Publications

Sodium hydrosulfide moderately alleviates the hallmark symptoms of Duchenne muscular dystrophy in mdx mice

Interactions Between the Circadian Clock and Heme Oxygenase in the Retina of Drosophila melanogaster

Heme Oxygenase-1 in Tumor Biology and Therapy

Heme oxygenase-1 (HO-1) degrades heme to carbon monoxide (CO), biliverdin, and ferrous iron. As HO-1 expression is highly increased by stressful conditions, the major role of the enzyme is the protection against oxidative injury. Additionally, it regulates cell proliferation, modulates inflammatory response and facilitates angiogenesis. Beneficial activities of HO-1 have been recognized in many pathological states e.g. atherosclerosis, diabetes, ischemia/reperfusion injury or organ transplantation. Interestingly HO-1 expression is very often boosted in tumor tissues and could be further elevated in response to radio-, chemo-, or photodynamic therapy. A growing body of evidence suggests that HO-1 may play a role in tumor induction and can potently improve the growth and spread of tumors. This review discusses the implications of HO-1 properties for tumor proliferation and cell death, differentiation, angiogenesis and metastasis, and tumor-related inflammation. Finally, it suggests that pharmacological agents that regulate HO activity or HO-1 gene silencing may become powerful tools for preventing the onset or progression of various cancers and sensitize them to anticancer therapies.

New strategies for application of plasmid and viral vectors in gene therapy

Pre-emptive hypoxia-regulated HO-1 gene therapy improves post-ischaemic limb perfusion and tissue regeneration in mice

Our results suggest the therapeutic potential of HO-1 for prevention of adverse effects in critical limb ischaemia.

Supplementary Figure 5 from Hypoxia-Regulated Overexpression of Soluble VEGFR2 Controls Angiogenesis and Inhibits Tumor Growth

<p>PDF - 3176KB, Characterization of B16F10 wt and msVEGFR2 expressing clones 13.3 and 16.4 tumors in vivo.</p>

OC19 EFFECT OF HYPOXIA AND HEME OXYGENASE-1 ON EXPRESSION OF ANGIOGENIC GENES IN HUMAN KERATINOCYTES

Cytosolic dsRNA sensors as regulators of transgene expression from AAV vectors in human iPSC-derived cardiomyocytes

Abstract Background Adeno-associated viral vectors (AAVs) are safe and efficient tools for delivery of therapeutic genes in numerous applications. Although they elicit only limited immune response in host cells, a recent study has demonstrated an association between decreasing transgene expression in the liver and induction of the immune response to cytoplasmic dsRNA. While AAVs are DNA vectors, it appears that their unique genome structure allows the generation of dsRNA in transduced cells. Since the expression cassette is flanked by inverted terminal repeats (ITRs), which can function as bidirectional promoters, assembly of sense and antisense transcripts can potentially result in formation of dsRNA structures, especially in cells exhibiting high transgene expression. Purpose In this study, we explored the mechanism of dsRNA recognition in cardiomyocytes, to unravel its role in the regulation of transgene expression from AAV vectors. Methods For this purpose, we utilised human iPSC-derived cardiomyocytes transduced with self-complementary AAV6 or AAV9 vectors encoding GFP. Since we focused on the long-term effects of transgene expression, all the analyses were performed 7–9 days after AAV transduction. Results We confirmed that ITR serves as a functional promoter in the cells, capable of maintaining approximately half of the fluorescence signal compared to the CMV promoter. Long-term transgene expression was associated with elevated expression of dsRNA sensors (MDA5, RIG-I, LGP2 and TLR3) and the appearance of small foci of dsRNA in the perinuclear region. Unlike an artificial ligand for dsRNA sensors, poly (I:C), dsRNA in AAV-transduced cells did not interact with the LGP2 protein. To induce dsRNA recognition, we stimulated the cells with IFNβ for 1 week, starting from day 1 after transduction. We found that transgene expression is significantly hampered in such conditions. Furthermore, IFNβ induced the direct interaction of dsRNA with the OAS1 protein in transduced cells. During viral infections, OAS1 and its effector nuclease RNAse L promote the degradation of mRNA to restrict the production of viral proteins. Since RNAse L is not specific for particular transcripts, we hypothesize that activation of the OAS1 pathway in transduced cells is responsible for downregulation of transgene expression. While IFNβ induced OAS1-dsRNA recognition, we did not observe such an event in cells transduced with AAV and maintained under standard conditions. In turn, AAV transduction resulted in a reduction in OAS1 expression at the mRNA and protein level, suggesting a possible reason for tolerance to dsRNA in transduced cardiomyocytes. Conclusions Our data present a novel mechanism of regulation of transgene expression in AAV-transduced cells involving OAS1–RNAse L pathway and provide deeper insight into the general signal transduction routes activated in response to cytosolic dsRNA in human iPSC-derived cardiomyocytes. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): National Science Centre - PRELUDUM grant 2019/33/N/NZ1/03066Jagiellonian University, Faculty of Biochemistry, Biophysics and Biotechnology - grant for young scientists MNS 6/2020

