Publications

Inhibition of cyclin-dependent kinase 5 affects early neuroinflammatory signalling in murine model of amyloid beta toxicity.

Wilkaniec A1, Gąssowska-Dobrowolska M1, Strawski M2, Adamczyk A1, Czapski GA3.

Author information:

1 Department of Cellular Signalling, Mossakowski Medical Research Centre Polish Academy of Sciences, Pawińskiego 5, 02-106, Warsaw, Poland.

2 Laboratory of Electrochemistry, Faculty of Chemistry, University of Warsaw, Pasteura 1, 02-093, Warsaw, Poland.

3 Department of Cellular Signalling, Mossakowski Medical Research Centre Polish Academy of Sciences, Pawińskiego 5, 02-106, Warsaw, Poland. This email address is being protected from spambots. You need JavaScript enabled to view it..

 

ABSTRACT:

BACKGROUND:

Cyclin-dependent kinase 5 (Cdk5) belongs to the family of proline-directed serine/threonine kinases and plays a critical role in neuronal differentiation, migration, synaptogenesis, plasticity, neurotransmission and apoptosis. The deregulation of Cdk5 activity was observed in post mortem analysis of brain tissue of Alzheimer's disease (AD) patients, suggesting the involvement of Cdk5 in the pathomechanism of this neurodegenerative disease. However, our recent study demonstrated the important function of Cdk5 in regulating inflammatory reaction.

 

METHODS:

Since the role of Cdk5 in regulation of inflammatory signalling in AD is unknown, we investigated the involvement of Cdk5 in neuroinflammation induced by single intracerebroventricular (icv) injection of amyloid beta protein (Aβ) oligomers in mouse. The brain tissue was analysed up to 35 days post injection. Roscovitine (intraperitoneal administration) was used as a potent Cdk5 inhibitor. The experiments were also performed on human neuroblastoma SH-SY5Y as well as mouse BV2 cell lines treated with exogenous oligomeric Aβ.

 

RESULTS:

Our results demonstrated that single injection of Aβ oligomers induces long-lasting activation of microglia and astrocytes in the hippocampus. We observed also profound, early inflammatory response in the mice hippocampus, leading to the significant elevation of pro-inflammatory cytokines expression (e.g. TNF-α, IL-1β, IL-6). Moreover, Aβ oligomers elevated the formation of truncated protein p25 in mouse hippocampus and induced overactivation of Cdk5 in neuronal cells. Importantly, administration of roscovitine reduced the inflammatory processes evoked by Aβ in the hippocampus, leading to the significant decrease of cytokines level.

 

CONCLUSIONS:

These studies clearly show the involvement of Cdk5 in modulation of brain inflammatory response induced by Aβ and may indicate this kinase as a novel target for pharmacological intervention in AD.

 

KEYWORDS:

Alzheimer’s disease; Amyloid beta; Cdk5; Cytokines; Gene expression; Neuroinflammation

Extracellular Alpha-Synuclein Oligomers Induce Parkin S-Nitrosylation: Relevance to Sporadic Parkinson's Disease Etiopathology.

Mol Neurobiol. 2018 Apr 21. doi: 10.1007/s12035-018-1082-0. [Epub ahead of print]

Wilkaniec A1, Lenkiewicz AM1, Czapski GA1, Jęśko HM1, Hilgier W2, Brodzik R3, Gąssowska-Dobrowolska M1, Culmsee C4, Adamczyk A5.

Author information:

1 Department of Cellular Signalling, Mossakowski Medical Research Centre, Polish Academy of Sciences, Pawińskiego 5 Street, 02-106, Warsaw, Poland.

2 Department of Neurotoxicology, Mossakowski Medical Research Centre, Polish Academy of Sciences, Pawińskiego 5 Street, 02-106, Warsaw, Poland.

3 BLIRT S.A., Trzy Lipy 3/1.38, 80-172, Gdańsk, Poland.

4 Institute of Pharmacology and Clinical Pharmacy, University of Marburg, 35043, Marburg, Germany.

5 Department of Cellular Signalling, Mossakowski Medical Research Centre, Polish Academy of Sciences, Pawińskiego 5 Street, 02-106, Warsaw, Poland. This email address is being protected from spambots. You need JavaScript enabled to view it..

