Targeted Multiple Reaction Monitoring Analysis of CSF Identifies UCHL1 and GPNMB as Candidate Biomarkers for ALS

Abstract

The neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and Parkinson’s disease (PD) share some common molecular deficits including disruption of protein homeostasis leading to disease-specific protein aggregation. While insoluble protein aggregates are the defining pathological confirmation of diagnosis, patient stratification based on early molecular etiologies may identify distinct subgroups within a clinical diagnosis that would respond differently in therapeutic development programs. We are developing targeted multiple reaction monitoring (MRM) mass spectrometry methods to rigorously quantify CSF proteins from known disease genes involved in lysosomal, ubiquitin-proteasomal, and autophagy pathways. Analysis of CSF from 21 PD, 21 ALS, and 25 control patients, rigorously matched for gender, age, and age of sample, revealed significant changes in peptide levels between PD, ALS, and control. In patients with PD, levels of two peptides for chromogranin B (CHGB, secretogranin 1) were significantly reduced. In CSF of patients with ALS, levels of two peptides from ubiquitin carboxy-terminal hydrolase like protein 1 (UCHL1) and one peptide each for glycoprotein non-metastatic melanoma protein B (GPNMB) and cathepsin D (CTSD) were all increased. Analysis of patients with ALS separated into two groups based on length of survival after CSF sampling revealed that the increases in GPNMB and UCHL1 were specific for short-lived ALS patients. While analysis of additional cohorts is required to validate these candidate biomarkers, this study suggests methods for stratification of ALS patients for clinical trials and identifies targets for drug efficacy measurements during therapeutic development.Electronic supplementary materialThe online version of this article (10.1007/s12031-019-01411-y) contains supplementary material, which is available to authorized users.

UCHL1 from serum and CSF is a candidate biomarker for amyotrophic lateral sclerosis

“…Our results indicated that CSF UCHL1 is a promising biomarker for ALS diagnosis, consistent with some previous studies 14,16 . Specifically, we found significantly elevated CSF UCHL1 in both discovery and validation cohorts.…”

“…Summarily, we selected a panel of candidate neurodegenerative biomarkers, based on our previous research and other published studies. These included Glial fibrillary acidic protein (GFAP), 8,9 DJ1, 10 S100B, 9,11 Amyloid beta 40 (Abeta 40), Amyloid beta 42 (Abeta 42), 9 Transglutaminase 2 (TGM2), 12 α‐synulein, 13 Ubiquitin C‐terminal hydrolase L1 (UCHL1), 14 and Neuron specific enolase (NSE) 15 . We recruited a discovery cohort, comprising 20 ALS patients and 20 patients with other neurological diseases, and screened it for possible biomarkers using the Luminex platform.…”

Discovery of Biomarkers for Amyotrophic Lateral Sclerosis and Frontotemporal Lobar Degeneration From Human Cerebrospinal Fluid Using Mass Spectrometry-Based Proteomics

“…UCHL1 is a ubiquitin-protein hydrolase important for protein homeostasis that is expressed in neurons and neuroendocrine cells, whereas APOB is associated with metabolic changes in ALS [30,35]. Consistent with previous studies, we found that all of these candidate proteins were elevated in CSF samples from patients with ALS [17,22,23,[36][37][38][39]. This observation independently strengthens the potential of these proteins as reliable biomarkers for ALS, and importantly, validates our approach as a method to identify novel candidate biomarkers.…”

Characterization of Fluid Biomarkers Reveals Lysosome Dysfunction and Neurodegeneration in Neuronopathic MPS II Patients

“…In addition to the classical fluid-based biomarkers of neurodegeneration such as Aβ 42 and Tau, markers of neuronal health and astrogliosis such as Ubiquitin C-Terminal hydroxylase (UCH-L1) and glial fibrillary acidic protein (GFAP) are also seen to be increased in neurodegenerative disorders such as Alzheimer’s disease, ALS, and frontotemporal dementia [ 45 , 46 , 47 , 48 ]. Our data show that a subset of these markers is also elevated in CSF of nMPS II patients relative to the control group.…”