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fibroblast secretome analysis

Jake Cosme, Hongbo Guo, Sina Hadipour-Lakmehsari, Andrew Emili, Anthony O Gramolini
Cardiac fibroblasts (CF) represent a major subpopulation of cells in the developing and adult heart. Cardiomyocyte (CM) and CF intercellular communication occurs through paracrine interactions and modulate myocyte development and stress response. Detailed proteomic analysis of the CF secretome in normal and stressed conditions may offer insights into the role of CF in heart development and disease. Primary neonatal mouse CF were isolated and cultured for 24h in 21% (normoxic) or 2% (hypoxic) O2. Conditioned media was separated to obtain EXO and EXO-depleted secretome fractions...
June 22, 2017: Journal of Proteome Research
Jennifer Schulze, Odett Kaiser, Gerrit Paasche, Hans Lamm, Andreas Pich, Andrea Hoffmann, Thomas Lenarz, Athanasia Warnecke
Hyperbaric oxygen therapy (HBOT) is a noninvasive widely applied treatment that increases the oxygen pressure in tissues. In cochlear implant (CI) research, intracochlear application of neurotrophic factors (NTFs) is able to improve survival of spiral ganglion neurons (SGN) after deafness. Cell-based delivery of NTFs such as brain-derived neurotrophic factor (BDNF) may be realized by cell-coating of the surface of the CI electrode. Human mesenchymal stem cells (MSC) secrete a variety of different neurotrophic factors and may be used for the development of a biohybrid electrode in order to release endogenously-derived neuroprotective factors for the protection of residual SGN and for a guided outgrowth of dendrites in the direction of the CI electrode...
2017: PloS One
Massimiliano Gaetani, Cinzia Maria Chinnici, Anna Paola Carreca, Claudia Di Pasquale, Giandomenico Amico, Pier Giulio Conaldi
Scarless wound healing and functional regeneration are typical processes of the fetus, gradually lost during post-natal life, and maximally attributed to fetal skin tissue and induced by fetal skin fibroblasts. The latter have been successfully applied to post-natal wounds, with clear advantages compared with autologous dermis grafts or adult fibroblast applications. Our goal was to functionally identify and uncover key factors and mechanisms through the analysis of secretomes, the principal players in all cell therapies based on mesenchymal stromal cells (MSCs)...
January 19, 2017: Journal of Tissue Engineering and Regenerative Medicine
Neri Minsky, Robert G Roeder
Secreted proteins serve pivotal roles in the development of multicellular organisms, acting as structural matrix, extracellular enzymes, and signal molecules. However, how the secretome is regulated remains incompletely understood. Here we demonstrate, unexpectedly, that peroxisome proliferator-activated receptor γ coactivator 1-α (PGC-1α), a critical transcriptional co-activator of metabolic gene expression, functions to down-regulate the expression of diverse genes encoding secreted molecules and extracellular matrix components to modulate the secretome...
January 6, 2017: Journal of Biological Chemistry
G M Sizemore, S Balakrishnan, A M Hammer, K A Thies, A J Trimboli, J A Wallace, S T Sizemore, R D Kladney, S A Woelke, L Yu, S A Fernandez, A Chakravarti, G Leone, M C Ostrowski
Fibroblasts within the mammary tumor microenvironment are active participants in carcinogenesis mediating both tumor initiation and progression. Our group has previously demonstrated that genetic loss of phosphatase and tensin homolog (PTEN) in mammary fibroblasts induces an oncogenic secretome that remodels the extracellular milieu accelerating ErbB2-driven mammary tumor progression. While these prior studies highlighted a tumor suppressive role for stromal PTEN, how the adjacent normal epithelium transforms in response to PTEN loss was not previously addressed...
April 20, 2017: Oncogene
Aihua Hou, Kai Pong Law, Min Qi Tin, Yoon Pin Lim, Louis Tong
Pterygium is a triangular shaped ocular fibrous surface lesion growing from conjunctiva towards central cornea, causing ocular irritation, astigmatism, and visual disturbance. The condition is characterized by epithelial proliferation, fibrovascular growth, chronic inflammation, and prominent extracellular matrix remodeling. Studies have suggested that aberrant extracellular proteins secreted by fibroblasts lead to abnormal matrix production and tissue invasion contributing to the development of the disease...
December 2016: Experimental Eye Research
Lucia Berti, Sonja Hartwig, Martin Irmler, Bernhard Rädle, Dorothea Siegel-Axel, Johannes Beckers, Stefan Lehr, Hadi Al-Hasani, Hans-Ulrich Häring, Martin Hrabě de Angelis, Harald Staiger
CONTEXT: Perivascular adipose tissue (PVAT) is suggested to impact on vascular cells via humoral factors, possibly contributing to endothelial dysfunction and atherosclerosis. OBJECTIVE: To address whether the hepatokine fibroblast growth factor (FGF) 21 affects the PVAT secretome. METHODS: Human perivascular (pre)adipocytes were subjected to targeted proteomics and whole-genome gene expression analysis. RESULTS: Preadipocytes, as compared to adipocytes, secreted higher amounts of inflammatory cytokines and chemokines...
