Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Jul;13(4):e1800105.
doi: 10.1002/prca.201800105. Epub 2019 Jan 25.

Quantitative Proteomic Profiling of Cerebrospinal Fluid to Identify Candidate Biomarkers for Alzheimer's Disease

Gajanan Sathe  1   2   3   4 , Chan Hyun Na  3   5   6 , Santosh Renuse  3 , Anil K Madugundu  1   2   3   4 , Marilyn Albert  5 , Abhay Moghekar  5 , Akhilesh Pandey  1   2   6   7
Affiliations

Quantitative Proteomic Profiling of Cerebrospinal Fluid to Identify Candidate Biomarkers for Alzheimer's Disease

Gajanan Sathe et al. Proteomics Clin Appl. 2019 Jul.

Abstract

Purpose: The aim of this study is to identify the potential cerebrospinal fluid (CSF) biomarkers for Alzheimer's disease and to evaluate these markers on independent CSF samples using parallel reaction monitoring (PRM) assays.

Experimental design: High-Resolution mass spectrometry and tandem mass tag (TMT) multiplexing technology are employed to identify potential biomarkers for Alzheimer's disease. Some of the identified potential biomarkers are validated using PRM assays.

Results: A total of 2327 proteins are identified in the CSF of which 139 are observed to be significantly altered in the CSF of AD patients. The proteins altered in AD includes a number of known AD marker such as MAPT, NPTX2, VGF, GFAP, and NCAM1 as well as novel biomarkers such as PKM and YWHAG. These findings are validated in a separate set of CSF specimens from AD dementia patients and controls. NPTX2, in combination with PKM or YWHAG, leads to the best results with AUCs of 0.935 and 0.933, respectively.

Conclusions and clinical relevance: The proteins that are found to be altered in the CSF of patients with AD could be used for monitoring disease progression and therapeutic response and perhaps also for early detection once they are validated in larger studies.

Keywords: Alzheimer's disease; TMT; biomarker; biomedical applications; cerebrospinal fluid; mass spectrometry; parallel reaction monitoring; proteome.

PubMed Disclaimer

Conflict of interest statement

Competing financial interests

The authors declare no competing financial interests.

Figures

Figure 1
Figure 1
A schematic of the workflow used to study the proteomic changes in CSF of AD patients. The CSF samples were depleted of high abundance proteins using an antibody-based MARS-14 depletion column. The proteome of CSF from cognitively normal individuals was compared with that of AD patients using a TMT-based quantitative proteomics approach in the discovery step. Validation was carried out using parallel reaction monitoring (PRM) assays for a subset of molecules that were found to be significantly altered in abundance in AD.
Figure 2
Figure 2
Summary of TMT-based discovery experiments (A) Distribution of fold-change values (log2 ratios) (AD/Controls) for proteins identified in the study (B) Volcano plot of identified proteins (c) Localization of proteins that are differentially abundant in CSF of AD patients, Representative MS/MS spectra of peptides identified from D) Microtubule associated protein tau (MAPT) (E) 14-3-3 gamma (YWHAG) (F) Pyruvate kinase (PKM)
Figure 3
Figure 3
Validation of known candidate proteins by parallel reaction monitoring assays: (A) Neuronal pentraxin 2 (NPTX2); (B) VGF nerve growth factor inducible (VGF); (C) Secretogranin 2 (SCG2). Peptide sequences and p-values are shown.
Figure 4
Figure 4
Validation of novel candidate proteins by parallel reaction monitoring assays: (A) 14-3-3 gamma (YWHAG); (B) Pyruvate kinase (PKM); (C) Gelsolin (GSN). Peptide sequences and p-values are shown.
Figure 5
Figure 5
ROC analysis for selected proteins to evaluate the utility of the markers: (A) ROC curve for individual markers (B) ROC curve with combination of markers; The area under the curve (AUC) is indicated.
See this image and copyright information in PMC

References

    1. Scheltens P, Blennow K, Breteler MM, de Strooper B, et al. Alzheimer's disease. Lancet. 2016;388:505–517. - PubMed
    1. Glenner GG, Wong CW. Alzheimer's disease: initial report of the purification and characterization of a novel cerebrovascular amyloid protein. Biochem Biophys Res Commun. 1984;120:885–890. - PubMed
    1. Lee VM, Balin BJ, Otvos L, Jr, Trojanowski JQ. A68: a major subunit of paired helical filaments and derivatized forms of normal Tau. Science. 1991;251:675–678. - PubMed
    1. Lin W, Zhang J, Liu Y, Wu R, et al. Studies on diagnostic biomarkers and therapeutic mechanism of Alzheimer's disease through metabolomics and hippocampal proteomics. Eur J Pharm Sci. 2017;105:119–126. - PubMed
    1. van Waalwijk van Doorn LJ, Gispert JD, Kuiperij HB, Claassen JA, et al. Improved Cerebrospinal Fluid-Based Discrimination between Alzheimer's Disease Patients and Controls after Correction for Ventricular Volumes. J Alzheimers Dis. 2017;56:543–555. - PubMed

Publication types