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Role of citrullination in diagnosing diseases

A citrullination, disease technology, applied in the field of the role of citrullination in diagnosing diseases

Inactive Publication Date: 2018-04-17
CEDARS SINAI MEDICAL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Challenges in identifying citrullinated proteins and modified amino acid residues

Method used

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  • Role of citrullination in diagnosing diseases
  • Role of citrullination in diagnosing diseases
  • Role of citrullination in diagnosing diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Embodiment 1: experimental method

[0092] Reagents and Materials

[0093] The following reagents were obtained: rabbit skeletal muscle PAD mix (PAD) (SignalChem); PAD2 (Sigma), heavy enzymatic myosin (HMM), tropomyosin (TM) (Sigma); F-actin (Cytoskeleton. Inc), cardiac troponin (TnI) (Abcam), anti-modified citrulline antibody (Millipore); sequencing grade Lys-C protease (WAKO), and protease inhibitor cocktail (Roche).

[0094] human heart tissue

[0095] Left ventricular tissue samples were obtained from Cris Dos Remedios, University of Sydney, Australia, after informed consent and approval by the local ethics committee. Samples were obtained from patients with HF (ischemic heart disease (ISHD) and idiopathic cardiomyopathy (IDCM), n=10 for each group) and non-heart failure donors during heart transplant surgery as previously described Heart (n=10) [Zhang P et al., Multiple reaction monitoring to identify site-specific troponin I phosphorylated residues in the faili...

Embodiment 2

[0136] Identification of cardiac citrullinated proteins.

[0137] To identify citrullination targets in the heart, we assessed the citrullinome in three groups ISHD, IDCM and non-failing donor hearts (n=10 each) using SWATH-MS [34]. SWATH-MS allows for quantification. Compared to non-heart failure controls, 53 citrullination sites were altered with HF (p<0.05) and are listed in Table 1A.

[0138] Table 1A: List of citrullinated proteins (p=0.05) with citrullinated peptide sequences. "Dea" is citrullination or deamination. "CAM" is carbamide methylation. Citrullinated proteins are grouped by cellular components. UP identifier = universal protein (UP) resource protein ID; p_kw = p-value of Kruskal-Wallis test (statistic)

[0139]

[0140]

[0141]

[0142] Table 1B shows all citrullinated peptides present in this study. These peptides are proteins with diverse cellular functions, including regulators of transcription and chromatin structure, cytoskeleton and contr...

Embodiment 3

[0182] Our experimental results characterize citrullinated proteins in normal and HF myocardium. Analysis revealed that citrullination was enriched in the mitochondrial and sarcomere subproteomes. Protein citrullination has a broad cellular distribution (Fig. 1), but is highly enriched in the mitochondrial and sarcomere subproteomes. The close association between these two subproteomes is not unexpected due to the high energy requirements of sarcomeres. We speculate that citrullination, similar to phosphorylation and acetylation, may potentially regulate muscle contractile proteins in a coordinated manner. Interestingly, multiple enzymes involved in metabolic pathways / metabolism were upregulated in ischemia but downregulated in IDCM ( Figure 6 ). To understand the topology and functional annotation of citrullinated protein-protein interactions in the cardiac system, protein networks were constructed using the STRING database ( Figure 2B ). The entire protein network con...

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Abstract

Provided herein are methods and markers for diagnosing cardiovascular disease and / or neurodegenearative diseases in a subject. The methods include obtaining a biological sample from a subject in needof diagnosis and detecting the amount of a citrullinated protein or a citrullinated peptide in the biological sample obtained from said subject.

Description

[0001] Statement Regarding Federally Funded Research [0002] This invention was made with Government support under Grant Numbers HL112586-01 and HHSN268201000032C awarded by the National Institutes of Health. The government has certain rights in this invention. Background technique [0003] All publications herein are incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. The following description includes information that may be useful in understanding the present invention. It is not an admission that any of the information presented herein is prior art or relevant to the presently claimed invention, or that any publication cited is prior art, either explicitly or implicitly. [0004] Citrullination, an irreversible post-translational modification (PTM) involving the conversion of arginine to citrulline by the peptidylarginine deiminase (PAD) family o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N33/6896G01N2440/18G01N2800/28G01N2800/2814G01N2800/2835G01N2800/32
Inventor J·凡艾克J·福特博伯E·科荣格R·侯维斯凯本杰明·菲利普·伯曼
Owner CEDARS SINAI MEDICAL CENT
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