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Article ; Online: Growth factor dependent changes in nanoscale architecture of focal adhesions.

Legerstee, Karin / Abraham, Tsion E / van Cappellen, Wiggert A / Nigg, Alex L / Slotman, Johan A / Houtsmuller, Adriaan B

Scientific reports

2021 Volume 11, Issue 1, Page(s) 2315

Abstract: Focal adhesions (FAs) are flat elongated structures that mediate cell migration and link the cytoskeleton to the extracellular matrix. Along the vertical axis FAs were shown to be composed of three layers. We used structured illumination microscopy to ... ...

Abstract	Focal adhesions (FAs) are flat elongated structures that mediate cell migration and link the cytoskeleton to the extracellular matrix. Along the vertical axis FAs were shown to be composed of three layers. We used structured illumination microscopy to examine the longitudinal distribution of four hallmark FA proteins, which we also used as markers for these layers. At the FA ends pointing towards the adherent membrane edge (heads), bottom layer protein paxillin protruded, while at the opposite ends (tails) intermediate layer protein vinculin and top layer proteins zyxin and VASP extended further. At the tail tips, only intermediate layer protein vinculin protruded. Importantly, head and tail compositions were altered during HGF-induced scattering with paxillin heads being shorter and zyxin tails longer. Additionally, FAs at protruding or retracting membrane edges had longer paxillin heads than FAs at static edges. These data suggest that redistribution of FA-proteins with respect to each other along FAs is involved in cell movement.
MeSH term(s)	Animals ; Cell Movement/immunology ; Cell Movement/physiology ; Cytoskeleton/genetics ; Cytoskeleton/metabolism ; Focal Adhesions/genetics ; Focal Adhesions/physiology ; Immunity, Cellular/genetics ; Immunity, Cellular/physiology ; Paxillin/genetics ; Paxillin/metabolism ; Vinculin/genetics ; Vinculin/metabolism ; Zyxin/genetics ; Zyxin/metabolism
Chemical Substances	Paxillin ; Zyxin ; Vinculin (125361-02-6)
Language	English
Publishing date	2021-01-27
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-81898-x
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Article ; Online: Compartmentalization of androgen receptors at endogenous genes in living cells.

Yavuz, Selçuk / Kabbech, Hélène / van Staalduinen, Jente / Linder, Simon / van Cappellen, Wiggert A / Nigg, Alex L / Abraham, Tsion E / Slotman, Johan A / Quevedo, Marti / Poot, Raymond A / Zwart, Wilbert / van Royen, Martin E / Grosveld, Frank G / Smal, Ihor / Houtsmuller, Adriaan B

Nucleic acids research

2023 Volume 51, Issue 20, Page(s) 10992–11009

Abstract: A wide range of nuclear proteins are involved in the spatio-temporal organization of the genome through diverse biological processes such as gene transcription and DNA replication. Upon stimulation by testosterone and translocation to the nucleus, ... ...

Abstract	A wide range of nuclear proteins are involved in the spatio-temporal organization of the genome through diverse biological processes such as gene transcription and DNA replication. Upon stimulation by testosterone and translocation to the nucleus, multiple androgen receptors (ARs) accumulate in microscopically discernable foci which are irregularly distributed in the nucleus. Here, we investigated the formation and physical nature of these foci, by combining novel fluorescent labeling techniques to visualize a defined chromatin locus of AR-regulated genes-PTPRN2 or BANP-simultaneously with either AR foci or individual AR molecules. Quantitative colocalization analysis showed evidence of AR foci formation induced by R1881 at both PTPRN2 and BANP loci. Furthermore, single-particle tracking (SPT) revealed three distinct subdiffusive fractional Brownian motion (fBm) states: immobilized ARs were observed near the labeled genes likely as a consequence of DNA-binding, while the intermediate confined state showed a similar spatial behavior but with larger displacements, suggesting compartmentalization by liquid-liquid phase separation (LLPS), while freely mobile ARs were diffusing in the nuclear environment. All together, we show for the first time in living cells the presence of AR-regulated genes in AR foci.
MeSH term(s)	Animals ; Cell Nucleus/genetics ; Cell Nucleus/metabolism ; Nuclear Proteins/metabolism ; Receptors, Androgen/metabolism ; Humans ; Mice ; Cell Line, Tumor
Chemical Substances	Nuclear Proteins ; Receptors, Androgen
Language	English
Publishing date	2023-10-04
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	186809-3
ISSN	1362-4962 ; 1362-4954 ; 0301-5610 ; 0305-1048
ISSN (online)	1362-4962 ; 1362-4954
ISSN	0301-5610 ; 0305-1048
DOI	10.1093/nar/gkad803
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Article: Identifying Molecular Changes in Early Cervical Cancer Samples of Patients That Developed Metastasis.

