Jozef Dulak

Jagiellonian University, Cracovia, Lesser Poland Voivodeship, Poland

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Publications (212)873.8 Total impact

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    ABSTRACT: Sarcoidosis is a multi-organ disease in which affected tissues are invaded with non-necrotizing granulomatous structures, mostly consisted of T helper 1 (Th1) cells and multinucleate giant cells. However, the etiology and pathogenesis of sarcoidosis is not known and the diagnosis is usually based on clinical examination involving radiography and histopathological analysis of biopsies of affected organs. Although the knowledge on the molecular background of sarcoidosis is limited, it seems that the important pathways involve transforming growth factor-β (TGF-β) and JAK/STAT, which may influence the interferon-γ (IFN-γ)-mediated signaling. Additionally, recently the role of microRNAs (miRNAs), the small non-coding RNA molecules, has been emphasized in different pathological conditions including autoimmune diseases. This review summarizes the current knowledge on the molecular pathways in the pathogenesis of sarcoidosis with a special emphasis on cytokines and miRNAs controlling immune cells proliferation and differentiation. Moreover, the possible role of T regulatory cells (CD4(+) CD25(+) FoxP3(+)) in this disease has been discussed.
    Journal of physiology and pharmacology: an official journal of the Polish Physiological Society 11/2015; 66(5):635-642. · 2.39 Impact Factor
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    ABSTRACT: Hypoxia-inducing pathologies as cancer develop pathologic and inefficient angiogenesis which rules tumor facilitating microenvironment, a key target for therapy. As such, the putative ability of endothelial precursor cells (EPCs) to specifically home to hypoxic sites of neovascularization prompted to design optimized, site-specific, cell-mediated, drug-/gene-targeting approach. Thus, EPC lines, were established from aorta-gonad-mesonephros (AGM) of murine 10.5dpc and 11.5dpc embryo, when endothelial repertoire is completed. Lines representing early endothelial differentiation steps were selected: MAgECs10.5, MagECs11.5. Distinct in maturation, they differently express VEGF receptors, VE-Cadherin and chemokine/receptors. MAgECs11.5, more differentiated than MAgEC10.5, displayed faster angiogenesis in vitro, different response to hypoxia and chemokines. Both MAgECs lines cooperated to tube-like formation with mature endothelial cells and invaded tumor spheroids through a vasculogenesis-like process. In vivo, both MAgEC-formed vessels established blood flow. Intravenously injected, both MAgECs invaded Matrigel(TM)-plugs and targeted tumors. Here we show that EPCs (MAgEC11.5) target tumor angiogenesis, allow local overexpression of hypoxia-driven soluble VEGF-receptor2 enabling drastic tumor growth reduction. We propose that such EPCs, able to target tumor angiogenesis, could act as therapeutic gene-vehicles, to inhibit tumor growth by vessel normalization resulting from tumor hypoxia alleviation.
    Cancer letters 11/2015; DOI:10.1016/j.canlet.2015.11.008 · 5.62 Impact Factor
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    ABSTRACT: The retina and the first optic neuropil (lamina) of Drosophila show circadian rhythms in various processes. To learn about the regulation of circadian rhythms in the retina and lamina and in two cell types, glial and the lamina L2 interneurons, we examined expression of the following clock genes; per, tim, clk, and cry and clock-controlled genes (ccgs); Atpα, nrv2, brp, Pdfr. We found that the expression of gene studied is specific for the retina and lamina. The rhythms of per and tim expression in the retina and glial cells are similar to that observed in the whole head and in clock neurons, while they differ in the lamina and L2 cells. In both the retina and lamina, CRY seems to be a repressor of clk expression. In L2 interneurons per expression is not cyclic indicating the other function of PER in those cells than in the circadian molecular clock. In contrast to per and tim, the pattern of clk and cry expression is similar in both the retina and lamina. The retina holds the autonomous oscillators but the expression of cry and ccgs, Atpα and nrv2, is also regulated by inputs from the pacemaker transmitted by PDF and ITP neuropeptides.
    Frontiers in Cellular Neuroscience 10/2015; 9. DOI:10.3389/fncel.2015.00353 · 4.29 Impact Factor

  • Circulation Research 10/2015; 117(9):e75-e76. DOI:10.1161/CIRCRESAHA.115.307450 · 11.02 Impact Factor
  • Agnieszka Loboda · Alicja Jozkowicz · Jozef Dulak ·

