Diabetes induced renal complications by leukocyte activation of nuclear factor κ-B and its regulated genes expression

Type 2 diabetes mellitus (T2D) is a metabolic disorder characterized by inappropriate insulin function. Despite wide progress in genome studies, defects in gene expression for diabetes prognosis still incompletely identified. Prolonged hyperglycemia activates NF-κB, which is a main player in vascular dysfunctions of diabetes. Activated NF-κB, triggers expression of various genes that promote inflammation and cell adhesion process. Alteration of pro-inflammatory and profibrotic gene expression contribute to the irreversible functional and structural changes in the kidney resulting in diabetic nephropathy (DN). To identify the effect of some important NF-κB related genes on mediation of DN progression, we divided our candidate genes on the basis of their function exerted in bloodstream into three categories (Proinflammatory; NF-κB, IL-1B, IL-6, TNF-α and VEGF); (Profibrotic; FN, ICAM-1, VCAM-1) and (Proliferative; MAPK-1 and EGF). We analyzed their expression profile in leukocytes of patients and explored their correlation to diabetic kidney injury features. Our data revealed the overexpression of both proinflammatory and profibrotic genes in DN group when compared to T2D group and were associated positively with each other in DN group indicating their possible role in DN progression. In DN patients, increased expression of proinflammatory genes correlated positively with glycemic control and inflammatory markers indicating their role in DN progression. Our data revealed that the persistent activation NF-κB and its related genes observed in hyperglycemia might contribute to DN progression and might be a good diagnostic and therapeutic target for DN progression. Large-scale studies are needed to evaluate the potential of these molecules to serve as disease biomarkers.     

Role of kidney biomarkers of chronic kidney disease: An update

Chronic kidney disease (CKD) is a progressive pathological condition marked by deteriorating renal function over time. Diagnostic of kidney disease depend on serum creatinine level and glomerular filtration rate which is detectable when kidney function become half. The detection of kidney damage in an early stage needs robust biomarkers. Biomarkers allow monitoring the disease progression at initial stages of disease. On the onset of impairment in cellular organization there is perturbation in signaling molecules which are either up-regulated or down-regulated and act as an indicator or biomarker of diseased stage. This review compiled the cell signaling of different kidney biomarkers associated with the onset of chronic kidney diseases. Delay in diagnosis of CKD will cause deterioration of nephron function which leads to End stage renal disease and at that point patients require dialysis or kidney transplant. Detailed information on the complex network in signaling pathway leading to a coordinated pattern of gene expression and regulation in CKD will undoubtedly provide important clues to develop novel prognostic and therapeutic strategies for CKD.     

Urinary proteomics and molecular determinants of chronic kidney disease: possible link to proteases

Chronic kidney disease (CKD) is the gradual decrease in renal function. Currently available biomarkers are effective only in detecting late stage CKD. Biomarkers of early stage CKD and prognostic biomarkers are required. We review the major findings in urinary proteomics in CKD during the last five years. Significant progress has been made and today urinary proteomics is applied in large randomized trials, and in patient management. Many of the biomarkers indicate altered protease activity. We therefore also review the literature on proteases associated with renal function loss. We anticipate in silico prediction tools of protease activity and additional system biology studies may contribute to biomarker discovery and elucidate the role of proteases in CKD development and progression. These approaches will enable the deciphering of the molecular pathophysiology of CKD, and hence definition of the most appropriate therapeutic targets in the future. Together with stable biomarker panels available today, this will significantly improve patient management.     

运动性急性肾损伤与新型肾功能生物标志物

大强度运动中,非创伤性急性肾损伤（acute kindey injury, AKI）经常发生,表现为血尿、蛋白尿、血红蛋白尿等。一般认为,中低程度的运动性急性肾损伤是可逆的,可完全恢复。但动物实验与人类研究均发现,严重的运动性肾损伤会导致“功能性”急性肾损伤发展为“结构性”急性肾损伤,并增加慢性肾病的风险。运动性急性肾损伤对机体的潜在健康威胁已引起国内外相关领域学者的广泛关注。血清肌酐 (serum creatinine, Scr)和尿量作为肾功能的传统经典标志物,不能特异性反映早期肾损伤,而新型肾损伤标志物可进一步明确损伤的位置及严重程度。在运动领域,利用新型生物标志物进行无创性检查,识别早期运动性急性肾损伤非常必要。本文综述了反映肾小球或肾小管损伤、细胞周期停滞和肾损伤修复的新型生物标志物,着重论述了尿中性粒细胞明胶酶相关脂质运载蛋白（NGAL)和肾损伤分子-1（KIM-1)与肾功能的关系,以及长时间耐力运动、急性运动和高强度间歇阻力运动3种运动形式对肾功能的影响,旨在引起重视,精准识别风险,及时进行早干预。     