de Geus, Vera / Ewing-Graham, Patricia C / de Koning, Willem / de Koning, Maurits N C / van den Bosch, Thierry P P / Nigg, Alex L / van Eijck, Casper H J / Jozwiak, Marta / van Beekhuizen, Heleen J / Mustafa, Dana A M

Frontiers in oncology

2022 Volume 11, Page(s) 715077

Abstract: Cervical cancer is one of the most common cancers in women worldwide. Patients diagnosed with early-stage cervical cancer have a good prognosis, however, 10-20% suffer from local or distant recurrent disease after primary treatment. Treatment options for ...

Abstract	Cervical cancer is one of the most common cancers in women worldwide. Patients diagnosed with early-stage cervical cancer have a good prognosis, however, 10-20% suffer from local or distant recurrent disease after primary treatment. Treatment options for recurrent cervical cancer are limited. Therefore, it is crucial to identify factors that can predict patients with an increased risk of recurrence to optimize treatment to prevent the recurrence of cervical cancer. We aimed to identify biomarkers in early-stage primary cervical cancer which recurred after surgery. Formalin-Fixed, Paraffin-Embedded surgical specimens of 34 patients with early-stage cervical cancer (FIGO 2009 stage 1B1) and 7 healthy controls were analyzed. Targeted gene expression profiling using the PanCancer IO 360 panel of NanoString Technology was performed. The findings were confirmed by performing immunohistochemistry stainings. Various genes, namely GLS, CD36, WNT5a, HRAS, DDB2, PIK3R2, and CDH2 were found to be differentially highly expressed in primary cervical cancer samples of patients who developed distant recurrence. In addition, The relative infiltration score of CD8+ T cells, CD80+CD86+ macrophages, CD163+MRC1+ macrophages, and FOXP3+IL2RA+ regulatory T cells were significantly higher in this group of samples. In contrast, no significant differences in gene expression and relative immune infiltration were found in samples of patients who developed local recurrence. The infiltration of CD8 and FOXP3 cells were validated by immunohistochemistry using all samples included in the study. We identified molecular alterations in primary cervical cancer samples from patients who developed recurrent disease. These findings can be utilized towards developing a molecular signature for the early detection of patients with a high risk to develop metastasis.
Language	English
Publishing date	2022-01-11
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2649216-7
ISSN	2234-943X
ISSN	2234-943X
DOI	10.3389/fonc.2021.715077
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Article ; Online: The deleted in oral cancer (DOC1 aka CDK2AP1) tumor suppressor gene is downregulated in oral squamous cell carcinoma by multiple microRNAs.