    Vascular Pharmacology 09/2015; DOI:10.1016/j.vph.2015.09.004 · 3.64 Impact Factor
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    ABSTRACT: Subcutaneous injection of the tumor cell suspension is a simple and commonly used tool for studying tumor development in vivo. However, subcutaneous models poorly resemble tumor complexity due to the fast growth not reflecting the natural course. Here, we describe an application of the new spheroid-plug model to combine the simplicity of subcutaneous injection with improved resemblance to natural tumor progression. Spheroid-plug model relies on in vitro formation of tumor spheroids, followed by injection of single tumor spheroid subcutaneously in Matrigel matrix. In spheroid-plug model, tumors grow slower in comparison to tumors formed by injection of cell suspension as assessed by 3D ultrasonography (USG) and in vivo bioluminescence measurements. The slower tumor growth rate in spheroid-plug model is accompanied by reduced necrosis. The spheroid-plug model ensures increased and more stable vascularization of tumor than classical subcutaneous tumor model as demonstrated by 3D USG Power Doppler examination. Flow cytometry analysis showed that tumors formed from spheroids have enhanced infiltration of endothelial cells as well as hematopoietic and progenitor cells with stem cell phenotype (c-Kit(+) and Sca-1(+)). They also contain more tumor cells expressing cancer stem cell marker CXCR4. Here, we show that spheroid-plug model allows investigating efficiency of anticancer drugs. Treatment of spheroid-plug tumors with known antiangiogenic agent axitinib decreased their size and viability. The antiangiogenic activity of axitinib was higher in spheroid-plug model than in classical model. Our results indicate that spheroid-plug model imitates natural tumor growth and can become a valuable tool for cancer research.
    Tumor Biology 09/2015; DOI:10.1007/s13277-015-4065-z · 3.61 Impact Factor

  • European Respiratory Journal 09/2015; 46(suppl 59):OA2932. DOI:10.1183/13993003.congress-2015.OA2932 · 7.64 Impact Factor
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    ABSTRACT: Lung mucoepidermoid carcinoma (MEC) is a very poorly characterized rare subtype of non-small-cell lung cancer (NSCLC) associated with more favorable prognoses than other forms of intrathoracic malignancies. We have previously identified that heme oxygenase-1 (HO-1, encoded by HMOX1) inhibits MEC tumor growth and modulates the transcriptome of microRNAs. Here we investigate the role of a major upstream regulator of HO-1 and a master regulator of cellular antioxidant responses, transcription factor Nrf2, in MEC biology. Nrf2 overexpression in the NCI-H292 MEC cell line mimicked the phenotype of HO-1 overexpressing cells, leading to inhibition of cell proliferation and migration and down-regulation of oncogenic miR-378. HMOX1 silencing identified HO-1 as a major mediator of Nrf2 action. Nrf2- and HO-1 overexpressing cells exhibited strongly diminished expression of multiple matrix metalloproteinases and inflammatory cytokine interleukin-1β, which was confirmed in an NCI-HO-1 xenograft model. Overexpression of HO-1 altered not only human MMP levels in tumor cells but also murine MMP levels within tumor microenvironments and metastatic niches. This could possibly contribute to decreased metastasis to the lungs and inhibitory effects of HO-1 on MEC tumor growth. Our profound transcriptome analysis and molecular characterization of the mucoepidermoid lung carcinoma helps to understand the specific clinical presentations of these tumors, emphasizing a unique antitumoral role of the Nrf2-HO-1 axis.
    Free Radical Biology and Medicine 09/2015; 89. DOI:10.1016/j.freeradbiomed.2015.08.004 · 5.74 Impact Factor
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    ABSTRACT: Stem cells are self-renewing cells that can differentiate into specialized cell type(s). Pluripotent stem cells, i.e. embryonic stem cells (ESC) or induced pluripotent stem cells (iPSC) differentiate into cells of all three embryonic lineages. Multipotent stem cells, like hematopoietic stem cells (HSC), can develop into multiple specialized cells in a specific tissue. Unipotent cells differentiate only into one cell type, like e.g. satellite cells of skeletal muscle. There are many examples of successful clinical applications of stem cells. Over million patients worldwide have benefited from bone marrow transplantations performed for treatment of leukemias, anemias or immunodeficiencies. Skin stem cells are used to heal severe burns, while limbal stem cells can regenerate the damaged cornea. Pluripotent stem cells, especially the patient-specific iPSC, have a tremendous therapeutic potential, but their clinical application will require overcoming numerous drawbacks. Therefore, the use of adult stem cells, which are multipotent or unipotent, can be at present a more achievable strategy. Noteworthy, some studies ascribed particular adult stem cells as pluripotent. However, despite efforts, the postulated pluripotency of such events like "spore-like cells", "very small embryonic-like stem cells" or "multipotent adult progenitor cells" have not been confirmed in stringent independent studies. Also plasticity of the bone marrow-derived cells which were suggested to differentiate e.g. into cardiomyocytes, has not been positively verified, and their therapeutic effect, if observed, results rather from the paracrine activity. Here we discuss the examples of recent studies on adult stem cells in the light of current understanding of stem cell biology.
    Acta biochimica Polonica 07/2015; 62(3). DOI:10.18388/abp.2015_1023 · 1.15 Impact Factor