December 2016: Archives of Physiology and Biochemistry
Paul Bălănescu, Anca Lădaru, Eugenia Bălănescu, Valer Pompilian, Daniela Gologanu, Simona Caraiola, Cristian Băicuş, Gheorghe A Dan
BACKGROUND: Proteomic candidate biomarkers for systemic sclerosis (Ssc) useful for appropriate patient evaluation and follow-up were identified in mass-spectrometry studies; however, most of these biomarkers were not evaluated and confirmed on independent patient samples. Up-regulation of reticulocalbin 1 (RCN1) and reticulocalbin 3 (RCN3) in the dermal fibroblast secretome originating from Ssc patients was previously described. The aim of the study was to evaluate circulating RCN1 and RCN3 as candidate biomarkers for Ssc clinical expression...
2016: Clinical Laboratory
M Strobel, H Pförtner, L Tuchscherr, U Völker, F Schmidt, N Kramko, H-J Schnittler, M J Fraunholz, B Löffler, G Peters, S Niemann
Host cell invasion is a major feature of Staphylococcus aureus and contributes to infection development. The intracellular metabolically active bacteria can induce host cell activation and death but they can also persist for long time periods. In this study a comparative analysis was performed of different well-characterized S. aureus strains in their interaction with a variety of host cell types. Staphylococcus aureus (strains 6850, USA300, LS1, SH1000, Cowan1) invasion was compared in different human cell types (epithelial and endothelial cells, keratinocytes, fibroblasts, osteoblasts)...
September 2016: Clinical Microbiology and Infection
Lusine Aghajanova, Signe Altmäe, Sergo Kasvandik, Andres Salumets, Anneli Stavreus-Evers, Linda C Giudice
OBJECTIVE: To determine expression of stanniocalcin-1 (STC1) in human endometrium with and without endometriosis and its regulation by steroid hormones. DESIGN: Laboratory study. SETTING: University. PATIENT(S): Nineteen women with endometriosis and 33 control women. INTERVENTION(S): Endometrial biopsy and fluid sampling. MAIN OUTCOME MEASURE(S): Analysis of early secretory (ESE) and midsecretory (MSE) endometrial secretomes from fertile women with the use of nano-liquid chromatography-dual mass spectrometry; real-time quantitative polymerase chain reaction, and immunohistochemistry for STC1 and its receptor calcium-sensing receptor (CASR) mRNA and proteins in endometrium with and without endometriosis; evaluation of STC1 and CASR mRNA expression in endometrial stromal fibroblasts (eSF) from women with and without endometriosis decidualized with the use of E2P or 8-bromo-cyclic adenosine monophosphate (cAMP)...
September 1, 2016: Fertility and Sterility
(no author information available yet)
[This corrects the article on p. 70 in vol. 10, PMID: 27064649.].
2016: Frontiers in Cellular Neuroscience
Bastien Paré, Lydia T Deschênes, Roxane Pouliot, Nicolas Dupré, Francois Gros-Louis
The proteins secreted by a particular type of cell, the secretome, play important roles in the regulation of many physiological processes via paracrine/autocrine mechanisms, and they are of increasing interest to help understanding rare diseases and to identify potential biomarkers and therapeutic targets. To facilitate ongoing research involving secreted proteins, we revisited cell culture protocols and whole secreted protein enrichment protocols. A reliable method for culturing and precipitating secreted protein from patient-derived fibroblast conditioned-medium was established...
2016: Frontiers in Cellular Neuroscience
Takayuki Ishige, Motoi Nishimura, Mamoru Satoh, Mai Fujimoto, Masaki Fukuyo, Toshihisa Semba, Sayaka Kado, Sachio Tsuchida, Setsu Sawai, Kazuyuki Matsushita, Akira Togawa, Hisahiro Matsubara, Atsushi Kaneda, Fumio Nomura
Gastric cancer is classified into two subtypes, diffuse and intestinal. The diffuse-type gastric cancer (DGC) has poorer prognosis, and the molecular pathology is not yet fully understood. The purpose of this study was to identify functional secreted molecules involved in DGC progression. We integrated the secretomics of six gastric cancer cell lines and gene expression analysis of gastric cancer tissues with publicly available microarray data. Hierarchical clustering revealed characteristic gene expression differences between diffuse- and intestinal-types...
February 19, 2016: Scientific Reports
Elizabete Bagordakis, Iris Sawazaki-Calone, Carolina Carneiro Soares Macedo, Carolina M Carnielli, Carine Ervolino de Oliveira, Priscila Campioni Rodrigues, Ana Lucia C A Rangel, Jean Nunes Dos Santos, Juha Risteli, Edgard Graner, Tuula Salo, Adriana Franco Paes Leme, Ricardo D Coletta
An important role has been attributed to cancer-associated fibroblasts (CAFs) in the tumorigenesis of oral squamous cell carcinoma (OSCC), the most common tumor of the oral cavity. Previous studies demonstrated that CAF-secreted molecules promote the proliferation and invasion of OSCC cells, inducing a more aggressive phenotype. In this study, we searched for differences in the secretome of CAFs and normal oral fibroblasts (NOF) using mass spectrometry-based proteomics and biological network analysis. Comparison of the secretome profiles revealed that upregulated proteins involved mainly in extracellular matrix organization and disassembly and collagen metabolism...