运动性急性肾损伤与新型肾功能生物标志物

大强度运动中,非创伤性急性肾损伤（acute kindey injury, AKI）经常发生,表现为血尿、蛋白尿、血红蛋白尿等。一般认为,中低程度的运动性急性肾损伤是可逆的,可完全恢复。但动物实验与人类研究均发现,严重的运动性肾损伤会导致“功能性”急性肾损伤发展为“结构性”急性肾损伤,并增加慢性肾病的风险。运动性急性肾损伤对机体的潜在健康威胁已引起国内外相关领域学者的广泛关注。血清肌酐 (serum creatinine, Scr)和尿量作为肾功能的传统经典标志物,不能特异性反映早期肾损伤,而新型肾损伤标志物可进一步明确损伤的位置及严重程度。在运动领域,利用新型生物标志物进行无创性检查,识别早期运动性急性肾损伤非常必要。本文综述了反映肾小球或肾小管损伤、细胞周期停滞和肾损伤修复的新型生物标志物,着重论述了尿中性粒细胞明胶酶相关脂质运载蛋白（NGAL)和肾损伤分子-1（KIM-1)与肾功能的关系,以及长时间耐力运动、急性运动和高强度间歇阻力运动3种运动形式对肾功能的影响,旨在引起重视,精准识别风险,及时进行早干预。     

Wnt signalling pathway in bladder cancer

Bladder cancer (BC) is one of the most common tumours of the urinary system and is also known as a highly malignant tumour. In addition to conventional diagnosis and treatment methods, recent research has focused on studying the molecular mechanisms related to BC, in the hope that new, less toxic and effective targeted anticancer drugs and new diagnostic markers can be discovered. It is known that the Wingless (Wnt) signalling pathway and its related genes, proteins and other substances are involved in multiple biological processes of various tumours. Clarifying the contribution of the Wnt signalling pathway in bladder tumours will help establish early diagnosis indicators, develop new therapeutic drugs and evaluate the prognosis for BC. This review aims to summarise previous studies related to BC and the Wnt signalling pathway, with a focus on exploring the participating substances and their mechanisms in the regulation of the Wnt signalling pathway to better determine how to promote new chemotherapeutic drugs, potential therapeutic targets and diagnostic biomarkers.     

The human urinary proteome reveals high similarity between kidney aging and chronic kidney disease

Aging induces morphological changes of the kidney and reduces renal function. We analyzed the low molecular weight urinary proteome of 324 healthy individuals from 2–73 years of age to gain insight on human renal aging. We observed age‐related modification of secretion of 325 out of over 5000 urinary peptides. The majority of these changes were associated with renal development before and during puberty, while 49 peptides were related to aging in adults. We therefore focussed the remainder of the study on these 49 peptides. The majority of these 49 peptides were also markers of chronic kidney disease, suggesting high similarity between aging and chronic kidney disease. Blinded evaluation of samples from healthy volunteers and diabetic nephropathy patients confirmed both the correlation of biomarkers with aging and with renal disease. Identification of a number of these aging‐related peptides led us to hypothesize that reduced proteolytic activity is involved in human renal aging. Finally, among the 324 supposedly healthy individuals, some had urinary aging‐related peptide excretion patterns typical of an individual significantly older than their actual age. In conclusion, these aging‐related biomarkers may allow noninvasive detection of renal lesions in healthy persons and show high resemblance between human aging and chronic kidney disease. This similarity has to be taken into account when searching for biomarkers of renal disease.     