  • Atherosclerosis 07/2015; 241(1):e84. DOI:10.1016/j.atherosclerosis.2015.04.295 · 3.99 Impact Factor
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    ABSTRACT: Endothelial progenitor cells (EPCs) have been extensively studied for almost 19 years now and were considered as a potential marker for endothelial regeneration ability. On the other hand, circulating endothelial cells (CEC) were studied as biomarker for endothelial injury. Yet, in the literature, there is also huge incoherency in regards to terminology and protocols used. This results in misleading conclusions on the role of so called "EPCs", especially in the clinical field. The discrepancies are mainly due to strong phenotypic overlap between EPCs and circulating angiogenic cells (CAC), therefore changes in "EPC" terminology have been suggested. Other factors leading to inconsistent results are varied definitions of the studied populations and the lack of universal data reporting, which could strongly affect data interpretation. The current review is focused on controversies concerning the use of "EPCs"/CAC and CEC as putative endothelial diagnostic markers. Copyright © 2015 Institute of Pharmacology, Polish Academy of Sciences. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.
    Pharmacological reports: PR 06/2015; 67(4). DOI:10.1016/j.pharep.2015.05.017 · 1.93 Impact Factor
  • A Loboda · A Józkowicz · J Dulak ·

    Thrombosis and Haemostasis 04/2015; 114(20150423). DOI:10.1160/TH15-01-0082 · 4.98 Impact Factor
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    ABSTRACT: Blood vessel formation is a fundamental process for the development of organism and tissue regeneration. Of importance, angiogenesis occurring during postnatal development is usually connected with inflammation. Here, we review how molecular and cellular mechanisms underlying inflammatory reactions regulate angiogenesis. Inflamed tissues are characterized by hypoxic conditions and immune cell infiltration. In this review, we describe an interplay of hypoxia-inducible factors (HIFs), HIF1 and HIF2, as well as NF-κB and nitric oxide in the regulation of angiogenesis. The mobilization of macrophages and the differential role of M1 and M2 macrophage subsets in angiogenesis are also discussed. Next, we present the current knowledge about microRNA regulation of inflammation in the context of new blood vessel formation. Finally, we describe how the mechanisms involved in inflammation influence tumor angiogenesis. We underlay and discuss the role of NF-E2-related factor 2/heme oxygenase-1 pathway as crucial in the regulation of inflammation-induced angiogenesis. © 2015 IUBMB Life, 2015. © 2015 International Union of Biochemistry and Molecular Biology.
    International Union of Biochemistry and Molecular Biology Life 04/2015; 67(3). DOI:10.1002/iub.1358 · 3.14 Impact Factor
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    ABSTRACT: Diabetes is associated with reduced expression of heme oxygenase-1 (HO-1), a heme-degrading enzyme of cytoprotective and proangiogenic properties. In myoblasts and satellite cells (mSCs) HO-1 improves survival, proliferation and production of proangiogenic growth factors. Induction of HO-1 in injured tissues facilitates neovascularization, the process impaired in diabetes. We aimed to examine whether conditioned media from the HO-1 overexpressing myoblast cell line can improve a blood-flow recovery in ischemic muscles of diabetic mice. Analysis of myogenic markers was performed at mRNA level in primary mSCs, isolated by a pre-plate technique from diabetic db/db and normoglycemic wild type mice, and then cultured under growth or differentiation conditions. Hind limb ischemia was performed by femoral artery ligation in db/db mice and blood recovery was monitored by Laser Doppler measurements. Mice were treated with a single intramuscular injection of conditioned media harvested from wild type C2C12 myoblast cell line, C2C12 cells stably transduced with HO-1 cDNA or with unconditioned media. Expression of HO-1 was lower in mSCs isolated from muscles of diabetic db/db mice when compared to their wild type counterparts, what was accompanied by increased levels of Myf5 or CXCR4, and decreased Mef2 or Pax7. Such cells displayed also diminished differentiation potential when cultured in vitro, as shown by less effective formation of myotubes and reduced expression of myogenic markers (myoD, myogenin, and myosin). Blood flow recovery after induction of severe hind limb ischemia was delayed in db/db mice compared to that in normoglycemic individuals. To improve muscle regeneration after ischemia, conditioned media collected from differentiating C2C12 cells (control and HO-1 overexpressing) were injected into hind limbs of diabetic mice. Analysis of blood flow revealed that media from HO-1 overexpressing cells accelerated blood-flow recovery, while immunohistochemical staining assessment of vessels density in injected muscle confirmed increased angiogenesis. The effect might be mediated by SDF-1α proangiogenic factor, as its secretion is elevated in HO-1 overexpressing cells. In conclusion, paracrine stimulation of angiogenesis in ischemic skeletal muscle using conditioned media may be a safe approach exploiting protective and proangiogenic properties of HO-1 in diabetes.
