ABSTRACT

Objectives

Renal amyloidosis (RA) is a rare protein misfolding disorder that prompts progressive renal insufficiency. This study aimed to decipher proteomic changes in human sera to understand the pathophysiology and molecular mechanisms underlying the disease development, hence assisting in the diagnosis of RA.

Methods

Serum proteomic analysis was performed using a gel-based approach followed by MALDI-TOF MS. RA patients with age and sex matched healthy volunteers were recruited from Max Super Speciality Hospital, New Delhi, India.

Results

Proteome profiles of serum revealed eight differentially expressed proteins namely, Zinc finger protein 624, Protein FAM183A, Calcium-binding mitochondrial carrier protein Scamc-3, V-type proton ATPase 116 kDa subunit A isoforms 2, Protein TXNRD3NB, ATP – dependent RNA helicase, Troponin C and Mitogen-activated protein kinase kinase kinase 7. These proteins were reported first time in RA. The increased levels of MAP3K7 and TROPONIN C were validated by bio-layer interferometry and their diagnostic accuracy was evaluated by ROC curve analysis. The differentially expressed proteins were predominantly associated with vesicular trafficking, transcriptional regulation, metabolic processes, apoptotic process and mitochondrial metabolism.

Conclusion

The results indicate that these proteomic signatures may be considered as potential molecular targets for RA diagnostics and therapeutics subject to validation on large sample size.

Abbreviations: AβP= Amyloid-beta protein, Aβ=Amyloid-beta, AL= Light chain amyloidosis, AA= Amyloid A, ALECT2= LECT2 amyloidosis, APS= Ammonium persulfate CKD= Chronic Kidney Diseases, EBRT= external beam radiation therapy, ESRD= End-Stage Kidney Disease, Glis2= Gli-similar 2, JNK= c-Jun NH 2-terminal kinase, MAPK= Mitogen-Activated Protein Kinase, MM=Multiple Myeloma, PHD= Prolyl hydroxylase, RA = Renal Amyloidosis, SAA= Serum Amyloid A, SD= Standard Deviation, Sepp= Selenoprotein, SCC= Squamous cell carcinoma, SDS= Sodium dodecyl sulfate, TEMED = tetramethyl ethylenediamine, TGF-Beta-1=Transforming growth factor- Beta-1, Trx = Thioredoxin, TrxR= Thioredoxin reductase

中文翻译：

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MAP3K7和肌钙蛋白C蛋白在肾淀粉样变性中的差异表达及相关扰动

摘要

目标

肾淀粉样变性 (RA) 是一种罕见的蛋白质错误折叠疾病，可导致进行性肾功能不全。本研究旨在破译人类血清中的蛋白质组学变化，以了解疾病发展的病理生理学和分子机制，从而协助 RA 的诊断。

方法

使用基于凝胶的方法和 MALDI-TOF MS 进行血清蛋白质组学分析。从印度新德里 Max Super 专科医院招募了年龄和性别匹配的健康志愿者的 RA 患者。

结果

血清的蛋白质组图谱揭示了八种差异表达的蛋白质，即锌指蛋白 624、蛋白质 FAM183A、钙结合线粒体载体蛋白 Scamc-3、V 型质子 ATPase 116 kDa 亚基 A 亚型 2、蛋白质 TXNRD3NB、ATP - 依赖的 RNA 解旋酶、肌钙蛋白 C 和丝裂原活化蛋白激酶激酶激酶 7。这些蛋白质是首次在 RA 中报道。MAP3K7 和肌钙蛋白 C 水平的增加通过生物层干涉测量法进行验证，并通过 ROC 曲线分析评估其诊断准确性。差异表达的蛋白质主要与囊泡运输、转录调控、代谢过程、凋亡过程和线粒体代谢有关。

结论

结果表明，这些蛋白质组学特征可以被认为是 RA 诊断和治疗的潜在分子靶点，需要在大样本量上进行验证。

缩写： AβP= 淀粉样蛋白-β，Aβ=淀粉样蛋白-β，AL= 轻链淀粉样变性，AA= 淀粉样蛋白 A，ALECT2= LECT2 淀粉样变性，APS= 过硫酸铵 CKD= 慢性肾病，EBRT= 外照射放射治疗，ESRD=终末期肾病，Glis2= Gli-similar 2，JNK= c-Jun NH 2-末端激酶，MAPK= 丝裂原活化蛋白激酶，MM=多发性骨髓瘤，PHD=脯氨酰羟化酶，RA=肾淀粉样变性，SAA=血清淀粉样蛋白 A，SD= 标准偏差，Sepp= 硒蛋白，SCC= 鳞状细胞癌，SDS= 十二烷基硫酸钠，TEMED = 四甲基乙二胺，TGF-Beta-1=转化生长因子 - Beta-1，Trx = 硫氧还蛋白，TrxR= 硫氧还蛋白还原酶