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Differences in protein levels between amyotrophic lateral sclerosis (ALS) patients and controls. Three antibodies targeting NEFM, RGS18, and SLC25A20 were found to reproducibly show significant differences between ALS patients (n = 367) and controls (n = 101) (P-values are shown below respective plot). NEFM, neurofilament medium polypeptide; RGS18, regulator of G-protein signaling 18; SLC25A20, solute carrier family 25 (carnitine/acylcarnitine translocase).
Source publication
Objective
Amyotrophic lateral sclerosis (ALS) is the most common adult motor neuron disease leading to muscular paralysis and death within 3–5 years from onset. Currently, there are no reliable and sensitive markers able to substantially shorten the diagnosis delay. The objective of the study was to analyze a large number of proteins in plasma from...
Similar publications
Objective: Amyotrophic lateral sclerosis (ALS) is the most common adult
motor neuron disease leading to muscular paralysis and death within 3–5 years
from onset. Currently, there are no reliable and sensitive markers able to substantially
shorten the diagnosis delay. The objective of the study was to analyze
a large number of proteins in plasma fro...
Citations
... It has been demonstrated that NEM, also known as NEFM, is linked to regulatory functions in dopaminergic neurotransmission (Kim et al., 2002) and is associated with the immune response (Barboni et al., 2014;Li et al., 2021). Increased levels of NEFM have been detected in various neurological diseases, such as brain damage (Martínez-Morillo et al., 2015), schizophrenia spectrum disorders (Runge et al., 2022), and amyotrophic lateral sclerosis (Häggmark et al., 2014). However, the association of NEFM with PD has not been reported previously and requires further investigation. ...
Background
Parkinson’s disease (PD) is Pengfei Zhang Liwen Zhao Pengfei Zhang Liwen Zhao a common neurological disorder involving a complex relationship with immune infiltration. Therefore, we aimed to explore PD immune infiltration patterns and identify novel immune-related diagnostic biomarkers.
Materials and methods
Three substantia nigra expression microarray datasets were integrated with elimination of batch effects. Differentially expressed genes (DEGs) were screened using the “limma” package, and functional enrichment was analyzed. Weighted gene co-expression network analysis (WGCNA) was performed to explore the key module most significantly associated with PD; the intersection of DEGs and the key module in WGCNA were considered common genes (CGs). The CG protein–protein interaction (PPI) network was constructed to identify candidate hub genes by cytoscape. Candidate hub genes were verified by another two datasets. Receiver operating characteristic curve analysis was used to evaluate the hub gene diagnostic ability, with further gene set enrichment analysis (GSEA). The immune infiltration level was evaluated by ssGSEA and CIBERSORT methods. Spearman correlation analysis was used to evaluate the hub genes association with immune cells. Finally, a nomogram model and microRNA-TF-mRNA network were constructed based on immune-related biomarkers.
Results
A total of 263 CGs were identified by the intersection of 319 DEGs and 1539 genes in the key turquoise module. Eleven candidate hub genes were screened by the R package “UpSet.” We verified the candidate hub genes based on two validation sets and identified six (SYT1, NEFM, NEFL, SNAP25, GAP43, and GRIA1) that distinguish the PD group from healthy controls. Both CIBERSORT and ssGSEA revealed a significantly increased proportion of neutrophils in the PD group. Correlation between immune cells and hub genes showed SYT1, NEFM, GAP43, and GRIA1 to be significantly related to immune cells. Moreover, the microRNA-TFs-mRNA network revealed that the microRNA-92a family targets all four immune-related genes in PD pathogenesis. Finally, a nomogram exhibited a reliable capability of predicting PD based on the four immune-related genes (AUC = 0.905).
Conclusion
By affecting immune infiltration, SYT1, NEFM, GAP43, and GRIA1, which are regulated by the microRNA-92a family, were identified as diagnostic biomarkers of PD. The correlation of these four genes with neutrophils and the microRNA-92a family in PD needs further investigation.
... To date, increased CSF levels of NFM have been reported for amyotrophic lateral sclerosis (ALS), 13 frontotemporal dementia, 46 and cerebral hemorrhage. 47 Elevated serum NFM levels have also been found in ALS 48 and traumatic brain injuries. 47 in patients of this study. ...