Stabile, Roberto / Cabezas, Mario Román / Verhagen, Mathijs P / Tucci, Francesco A / van den Bosch, Thierry P P / De Herdt, Maria J / van der Steen, Berdine / Nigg, Alex L / Chen, Meng / Ivan, Cristina / Shimizu, Masayoshi / Koljenović, Senada / Hardillo, Jose A / Verrijzer, C Peter / Baatenburg de Jong, Robert J / Calin, George A / Fodde, Riccardo

Cell death & disease

2023 Volume 14, Issue 5, Page(s) 337

Abstract: Cyclin-dependent kinase 2-associated protein 1 (CDK2AP1; also known as deleted in oral cancer or DOC1) is a tumor suppressor gene known to play functional roles in both cell cycle regulation and in the epigenetic control of embryonic stem cell ... ...

Abstract	Cyclin-dependent kinase 2-associated protein 1 (CDK2AP1; also known as deleted in oral cancer or DOC1) is a tumor suppressor gene known to play functional roles in both cell cycle regulation and in the epigenetic control of embryonic stem cell differentiation, the latter as a core subunit of the nucleosome remodeling and histone deacetylation (NuRD) complex. In the vast majority of oral squamous cell carcinomas (OSCC), expression of the CDK2AP1 protein is reduced or lost. Notwithstanding the latter (and the DOC1 acronym), mutations or deletions in its coding sequence are extremely rare. Accordingly, CDK2AP1 protein-deficient oral cancer cell lines express as much CDK2AP1 mRNA as proficient cell lines. Here, by combining in silico and in vitro approaches, and by taking advantage of patient-derived data and tumor material in the analysis of loss of CDK2AP1 expression, we identified a set of microRNAs, namely miR-21-5p, miR-23b-3p, miR-26b-5p, miR-93-5p, and miR-155-5p, which inhibit its translation in both cell lines and patient-derived OSCCs. Of note, no synergistic effects were observed of the different miRs on the CDK2AP1-3-UTR common target. We also developed a novel approach to the combined ISH/IF tissue microarray analysis to study the expression patterns of miRs and their target genes in the context of tumor architecture. Last, we show that CDK2AP1 loss, as the result of miRNA expression, correlates with overall survival, thus highlighting the clinical relevance of these processes for carcinomas of the oral cavity.
MeSH term(s)	Humans ; Cell Line, Tumor ; Cell Proliferation/genetics ; Cyclin-Dependent Kinase Inhibitor Proteins/metabolism ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; MicroRNAs/genetics ; MicroRNAs/metabolism ; Mouth Neoplasms/genetics ; Mouth Neoplasms/pathology ; Squamous Cell Carcinoma of Head and Neck/genetics ; Squamous Cell Carcinoma of Head and Neck/pathology ; Tumor Suppressor Proteins/genetics
Chemical Substances	Cyclin-Dependent Kinase Inhibitor Proteins ; MicroRNAs ; CDK2AP1 protein, human ; Tumor Suppressor Proteins
Language	English
Publishing date	2023-05-22
Publishing country	England
Document type	Journal Article
ZDB-ID	2541626-1
ISSN	2041-4889 ; 2041-4889
ISSN (online)	2041-4889
ISSN	2041-4889
DOI	10.1038/s41419-023-05857-2
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Article ; Online: Differentiating Cerebellar Impact on Thalamic Nuclei.

Gornati, Simona V / Schäfer, Carmen B / Eelkman Rooda, Oscar H J / Nigg, Alex L / De Zeeuw, Chris I / Hoebeek, Freek E

Cell reports

2018 Volume 23, Issue 9, Page(s) 2690–2704

Abstract: The cerebellum plays a role in coordination of movements and non-motor functions. Cerebellar nuclei (CN) axons connect to various parts of the thalamo-cortical network, but detailed information on the characteristics of cerebello-thalamic connections is ... ...

