bims-prodis 2019-01-27 papers

bims-prodis

Biomed News

on Proteomics in disease

Issue of 2019‒01‒27
twenty-five papers selected by
Nancy Gough
Bioserendipity

Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread.
Development of a Novel Diagnostic Biomarker Set for Rheumatoid Arthritis Using a Proteomics Approach.
Proteomics in kidney allograft transplantation - Application of molecular pathway analysis for kidney allograft disease phenotypic biomarker selection.
Differentially expressed plasma proteins of β-thalassemia/hemoglobin E patients in response to curcuminoids/vitamin E antioxidant cocktails.
Data-Independent Acquisition-Based Quantitative Proteomic Analysis Reveals Differences in Host Immune Response of Peripheral Blood Mononuclear Cells to Sepsis.
Quantitative Proteomics by SWATH-MS of Maternal Plasma Exosomes Determine Pathways Associated with Term and Preterm Birth.
A Strategy for Discovery and Verification of Candidate Biomarkers in Cerebrospinal Fluid of Preclinical Alzheimer's Disease.
Drug repositioning for dengue haemorrhagic fever by integrating multiple omics analyses.
Breast cancer-derived exosomes reflect the cell-of-origin phenotype.
Using a Targeted Proteomics Chip to Explore Pathophysiological Pathways for Incident Diabetes- The Malmö Preventive Project.
Dysregulated fatty acid metabolism in nasal polyp-derived eosinophils from patients with chronic rhinosinusitis.
High-resolution quantitative proteomics applied to the study of the specific protein signature in the sputum and saliva of active tuberculosis patients and their infected and uninfected contacts.
Discovery of Distinct Immune Phenotypes Using Machine Learning in Pulmonary Arterial Hypertension.
Serum protein signature of coronary artery disease in type 2 diabetes mellitus.
Proteomic Approach to the Potential Role of Angiotensin II in Barrett Dysplasia.
Mass spectrometry-based proteomics reveals the distinct nature of the skin proteomes of photoaged compared to intrinsically aged skin.
Is the placental proteome impaired in well-controlled gestational diabetes?
A Proteome Approach Reveals Differences between Fertile Women and Patients with Repeated Implantation Failure on Endometrial Level⁻Does hCG Render the Endometrium of RIF Patients?
The future of myeloma precision medicine: integrating the compendium of known drug resistance mechanisms with emerging tumor profiling technologies.
Integrating clinical, molecular, proteomic and histopathological data within the tissue context: Tissunomics.
Muscular proteomic profiling of deep pressure ulcers reveals myoprotective role of JAK2 in ischemia and reperfusion injury.
Targeting TNFα in severe psoriasis-mass spectrometry reveals a time-dependent specific inhibition of Factor H in responding patients.
Inter-Individual and Regional Variability in Drug Transporter Abundance at the Human Blood-Brain Barrier Measured by Quantitative Targeted Proteomics.
Plasma Peptidome Pattern of Breast Cancer Using Magnetic Beads-Based Plasma Fractionation and MALDI-TOF MS: A Case Control Study in Egypt
Injecting autologous platelet rich plasma solely into the knee joint is not adequate in treating geriatric patients with moderate to severe knee osteoarthritis.

Clin Proteomics. 2019 ;16 3

Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread.

Chandra Kirana, Lifeng Peng, Rose Miller, John P Keating, Corinne Glenn, Hongjun Shi, T William Jordan, Guy J Maddern, Richard S Stubbs.

  Biomarkers are urgently required to support current histological staging to provide additional accuracy in stratifying colorectal cancer (CRC) patients according to risk of spread to properly assign adjuvant chemotherapy after surgery. Chemotherapy is given to patients with stage III to reduce the risk of recurrence but is controversial in stage II patients. Up to 25% of stage II patients will relapse within 5 years after tumor removal and when this occurs cure is seldom possible. The aim of this study was to identify protein biomarkers to stratify risk of spread of CRC patients. Laser micro-dissection was used to isolate cancer cells from primary colorectal tumors of stage II patients which did or did not metastasize within 5 years after surgical resection. Protein expression differences between two groups of tumors were profiled by 2D-DIGE with saturation CyDye labeling and identified using MALDI-TOF mass spectrometry. Evaluation of protein candidates was conducted using tissue micro array (TMA) immunohistochemistry on 125 colorectal tumor tissue samples of different stages. A total of 55 differentially expressed proteins were identified. Ten protein biomarkers were chosen based on p value and ratio between non metastasized and metastazised groups and evaluated on 125 tissues using TMA immunohistochemistry. Expression of HLAB, protein 14-3-3β, LTBP3, ADAMTS2, JAG2 and NME2 on tumour cells was significantly associated with clinical parameters related to tumour progression, invasion and metastasis. Kaplan-Meier survival curve showed strong expression of six proteins was associated with good CRC specific survival. Expression of HLAB, ADAMTS2, LTBP3, JAG2 and NME2 on tumour cells, was associated with tumour progression and invasion, metastasis and CRC specific survival may serve as potential biomarkers to stratify CRC patients into low and high risk of tumour metastasis. Combined methods of laser microdissection, 2D DIGE with saturation labelling and MALDI-TOF MS proved to be resourceful techniques capable of identifying protein biomarkers to predict risk of spread of CRC to liver.

Keywords:  Biomarkers; Colorectal cancer; Liver metastasis; Prognosis; Proteomics

DOI:  https://doi.org/10.1186/s12014-019-9223-7
Biomed Res Int. 2018 ;2018 7490723

Development of a Novel Diagnostic Biomarker Set for Rheumatoid Arthritis Using a Proteomics Approach.

Sora Mun, Jiyeong Lee, Mi-Kyoung Lim, You-Rim Lee, Chunhwa Ihm, Seung Hoon Lee, Hee-Gyoo Kang.

Background: Rheumatoid arthritis (RA) is an autoimmune disease that starts with inflammation of the synovial membrane. Studies have been conducted to develop methods for efficient diagnosis of RA and to identify the mechanisms underlying RA development. Blood samples can be useful for detecting disturbance of homeostasis in patients with RA. Nanoliquid chromatography-tandem mass spectrometry (LC-MS/MS) is an efficient proteomics approach to analyze blood sample and quantify serum proteins.Methods: Serum samples of 18 healthy controls and 18 patients with RA were analyzed by LC-MS/MS. Selected candidate biomarkers were validated by enzyme-linked immunosorbent assay (ELISA) using sera from 43 healthy controls and 44 patients with RA.
Results: Thirty-eight proteins were significantly differentially expressed by more than 2-fold in healthy controls and patients with RA. Based on a literature survey, we selected six candidate RA biomarkers. ELISA was used to evaluate whether these proteins effectively allow distinguishing patients with RA from healthy controls and monitoring drug efficacy. SAA4, gelsolin, and vitamin D-binding protein were validated as potential biomarkers of RA for screening and drug efficacy monitoring of RA.
Conclusions: We identified a panel of three biomarkers for RA which has potential for application in RA diagnosis and drug efficacy monitoring. Further, our findings will aid in understanding the pathogenesis of RA.