... 55 For ALS and Alzheimer's dementia, dysregulated neurofilament phosphorylation with the accumulation of hyperphosphorylated forms has been described. 48,[56][57][58] Other Neurodegenerative Markers ...
Background
Schizophrenia spectrum disorders (SSD) can be associated with neurodegenerative processes causing disruption of neuronal, synaptic, or axonal integrity. Some previous studies have reported alterations of neurodegenerative markers (such as amyloid beta [Aβ], tau, or neurofilaments) in patients with SSD. However, the current state of research remains inconclusive. Therefore, the rationale of this study was to investigate established neurodegenerative markers in the cerebrospinal fluid (CSF) of a large group of patients with SSD.
Study Design
Measurements of Aβ1–40, Aß1–42, phospho- and total-tau in addition to neurofilament light (NFL), medium (NFM), and heavy (NFH) chains were performed in the CSF of 100 patients with SSD (60 F, 40 M; age 33.7 ± 12.0) and 39 controls with idiopathic intracranial hypertension (33 F, 6 M; age 34.6 ± 12.0) using enzyme-linked immunoassays.
Study Results
The NFM levels were significantly increased in SSD patients (P = .009), whereas phospho-tau levels were lower in comparison to the control group (P = .018). No other significant differences in total-tau, beta-amyloid-quotient (Aβ1–42/Aβ1–40), NFL, and NFH were identified.
Conclusions
The findings argue against a general tauopathy or amyloid pathology in patients with SSD. However, high levels of NFM, which has been linked to regulatory functions in dopaminergic neurotransmission, were associated with SSD. Therefore, NFM could be a promising candidate for further research on SSD.
... 88 Furthermore, in an earlier study, high levels of NfM were observed in amyotrophic lateral sclerosis plasma. 89 Further research is required to establish any potential utility of NfM as a biomarker for frontotemporal dementia and amyotrophic lateral sclerosis. ...
Frontotemporal dementia refers to a group of neurodegenerative disorders characterised by behaviour and language alterations and focal brain atrophy. Amyotrophic lateral sclerosis is a rapidly progressing neurodegenerative disease characterised by loss of motor neurons resulting in muscle wasting and paralysis. Frontotemporal dementia and amyotrophic lateral sclerosis are considered to exist on a disease spectrum given substantial overlap of genetic and molecular signatures. The predominant genetic abnormality in both frontotemporal dementia and amyotrophic lateral sclerosis is an expanded hexanucleotide repeat sequence in the C9ORF72 gene. In terms of brain pathology, abnormal aggregates of TAR-DNA binding protein-43 are predominantly present in frontotemporal dementia and amyotrophic lateral sclerosis patients. Currently, sensitive and specific diagnostic and disease surveillance biomarkers are lacking for both diseases. This has impeded the capacity to monitor disease progression during life and the development of targeted drug therapies for the two diseases. The purpose of this review is to examine the status of current biofluid biomarker discovery and development in frontotemporal dementia and amyotrophic lateral sclerosis. The major pathogenic proteins implicated in different frontotemporal dementia and amyotrophic lateral sclerosis molecular subtypes and proteins associated with neurodegeneration and the immune system will be discussed. Furthermore, the use of mass spectrometry-based proteomics as an emerging tool to identify new biomarkers in frontotemporal dementia and amyotrophic lateral sclerosis will be summarised.
... By contrast, full length (200 kDa) or oligomeric NfH were predominant in CSF and blood (Petzold et al., 2003;Shaw et al., 2005;Lewis et al., 2008). Recent studies also suggest full length (150 kDa) or trimeric NfM (450 kDa) in blood (Haggmark et al., 2014). A comprehensive list of widely used capture and detection antibodies to NfPs in ELISA is shown in Table 1. ...
... Different subunits might reflect different neurodegenerative processes. In addition to commonly used NfL and pNfH, some studies also found potential values of other Nf subunits, i.e., NfM (Hu et al., 2002;Haggmark et al., 2014;Martinez-Morillo et al., 2014;Zucchi et al., 2018;Remnestal et al., 2020), INA (Martinez-Morillo et al., 2014) and PRPH (Finderlater, 2010;Liang et al., 2019;Sabbatini et al., 2021) as biomarkers in CSF or serum in neurological diseases or injuries. ...