Abstract	The cerebellum plays a role in coordination of movements and non-motor functions. Cerebellar nuclei (CN) axons connect to various parts of the thalamo-cortical network, but detailed information on the characteristics of cerebello-thalamic connections is lacking. Here, we assessed the cerebellar input to the ventrolateral (VL), ventromedial (VM), and centrolateral (CL) thalamus. Confocal and electron microscopy showed an increased density and size of CN axon terminals in VL compared to VM or CL. Electrophysiological recordings in vitro revealed that optogenetic CN stimulation resulted in enhanced charge transfer and action potential firing in VL neurons compared to VM or CL neurons, despite that the paired-pulse ratio was not significantly different. Together, these findings indicate that the impact of CN input onto neurons of different thalamic nuclei varies substantially, which highlights the possibility that cerebellar output differentially controls various parts of the thalamo-cortical network.
MeSH term(s)	Animals ; Axons/metabolism ; Axons/ultrastructure ; Cerebellar Nuclei/physiology ; Cerebellar Nuclei/ultrastructure ; Cerebellum/physiology ; Cerebellum/ultrastructure ; Dendrites/physiology ; Electric Stimulation ; Excitatory Postsynaptic Potentials ; Female ; Male ; Mice, Inbred C57BL ; Receptors, Ionotropic Glutamate/antagonists & inhibitors ; Synapses/physiology ; Synapses/ultrastructure ; Synaptic Transmission ; Thalamic Nuclei/physiology
Chemical Substances	Receptors, Ionotropic Glutamate
Language	English
Publishing date	2018-06-27
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2649101-1
ISSN	2211-1247 ; 2211-1247
ISSN (online)	2211-1247
ISSN	2211-1247
DOI	10.1016/j.celrep.2018.04.098
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Article ; Online: Impact of early- and late-onset preeclampsia on features of placental and newborn vascular health.

Herzog, Emilie M / Eggink, Alex J / Reijnierse, Anniek / Kerkhof, Martina A M / de Krijger, Ronald R / Roks, Anton J M / Reiss, Irwin K M / Nigg, Alex L / Eilers, Paul H C / Steegers, Eric A P / Steegers-Theunissen, Régine P M

Placenta

2017 Volume 49, Page(s) 72–79

Abstract: Introduction: Offspring exposed to preeclampsia (PE) show an increased risk of cardiovascular disease in adulthood. We hypothesize that this is mediated by a disturbed vascular development of the placenta, umbilical cord and fetus. Therefore, we ... ...

Abstract	Introduction: Offspring exposed to preeclampsia (PE) show an increased risk of cardiovascular disease in adulthood. We hypothesize that this is mediated by a disturbed vascular development of the placenta, umbilical cord and fetus. Therefore, we investigated associations between early-onset PE (EOPE), late-onset PE (LOPE) and features of placental and newborn vascular health. Methods: We performed a nested case-control study in The Rotterdam Periconceptional Cohort, including 30 PE pregnancies (15 EOPE, 15 LOPE) and 218 control pregnancies (164 uncomplicated controls, 54 complicated controls including 28 fetal growth restriction, 26 preterm birth) and assessed macroscopic and histomorphometric outcomes of the placenta and umbilical cord. Results: A significant association was observed between PE and a smaller umbilical vein area and wall thickness, independent of gestational age and birth weight. In EOPE we observed significant associations with a lower weight, length and width of the placenta, length of the umbilical cord, and thickness and wall area of the umbilical vein and artery. These associations attenuated after gestational age and birth weight adjustment. In LOPE a significant association with a larger placental width and smaller umbilical vein wall thickness was shown, independent of gestational age and birth weight. Discussion: Our study suggests that PE is associated with a smaller umbilical cord vein area and wall thickness, independent of gestational age and birth weight, which may serve as a proxy of disturbed cardiovascular development in the newborn. Follow-up studies are needed to ultimately predict and lower the risk of cardiovascular disease in offspring exposed to PE.
MeSH term(s)	Birth Weight/physiology ; Case-Control Studies ; Female ; Fetal Development/physiology ; Fetal Growth Retardation/pathology ; Fetal Growth Retardation/physiopathology ; Humans ; Infant, Newborn ; Organ Size/physiology ; Placenta/blood supply ; Placenta/pathology ; Placenta/physiopathology ; Pre-Eclampsia/pathology ; Pre-Eclampsia/physiopathology ; Pregnancy
Language	English
Publishing date	2017
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	603951-0
ISSN	1532-3102 ; 0143-4004
ISSN (online)	1532-3102
ISSN	0143-4004
DOI	10.1016/j.placenta.2016.11.014
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Zs.A 1578: Show issues