Circulating biomarkers of protein oxidation for Alzheimer disease: expectations within limits

Alzheimer disease (AD), the most common dementing disorder, is a multifactorial disease with complex etiology. Among different hypotheses proposed for AD one of the most corroborated is the "oxidative stress hypothesis". Although recent studies extensively demonstrated the specific oxidative modification of selected proteins in the brain of AD patients and how their dysfunction possibly correlates with the pathology, there is still an urgent need to extend these findings to peripheral tissue. So far very few studies showed oxidative damage of proteins in peripheral tissues and current findings need to be replicated. Another limit in AD research is represented by the lack of highly specific diagnostic tools for early diagnosis. For a full screening and early diagnosis, biomarkers easily detectable in biological samples, such as blood, are needed. The search of reliable biomarkers for AD in peripheral blood is a great challenge. A few studies described a set of plasma markers that differentiated AD from controls and were shown to be useful in predicting conversion from mild cognitive impairment, which is considered a prodromal stage, to AD. We review the current state of knowledge on peripheral oxidative biomarkers for AD, including proteomics, which might be useful for early diagnosis and prognosis.     

Glycation and biomarkers of vascular complications of diabetes

Propensity to diabetic nephropathy (DN), retinopathy (DR), and cardiovascular disease (CVD) varies between individuals. Current biomarkers such as indicators of glycemia (HbA1c), retinal examinations, and albuminuria, cannot detect early tissue damage. HbAIc also doesn’t reflect most glycative and oxidative chemical pathways that cause complications, and studies of new biomarkers to measure their end-products are needed. This review proposes the study of advanced glycation end products (AGEs) and oxidation end-products (OPs) in long-term diabetes outcome studies. AGEs integrate the activity of glycation pathways that form dicarbonyls, while OPs reflect superoxides, hydroxyl radicals, and peroxides. We discuss using these biomarkers to predict risk of development and progression of DN, DR, and CVD, and to determine if they confer risk independently of the level of HbA1c. We also discuss methods and guidelines to document sample quality in such studies. These studies have the potential to validate unique biomarkers during the early stages of diabetes in those who are at high risk of diabetic complications. Information on basic mechanisms responsible for complications could also stimulate development of therapeutic approaches to delay or arrest them. The ultimate goal is to predict those requiring aggressive therapies during the earliest stages, when prevention or reversal of complications is still possible.     

Stage-specific quantitative changes in renal and urinary proteome during the progression and development of streptozotocin-induced diabetic nephropathy in rats

Diabetic nephropathy (DN) is a microvascular complication associated with diabetes causing slow deterioration of kidneys leading to end-stage renal disease. Timely intervention and diagnosis are crucial in order to ameliorate and halt the progression of DN. Current diagnosis of DN consists of urine assays for detection of microalbuminuria, which have inadequate specificity and sensitivity. Hence, there arises a need to discover stage-specific biomarkers which can aid in the early detection of DN and also in identifying the mechanisms underlying pathogenesis of DN. Therefore the present study was undertaken to identify the differentially expressed proteins in the urine and to examine the pattern of proteomic changes occurring in the rat kidneys during the course of progression of streptozotocin-induced model of DN in rats. Two-dimensional gel electrophoresis coupled to MALDI-TOF mass spectrometry was employed to identify the differentially expressed proteins under diabetic conditions. Among the identified proteins Calgranulin A and Calgranulin B appeared in the urinary proteome at the fourth week of induction of diabetes while we recorded a time-dependent decrease in the expression of major urinary protein (alpha 2u globulin) in the urine as well as kidneys of diabetic rats. Parallel monitoring of targeted proteomic changes in the renal proteome revealed an increase in histone H2B phosphorylation at serine14 along with a gradual decrease in Bcl-2 and MMP-13 expression during the course of progression and development of streptozotocin-induced DN.     

Urinary biomarkers for monitoring disease progression in the Han:SPRD-cy rat model of autosomal-dominant polycystic kidney disease