    Stem Cell Research & Therapy 04/2015; 6(1):61. DOI:10.1186/s13287-015-0063-8 · 3.37 Impact Factor
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    ABSTRACT: Heme oxygenase-1 (HO-1) mitigates cellular injury by antioxidant, anti-apoptotic, anti-inflammatory and proangiogenic effects. Vascular endothelial growth factor (VEGF) is a critical regulator of blood vessels growth. Their coordinated action was analyzed in a model of femoral artery ligation (FAL) in mice lacking HO-1 gene (HO-1 KO). Gastrocnemius skeletal muscles of HO-1 KO mice were preemptively injected with plasmids containing hypoxia-response element (HRE) driving the expression of only HO-1 (pHRE-HO1) or both HO-1 and VEGF (pHRE-HO1-VEGF). At day 14th the pHRE-HO1 vector increased an impaired post-ischemic blood flow recovery in HO-1 KO mice to the level observed in wild-type (WT) mice subjected to FAL and pHRE-HO1-VEGF restored it already at day 7. The pHRE-HO1 gene therapy diminished, when compared to control pHRE-empty-treated HO-1 KO mice, the expression of toll-like receptors (TLR4 and TLR9) and inflammatory cytokines (IL-1β, IL-6 and TNFα) at day 3, whereas opposite effects were observed following concomitant HO-1 and VEGF gene transfer. Moreover, HO-1 diminished ischemia-induced expression of MyoD involved in satellite cell differentiation in HO-1 KO mice. Our results confirm the therapeutic potential of HO-1 and VEGF against critical limb ischemia although, their concomitant delivery may have contradictory actions on the resolution of inflammation. Copyright © 2015. Published by Elsevier Inc.
    Vascular Pharmacology 04/2015; 71. DOI:10.1016/j.vph.2015.02.011 · 3.64 Impact Factor
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    ABSTRACT: C57BL/6 is the most often used laboratory mouse strain. However, sometimes it is beneficial to cross the transgenic mice on the C57BL/6 background to the other strain, such as FVB. Although this is a common strategy, the influence of crossing these different strains on homeostatic expression of cytokines is not known. Here we have investigated the differences in the expression of selected cytokines between C57BL/6J and C57BL/6JxFVB mice in serum and skeletal muscle. We have found that only few cytokines were altered by crossing of the strains. Concentrations of IL5, IL7, LIF, MIP-2, and IP-10 were higher in serum of C57BL/6J mice than in C57BL/6JxFVB mice, whereas concentration of G-CSF was lower in C57BL/6J. In the skeletal muscle only the concentration of VEGF was higher in C57BL/6J mice than in C57BL/6JxFVB mice. Concluding, the differences in cytokine expression upon crossing C57BL/6 and FVB strain in basal conditions are not profound.
    Mediators of Inflammation 04/2015; 2015:762419. DOI:10.1155/2015/762419 · 3.24 Impact Factor
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    Scientific Reports 02/2015; · 5.58 Impact Factor
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    ABSTRACT: Mouse and human induced pluripotent stem cells (iPSCs) may represent a novel approach for modeling diabetes. Taking this into consideration, the aim of this study was to generate and evaluate differentiation potential of iPSCs from lep(db/db) (db/db) mice, the model of diabetes type 2 as well as from patients with Maturity Onset Diabetes of the Young 3 (HNF1A MODY). Murine iPSC colonies from both wild type and db/db mice were positive for markers of pluripotency: Oct3/4A, Nanog, SSEA1, CDy1 and alkaline phosphatase and differentiated in vitro and in vivo into cells originating from three germ layers. However, our results suggest impaired differentiation of db/db cells into endothelial progenitor-like cells expressing CD34 and Tie2 markers and their reduced angiogenic potential. Human control and HNF1A MODY reprogrammed cells also expressed pluripotency markers: OCT3/4A, SSEA4, TRA-1-60, TRA-1-81, formed embryoid bodies (EBs) and differentiated into cells of three germ layers. Additionally, insulin expressing cells were obtained from those partially reprogrammed cells with direct as well as EB-mediated differentiation method. Our findings indicate that disease-specific iPSCs may help to better understand the mechanisms responsible for defective insulin production or vascular dysfunction upon differentiation toward cell types affected by diabetes.
    Scientific Reports 02/2015; 5:8597. DOI:10.1038/srep08597 · 5.58 Impact Factor
  • Agnieszka Loboda · Alicja Jozkowicz · Jozef Dulak ·