Biomarkers of neurodegeneration and neuronal injury have the potential to improve diagnostic accuracy, disease monitoring, prognosis, and measure treatment efficacy. Neurofilament proteins (NfPs) are well suited as biomarkers in these contexts because they are major neuron-specific components that maintain structural integrity and are sensitive to neurodegeneration and neuronal injury across a wide range of neurologic diseases. Low levels of NfPs are constantly released from neurons into the extracellular space and ultimately reach the cerebrospinal fluid (CSF) and blood under physiological conditions throughout normal brain development, maturation, and aging. NfP levels in CSF and blood rise above normal in response to neuronal injury and neurodegeneration independently of cause. NfPs in CSF measured by lumbar puncture are about 40-fold more concentrated than in blood in healthy individuals. New ultra-sensitive methods now allow minimally invasive measurement of these low levels of NfPs in serum or plasma to track disease onset and progression in neurological disorders or nervous system injury and assess responses to therapeutic interventions. Any of the five Nf subunits – neurofilament light chain (NfL), neurofilament medium chain (NfM), neurofilament heavy chain (NfH), alpha-internexin (INA) and peripherin (PRPH) may be altered in a given neuropathological condition. In familial and sporadic Alzheimer’s disease (AD), plasma NfL levels may rise as early as 22 years before clinical onset in familial AD and 10 years before sporadic AD. The major determinants of elevated levels of NfPs and degradation fragments in CSF and blood are the magnitude of damaged or degenerating axons of fiber tracks, the affected axon caliber sizes and the rate of release of NfP and fragments at different stages of a given neurological disease or condition directly or indirectly affecting central nervous system (CNS) and/or peripheral nervous system (PNS). NfPs are rapidly emerging as transformative blood biomarkers in neurology providing novel insights into a wide range of neurological diseases and advancing clinical trials. Here we summarize the current understanding of intracellular NfP physiology, pathophysiology and extracellular kinetics of NfPs in biofluids and review the value and limitations of NfPs and degradation fragments as biomarkers of neurodegeneration and neuronal injury.
... 52 In another study, we identified increased levels in plasma from ALS patients compared with controls. 53 Further studies will elucidate the potential added value of NEFM in relation to NEFL and NEFH. Apart from being brain-enriched, another characteristic that several of the studied proteins have in common is that they have calmodulin-binding properties. ...
Objective
Decreased amyloid beta (Aβ) 42 together with increased tau and phospho-tau in cerebrospinal fluid (CSF) is indicative of Alzheimer’s disease (AD). However, the molecular pathophysiology underlying the slowly progressive cognitive decline observed in AD is not fully understood and it is not known what other CSF biomarkers may be altered in early disease stages.
Methods
We utilized an antibody-based suspension bead array to analyze levels of 216 proteins in CSF from AD patients, patients with mild cognitive impairment (MCI), and controls from two independent cohorts collected within the AETIONOMY consortium. Two additional cohorts from Sweden were used for biological verification.
Results
Six proteins, amphiphysin (AMPH), aquaporin 4 (AQP4), cAMP-regulated phosphoprotein 21 (ARPP21), growth-associated protein 43 (GAP43), neurofilament medium polypeptide (NEFM), and synuclein beta (SNCB) were found at increased levels in CSF from AD patients compared with controls. Next, we used CSF levels of Aβ42 and tau for the stratification of the MCI patients and observed increased levels of AMPH, AQP4, ARPP21, GAP43, and SNCB in the MCI subgroups with abnormal tau levels compared with controls. Further characterization revealed strong to moderate correlations between these five proteins and tau concentrations.
Interpretation
In conclusion, we report six extensively replicated candidate biomarkers with the potential to reflect disease development. Continued evaluation of these proteins will determine to what extent they can aid in the discrimination of MCI patients with and without an underlying AD etiology, and if they have the potential to contribute to a better understanding of the AD continuum.
... Suspension bead array. The procedure for suspension bead arrays was performed as described previously 54,55 . In short, samples were distributed in 96-well microtiter plates, diluted 1:10 in PBS and the protein content was directly labeled with biotin. ...