July 2016: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
M M Koczorowska, S Tholen, F Bucher, L Lutz, J N Kizhakkedathu, O De Wever, U F Wellner, M L Biniossek, A Stahl, S Lassmann, O Schilling
Cancer associated fibroblasts (CAFs) constitute an abundant stromal component of most solid tumors. Fibroblast activation protein (FAP) α is a cell surface protease that is expressed by CAFs. We corroborate this expression profile by immunohistochemical analysis of colorectal cancer specimens. To better understand the tumor-contextual role of FAPα, we investigate how FAPα shapes functional and proteomic features of CAFs using loss- and gain-of function cellular model systems. FAPα activity has a strong impact on the secreted CAF proteome ("secretome"), including reduced levels of anti-angiogenic factors, elevated levels of transforming growth factor (TGF) β, and an impact on matrix processing enzymes...
January 2016: Molecular Oncology
Sun-Xia Chen, Xiao-En Xu, Xiao-Qing Wang, Shu-Jian Cui, Lei-Lei Xu, Ying-Hua Jiang, Yang Zhang, Hai-Bo Yan, Qian Zhang, Jie Qiao, Peng-Yuan Yang, Feng Liu
The tumor cell proliferation, migration and invasion were influenced by the interaction between the cancer cells and their microenvironment. In current study, we established two pairs of the primary fibroblast cultures from colorectal adenocarcinoma tissues and the normal counterparts and identified 227 proteins in the colonic fibroblast secretomes; half of these proteins were novel. The mass spectrometry data and analyzed results presented here provide novel insights into the molecular characteristics and modulatory role of colon cancer associated fibroblasts...
December 2014: Data in Brief
Radu Albulescu, Cristiana Tanase, Elena Codrici, Daniela I Popescu, Sanda M Cretoiu, Laurentiu M Popescu
Telocytes (TCs) are interstitial cells that are present in numerous organs, including the heart interstitial space and cardiac stem cell niche. TCs are completely different from fibroblasts. TCs release extracellular vesicles that may interact with cardiac stem cells (CSCs) via paracrine effects. Data on the secretory profile of TCs and the bidirectional shuttle vesicular signalling mechanism between TCs and CSCs are scarce. We aimed to characterize and understand the in vitro effect of the TC secretome on CSC fate...
August 2015: Journal of Cellular and Molecular Medicine
Pascal Schlage, Tobias Kockmann, Jayachandran N Kizhakkedathu, Ulrich auf dem Keller
Secreted proteases act on interstitial tissue secretomes released from multiple cell types. Thus, substrate proteins might be part of higher molecular complexes constituted by many proteins with diverse and potentially unknown cellular origin. In cell culture, these may be reconstituted by mixing native secretomes from different cell types prior to incubation with a test protease. Although current degradomics techniques could identify novel substrate proteins in these complexes, all information on the cellular origin is lost...
July 2015: Proteomics
Daniel M Waldera Lupa, Faiza Kalfalah, Kai Safferling, Petra Boukamp, Gereon Poschmann, Elena Volpi, Christine Götz-Rösch, Francoise Bernerd, Laura Haag, Ulrike Huebenthal, Ellen Fritsche, Fritz Boege, Niels Grabe, Julia Tigges, Kai Stühler, Jean Krutmann
Most molecular hallmarks of cellular senescence have been identified in studies of cells aged in vitro by driving them into replicative or stress-induced senescence. Comparatively, less is known about the characteristic features of cells that have aged in vivo. Here we provide a systematic molecular analysis of normal human dermal fibroblasts (NHDFs) that were isolated from intrinsically aged human skin of young versus middle aged versus old donors. Intrinsically aged NHDFs in culture exhibited more frequently nuclear foci positive for p53 binding protein 1 and promyelocytic leukemia protein reminiscent of 'DNA segments with chromatin alterations reinforcing senescence (DNA-SCARS)'...
August 2015: Journal of Investigative Dermatology
Pramod Kumar, Abhigyan Satyam, Xingliang Fan, Yury Rochev, Brian J Rodriguez, Alexander Gorelov, Lokesh Joshi, Michael Raghunath, Abhay Pandit, Dimitrios I Zeugolis
Tissue engineering by self-assembly uses the cells' secretome as a regeneration template and biological factory of trophic factors. Despite the several advantages that have been witnessed in preclinical and clinical settings, the major obstacle for wide acceptance of this technology remains the tardy extracellular matrix formation. In this study, we assessed the influence of macromolecular crowding (MMC)/excluding volume effect, a biophysical phenomenon that accelerates thermodynamic activities and biological processes by several orders of magnitude, in human corneal fibroblast (HCF) culture...
July 2015: Tissue Engineering. Part C, Methods
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