DOI: https://doi.org/10.1155/2018/7490723
Proteomics Clin Appl. 2019 Jan 25. e1800091

Proteomics in kidney allograft transplantation - Application of molecular pathway analysis for kidney allograft disease phenotypic biomarker selection.

David Marx, Jochen Metzger, Jérôme Olagne, Iwona Belczacka, Stanislas Faguer, Magali Colombat, Holger Husi, William Mullen, Wilfried Gwinner, Sophie Caillard.

  PURPOSE: There is a need for accurate, robust, non-invasive methods to provide early diagnosis of graft lesions after kidney transplantation. A multitude of proteomic biomarkers for the major kidney allograft disease phenotypes defined by the BANFF classification criteria have been described in literature. None of these biomarkers have been established in the clinic. A key reason for this is the lack of clinical validation which is difficult, as even the gold standard of diagnosis, kidney biopsy, is often ambiguous.EXPERIMENTAL DESIGN: We evaluated semantic clustering by ReviGO on top of transcriptomic pathway analysis to connect histological and transcriptomic kidney allograft disease characteristics with proteomic biomarker qualification.
RESULTS: By using public data generated in microarray studies of kidney allograft tissue, we were able to identify biological processes and key molecules specifically associated with the different kidney allograft disease phenotypes.
CONCLUSIONS AND CLINICAL RELEVANCE: Semantic clustering holds the promise to guide adaptation of proteomic marker panels to molecular pathology. This can support the development of noninvasive tests (e.g. in urine, by capillary electrophoresis mass spectrometry) that simultaneously detect diverse kidney allograft phenotypes with high accuracy and sensitivity. This article is protected by copyright. All rights reserved.

Keywords:  allograft biopsy; biomolecular pathways; kidney transplantation; protein marker selection; proteomics

DOI:  https://doi.org/10.1002/prca.201800091
Hematology. 2019 Dec;24(1): 300-307

Differentially expressed plasma proteins of β-thalassemia/hemoglobin E patients in response to curcuminoids/vitamin E antioxidant cocktails.

Jirawan Panachan, Daranee Chokchaichamnankit, Churat Weeraphan, Chantragan Srisomsap, Patarabutr Masaratana, Suneerat Hatairaktham, Narumol Panichkul, Jisnuson Svasti, Ruchaneekorn W Kalpravidh.

  OBJECTIVE: Iron overload and oxidative stress are the major causes of serious complications and mortality in thalassemic patients. Our previous work supports the synergistic effects of antioxidant cocktails (curcuminoids or vitamin E, N-acetylcysteine, and deferiprone) in treatment of β-thalassemia/Hb E patients. This further 2-DE-based proteomic study aimed to identify the plasma proteins that expressed differentially in response to antioxidant cocktails.METHODS: Frozen plasma samples of ten normal subjects and ten β-thalassemia/Hb E patients at three-time points (baseline, month 6, and month 12) were reduced the dynamic range of proteome using ProteoMiner kit and separated proteins by two-dimensional gel electrophoresis. Differentially expressed proteins were identified using tandem mass spectrometry. Several plasma proteins were validated by ELISA and Western blot analysis.
RESULTS: Thirteen and 11 proteins were identified with altered expression levels in the curcuminoids- and vitamin E cocktail groups, respectively. The associations between vitronectin (VTN) expression and total bilirubin levels, as well as between serum paraoxonase/arylesterase 1 (PON1) expression and blood reactive oxygen species were observed. Validation results were consistent with proteomics results.
DISCUSSION AND CONCLUSIONS: These plasma proteins may provide better understanding of the mechanisms underlying the therapeutic effects of antioxidant cocktails in thalassemic patients.

Keywords:  N-acetylcysteine; antioxidant; curcuminoids; deferiprone; plasma proteomics; thalassemia; vitamin E; β-thalassemia/Hb E

DOI:  https://doi.org/10.1080/16078454.2019.1568354
Scand J Immunol. 2019 Jan 22. e12748

Data-Independent Acquisition-Based Quantitative Proteomic Analysis Reveals Differences in Host Immune Response of Peripheral Blood Mononuclear Cells to Sepsis.

Yiqing Tong, Xin Ku, Chunrong Wu, Jianjun Liu, Chunhui Yang, Wenzhi Tang, Wei Yan, Jianguo Tang.

  This study was aimed to uncover proteins that are differentially expressed in sepsis. Data Independent Acquisition (DIA) was used for analysis to identify differentially expressed proteins in peripheral blood mononuclear cells (PBMCs) of patients. A total of 24 non-septic Intensive Care Unit (ICU) patients, 11 septic shock patients and 27 patients diagnosed with sepsis were recruited for the Mass Spectrometry (MS) discovery. PBMCs were isolated from routine blood samples and digested into peptides. A DIA workflow was developed using a quadrupole-Orbitrap Liquid Chromatography (LC)-MS system, and mass spectra peaks were extracted by Skyline software. Orthogonal partial least squares discriminant analysis (OPLS-DA) and partial least squares discriminant analysis (PLS-DA) were applied to distinguish the patient groups at the level of fragment ion and peptide. Differentially expressed proteins in the patient groups were verified by Enzyme linked immunosorbent assay (ELISA). Receiver operating characteristic (ROC) curves were used to evaluate the protein expression. A total of 1,062 fragment ions and 122 proteins were identified in the MS-DIA analysis conducted by Skyline software. Using gene ontology clustering analysis, we discovered that 51 of the 122 identified proteins were associated with biological processes, including carbon metabolism, biosynthesis of antibiotics, platelet activation, bacterial invasion of epithelial cells and complement, and coagulation cascades. Among them, five proteins (High-mobility group box1 (HMGB1), Matrix metalloproteinase 8 (MMP8), Neutrophil gelatinase-associated lipocalin (NGAL), Lactotransferrin (LTF), and Grancalcin (GCA)) were identified by ELISA as closely related to the development of sepsis. The ROC curves displayed good sensitivity and specificity. This article is protected by copyright. All rights reserved.