Apart from well-defined factors in neuronal cells¹, only a few reports consider that the variability of sporadic amyotrophic lateral sclerosis (ALS) progression can depend on less-defined contributions from glia2,3 and blood vessels⁴. In this study we use an expression-weighted cell-type enrichment method to infer cell activity in spinal cord samples from patients with sporadic ALS and mouse models of this disease. Here we report that patients with sporadic ALS present cell activity patterns consistent with two mouse models in which enrichments of vascular cell genes preceded microglial response. Notably, during the presymptomatic stage, perivascular fibroblast cells showed the strongest gene enrichments, and their marker proteins SPP1 and COL6A1 accumulated in enlarged perivascular spaces in patients with sporadic ALS. Moreover, in plasma of 574 patients with ALS from four independent cohorts, increased levels of SPP1 at disease diagnosis repeatedly predicted shorter survival with stronger effect than the established risk factors of bulbar onset or neurofilament levels in cerebrospinal fluid. We propose that the activity of the recently discovered perivascular fibroblast can predict survival of patients with ALS and provide a new conceptual framework to re-evaluate definitions of ALS etiology.
... As the method only consumes a few microliters of sample, it is especially useful in settings such as those involving infants and young children, where limited sample volumes may be collected. This method has previously been applied in both pediatric and adult cohorts in other contexts such as childhood asthma and malaria, [12,30] liver and kidney disease, [11,31] nervous system/neurodegenerative disease, [32,33] and cancer. [34] Although our study included a large number of children, the number of children per lifestyle group was too small to allow for analysis stratified by exposure, such as vaccination and breast feeding or outcome such as sensitization to allergens. ...
Purpose
Little is known about the longitudinal development of different plasma protein levels during early childhood and particularly in relation to lifestyle factors. This study aimed to monitor the plasma proteome early in life and the influence of different lifestyles.
Experimental design
A multiplex bead‐based immunoassay was used to analyze plasma levels of 97 proteins in 280 blood samples longitudinally collected in children at 6, 12, 24 and 60 months of age living in families with an anthroposophic (n = 15), partly anthroposophic (n = 27) or non‐anthroposophic (n = 28) lifestyle.
Results
A total of 68 proteins (70%) showed significantly altered plasma levels between 6 months and 5 years of age. In lifestyle stratified analysis, 59 of 97 (61%) proteins were altered over time within one or more of the three lifestyle groups. Nearly half of these proteins (28 out of 59) changed irrespective of lifestyle. The temporal changes represented four longitudinal trends of the plasma proteins during development, also following stratification of lifestyle.
Conclusions and clinical relevance
Our findings contribute to understand the development of the plasma proteome under the influence of lifestyle exposures in early childhood.
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... Although CSF and serum NF-L is known to increase in several neurodegenerative diseases [49,50] including FTD, NF-M has to our knowledge not been investigated in the context of FTD before. However, high CSF levels of NF-M has previously been reported in stroke patients [51] and using the same NF-M antibodies as in this study, high levels of NF-M were observed in plasma of ALS patients [52]. ...
... The semi-quantitative nature of the SBA assay could be viewed as a limitation since no definite concentrations are obtained. However, it has previously been shown that the protein levels acquired by the use of suspension bead arrays can be reproduced using other methods [28,52,66,67]. ...
Background:
The clinical presentations of frontotemporal dementia (FTD) are diverse and overlap with other neurological disorders. There are, as of today, no biomarkers in clinical practice for diagnosing the disorders. Here, we aimed to find protein markers in cerebrospinal fluid (CSF) from patients with FTD, presymptomatic mutation carriers and non-carriers.
Methods:
Antibody suspension bead arrays were used to analyse 328 proteins in CSF from patients with behavioural variant FTD (bvFTD, n = 16) and progressive primary aphasia (PPA, n = 13), as well as presymptomatic mutation carriers (PMC, n = 16) and non-carriers (NC, n = 8). A total of 492 antibodies were used to measure protein levels by direct labelling of the CSF samples. The findings were further examined in an independent cohort including 13 FTD patients, 79 patients with Alzheimer's disease and 18 healthy controls.