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Article: Immunomics of Renal Allograft Acute T Cell-Mediated Rejection Biopsies of Tacrolimus- and Belatacept-Treated Patients.

van der Zwan, Marieke / Baan, Carla C / Colvin, Robert B / Smith, Rex N / White, Rebecca A / Ndishabandi, Dorothy / Nigg, Alex L / van den Bosch, Thierry P P / de Graav, Gretchen N / Clahsen-van Groningen, Marian C / Hesselink, Dennis A

Transplantation direct

2018 Volume 5, Issue 1, Page(s) e418

Abstract: Background: Belatacept-based therapy in kidney transplant recipient has been shown to increase long-term renal allograft and patient survival compared with calcineurin inhibitor-based therapy, however, with an increased risk of acute T cell-mediated ... ...

Abstract	Background: Belatacept-based therapy in kidney transplant recipient has been shown to increase long-term renal allograft and patient survival compared with calcineurin inhibitor-based therapy, however, with an increased risk of acute T cell-mediated rejection (aTCMR). An improved understanding of costimulation blockade-resistant rejections could lead to a more personalized approach to belatacept therapy. Here, immunomic profiles of aTCMR biopsies of patients treated with either tacrolimus or belatacept were compared. Methods: Formalin-fixed paraffin-embedded renal transplant biopsies were used for immunohistochemistry and gene expression analysis using the innovative NanoString technique. To validate NanoString, transcriptomic profiles of patients with and without biopsy-proven aTCMR were compared. Biopsies from 31 patients were studied: 14 tacrolimus-treated patients with aTCMR, 11 belatacept-treated patients with aTCMR, and 6 controls without rejection. Results: A distinct pattern was seen in biopsies with aTCMR compared to negative controls: 78 genes had a higher expression in the aTCMR group (false discovery rate Conclusions: In this small pilot study, no difference was found in immunomics of aTCMR biopsies of tacrolimus- and belatacept-treated patients. This suggests that clinically diagnosed aTCMR reflects a final common pathway of allorecognition which is unaffected by the type of immunosuppressive therapy.
Language	English
Publishing date	2018-12-20
Publishing country	United States
Document type	Journal Article
ISSN	2373-8731
ISSN	2373-8731
DOI	10.1097/TXD.0000000000000857
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Article ; Online: Prognostic value of intra-tumoral CD8

Sideras, Kostandinos / Galjart, Boris / Vasaturo, Angela / Pedroza-Gonzalez, Alexander / Biermann, Katharina / Mancham, Shanta / Nigg, Alex L / Hansen, Bettina E / Stoop, Hans A / Zhou, Guoying / Verhoef, Cornelis / Sleijfer, Stefan / Sprengers, Dave / Kwekkeboom, Jaap / Bruno, Marco J

Journal of surgical oncology

2018 Volume 118, Issue 1, Page(s) 68–76

Abstract: Background and objectives: Patients with isolated colorectal-cancer-liver-metastases (CRCLM) frequently undergo metastatectomy. Tumor-infiltrating-lymphocytes (TILs) have prognostic potential in the setting of primary colorectal cancer, however, their ... ...

