University of Colorado
Neurology

Author Of 2 Presentations

Biomarkers and Bioinformatics Poster Presentation

P0061 - Determining the effect of blood anticoagulants on the detection level of neurobiomarkers in headache and control patients on the SIMOA platform (ID 1833)

Speakers
Presentation Number
P0061
Presentation Topic
Biomarkers and Bioinformatics

Abstract

Background

In the development of the sensitive single molecule array (SIMOA) technology, serum and blood plasma from EDTA collection tubes were used to verify and validate blood levels of NFL, GFAP, Tau and UCHL1 across healthy controls[1]. As the need arises to establish extensive baseline levels of these neurobiomarkers in order to better evaluate neurodegeneration in a wide variety of patients including multiple sclerosis (MS) patients, a validation study across a variety of blood collection anticoagulants is necessary to ensure there are no significant differences in blood levels due to the additives themselves.

Objectives

To determine whether SIMOA results are reliable and comparable within patients and between cohorts, we measured blood levels of the neurobiomarkers neurofilament (NFL), glial fibrillary acidic protein (GFAP), tau and ubiquitin C-terminal hydrolase L1 (UCHL1) in samples collected with different blood collection additives.

Methods

Serum and blood plasma were collected voluntarily from either headache or healthy control patients in one of four different blood collection tubes representing the 4 most common types of anticoagulant: none (serum), EDTA, sodium heparin (hep) and sodium citrate (NaC). Plasma and serum were isolated from whole blood by centrifugation at 1500 x g for 20 minutes. Plasma was centrifuged at 400 x g for 10 minutes to further deplete cells. Biomarker levels of NFL, GFAP, Tau and UCHL1 were measured via SIMOA with the Neuro4Plex A Advantage kit in the Quanterix SR-X machine according to manufacturer instructions. Statistical comparisons were made using GraphPad Prism analysis software.

Results

No statistically significant differences in blood concentrations were found between the anticoagulants for the NFL, GFAP or UCHL1 biomarkers in either the headache patients or the healthy control patients. However, Tau levels were significantly lower in all serum samples (p value <0.0001) from both patient cohorts with average levels 65-80% lower than plasma.

Conclusions

Use of serum should be avoided when establishing baseline blood levels of the neurobiomarker Tau on the SIMOA platform.

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Biomarkers and Bioinformatics Poster Presentation

P0111 - Monitoring Multiple Sclerosis Treatment with Plasma Biomarkers: NfL, GFAP, UCH-L1, and Tau (ID 909)

Speakers
Presentation Number
P0111
Presentation Topic
Biomarkers and Bioinformatics

Abstract

Background

Blood neurofilament light (NfL) levels have been linked to multiple sclerosis (MS) activity and progression but are affected by factors such as age and body mass index (BMI). Less is known about what factors affect blood levels of Glial Fibrillary Acid Protein (GFAP), Ubiquitin Carboxy-Terminal Hydrolase L1 (UCH-L1), and Tau. Single Molecule Array (SIMOA) platforms allow multiplex measurement of these biomarkers with high sensitivity.

Objectives

To evaluate factors associated with higher levels of four plasma biomarkers—NfL, GFAP, UCH-L1, and Tau—in individuals with MS on immunotherapy.

Methods

Subjects with MS between 18-65 years taking dimethyl fumarate (n=40), fingolimod (n=37), natalizumab (n=47), or rituximab (n=90) for at least 1 year were identified from Rocky Mountain MS Center Biorepository. Neuro 4-plex A plasma assays were conducted on the Quanterix SR-X SIMOA platform. Biomarker concentrations were log transformed. For each biomarker, summary statistics were generated, and logistic regressions on the probability of having a level in the top quartile, adjusting for age, were performed with different explanatory variables including gender, BMI, disease duration, length on disease modifying therapy (DMT), DMT type, and MS subtype (relapsing MS [RMS] or progressive MS [PMS]). All statistics were generated in SAS.

Results

Included were 214 subjects (194 RMS, 20 PMS; 70.3% female; 86.9% Caucasian) with mean age of 44.1(SD 9.8). Mean disease and treatment durations were 150.7(SD 95.7) and 49.7(SD 33.5) months, respectively. Means (IQR) for NfL, GFAP, UCH-L1 and Tau were 6.6(3.9-7.1), 66.9(45.5-81.4), 11.5(7.1-13.8), and 1.2(0.8-1.5) pg/ml, respectively. NfL, GFAP, and UCH-L1 increased with age. (Remainder of results are given age-adjusted.) PMS was more likely than RMS to be in the top quartile for NfL (OR 3.5,p=0.01), GFAP (OR 2.6,p=0.06), and UCH-L1 (OR 2.8,p=0.04). Longer disease duration (5 years) increased the likelihood of elevated NfL (OR 1.3,p=0.02) and elevated GFAP (OR 1.3,p=0.03). Higher BMI (5 units) decreased the likelihood of elevated NfL (OR 0.6,p=0.0007) and GFAP (OR 0.8,p=0.03) but increased the likelihood of having an elevated Tau (OR 1.4,p=0.002). Ethnicity and treatment duration had no effect, but men were more likely to have elevated UCH-L1 (OR 2.1,p=0.03) and lower Tau (OR 0.2,p=0.0004). Comparing DMTs, no biomarker differences were observed except subjects on rituximab and dimethyl fumarate were less likely to have elevated Tau.

Conclusions

Plasma NfL, GFAP, and UCH-L1 are promising biomarkers to differentiate relapsing from progressive MS. Age and BMI should be incorporated into biomarker models to determine normal thresholds. No differences were observed between treatments for NFL, GFAP, or UCH-L1, but subjects on dimethyl fumarate and rituximab were less likely to have elevated Tau. Lack of randomization or repeated measures limited comparative effectiveness analyses.

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