Keywords:  Data-Independent Acquisition; Sepsis; peripheral blood mononuclear cells; proteomics

DOI:  https://doi.org/10.1111/sji.12748
Endocrinology. 2019 Jan 21.

Quantitative Proteomics by SWATH-MS of Maternal Plasma Exosomes Determine Pathways Associated with Term and Preterm Birth.

Ramkumar Menon, Christopher Luke Dixon, Samantha Sheller-Miller, Stephen J Fortunato, George R Saade, Carlos Palma, Andrew Lai, Dominic Guanzon, Carlos Salomon.

Introduction: Exosomes are membrane-bound nanovesicles that transport molecular signals between cells. This study determined changes in maternal plasma exosome proteomics contents in term and preterm births.Methods: Maternal plasma (MP) samples were collected from group 1: term not-in labor (TNIL, n=13); group 2: term in labor (TL, n= 11), group 3: preterm premature rupture of membranes (pPROM, n=8); and group 4: preterm birth (PTB, n=13). Exosomes isolated from plasma by differential density centrifugation followed by size exclusion chromatography were characterised by morphology (electron microscopy), quantity and size (nanoparticle tracking analysis and markers (western blot). A quantitative, information-independent acquisition (Sequential Windowed Acquisition of All Theoretical Mass Spectra [SWATH]) approach was used to determine the protein profile in exosomes. Ingenuity Pathway Analysis (IPA) determined pathways associated with the protein profile identified in exosomes.
Results: MP exosomes were spherical, had a mean diameter of 120 nm and positive for exosomal proteins CD63 and TSG101 irrespective of pregnancy status. No significant changes in exosome quantities were seen in maternal circulation across the groups. SWATH identified 72 statistically significant proteins across the groups studied. Bioinformatics analysis showed the proteins within the exosomes in TNIL, TL, pPROM, and PTB target pathways mainly associated with inflammatory and metabolic signals.
Conclusions: Exosomal data suggest that homeostatic imbalances, specifically inflammatory and endocrine signaling might disrupt pregnancy maintenance resulting in labor-related changes both at term and preterm. Reflection of physiologic changes in exosomes is suggestive of its usefulness as biomarkers and cellular function indicators.

DOI: https://doi.org/10.1210/en.2018-00820
Front Mol Neurosci. 2018 ;11 483

A Strategy for Discovery and Verification of Candidate Biomarkers in Cerebrospinal Fluid of Preclinical Alzheimer's Disease.

Xiaofang Zhong, Jingxin Wang, Cynthia Carlsson, Ozioma Okonkwo, Henrik Zetterberg, Lingjun Li.

  Alzheimer's disease (AD), a progressive neurodegenerative disease, is characterized by the accumulation of senile plaques, neurofibrillary tangles, and loss of synapses and neurons in the brain. The pathophysiological process of AD begins with a long asymptomatic phase, which provides a potential opportunity for early therapeutic intervention. Therefore, it is crucial to define putative biomarkers via reliable and validated methods for early diagnosis of AD. Here, we characterized candidate biomarkers by discovery proteomics analysis of cerebrospinal fluid (CSF), revealing that 732 and 704 proteins with more than one unique peptide were identified in healthy controls and preclinical AD patients, respectively. Among them, 79 and 98 proteins were significantly altered in preclinical AD for women and men, respectively, many of which have been demonstrated with consistent regulation pattern in patients with mild cognitive impairment or AD dementia. In-house developed 5-plex isotopic N,N-dimethyl leucine (iDiLeu) tags were further utilized to verify candidate biomarkers, neurosecretory protein VGF (VGF) and apolipoprotein E (apoE). By labeling peptide standards with different iDiLeu tags, a four-point internal calibration curve was constructed to allow for determination of the absolute amount of target analytes in CSF through a single liquid chromatography-mass spectrometry run.

Keywords:  Alzheimer’s disease; biomarker; cerebrospinal fluid; iDiLeu; isotopic labeling for quantitation; label-free quantitation; targeted quantitative proteomics

DOI:  https://doi.org/10.3389/fnmol.2018.00483
Sci Rep. 2019 Jan 24. 9(1): 523

Drug repositioning for dengue haemorrhagic fever by integrating multiple omics analyses.

Takayuki Amemiya, M Michael Gromiha, Katsuhisa Horimoto, Kazuhiko Fukui.

To detect drug candidates for dengue haemorrhagic fever (DHF), we employed a computational drug repositioning method to perform an integrated multiple omics analysis based on transcriptomic, proteomic, and interactomic data. We identified 3,892 significant genes, 389 proteins, and 221 human proteins by transcriptomic analysis, proteomic analysis, and human-dengue virus protein-protein interactions, respectively. The drug candidates were selected using gene expression profiles for inverse drug-disease relationships compared with DHF patients and healthy controls as well as interactomic relationships between the signature proteins and chemical compounds. Integrating the results of the multiple omics analysis, we identified eight candidates for drug repositioning to treat DHF that targeted five proteins (ACTG1, CALR, ERC1, HSPA5, SYNE2) involved in human-dengue virus protein-protein interactions, and the signature proteins in the proteomic analysis mapped to significant pathways. Interestingly, five of these drug candidates, valparoic acid, sirolimus, resveratrol, vorinostat, and Y-27632, have been reported previously as effective treatments for flavivirus-induced diseases. The computational approach using multiple omics data for drug repositioning described in this study can be used effectively to identify novel drug candidates.

DOI: https://doi.org/10.1038/s41598-018-36636-1
Proteomics. 2019 Jan 22. e1800180

Breast cancer-derived exosomes reflect the cell-of-origin phenotype.

Shu Wen Wen, Luize G Lima, Richard J Lobb, Emma L Norris, Marcus L Hastie, Sophie Krumeich, Andreas Möller.

  A manner in which cells can communicate with each other is via secreted nanoparticles termed exosomes. These vesicles contain lipids, nucleic acids and proteins, and are said to reflect the cell-of-origin. However, for the exosomal protein content, there is limited evidence in the literature to verify this statement. Here, we use proteomic assessment combined with pathway-enrichment analysis to demonstrate that the protein cargo of exosomes reflects the epithelial/mesenchymal phenotype of secreting breast cancer cells. Given that epithelial-mesenchymal plasticity is known to implicate various stages of cancer progression, our results suggest that breast cancer subtypes with distinct epithelial and mesenchymal phenotypes may be distinguished by directly assessing the protein content of exosomes. Additionally, our work is a substantial step towards verifying the statement that cell-derived exosomes reflect the phenotype of the cells-of-origin. This article is protected by copyright. All rights reserved.