Abstract	Background and objectives: Patients with isolated colorectal-cancer-liver-metastases (CRCLM) frequently undergo metastatectomy. Tumor-infiltrating-lymphocytes (TILs) have prognostic potential in the setting of primary colorectal cancer, however, their role in CRCLM is less studied. We aimed to study the spatial distribution and prognostic role of tumor-infiltrating CD8 Methods: TILs were isolated from fresh metastatic tissues of 47 patients with CRCLM. Archived paraffin-embedded tissue, from the same patients, was retrieved. CD8 Results: By immunohistochemistry, individual densities of intra-tumoral or peri-tumoral CD8 Conclusions: The ratio of cytotoxic (CD8
MeSH term(s)	Adult ; Aged ; Aged, 80 and over ; CD8-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/pathology ; Cohort Studies ; Colorectal Neoplasms/immunology ; Colorectal Neoplasms/secondary ; Female ; Forkhead Transcription Factors/immunology ; Humans ; Immunohistochemistry ; Liver Neoplasms/immunology ; Liver Neoplasms/pathology ; Lymphocytes, Tumor-Infiltrating/immunology ; Male ; Middle Aged ; Prognosis ; T-Lymphocyte Subsets/immunology ; T-Lymphocytes, Regulatory/immunology
Chemical Substances	FOXP3 protein, human ; Forkhead Transcription Factors
Language	English
Publishing date	2018-06-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	82063-5
ISSN	1096-9098 ; 0022-4790
ISSN (online)	1096-9098
ISSN	0022-4790
DOI	10.1002/jso.25091
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Zs.A 706: Show issues

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Article ; Online: T lymphocytes facilitate brain metastasis of breast cancer by inducing Guanylate-Binding Protein 1 expression.

Mustafa, Dana A M / Pedrosa, Rute M S M / Smid, Marcel / van der Weiden, Marcel / de Weerd, Vanja / Nigg, Alex L / Berrevoets, Cor / Zeneyedpour, Lona / Priego, Neibla / Valiente, Manuel / Luider, Theo M / Debets, Reno / Martens, John W M / Foekens, John A / Sieuwerts, Anieta M / Kros, Johan M

Acta neuropathologica

2018 Volume 135, Issue 4, Page(s) 581–599

Abstract: The discovery of genes and molecular pathways involved in the formation of brain metastasis would direct the development of therapeutic strategies to prevent this deadly complication of cancer. By comparing gene expression profiles of Estrogen Receptor ... ...

Abstract	The discovery of genes and molecular pathways involved in the formation of brain metastasis would direct the development of therapeutic strategies to prevent this deadly complication of cancer. By comparing gene expression profiles of Estrogen Receptor negative (ER-) primary breast tumors between patients who developed metastasis to brain and to organs other than brain, we found that T lymphocytes promote the formation of brain metastases. To functionally test the ability of T cells to promote brain metastasis, we used an in vitro blood-brain barrier (BBB) model. By co-culturing T lymphocytes with breast cancer cells, we confirmed that T cells increase the ability of breast cancer cells to cross the BBB. Proteomics analysis of the tumor cells revealed Guanylate-Binding Protein 1 (GBP1) as a key T lymphocyte-induced protein that enables breast cancer cells to cross the BBB. The GBP1 gene appeared to be up-regulated in breast cancer of patients who developed brain metastasis. Silencing of GBP1 reduced the ability of breast cancer cells to cross the in vitro BBB model. In addition, the findings were confirmed in vivo in an immunocompetent syngeneic mouse model. Co-culturing of ErbB2 tumor cells with activated T cells induced a significant increase in Gbp1 expression by the cancer cells. Intracardial inoculation of the co-cultured tumor cells resulted in preferential seeding to brain. Moreover, intracerebral outgrowth of the tumor cells was demonstrated. The findings point to a role of T cells in the formation of brain metastases in ER- breast cancers, and provide potential targets for intervention to prevent the development of cerebral metastases.
MeSH term(s)	Adult ; Aged ; Animals ; Blood-Brain Barrier/metabolism ; Blood-Brain Barrier/pathology ; Brain Neoplasms/metabolism ; Brain Neoplasms/secondary ; Breast Neoplasms/metabolism ; Breast Neoplasms/pathology ; Cells, Cultured ; Coculture Techniques ; Female ; GTP-Binding Proteins/genetics ; GTP-Binding Proteins/metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Mice ; Middle Aged ; Neoplasm Metastasis/physiopathology ; Neoplasm Transplantation ; Proteome ; RNA, Messenger/metabolism ; T-Lymphocytes/metabolism
Chemical Substances	GBP1 protein, human ; Proteome ; RNA, Messenger ; GTP-Binding Proteins (EC 3.6.1.-)
Language	English
Publishing date	2018-01-19
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1079-0
ISSN	1432-0533 ; 0001-6322
ISSN (online)	1432-0533
ISSN	0001-6322
DOI	10.1007/s00401-018-1806-2
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Article ; Online: Proteomic signatures of extracellular vesicles secreted by nonmineralizing and mineralizing human osteoblasts and stimulation of tumor cell growth.