    Toxicology 12/2014; 328. DOI:10.1016/j.tox.2014.12.002 · 3.62 Impact Factor
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    ABSTRACT: Background Peroxisome proliferator-activated receptor-¿ (PPAR¿) agonists, which have been used as insulin sensitizers in diabetic patients, may improve functions of endothelial cells (ECs). We investigated the effect of PPAR¿ on angiogenic activities of murine ECs and bone marrow-derived proangiogenic cells (PACs).MethodsPACs were isolated from bone marrow of 10¿12 weeks old, wild type, db/db and PPAR¿ heterozygous animals. Cells were cultured on fibronectin and gelatin coated dishes in EGM-2MV medium. For in vitro stimulations, rosiglitazone (10 ¿mol/L) or GW9662 (10 ¿mol/L) were added to 80% confluent cell cultures for 24 hours. Angiogenic potential of PACs and ECs was tested in vitro and in vivo in wound healing assay and hind limb ischemia model.ResultsECs and PACs isolated from diabetic db/db mice displayed a reduced angiogenic potential in ex vivo and in vitro assays, the effect partially rescued by incubation of cells with rosiglitazone (PPAR¿ activator). Correction of diabetes by administration of rosiglitazone in vivo did not improve angiogenic potential of isolated PACs or ECs. In a hind limb ischemia model we demonstrated that local injection of conditioned media harvested from wild type PACs improved the blood flow restoration in db/db mice, confirming the importance of paracrine action of the bone marrow-derived cells.Transcriptome analysis showed an upregulation of prooxidative and proinflammatory pathways, and downregulation of several proangiogenic genes in db/db PACs. Interestingly, db/db PACs had also a decreased level of PPAR¿ and changed expression of PPAR¿-regulated genes. Using normoglycemic PPAR¿+/¿ mice we demonstrated that reduced expression of PPAR¿ does not influence neovascularization either in wound healing or in hind limb ischemia models.Conclusions In summary, activation of PPAR¿ by rosiglitazone improves angiogenic potential of diabetic ECs and PACs, but decreased expression of PPAR¿ in diabetes does not impair angiogenesis.
    Cardiovascular Diabetology 11/2014; 13(1):150. DOI:10.1186/PREACCEPT-1453357592137930 · 4.02 Impact Factor

Publication Stats

5k Citations
873.80 Total Impact Points


  • 1998-2015
    • Jagiellonian University
      • • Department of Medical Biotechnology
      • • Department of Cell Biochemistry
      • • Department of Clinical Biochemistry
      Cracovia, Lesser Poland Voivodeship, Poland
  • 2006
    • Kyushu University
      Hukuoka, Fukuoka, Japan
  • 1999-2005
    • University of Innsbruck
      • Institute of Biochemistry
      Innsbruck, Tyrol, Austria
    • Stanford University
      • Division of Cardiovascular Medicine
      Palo Alto, California, United States
  • 2001
    • Medical College of Wisconsin
      • Department of Anesthesiology
      Milwaukee, Wisconsin, United States
  • 1995-1999
    • Collegium Medicum of the Jagiellonian University
      • Chair of Clinical Biochemistry
      Cracovia, Lesser Poland Voivodeship, Poland