Keywords:  Breast cancer; epithelial-to-mesenchymal transition; exosomes; proteomic

DOI:  https://doi.org/10.1002/pmic.201800180
Sci Rep. 2019 Jan 22. 9(1): 272

Using a Targeted Proteomics Chip to Explore Pathophysiological Pathways for Incident Diabetes- The Malmö Preventive Project.

John Molvin, Manan Pareek, Amra Jujic, Olle Melander, Lennart Råstam, Ulf Lindblad, Bledar Daka, Margrét Leósdóttir, Peter M Nilsson, Michael H Olsen, Martin Magnusson.

Multiplex proteomic platforms provide excellent tools for investigating associations between multiple proteins and disease (e.g., diabetes) with possible prognostic, diagnostic, and therapeutic implications. In this study our aim was to explore novel pathophysiological pathways by examining 92 proteins and their association with incident diabetes in a population-based cohort (146 cases of diabetes versus 880 controls) followed over 8 years. After adjusting for traditional risk factors, we identified seven proteins associated with incident diabetes. Four proteins (Scavenger receptor cysteine rich type 1 protein M130, Fatty acid binding protein 4, Plasminogen activator inhibitor 1 and Insulin-like growth factor-binding protein 2) with a previously established association with incident diabetes and 3 proteins (Cathepsin D, Galectin-4, Paraoxonase type 3) with a novel association with incident diabetes. Galectin-4, with an increased risk of diabetes, and Paraoxonase type 3, with a decreased risk of diabetes, remained significantly associated with incident diabetes after adjusting for plasma glucose, implying a glucose independent association with diabetes.

DOI: https://doi.org/10.1038/s41598-018-36512-y
Allergy. 2019 Jan 22.

Dysregulated fatty acid metabolism in nasal polyp-derived eosinophils from patients with chronic rhinosinusitis.

Jun Miyata, Koichi Fukunaga, Yusuke Kawashima, Takashi Watanabe, Akina Saitoh, Tomomi Hirosaki, Yasutomo Araki, Toru Kikawada, Tomoko Betsuyaku, Osamu Ohara, Makoto Arita.

  BACKGROUND: Eosinophils are multifunctional granulocytes capable of releasing various cytokines, chemokines, and lipid mediators. We previously reported dysregulated fatty acid metabolism in peripheral blood-derived eosinophils from patients with severe asthma. However, functional characteristics of eosinophils present in allergic inflammatory tissues remains largely uncharacterized.METHODS: We established a method for isolating CD69hi CCR3low CXCR4- siglec-8int eosinophils from nasal polyps of patients with eosinophilic rhinosinusitis (NP-EOS). Multi-omics analysis including lipidomics, proteomics, and transcriptomics was performed to analyze NP-EOS as compared with peripheral blood-derived eosinophils from healthy subjects (PB-EOS).
RESULTS: Lipidomic analysis revealed impaired synthesis of prostaglandins and 15-lipoxygenase (15-LOX)-derived mediators, and selective upregulation of leukotriene D4 production. Furthermore, proteomics and transcriptomics revealed changes in the expression of specific enzymes (GGT5, DPEP2, and 15-LOX) responsible for dysregulated lipid metabolism. Ingenuity pathway analysis indicated the importance of type 2 cytokines and pattern recognition receptor pathways. Stimulation of PB-EOS with eosinophil activators IL-5, GM-CSF, and agonists of TLR2 and NOD2 mimicked the observed changes in lipid metabolism.
CONCLUSION: Inflammatory tissue-derived eosinophils possess a specific phenotype with dysregulated fatty acid metabolism that may be targeted therapeutically to control eosinophilic inflammatory diseases. This article is protected by copyright. All rights reserved.

Keywords:  GGT5; chronic rhinosinusitis; human eosinophil; lipid mediator; multi-omics

DOI:  https://doi.org/10.1111/all.13726
J Proteomics. 2019 Jan 17. pii: S1874-3919(19)30018-1. [Epub ahead of print]

High-resolution quantitative proteomics applied to the study of the specific protein signature in the sputum and saliva of active tuberculosis patients and their infected and uninfected contacts.

Jesús Mateos, Olivia Estévez, África González-Fernández, Luis Anibarro, Ángeles Pallarés, Rajko Reljic, José M Gallardo, Isabel Medina, Mónica Carrera.

  Our goal was to establish panels of protein biomarkers that are characteristic of patients with microbiologically confirmed pulmonary tuberculosis (TB) and their contacts, including latent TB-infected (LTBI) and uninfected patients. Since the first pathogen-host contact occurs in the oral and nasal passages the saliva and sputum were chosen as the biological fluids to be studied. Quantitative shotgun proteomics was performed using a LTQ-Orbitrap-Elite platform. For active TB patients, both fluids exhibited a specific accumulation of proteins that were related to complement activation, inflammation and modulation of immune response. In the saliva of TB patients, a decrease of in proteins related to glucose and lipid metabolism was detected. In contrast, the sputum of uninfected contacts presented a specific proteomic signature that was composed of proteins involved in the perception of bitter taste, defense against pathogens and innate immune response, suggesting that those are key events during the initial entry of the pathogen in the host. SIGNIFICANCE: This is the first study to compare the saliva and sputum from active TB patients and their contacts. Our findings strongly suggest that TB patients show not only an activation of processes that are related to complement activation and modulation of inflammation but also an imbalance in carbohydrate and lipid metabolism. In addition, those individuals who do not get infected after direct exposure to the pathogen display a typical proteomic signature in the sputum, which is a reflection of the secretion from the nasal and oral mucosa, the first immunological barriers that M. tuberculosis encounters in the host. Thus, this result indicates the importance of the processes related to the innate immune response in fighting the initial events of the infection.

Keywords:  Acute-phase response; Biomarker discovery; Innate immune response; Shotgun proteomics; Tuberculosis

DOI:  https://doi.org/10.1016/j.jprot.2019.01.010
Circ Res. 2019 Jan 21.

Discovery of Distinct Immune Phenotypes Using Machine Learning in Pulmonary Arterial Hypertension.

Andrew J Sweatt, Haley K Hedlin, Vidhya Balasubramanian, Andrew Hsi, Lisa K Blum, William H Robinson, Francois Haddad, Peter M Hickey, Robin A Condliffe, Allan Lawrie, Mark R Nicolls, Marlene Rabinovitch, Purvesh Khatri, Roham T Zamanian.