Morhayim, Jess / van de Peppel, Jeroen / Demmers, Jeroen A A / Kocer, Gulistan / Nigg, Alex L / van Driel, Marjolein / Chiba, Hideki / van Leeuwen, Johannes P

FASEB journal : official publication of the Federation of American Societies for Experimental Biology

2015 Volume 29, Issue 1, Page(s) 274–285

Abstract: Beyond forming bone, osteoblasts play pivotal roles in various biologic processes, including hematopoiesis and bone metastasis. Extracellular vesicles (EVs) have been implicated in intercellular communication via transfer of proteins and nucleic acids ... ...

Abstract	Beyond forming bone, osteoblasts play pivotal roles in various biologic processes, including hematopoiesis and bone metastasis. Extracellular vesicles (EVs) have been implicated in intercellular communication via transfer of proteins and nucleic acids between cells. We focused on the proteomic characterization of nonmineralizing (NMOBs) and mineralizing (MOBs) human osteoblast (SV-HFOs) EVs and investigated their effect on human prostate cancer (PC3) cells by microscopic, proteomic, and gene expression analyses. Proteomic analysis showed that 97% of the proteins were shared among NMOB and MOB EVs, and 30% were novel osteoblast-specific EV proteins. Label-free quantification demonstrated mineralization stage-dependent 5-fold enrichment of 59 and 451 EV proteins in NMOBs and MOBs, respectively. Interestingly, bioinformatic analyses of the osteoblast EV proteomes and EV-regulated prostate cancer gene expression profiles showed that they converged on pathways involved in cell survival and growth. This was verified by in vitro proliferation assays where osteoblast EV uptake led to 2-fold increase in PC3 cell growth compared to cell-free culture medium-derived vesicle controls. Our findings elucidate the mineralization stage-specific protein content of osteoblast-secreted EVs, show a novel way by which osteoblasts communicate with prostate cancer, and open up innovative avenues for therapeutic intervention.
MeSH term(s)	Calcification, Physiologic/genetics ; Calcification, Physiologic/physiology ; Cell Communication/genetics ; Cell Communication/physiology ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Extracellular Matrix/metabolism ; Extracellular Matrix/pathology ; Extracellular Matrix Proteins/genetics ; Extracellular Matrix Proteins/metabolism ; Gene Expression Profiling ; Humans ; Male ; Neoplasm Proteins/genetics ; Neoplasm Proteins/metabolism ; Osteoblasts/metabolism ; Osteoblasts/pathology ; Prostatic Neoplasms/genetics ; Prostatic Neoplasms/metabolism ; Prostatic Neoplasms/pathology ; Proteomics ; Tumor Microenvironment
Chemical Substances	Extracellular Matrix Proteins ; Neoplasm Proteins
Language	English
Publishing date	2015-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	639186-2
ISSN	1530-6860 ; 0892-6638
ISSN (online)	1530-6860
ISSN	0892-6638
DOI	10.1096/fj.14-261404
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