The Han:SRPD-cy rat is a well-recognized model of human autosomal-dominant polycystic kidney disease. The disease is characterized by the development of progressive renal cysts, leading to declining renal function. Disease progression typically is monitored by measurement of plasma urea concentration. Although plasma urea may be an adequate measure of overall renal function, urinary biomarkers capable of accurately monitoring disease progression may be equally useful. The goal of this study was to assess several urinary biomarkers as potential markers of disease progression in male and female Han:SPRD-cy rats. These biomarkers were compared with changes in plasma urea concentration and morphometric changes as the disease progressed. Urinary activity of N-acetyl-β-D-glucosaminidase and concentration of α-glutathione S-transferase were measured as markers of proximal tubular dysfunction, glutathione S-transferase Yb1 as a distal tubular marker, and collagen IV as a biomarker for glomerular lesions. Urinary albumin was used as biomarker of glomerular or proximal tubular lesions. Albuminuria increased in male rats as the disease progressed, correlating with increasing plasma urea and morphologic changes. Urine concentrations of α-glutathione S-transferase decreased significantly in the male heterozygotic compared with wildtype rats in the later stages of the disease. Urinary concentrations of glutathione S-transferase Yb1 and collagen IV and activity of N-acetyl-β-D-glucosaminidase did not change during disease progression. Measurement of urinary albumin and concentrations of α-glutathione S-transferase may be useful for monitoring disease progression in the male Han:SPRD-cy rat model in future experiments.     

Biosensors for cancer markers diagnosis

Point-of-care diagnostic devices present a viable option for the rapid and sensitive detection and analysis of cancer markers. With the growing number of cancer cases being diagnosed worldwide and the increased number of fatalities due to late disease detection, biosensors can play an important role in the early diagnosis of cancer. Molecular profiles of patients are being increasingly studied using new molecular tools such as genomic and proteomic techniques. These methods combined with bioinformatics tools are generating new data which is being employed in the elucidation of new disease biomarkers. As with many disease conditions finding specific and sensitive markers that are associated with only one type of the disease can be difficult. In addition to this, the level of the biomarkers in biological fluids can vary depending on different disease conditions and stages. A number of molecular markers are therefore usually evaluated for cancer diagnosis and these can include proteins, peptides, over/under expression of gene markers and gene mutations. This review provides an overview of the biosensor technology available today, areas which are currently being developed and researched for cancer markers diagnosis—and a consideration of future prospects for the technology.     

胆管癌诊断的分子生物学标志研究进展

胆管癌(cholangiocarcinoma, CCA)是第二常见的原发性肝脏恶性肿瘤。由于该病症隐匿,早期诊断困难,大多数患者在晚期才被发现且患者预后较差,因此早期诊断成为胆管癌治疗的关键。分子生物学标志可以标记系统、器官、组织、细胞及亚细胞结构及其功能的改变。大量研究表明,某些分子生物学标记物的检测有助于CCA的早期诊断,故本文旨在总结与CCA诊断相关的分子生物学标记物及其研究进展。     

Integrated gene networks in breast cancer development

Breast cancer is a complex and heterogenous disease. Classical molecular medical approaches cannot fully understand and comprehend its pathogenesis. In this review, the development of new biological markers for the early detection and creation of guided and specific therapy of breast cancer are discussed in light of the rapid advances in the “omics”. Results of cancer research in combination with large-scale methods that examine the expression status of genes and proteins have identified a large number of new biomarkers as well as confirmed the human growth hormone as an important player in the pathogenesis of this disease through its autocrine regulation where it influences the activation of Pax5 and HOXA1 gene networks.     

Urinary proteomics: a tool to discover biomarkers of kidney diseases

There is intense interest in applying proteomics to urine analysis in order to promote a better understanding of kidney disease processes, develop new biomarkers for diagnosis and detect early factors that contribute to end-stage renal diseases. This interest creates numerous opportunities as well as challenges. To fulfill this task, proteomics requires, in its different stages of realization, various technological platforms with high sensitivity, high throughput and large automation ability. In this review, we will give an overview of promising proteomic methods that can be used for analyzing urinary proteome and detecting biomarkers for different kidney diseases. Furthermore, we will focus on the current status and future directions in investigating kidney diseases using urinary proteomics.     

Applications of urinary proteomics in biomarker discovery

Urine is an important source of biomarkers. This article reviews current advances, major challenges, and future prospects in the field of urinary proteomics. Because the practical clinical problem is to distinguish diseases with similar symptoms, merely comparing samples from patients of a particular disease to those of healthy individuals is inadequate for finding biomarkers with sufficient diagnostic power. In addition, the variation of expression levels of urinary proteins among healthy individuals and individuals under different physiological conditions adds to the difficulty in identifying biomarkers. We propose that establishing the natural variation in urinary protein expression among a healthy population can serve as a reference to help identify protein abundance changes that are caused by disease, not by individual variations or physiological changes. We also discuss that comparing protein expression levels between urine and plasma may reveal the physiological function of the kidney and that may facilitate biomarker discovery. Finally, we propose that establishing a data-sharing platform for data collection and integrating results from all urinary biomarker studies will help promote the development of urinary proteomics.     