Analysis of Cell Cycle Status of Murine Hematopoietic Stem Cells

Introduction to the special issue dedicated to the founding Editor, Dr. Waclaw Szybalski

Role of Heme Oxygenase-1 in Postnatal Differentiation of Stem Cells: A Possible Cross-Talk with MicroRNAs

It seems that interplay between HO-1 and miRNAs may be important in regulating fates of stem and progenitor cells and needs further intensive studies.

HMG-CoA Reductase Inhibitors Regulate Inflammatory Transcription Factors in Human Endothelial and Vascular Smooth Muscle Cells

Objective— Pleiotropic atheroprotective effects of HMG-CoA reductase inhibitors may be mediated on the level of vascular gene transcription. The aim of this study was to characterize the effects of statins on the activation of transcription factors known to regulate inflammation and cell proliferation/differentiation. Methods and Results— Simvastatin, atorvastatin, and lovastatin (0.1 to 10 μmol/L) inhibited the binding of nuclear proteins to both the nuclear factor-kappa B (NF-κB) and activator protein-1 (AP-1) DNA consensus oligonucleotides in human endothelial and vascular smooth muscle cells as assessed by electrophoretic mobility shift assay (EMSA). The inhibitory effects of statins on NF-κB or AP-1–dependent transcriptional activity were examined by transient transfection studies. HMG-CoA reductase inhibitors upregulated IκB-α protein levels in endothelial cells and decreased c-Jun mRNA expression in smooth muscle cells as analyzed by Western and Northern blotting, respectively. Furthermore, statins inhibited DNA binding of hypoxia-inducible factor-1α. Downstream effects of statins included inhibition of plasminogen activator inhibitor-1 and vascular endothelial growth factor-A mRNA levels in endothelial cells. Conclusions— HMG-CoA reductase inhibitors downregulate the activation of transcription factors NF-κB, AP-1, and hypoxia-inducible factor-1α. These findings support the concept that statins have antiinflammatory and antiproliferative effects that are relevant in the treatment of atherosclerotic diseases.

Supplementary materials and methods from Heme Oxygenase-1 Controls an HDAC4-miR-206 Pathway of Oxidative Stress in Rhabdomyosarcoma

<p>Detailed description of methods used in the study with Supplementary Figures legends</p>

Vascular endothelial growth factor: angiogenesis, atherogenesis or both?

Gel Electrophoresis of DNA --- New Measurements and the Repton Model at High Fields

Faculty of Biotechnology, Jagellonian UniversityGronostajowa 7, 30-387 Cracow, Poland(Received December 7, 2004)The paper is dedicated to Prof. Andrzej Fulinski for his 70th birthdayNew experimental data are presented on the gel electrophoresisofDNA.Experiment was made for molecules of length 173kbp, in 1 percent agarosegel, in TAE 1× buffer and the field intensity between 5 and 9V/cm. Theresults are compared with our computer simulations, performed within therepton model of Duke and Rubinstein. The ranges of field and moleculelength are determined, where the geometration effect appears. We investi-gate also the field dependence of the velocity and the diffusion coefficientat the border of the geometration regime.PACS numbers: 87.15.Tt, 05.10.Ln, 83.10.Kn

Expression of β-galactosidase gene and endothelial nitric oxide synthase gene in rat vascular smooth muscle cells after in vitro lipotransfection

Unproven cell interventions in Poland and the exploitation of European Union law on advanced therapy medicinal products

Hypoxic angiogenic transcriptome in human keratinocytes and microvascular endothelial cells: Macroarray and real-time PCR analysis.