Dataset Information

Isotopic N,N-dimethyl leucine tags for absolute quantification of clusterin and apolipoprotein E in Alzheimer's disease

ABSTRACT: Alzheimer’s disease (AD) is the most common form of dementia and one of the leading causes of death in the United States. In past decades, extensive efforts have been devoted to improving our understanding of AD pathogenesis and accelerating biomarker discovery for early diagnosis and clinical treatment of AD. Herein, this study aims to quantify clusterin (CLU) and apolipoprotein E (APOE) in blood samples from AD patients and evaluate these two proteins as potential biomarkers in AD diagnosis. In-house synthesized 5-plex isotopic N,N-dimethyl leucine (iDiLeu) tags were used to label target peptide standards at different concentrations to construct standard curves. Our study reveals that the levels of CLU and APOE exhibit clear disparities in male vs. female AD groups but not in male vs. female non-AD groups, while the levels of serum CLU and APOE do not show statistical differences in the AD groups and non-AD groups. Principal component analysis (PCA) with CLU and APOE showed some separation between the AD and non-AD participants. Significance: Dissecting CLU and APOE heterogeneity in AD pathogenesis may therefore facilitate delineating the pathological relevance for specific pathways between different genders, leading to personalized medicine in the future. Collectively, this study introduces a cost-effective absolute quantitative proteomics strategy for target protein quantitation and lays the foundation for future investigation of CLU and APOE as potential biomarkers for AD.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Serum

DISEASE(S): Alzheimer's Disease

SUBMITTER: Yuan Liu

LAB HEAD: Lingjun Li

PROVIDER: PXD030724 | Pride | 2022-10-05

REPOSITORIES: Pride

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Isotopic N,N-dimethyl leucine tags for absolute quantification of clusterin and apolipoprotein E in Alzheimer's disease.

Liu Yuan Y Zhang Hua H Zhong Xiaofang X Li Zihui Z Zetterberg Henrik H Li Lingjun L

Journal of proteomics 20220203

Alzheimer's disease (AD) is the most common form of dementia and one of the leading causes of death in the United States. In the past decades, extensive efforts have been devoted to biomarker discovery for early diagnosis and treatment of AD. Herein, this study aims to quantify clusterin (CLU) and apolipoprotein E (APOE) in blood samples from AD patients and evaluate these two proteins as potential biomarkers in AD diagnosis. In-house synthesized 5-plex isotopic N,N-dimethyl leucine (iDiLeu) tag ...[more]

PMID: 35124278

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Project description:Microglia are innate immune cells of the brain that perform phagocytic and inflammatory functions in disease conditions. Transcriptomic studies of acutely-isolated microglia have provided novel insights into their molecular and functional diversity in homeostatic and neurodegenerative disease states. State-of-the-art mass spectrometric methods can comprehensively characterize proteomic alterations in microglia in neurodegenerative disorders, potentially providing novel functionally-relevant molecular insights that are not provided by transcriptomics. However, proteomic profiling of adult primary microglia in neurodegenerative disease conditions has not been performed. We performed quantitative proteomic analyses of purified CD11b+ acutely-isolated microglia adult mice in normal, acute neuroinflammatory (LPS-treatment) and chronic neurodegenerative states (5xFAD model of Alzheimer’s disease [AD]) using tandem mass tag mass spectrometry. Differential expression analyses were performed to characterize specific microglial proteomic changes in 5xFAD mice and identify overlap with LPS-induced pro-inflammatory changes. Our results were also contrasted with existing proteomic data from wild-type mouse microglia and from existing microglial transcriptomic data from wild-type and 5xFAD mice. Neuropathological validation studies of select proteins were performed in human AD and 5xFAD brains. Of 4,133 proteins identified, 187 microglial proteins were differentially expressed in the 5xFAD mouse model of AD pathology, including proteins with previously known (Apoe, Clu and Htra1) as well as previously unreported relevance to AD biology (Cotl1 and Hexb). Proteins upregulated in 5xFAD microglia shared significant overlap with pro-inflammatory changes observed in LPS-treated mice. Several proteins increased in human AD brain were also upregulated by 5xFAD microglia (Aβ peptide, Apoe, Htra1, Cotl1 and Clu). Cotl1 was identified as a novel microglia-specific marker with increased expression and strong association with AD neuropathology. Apoe protein was also detected within plaque-associated microglia in which Apoe and Aβ were highly co-localized suggesting a role for Apoe in phagocytic clearance of Aβ. We report the first comprehensive comparative proteomic study of adult mouse microglia derived from acute neuroinflammatory and AD models, representing a valuable resource to the neuroscience research community. We highlight shared and unique microglial proteomic changes in acute neuroinflammatory, aging and AD mouse models in addition to identifying novel roles for microglial proteins in human neurodegeneration.

Project description:Background: Preclinical studies showed thatmesenchymal stem cells (MSCs) ameliorate tau phosphorylation, amyloid-betaaccumulation, and inflammation in Alzheimer’s disease (AD) mouse models viasecretion of neurotrophic factors and cytokines. We aimed to identify CSFbiomarkers that can be used to predict or monitor the response to MSCs inpatients with AD. Methods: AD patients were injected withhuman umbilical cord blood-MSCs (n=22) or placebo (n=12). The cerebrospinalfluid (CSF) samples were collected at baseline, one day after the firstinjection, and one day after the third injection. The patients injected withMSCs were classified into good responder (GR) or poor responder (PR) groupsbased on the rate of changes in the ratio of total-tau and phosphorylated-tauin the CSF. We selected three typical participants in each group, and their CSFprotein levels were analyzed using liquid chromatography-tandem massspectrometry (LC-MS/MS). Results: In the LC-MS/MS analysis 1,667proteins were identified. 11 proteins showed significant differences betweenthe typical GR and PR at baseline. Based on their significance level and knownfunctions two proteins, reticulocalbin-3 (RCN3) and follistatin-related protein3 (FSTL3), were selected as potential biomarkers to predict MSC response. 173proteins showed significant change one day after the third injection comparedto the baseline in typical GR. We excluded 45 proteins that showed significantchange after the third injection compared to the baseline in the typical PR.Based on their significance level and known function, four proteins,scrapie-responsive protein 1 (SCRG1), neural proliferation and differentiationcontrol protein (NPDC1), apolipoprotein E (ApoE), and cystatin C (CysC), wereselected as potential biomarker to monitor MSC response. Additionally,functional analysis revealed that the increased CSF proteins after the thirdinjection compared to the baseline in the typical GR were associated withsynaptogenesis. Conclusions: This study identified twoproteins (RCN3 and FSTL3) that may be potential biomarkers for predicting MSCresponse and four proteins (SCRG1, NPDC1, ApoE, CysC) that may be potentialbiomarkers for monitoring MSC response in patients with AD. Further studies areneeded to validate our results.

			Action	DRS
	AD10_peptide.csv	Csv
	AD1_peptide.csv	Csv
	AD2_peptide.csv	Csv
	AD3_peptide.csv	Csv
	AD4_peptide.csv	Csv