Results:
We found significantly altered protein levels in CSF from FTD patients compared to unaffected individuals (PMC and NC) for 26 proteins. The analysis show patterns of separation between unaffected individuals and FTD patients, especially for those with a clinical diagnosis of bvFTD. The most statistically significant differences in protein levels were found for VGF, TN-R, NPTXR, TMEM132D, PDYN and NF-M. Patients with FTD were found to have higher levels of TN-R and NF-M, and lower levels of VGF, NPTXR, TMEM132D and PDYN, compared to unaffected individuals. The main findings were reproduced in the independent cohort.
Conclusion:
In this pilot study, we show a separation of FTD patients from unaffected individuals based on protein levels in CSF. Further investigation is required to explore the CSF profiles in larger cohorts, but the results presented here has the potential to enable future clinical utilization of these potential biomarkers within FTD.
... Neurofilament medium (NEFM) is relevant to the elongation of neuronal structures (Pezzini et al., 2017). In a larger cohort with 367 amyotrophic lateral sclerosis (ALS) patients and 101 controls, plasma NEFM levels were significantly elevated in ALS patients compared with controls (Häggmark et al., 2014). NEFM was also down-regulated in ZF-like aldosteroneproducing adenomas and contributed to a D1R/D2R imbalance (Maniero et al., 2017). ...
Background : The pathogenesis of Alzheimer’s disease (AD) remains to be elucidated. This study aimed to identify the hub genes in AD pathogenesis and determine their functions and pathways.
Methods : A co-expression network for an AD gene dataset with 401 samples was constructed, and the AD status-related genes were screened. The hub genes of the network were identified and validated by an independent cohort. The functional pathways of hub genes were analyzed.
Results : The co-expression network revealed a module that related to the AD status, and 101 status-related genes were screened from the trait-related module. Gene enrichment analysis indicated that these status-related genes are involved in synaptic processes and pathways. Four hub genes ( ENO2 , ELAVL4 , SNAP91 , and NEFM ) were identified from the module, and these hub genes all participated in AD-related pathways, but the associations of each gene with clinical features were variable. An independent dataset confirmed the different expression of hub genes between AD and controls.
Conclusions : Four novel genes associated with AD pathogenesis were identified and validated, which provided novel therapeutic targets for AD.
... Molecular indicators of glutathione excitotoxicity, impaired axonal transport, oxidative stress, mitochondrial dysfunction, growth factor deficiency, protein aggregation, abnormal RNA metabolism, and apoptosis were observed in the human brain, spinal cord, and transgenic disease models long before the onset of symptoms. Excessive glutamate leading to neuronal degeneration is thought to be one of the pathogenesis of ALS, a fatal neurodegenerative disorder caused by selective death of motor neurons in the brain and spinal cord [50,51]. Riluzol, a drug that is used to treat ALS, inhibits the activation of postsynaptic neurons that prevent glutamate release and reduce glutamate-induced sodium ecototoxicity [50]. ...
... Excessive glutamate leading to neuronal degeneration is thought to be one of the pathogenesis of ALS, a fatal neurodegenerative disorder caused by selective death of motor neurons in the brain and spinal cord [50,51]. Riluzol, a drug that is used to treat ALS, inhibits the activation of postsynaptic neurons that prevent glutamate release and reduce glutamate-induced sodium ecototoxicity [50]. Loss of EAAT1 was observed in patients with ALS. ...
The solute carrier (SLC) superfamily is one of the major sub-groups of membrane proteins in mammalian cells. The solute carrier proteins include more than 400 different membrane-spanning solute carriers organized with 65 families in the human. In solute carrier family neurons, neurotransmitter is considered to be a pharmacological target of neuropsychiatric drugs because of their important role in the recovery of neurotransmitters such as GABA, glutamate, serotonin, dopamine and noradrenaline and regulation of their concentration in synaptic regions. Therefore, solute carrier transporters play vital and different roles in neurodegenerative disorders. In this article, the role of solute carrier transporters in neurodegenerative disorders such as Alzheimer disease, amyotrophic lateral sclerosis, Huntington disease, Parkinson's diseases, depression, post-traumatic stress disorder, dementia, schizophrenia, and Epilepsy reviewed and discussed to see how defects or absences in SLC transporter cause neurodegenerative disorders. In this review, we try to summarize what is known about solute carriers with respect to brain distribution and expression. The review summarizes current knowledge on the roles of solute carrier transporters in neurodegenerative disorders.