Proteomic Analysis of Urine to Identify Breast Cancer Biomarker Candidates Using a Label-Free LC-MS/MS Approach

Introduction

Breast cancer is a complex heterogeneous disease and is a leading cause of death in women. Early diagnosis and monitoring progression of breast cancer are important for improving prognosis. The aim of this study was to identify protein biomarkers in urine for early screening detection and monitoring invasive breast cancer progression.

Method

We performed a comparative proteomic analysis using ion count relative quantification label free LC-MS/MS analysis of urine from breast cancer patients (n = 20) and healthy control women (n = 20).

Results

Unbiased label free LC-MS/MS-based proteomics was used to provide a profile of abundant proteins in the biological system of breast cancer patients. Data analysis revealed 59 urinary proteins that were significantly different in breast cancer patients compared to the normal control subjects (p<0.05, fold change >3). Thirty-six urinary proteins were exclusively found in specific breast cancer stages, with 24 increasing and 12 decreasing in their abundance. Amongst the 59 significant urinary proteins identified, a list of 13 novel up-regulated proteins were revealed that may be used to detect breast cancer. These include stage specific markers associated with pre-invasive breast cancer in the ductal carcinoma in-situ (DCIS) samples (Leucine LRC36, MAST4 and Uncharacterized protein CI131), early invasive breast cancer (DYH8, HBA, PEPA, uncharacterized protein C4orf14 (CD014), filaggrin and MMRN2) and metastatic breast cancer (AGRIN, NEGR1, FIBA and Keratin KIC10). Preliminary validation of 3 potential markers (ECM1, MAST4 and filaggrin) identified was performed in breast cancer cell lines by Western blotting. One potential marker MAST4 was further validated in human breast cancer tissues as well as individual human breast cancer urine samples with immunohistochemistry and Western blotting, respectively.

Conclusions

Our results indicate that urine is a useful non-invasive source of biomarkers and the profile patterns (biomarkers) identified, have potential for clinical use in the detection of BC. Validation with a larger independent cohort of patients is required in the following study.     

Advances in ovarian cancer proteomics: the quest for biomarkers and improved therapeutic interventions

Epithelial ovarian cancer is the leading cause of cancer-related death among gynecological cancers due to the asymptomatic nature of the disease, a lack of early detection markers and the development of resistance to current chemotherapeutic agents. Currently available tests (CA-125, transvaginal ultrasound or combination of both) lack the sensitivity and specificity to be useful as an efficient screening tool for surveillance of the general population. Thus, there is an urgent need for the development and validation of new molecular markers that would be both specific and sensitive indicators of disease onset, as well as progression. Proteomic profiling has emerged as a powerful tool to study ovarian cancer in an unbiased way at the molecular level, to monitor the effects of given treatment options and for the discovery of biomarkers. In this review we discuss the challenges associated with proteomics-based biomarker discovery and some recent concepts to potentially overcome these hurdles. Recent proteomics work on ovarian cancer cells and tissues will be discussed in light of obtaining new insights into fundamental biological processes, as well as their potential integration with ongoing biomarker discovery pipelines.     

Advances in ovarian cancer proteomics: the quest for biomarkers and improved therapeutic interventions

Epithelial ovarian cancer is the leading cause of cancer-related death among gynecological cancers due to the asymptomatic nature of the disease, a lack of early detection markers and the development of resistance to current chemotherapeutic agents. Currently available tests (CA-125, transvaginal ultrasound or combination of both) lack the sensitivity and specificity to be useful as an efficient screening tool for surveillance of the general population. Thus, there is an urgent need for the development and validation of new molecular markers that would be both specific and sensitive indicators of disease onset, as well as progression. Proteomic profiling has emerged as a powerful tool to study ovarian cancer in an unbiased way at the molecular level, to monitor the effects of given treatment options and for the discovery of biomarkers. In this review we discuss the challenges associated with proteomics-based biomarker discovery and some recent concepts to potentially overcome these hurdles. Recent proteomics work on ovarian cancer cells and tissues will be discussed in light of obtaining new insights into fundamental biological processes, as well as their potential integration with ongoing biomarker discovery pipelines.