  RATIONALE: Accumulating evidence implicates inflammation in pulmonary arterial hypertension (PAH) and therapies targeting immunity are under investigation, though it remains unknown if distinct immune phenotypes exist.OBJECTIVE: Identify PAH immune phenotypes based on unsupervised analysis of blood proteomic profiles.
METHODS AND RESULTS: In a prospective observational study of Group 1 PAH patients evaluated at Stanford University (discovery cohort, n=281) and University of Sheffield (validation cohort, n=104) between 2008-2014, we measured a circulating proteomic panel of 48 cytokines, chemokines, and factors using multiplex immunoassay. Unsupervised machine learning (consensus clustering) was applied in both cohorts independently to classify patients into proteomic immune clusters, without guidance from clinical features. To identify central proteins in each cluster, we performed partial correlation network analysis. Clinical characteristics and outcomes were subsequently compared across clusters. Four PAH clusters with distinct proteomic immune profiles were identified in the discovery cohort. Cluster 2 (n=109) had low cytokine levels similar to controls. Other clusters had unique sets of upregulated proteins central to immune networks- cluster 1 (n=58)(TRAIL, CCL5, CCL7, CCL4, MIF), cluster 3 (n=77)(IL-12, IL-17, IL-10, IL-7, VEGF), and cluster 4 (n=37)(IL-8, IL-4, PDGF-β, IL-6, CCL11). Demographics, PAH etiologies, comorbidities, and medications were similar across clusters. Non-invasive and hemodynamic surrogates of clinical risk identified cluster 1 as high-risk and cluster 3 as low-risk groups. Five-year transplant-free survival rates were unfavorable for cluster 1 (47.6%, CI 35.4-64.1%) and favorable for cluster 3 (82.4%, CI 72.0-94.3%)(across-cluster p<0.001). Findings were replicated in the validation cohort, where machine learning classified four immune clusters with comparable proteomic, clinical, and prognostic features.
CONCLUSIONS: Blood cytokine profiles distinguish PAH immune phenotypes with differing clinical risk that are independent of World Health Organization Group 1 subtypes. These phenotypes could inform mechanistic studies of disease pathobiology and provide a framework to examine patient responses to emerging therapies targeting immunity.

Keywords:  cytokines and growth factors; machine learning; phenotype; proteomics

DOI:  https://doi.org/10.1161/CIRCRESAHA.118.313911
J Transl Med. 2019 Jan 24. 17(1): 17

Serum protein signature of coronary artery disease in type 2 diabetes mellitus.

Ramu Adela, Podduturu Naveen Chander Reddy, Tarini Shankar Ghosh, Suruchi Aggarwal, Amit Kumar Yadav, Bhabatosh Das, Sanjay K Banerjee.

  BACKGROUND: Coronary artery disease (CAD) is the leading cause of morbidity and mortality in patients with type 2 diabetes mellitus (T2DM). The purpose of the present study was to discriminate the Indian CAD patients with or without T2DM by using multiple pathophysiological biomarkers.METHODS: Using sensitive multiplex protein assays, we assessed 46 protein markers including cytokines/chemokines, metabolic hormones, adipokines and apolipoproteins for evaluating different pathophysiological conditions of control, T2DM, CAD and T2DM with CAD patients (T2DM_CAD). Network analysis was performed to create protein-protein interaction networks by using significantly (p < 0.05) altered protein markers in each disease using STRING 10.5 database. We used two supervised analysis methods i.e., between class analysis (BCA) and principal component analysis (PCA) to reveals distinct biomarkers profiles. Further, random forest classification (RF) was used to classify the diseases by the panel of markers.
RESULTS: Our two supervised analysis methods BCA and PCA revealed a distinct biomarker profiles and high degree of variability in the marker profiles for T2DM_CAD and CAD. Thereafter, the present study identified multiple potential biomarkers to differentiate T2DM, CAD, and T2DM_CAD patients based on their relative abundance in serum. RF classified T2DM based on the abundance patterns of nine markers i.e., IL-1β, GM-CSF, glucagon, PAI-I, rantes, IP-10, resistin, GIP and Apo-B; CAD by 14 markers i.e., resistin, PDGF-BB, PAI-1, lipocalin-2, leptin, IL-13, eotaxin, GM-CSF, Apo-E, ghrelin, adipsin, GIP, Apo-CII and IP-10; and T2DM _CAD by 12 markers i.e., insulin, resistin, PAI-1, adiponectin, lipocalin-2, GM-CSF, adipsin, leptin, Apo-AII, rantes, IL-6 and ghrelin with respect to the control subjects. Using network analysis, we have identified several cellular network proteins like PTPN1, AKT1, INSR, LEPR, IRS1, IRS2, IL1R2, IL6R, PCSK9 and MYD88, which are responsible for regulating inflammation, insulin resistance, and atherosclerosis.
CONCLUSION: We have identified three distinct sets of serum markers for diabetes, CAD and diabetes associated with CAD in Indian patients using nonparametric-based machine learning approach. These multiple marker classifiers may be useful for monitoring progression from a healthy person to T2DM and T2DM to T2DM_CAD. However, these findings need to be further confirmed in the future studies with large number of samples.

Keywords:  Adipokines; Apolipoproteins; Coronary artery diseases; Cytokines/chemokines; Metabolic hormones and biomarkers; Type 2 diabetes mellitus

DOI:  https://doi.org/10.1186/s12967-018-1755-5
Proteomics Clin Appl. 2019 Jan 21. e1800102

Proteomic Approach to the Potential Role of Angiotensin II in Barrett Dysplasia.

Svein Olav Bratlie, Ville Wallenius, Anders Edebo, Lars Fändriks, Anna Casselbrant.

  BACKGROUND AND AIM: Dysplasia in Barrett's esophagus (BE) is regarded as a pre-neoplastic lesion. The renin-angiotensin system (RAS), which is known for its role in electrolyte homeostasis and hemodynamics, has also been shown to have tissue-based features linked to proliferation, inflammation, and cancer. RAS has been found to be associated with BE dysplasia. The aim of this study was to investigate possible effects of the RAS in BE dysplasia by using RAS-interfering pharmaceutical agents and by assessment of global protein expression in esophageal mucosal biopsies.METHODS: Endoscopic biopsies were taken from 18 BE in patients with low-grade dysplasia (LGD) before and after three weeks of treatment with either angiotensin-converting enzyme (ACE) inhibitors (enalapril 5 mg; n = 6) or angiotensin II receptor type 1 (AT1R) blockers (candesartan 8 mg; n = 6), or no-treatment (n = 6). A global proteomics analysis by 2-D gel electrophoresis and mass spectrometry (MS) was then performed to identify proteins that were regulated after interference with RAS. Criteria-based selection of the proteins of particular interest was performed in two steps.
RESULTS: Three proteins were identified to show significant modulation of expression after three weeks of treatment: 60 kDa heat shock protein (downregulated), protein disulphide isomerase A3 (downregulated), and inorganic pyrophosphatase (upregulated).
CONCLUSION: We detected three proteins with no previously known links to esophageal RAS, but with possible relevance for the development of EAC. The fact that expression of these proteins was influenced by interference with the RAS suggests an involvement of AngII in the development of EAC in BE. This article is protected by copyright. All rights reserved.

