeneity, it requires a long scan time and is computationally expensive. This approach acquires more data than is required for the basic acquisition of MWF. Vidarsson et al. (2005) describe a 3-echo sequence which enables selective suppression of structures with low signal fraction of short T2 components. They conceptualize the imaging process as a lter design problem, in which images comprising a linear combination of magnitude data are accordingly weighed. To facilitate consistent discussion of mathematics, the theory from Vidarsson et al. (2005) has been adapted to suit notation used in earlier sections of this paper. Multiexponential decay can be described as a general phenomenon in the form y = As + Noise, where y, A, and s represent the same quantities as earlier described. Approximation of s involves the construction of a pseudo-inverse matrix A via least squares estimation such that s(e) = A y, where s(e) denotes an estimate of the signal vector. The short fraction, sshort , is found by summing the short-T2 spectrum as in sshort = [1 . . . 1 0 . . . 0]s(e) . = aT y , where aT = [1 . . . 1 0 . . . 0]A . a is therefore a vector of the weights of dierent measurements, and can be determined by optimization, which avoids calculation of s(e) . Based on user-specied inputs to dene the degree to which longer T2 signal fractions are to be suppressed, an algorithm can optimize a to maximize the signal to noise ratio for the MWF. This optimization is solved in the form of a second order cone program, and determines the necessary scan parameters. This approach sacrices robustness against noise for faster scan and reconstruction times.
(e) (e)

Histopathology Studies of Myelin Water Imaging

The validation of myelin water imaging is possible by studying samples of formalin xed brains. It has been demonstrated that MRI of formalin xed brains is highly representative of in vivo scanning (Nagara et al., 1987). In addition, T2 relaxation components obtained from these brains are preserved with a slight shift towards faster relaxation times. This motivated studies to elucidate the relationship between the presence of myelin in histological samples and the localization and intensity of the short T2 signal fraction. Moore et al. (2000) performed 32-echo MRI scans on two formalin xed brains with a 5 mm resolution in the slice selective direction. Each brain was then sectioned along the centres of the imaged slices and each section was photographed, dehydrated, and embedded with paran. Luxol fast blue (LFB) is a copper phthalocyanine dye which has demonstrated specic binding to lipoproteins of the myelin sheath. It has been found to be useful in the study of cell membrane 8

structures due to its adherence to phospholipids and is a common dye used for the staining and visualization of myelin in central nervous system histology. Moore et al. (2000) stained paran embedded formalin xed slices using LFB by the Bielschowsky technique. The distribution and amplitude of dye were compared qualitatively with the amplitude of the short T2 fraction detected by the 32-echo MRI scan. They found that myelin distribution corresponded with areas of increased MWF. In addition, areas found to have demyelinated plaques as a result of MS lack the short T2 relaxation component. Further work pursued the quantitative investigation of MWF correlations with histological myelin density. Laule et al. (2006) performed a similar protocol of formalin xing and sectioning brain samples from 13 MS subjects. Brain sections stained with LFB were scanned for the optical density of dye. These digital histopathy images were registered to MRI images that involved manual placement of 10 matching anchor points on the two images followed by computational analysis. Regions of interest were selected in the resulting registered images and LFB optical density and MWF were measured. MWF was found to correlate highly with LFB optical density (R2 0.67). In addition, MWF and LFB optical density values in both lesion and gray matter tissue were found to be clearly lower than those detected in white matter (Figure 4). This work was repeated at 7 T revealing an increased mean R2 value of 0.78, which was attributed to better attribution of MRI values with histopathology values due to increased SNR and better comparative specicity due to decreased slice-selected thickness (Laule et al., 2008).

Research Applications of Myelin Water Imaging

Longitudinal study of MS has found that MWF can be used to monitor the myelination state of lesions over time (Vavasour et al., 2009). Although nuclear relaxation measures have demonstrated short term (6 month) stability in relapsing remitting MS normal appearing white matter, it is hypothesized that they vary over the long term (> 2 years) given the characteristics of pathological progression (Liang et al., 2012). Based on these ndings, quantication of MWF may serve as a biomarker of subclinical MS development and aid in its diagnosis and prognosis. Additionally, the use of myelin water imaging may be benecial in assessing pharmacological treatments of MS. For example, glatiramer acetate (GA) is a copolymer which has had demonstrated neuroprotective eects both in relieving inammation and in facilitating remyelination (Aharoni, 2012). Current research in progress at the UBC MRI Research Centre involves the use of transcranial magnetic stimulation in combination with myelin water imaging to establish neurophysiological correlates of increased or decreased MWF. Early work in this area has shown a negative correlation between the degree of transcallosal inhibition, an evoked phenomenon dependent on intercortical bres, with MWF in select regions of the posterior corpus callosum. Characterizing this relationship may be useful in order to provide bimodal assessment of functional decline or recovery when using treatment options such as GA. Relaxation based imaging has also been applied to the study of other diseases such as Alzheimers disease. Increased T2 decay found in Alzheimers patients has been attributed to a decrease in local iron concentrations as a consequence of oligodendrocyte and Schwann cell loss. Another area is in the study of phenylketonuria (PKU), a congenital disease characterized by decient phenylalanine metabolism. One of its common symptoms is severe mental retardation due to the excessive buildup 9

d
Figure 4: Qualitative and quantitative correlations of myelin water imaging with histopathology. 32-echo structural MRI was acquired in formalin xed brains from 13 MS subjects (a) and was subject to NNLS analysis to decompose the multiple exponential components of the T2 decay curves. The signal fraction of short T2 decay was quantied and used to construct a myelin water map (b). The brain samples were sectioned and paran embedded along the centres of the slices acquired by MRI and stained with Luxol fast blue to visualize myelin (c). Myelin visualized by histopathology was found to correlate both qualitatively and quantitatively (d) with myelin water fraction, and was denser in white matter than in gray matter and lesions. Figure adapted from Laule et al. (2006).

10

of phenylalanine in the brain which results in progressive brain damage. MWF in the NAWM of PKU patients was signicantly reduced by up to 56% when compared to normal control.

Conclusion

Techniques for the decomposition of multiexponential decay curves have enabled new modes of contrast in MRI unattainable by conventional methods. The development of these techniques has progressed through theory, practice, and validation. Fractional contributions to T1 and T2 relaxation have been shown to represent protons of water molecules in dierent environments. In particular, the short T2 signal fraction correlates with myelin composition in a qualitative and quantitative fashion. Myelin plays a pivotal role in the coordinated conduction of nerve signals in the central and peripheral nervous systems. As such, methods capable of in vivo assessment of myelination promise to deliver diagnostic and therapeutic benets to patients with neurological disease. They may also oer future capabilities in elucidating functions of myelin and its role in the central nervous system, particularly in cases of pathogenesis. All considered, myelin water imaging can be considered a past and future success of MRI, highlighting the strengths of this imaging modality. It harnesses the multimodal and quantitative natures of MRI to provide direct information about biological structure and function.

11

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