A selective detection of lysophosphatidylcholine in dried blood spots for diagnosis of adrenoleukodystrophy by LC-MS/MS

Abstract

X-linked adrenoleukodystrophy (X-ALD) is a rare inherited metabolic disorder characterized by an impaired beta-oxidation of very long chain fatty acids in the peroxisomes. Recent studies have suggested that 1-hexacosanoyl-2-hydroxy-sn-glycero-3-phosphocholine (Lyso-PC 26:0) can be a sensitive biomarker for X-ALD. Although approximately 10-fold increase in the concentration of Lyso-PC 26:0 in DBSs from X-ALD-affected individuals were reported, whether the carriers might be distinguished from the healthy controls remained unclear. To address this question, we have validated previously developed LC-MS/MS-based analytical procedures using QC DBS. We found that the recovery of Lyso-PC 26:0 from the QC DBSs was 73.6 ± 0.3% when 2 μM of Lyso-PC 26:0 was spiked into the blood. Based on this result, the amounts of Lyso-PC 26:0 in the controls and ALD-affected individuals were 0.090 ± 0.004 (n = 11) and 1.078 ± 0.217 (n = 4) pmol/DBS, respectively. Interestingly, the concentration of Lyso-PC 26:0 in the carriers were 0.548 ± 0.095 pmol/DBS (n = 3), indicating that the carriers and the healthy controls can be distinguished. These results suggest that LC-MS/MS-based technique can be used for the detection of asymptomatic carriers and X-ALD-affected subjects in the newborn screening.

Application of a diagnostic methodology by quantification of 26:0 lysophosphatidylcholine in dried blood spots for Japanese newborn screening of X-linked adrenoleukodystrophy

“…1) in keeping with previous studies on LysoPCs [13], [14], [17]. The heterogynous female (red dot) in our study did not present obvious differentiation of newborns but did differentiate healthy adults (Fig.…”

“…As new technologies for high throughput screening of X-ALD become feasible [13] , [14] , patients can be diagnosed even at birth and treated at a very early stage. Studies on quantification of lysophosphatidylcholines (LysoPCs) in dried blood spots (DBS) for newborn screening have presented satisfactory results, including discrimination of heterogynous females [13] , [14] , [15] , [16] , [17] . Newborn screening of X-ALD has not been implemented in Japan to date.…”

Comprehensive Evaluation of the NeoBase 2 Non-derivatized MSMS Assay and Exploration of Analytes With Significantly Different Concentrations Between Term and Preterm Neonates

“…The diagnosis of ADA-SCID can be supported by lymphopenia and low Ig concentrations [ 36 ]. The C20:0-LPC concentration does not significantly differ between healthy controls and ALD [ 37 , 38 ], and increases in C24:0-LPC, C26:0-LPC, and the ratio of these markers to C20:0-LPC or C22:0-LPC are used for diagnosis [ 37 - 39 ].…”

Newborn Screening for X-Linked Adrenoleukodystrophy: Past, Present, and Future

“…Newborn screening for X-linked adrenoleukodystrophy (ALD) was added to the Recommended Uniform Screening Panel in February 2016. The implementation of this screening continues to expand in the United States and is being considered in other countries [1][2][3][4][5][6][7][8][9]. It is an opportune time to evaluate the development, performance, and the future of the endeavor.…”