Keywords:  Barrett's esophagus; cancer; endoscopy; low-grade dysplasia; proteomics; renin-angiotensin system

DOI:  https://doi.org/10.1002/prca.201800102
Int J Cosmet Sci. 2019 Jan 20.

Mass spectrometry-based proteomics reveals the distinct nature of the skin proteomes of photoaged compared to intrinsically aged skin.

V L Newton, I Riba-Garcia, C E M Griffiths, A V Rawlings, R Voegeli, R D Unwin, M J Sherratt, R E B Watson.

  OBJECTIVE: With increasing age skin is subject to alterations in its organisation, which impacts on its function as well as having clinical consequences. Proteomics is a useful tool for non-targeted, semi-quantitative simultaneous investigation of high numbers of proteins. In the current study we utilise proteomics to characterise and contrast age-associated differences in photoexposed and photoprotected skin, with a focus on the epidermis, dermal-epidermal junction and papillary dermis.METHODS: Skin biopsies from buttock (photoprotected) and forearm (photoexposed) of healthy volunteers (aged 18-30 or ≥65 years) were transversely sectioned from the stratum corneum to a depth of 250 μm. Following SDS-PAGE, each sample lane was segmented prior to analysis by liquid chromatography-mass spectrometry/mass spectrometry. Pathways analysis was carried out using Ingenuity IPA.
RESULTS: Comparison of skin proteomes at buttock and forearm sites revealed differences in relative protein abundance. Ageing in skin on the photoexposed forearm resulted in 80% of the altered proteins being increased with age, in contrast to the photoprotected buttock where 74% of altered proteins with age were reduced. Functionally, age-altered proteins in the photoexposed forearm were associated with conferring structure, energy and metabolism. In the photoprotected buttock proteins associated with gene expression, free-radical scavenging, protein synthesis and protein degradation were most frequently altered.
CONCLUSION: This study highlights the necessity of not considering photoageing as an accelerated intrinsic ageing, but as a distinct physiological process. This article is protected by copyright. All rights reserved.

Keywords:  Cell culture; Dermal-epidermal junction; Dermis; Elisa; Epidermis; Genomics; Proteomics; Skin ageing; Skin physiology; Structure

DOI:  https://doi.org/10.1111/ics.12513
J Mass Spectrom. 2019 Jan 24.

Is the placental proteome impaired in well-controlled gestational diabetes?

Silvia Burlina, Cristina Banfi, Maura Brioschi, Silvia Visentin, Maria Grazia Dalfrà, Pietro Traldi, Annunziata Lapolla.

  In pregnancy complicated by gestational diabetes (GDM), the human placenta shows several pathological functional and structural changes, but the extent to which maternal glycemic control contributes to placental abnormalities remains unclear. The aim of this study was to profile and compare the proteome of placentas from healthy pregnant women and those with GDM, to investigate the placenta-specific protein composition and possible changes of its function in presence of GDM. Quantitative proteomic analysis, based on LC-MSE approach, revealed that higher (~15% increase) levels of galectin 1 and collagen alpha-1 XIV chain (although the difference regarding the latter was at the limit of significance) were present in GDM samples, while heat shock 70 kDa protein 1A/1B was less abundant in GDM placental tissue. These data seem to indicate that GDM, when well controlled, did not markedly affect the placental proteome.

Keywords:  gestational diabetes; human placenta; label-free LC-MSE analysis; proteomic analysis

DOI:  https://doi.org/10.1002/jms.4336
Int J Mol Sci. 2019 Jan 19. pii: E425. [Epub ahead of print]20(2):

A Proteome Approach Reveals Differences between Fertile Women and Patients with Repeated Implantation Failure on Endometrial Level⁻Does hCG Render the Endometrium of RIF Patients?

Alexandra P Bielfeld, Sarah Jean Pour, Gereon Poschmann, Kai Stühler, Jan-Steffen Krüssel, Dunja M Baston-Büst.

  BACKGROUND: The molecular signature of endometrial receptivity still remains barely understood, especially when focused on the possible benefit of therapeutical interventions and implantation-related pathologies. Therefore, the protein composition of tissue and isolated primary cells (endometrial stromal cells, ESCs) from endometrial scratchings of ART (Assisted Reproductive Techniques) patients with repeated implantation failure (RIF) was compared to volunteers with proven fertility during the time of embryo implantation (LH + 7). Furthermore, an analysis of the endometrial tissue of fertile women infused with human chorionic gonadotropin (hCG) was conducted.METHODS: Endometrial samples (n = 6 RIF, n = 10 fertile controls) were split into 3 pieces: 1/3 each was frozen in liquid nitrogen, 1/3 fixed in PFA and 1/3 cultured. Protein lysates prepared from fresh frozen tissue were processed for mass spectrometric analysis.
RESULTS: Three proteins (EPPK1, BCLAF1 and PTMA) showed a statistically altered abundance in the endometrial tissue of RIF patients. Furthermore, pathways like metabolism, immune system, ferroptosis and the endoplasmic reticulum were altered in RIF patients. Remarkably, endometrial tissues of RIF patients showed a significantly higher (p-value = 9 × 10-8) protein intensity correlation (Pearson's correlation coefficient = 0.95) compared to fertile women (Pearson's correlation coefficient = 0.88). The in vivo infusion of hCG stimulated proteins of endocytosis, HIF1 signalling and chemokine production. Notably, patients suffering from RIF had a clinical pregnancy rate of 19% after the intrauterine infusion of hCG before embryo transfer (ET) compared to their failed previous cycles.
CONCLUSION: Our study showed for the first time that the endometrial proteome composition of RIF patients differs from fertile controls during the window of implantation. The intrauterine infusion of hCG prior to an embryo transfer might improve the chemokine triggered embryo-endometrial dialogue and intensify the angiogenesis and immune response. From a clinical point of view, the hCG infusion prior to an embryo transfer might increase the pregnancy rate of RIF patients.

Keywords:  embryo transfer; endometrial receptivity; human chorionic gonadotropin; window of implantation

DOI:  https://doi.org/10.3390/ijms20020425
Leukemia. 2019 Jan 25.

The future of myeloma precision medicine: integrating the compendium of known drug resistance mechanisms with emerging tumor profiling technologies.

Taylor Harding, Linda Baughn, Shaji Kumar, Brian Van Ness.

Multiple myeloma (MM) is a hematologic malignancy that is considered mostly incurable in large part due to the inability of standard of care therapies to overcome refractory disease and inevitable drug-resistant relapse. The post-genomic era has been a productive period of discovery where modern sequencing methods have been applied to large MM patient cohorts to modernize our current perception of myeloma pathobiology and establish an appreciation for the vast heterogeneity that exists between and within MM patients. Numerous pre-clinical studies conducted in the last two decades have unveiled a compendium of mechanisms by which malignant plasma cells can escape standard therapies, many of which have potentially quantifiable biomarkers. Exhaustive pre-clinical efforts have evaluated countless putative anti-MM therapeutic agents and many of these have begun to enter clinical trial evaluation. While the palette of available anti-MM therapies is continuing to expand it is also clear that malignant plasma cells still have mechanistic avenues by which they can evade even the most promising new therapies. It is therefore becoming increasingly clear that there is an outstanding need to develop and employ precision medicine strategies in MM management that harness emerging tumor profiling technologies to identify biomarkers that predict efficacy or resistance within an individual's sub-clonally heterogeneous tumor. In this review we present an updated overview of broad classes of therapeutic resistance mechanisms and describe selected examples of putative biomarkers. We also outline several emerging tumor profiling technologies that have the potential to accurately quantify biomarkers for therapeutic sensitivity and resistance at genomic, transcriptomic and proteomic levels. Finally, we comment on the future of implementation for precision medicine strategies in MM and the clear need for a paradigm shift in clinical trial design and disease management.

DOI: https://doi.org/10.1038/s41375-018-0362-z
Histopathology. 2019 Jan 22.

Integrating clinical, molecular, proteomic and histopathological data within the tissue context: Tissunomics.

S Ramón Y Cajal, S Hümmer, V Peg, X Matias Guiu, I De Torres, J Castellvi, E Martinez-Saez, J Hernandez-Losa.

  Malignant tumors show a marked degree of morphological, molecular and proteomic heterogeneity. This variability is closely related to microenvironmental factors and the location of the tumor. The activation of genetic alterations is very tissue dependent and only few tumors have distinct genetic alterations. Importantly, the activation state of proteins and signaling factors is heterogeneous in the primary tumor and in metastases and recurrences. The molecular diagnosis based only in genetic alterations can drive to treatments with unpredictable responses depending on the tumor location such as the tumor response in melanomas vs. colon carcinomas with BRAF mutations. Therefore, we understand that the correct evaluation of tumors requires a system that integrates both morphological, molecular and protein information in a clinical and pathological context, where intratumoral heterogeneity can be assessed. Then, we propose the term tissunomics where the diagnosis will be contextualized in each tumor based on the complementation of the pathological, molecular, protein expression, environmental cells and clinical data. 1.General background of tumour pathology. 2.Current situation and challenges. Why are malignant tumours heterogeneous?. Some oncogenic alterations are tissue-specific. Some histopathological features suggest oncogenic alterations. 3.Problems and barriers. Bias and limitations of molecular classification and somatic theory. Gene expression heterogeneity and environmental cells. 4.The solution: to integrate clinical, radiological, molecular and expression data in a tissue context. 5.Conclusions. This article is protected by copyright. All rights reserved.

Keywords:  Tumour heterogeneity; genomics; histopathology; proteomics

DOI:  https://doi.org/10.1111/his.13828
Am J Transl Res. 2018 ;10(11): 3413-3429

Muscular proteomic profiling of deep pressure ulcers reveals myoprotective role of JAK2 in ischemia and reperfusion injury.

Zan Liu, Licheng Ren, Xu Cui, Le Guo, Bimei Jiang, Jie Zhou, Pengfei Liang, Jizhang Zeng, Zhiyou He, Pihong Zhang.

Pressure ulcers (PUs) are a complex and serious clinical problem. Deep tissue injury (DTI) is either the outcome or the trigger of deep PUs. However, the cellular and molecular mechanisms that contribute to the pathogenesis of deep PUs remain unclear. In this study, the degeneration characteristics and increased autophagy and apoptosis were observed in deep PU muscle tissues. Muscular proteome of deep PU revealed that a total of 520 proteins were differentially expressed, particularly, JAK2 was down-regulated. Intriguingly, expression of JAK2 in C2C12 myoblasts exposed to oxygen-glucose deprivation and reoxygenation (OGD/R) insult was also distinctly reduced. Ex vivo, we transfected C2C12 myoblasts with lentivirus carrying the JAK2 plasmid and found that JAK2-overexpressed myoblasts exhibited a decrease in autophagy and apoptosis after OGD/R treatment, as well as less cell death. Finally, Western blot analysis determined that p-JAK2, p-AKT, p-mTOR and p-ERK1/2 levels were significantly elevated, accompanied by JAK2 overexpression but without p-STAT3, and inhibition of the AKT and ERK1/2 pathway resulted in elevated apoptosis and/or autophagy. These results demonstrated that JAK2 may play an important protective role in muscular ischemia and reperfusion injury during DTI development by inhibition of autophagy and apoptosis through the AKT and ERK1/2 pathways.

Keywords: Deep pressure ulcers; JAK2; apoptosis; autophagy; ischemia/reperfusion injury
Am J Transl Res. 2018 ;10(12): 4338-4349

Targeting TNFα in severe psoriasis-mass spectrometry reveals a time-dependent specific inhibition of Factor H in responding patients.

Patrice Morlière, Joana F Antunes, Virginie Nourry, Francisco M Amado, Pedro M Domingues, Ana C Guerreiro, João A Ferreira, João N Maia-Silva, Jean-Claude Mazière, René C Santus, Paulo L Filipe.

Drugs targeting TNFα (eg, Etanercept®) provide effective control of severe psoriasis. In absence of validated biological parameters of inflammation in psoriasis most decisions on therapeutics have relied mostly on clinical criteria, namely the "Psoriasis Area and Severity Index" (PASI). The purpose of this study was to assess by mass spectrometry alterations in concentrations of serum proteins that specifically correlated with effectiveness of Etanercept treatment. This prospective study enrolled 10 patients suffering from moderate to severe psoriasis (PASI score > 10 and < 17) and treated with Etanercept over a period of 24 weeks; 10 healthy, age-matched volunteers provided controls. Serum proteins sensitive to Etanercept treatment were identified using SELDI-TOF (surface-enhanced laser desorption and ionization - time of flight) coupled to nano LC-ESI/MS (nano liquid chromatography-electrospray ionization/tandem mass spectrometry) technologies. For comparisons between groups of individuals p-values (considered significant when < 0.01) were estimated with non-parametric tests, namely Mann-Whitney (for unpaired data) and Wilcoxon signed-rank (for paired data). In responding patients it could be shown using SELDI-TOF spectrometry that two proteins (134 kDa and 4.3 kDa) return to control levels by 24 weeks of treatment. Using nano LC-ESI/MS the 134 kDa species was identified as complement Factor H. These observations deserve further analyses utilizing larger cohorts of patients. Determination of Factor H levels may become a complementary tool to follow remission or predict the onset of relapse in the follow-up of patients under treatment with Etanercept.

Keywords: Etanercept®; TNFα; complement factor H; mass spectrometry; psoriasis; serum
Clin Pharmacol Ther. 2019 Jan 23.

Inter-Individual and Regional Variability in Drug Transporter Abundance at the Human Blood-Brain Barrier Measured by Quantitative Targeted Proteomics.

Sarah Billington, Laurent Salphati, Cornelis E C A Hop, Xiaoyan Chu, Raymond Evers, Doug Burdette, Christopher Rowbottom, Yurong Lai, Guangqing Xiao, W Griffith Humphreys, Tot Bui Nguyen, Bhagwat Prasad, Jashvant D Unadkat.

  For in-vitro to in-vivo extrapolation (IVIVE) of brain distribution of drugs that are transported at the human blood-brain barrier (BBB), it's important to quantify the inter-individual and regional variability of drug transporter abundance at this barrier. Therefore, using quantitative targeted proteomics, we compared the abundance of ABC and SLC transporters in brain microvascular endothelial cells (BMEC) isolated from post-mortem specimens of two matched brain regions, the occipital (BA17) and parietal (BA39) lobe, from 30 adults. Of the quantifiable transporters, the abundance ranked: GLUT1>BCRP>P-gp>ENT1>OATP2B1. The abundance of MRP1/2/3/4, OATP1A2, OAT3, OCT1/2, OCTN1/2, or ENT2 was below the limit of quantification. Transporter abundance per g of tissue (scaled using GLUT1 abundance in BMEC vs. brain homogenate) in BA17 was 30-42% higher than BA39. The inter-individual variability in transporter abundance (%CV) was 35-57% (BA17) and 27-46% (BA39). These data can be used in proteomics-informed bottom-up IVIVE to predict human brain drug distribution. This article is protected by copyright. All rights reserved.

Keywords:  blood-brain barrier; in-vitro to in-vivo correlation; proteomics; transporters

DOI:  https://doi.org/10.1002/cpt.1373
Asian Pac J Cancer Prev. 2019 Jan 25. 20(1): 175-184

Plasma Peptidome Pattern of Breast Cancer Using Magnetic Beads-Based Plasma Fractionation and MALDI-TOF MS: A Case Control Study in Egypt

Ahmad Zaki, Ragaa Abdelkader Ramadan, Pacint Moez, Hala Ghareeb, Ahmad Elkarmouty.

Objective: The present study aimed to determine peptidome patterns in breast cancer (BC). Methods: We analyzed the plasma proteomic proﬁling of 80 BC patients and 50 healthy controls, using hydrophobic interaction chromatography magnetic beads (MB-HIC8) separation followed by Matrix assisted laser desorption ionization/ time of flight mass spectrometry (MALDI-TOF MS). Results: ClinProTools software identiﬁed 92 peaks that differed among the analyzed groups, 33 peaks were signiﬁcantly different (P < 0.05). Of those, 22 peaks were up-regulated while 11 peaks were down-regulated in BC patients compared with the healthy controls. Three peptide ion signatures (m/z 1,570.31, 1,897.4 and 2,568.17) were provided by the Quick Classifier model to discriminate BC patients from healthy control subjects with 96.4% accuracy. External validation was performed by an independent group and this achieved a sensitivity of 100% and a specificity of 76.9%. Conclusion: MALDI-TOF MS has good analytical performance in distinguishing BC patients from healthy controls.

Keywords: Matrix assisted laser desorption ionization/ time of flight mass spectrometry; magnetic beads; ClinProTools
Exp Gerontol. 2019 Jan 19. pii: S0531-5565(18)30517-5. [Epub ahead of print]119 1-6

Injecting autologous platelet rich plasma solely into the knee joint is not adequate in treating geriatric patients with moderate to severe knee osteoarthritis.

Carl P C Chen, Jean-Lon Chen, Chih-Chin Hsu, Yu-Cheng Pei, Wei-Han Chang, Hsueh-Chih Lu.

Knee pain caused by osteoarthritis (OA) is commonly seen in geriatric patients. Patients with knee OA are often complicated with joint pain, soreness, and weakness. The injection of autologous platelet rich plasma (aPRP) has been proven to be effective in treating mild knee OA. The effect of injecting aPRP in treating moderate to severe degrees of knee OA remains controversial. This study aimed to evaluate the effectiveness of aPRP in treating patients with at least grade 2 on the Kellgren and Lawrence system for the classification of knee OA using a proteomic approach and clinical evaluation tool of Lequesne index. Musculoskeletal ultrasound was used for accurate needle placement into the knee joint, and to the perimeniscal soft tissue for the injection of aPRP. Three monthly aPRP injections were performed. Group 1 patients received intra-articular (IA) injection only, while group 2 received simultaneous IA and pes anserinus aPRP injections. After two monthly aPRP injections, both groups revealed significant drops in average SF total protein concentrations, and increases in the protein concentrations associated with chelation and anti-aging (eg/transthyretin, matrilin, and complement). However, it is group 2 that revealed significant decreases in the protein concentrations associated with inflammation (eg/immunoglobulin and apolipoprotein), and improved knee functional status. SF appeared to become less susceptible to degeneration after aPRP injections in group 2. As a result, at least 2 monthly injection of IA aPRP in conjunction with accurate injection of aPRP to the perimeniscal soft tissue structure such as the pes anserinus may be a viable option in treating patients with moderate to severe degrees of knee OA.

DOI: https://doi.org/10.1016/j.exger.2019.01.018