 

ABSTRACT:

α-Synuclein (ASN) and parkin, a multifunctional E3 ubiquitin ligase, are two proteins that are associated with the pathophysiology of Parkinson's disease (PD). Excessive release of ASN, its oligomerization, aggregation, and deposition in the cytoplasm contribute to neuronal injury and cell death through oxidative-nitrosative stress induction, mitochondrial impairment, and synaptic dysfunction. In contrast, overexpression of parkin provides protection against cellular stresses and prevents dopaminergic neural cell loss in several animal models of PD. However, the influence of ASN on the function of parkin is largely unknown. Therefore, the aim of this study was to investigate the effect of extracellular ASN oligomers on parkin expression, S-nitrosylation, as well as its activity. For these investigations, we used rat pheochromocytoma (PC12) cell line treated with exogenous oligomeric ASN as well as PC12 cells with parkin overexpression and parkin knock-down. The experiments were performed using spectrophotometric, spectrofluorometric, and immunochemical methods. We found that exogenous ASN oligomers induce oxidative/nitrosative stress leading to parkin S-nitrosylation. Moreover, this posttranslational modification induced the elevation of parkin autoubiquitination and degradation of the protein. The decreased parkin levels resulted in significant cell death, whereas parkin overexpression protected against toxicity induced by extracellular ASN oligomers. We conclude that lowering parkin levels by extracellular ASN may significantly contribute to the propagation of neurodegeneration in PD pathology through accumulation of defective proteins as a consequence of parkin degradation.

KEYWORDS:

Alpha-synuclein; Parkin; Parkinson’s disease; S-nitrosylation

Neuroprotective Potential and Paracrine Activity of Stromal Vs. Culture-Expanded hMSC Derived from Wharton Jelly under Co-Cultured with Hippocampal Organotypic Slices.

Mol Neurobiol. 2018 Jul;55(7):6021-6036. doi: 10.1007/s12035-017-0802-1. Epub 2017 Nov 13.

Dabrowska S 1, Sypecka J 1, Jablonska A 1, Strojek L 1, Wielgos M 2, Domanska-Janik K 1,
Sarnowska A 3.

1 Mossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, Warsaw, Poland.

2 1st Department of Obstetrics and Gynecology, Medical University of Warsaw, Warsaw, Poland.

3 Mossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, Warsaw, Poland. This email address is being protected from spambots. You need JavaScript enabled to view it..

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Intra-arterial delivery of cell therapies for stroke.

Stroke. 2018 May;49(5):1075-1082. doi: 10.1161/STROKEAHA.117.018288. Epub 2018 Apr 18.

Guzman R 1, Janowski M 2,3,4,5, Walczak P 6,3,7.

1 From the Department of Neurosurgery and Biomedicine, University Hospital Basel, University of Basel, Switzerland (R.G.).

2 Russell H. Morgan Department of Radiology and Radiological Science, the Johns Hopkins University School of Medicine, Baltimore, MD (M.J., P.W.).

3 Cellular Imaging Section and Vascular Biology Program, Institute for Cell Engineering, the Johns Hopkins University School of Medicine, Baltimore, MD (M.J., P.W.).

4 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences (M.J.).

5 Department of Neurosurgery, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland (M.J.).

6 Russell H. Morgan Department of Radiology and Radiological Science, the Johns Hopkins University School of Medicine, Baltimore, MD (M.J., P.W.) This email address is being protected from spambots. You need JavaScript enabled to view it..

7 Department of Neurology and Neurosurgery, Faculty of Medical Sciences, University of Warmia and Mazury, Olsztyn, Poland (P.W.).

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Hydrogel-based scaffolds to support intrathecal stem cell transplantation as a gateway to the spinal cord: clinical needs, biomaterials, and imaging technologies.

NPJ Regen Med. 2018 Apr 4;3:8. doi: 10.1038/s41536-018-0046-3. eCollection 2018.

Oliveira JM1,2,3, Carvalho L1,2, Silva-Correia J1,2, Vieira S1,2, Majchrzak M4, Lukomska B4, Stanaszek L4, Strymecka P4, Malysz-Cymborska I5, Golubczyk D5, Kalkowski L5, Reis RL1,2,3, Janowski M4,6,7, Walczak P5,6,7.

1 3B´s Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence, Tissue Engineering and Regenerative Medicine, AvePark, Zona Industrial da Gandra, 4805-017 Barco, Guimarães Portugal.

2 2ICVS/3B's - PT Government Associate Laboratory, Braga, Portugal.

3 3The Discoveries Centre for Regenerative and Precision Medicine, Headquarters at University of Minho, Avepark, 4805-017 Barco, Guimarães Portugal.

4 4NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

5 5Department of Neurology and Neurosurgery, School of Medicine, Collegium Medicum, University of Warmia and Mazury, Olsztyn, Poland.

6 6Russel H, Morgan Department of Radiology and Radiological Science, Johns Hopkins University, Baltimore, MD USA.

7 7Vascular Biology Program, Institute for Cell Engineering, Johns Hopkins University, Baltimore, MD USA.

Hydrogel-based scaffolds to support intrathecal stem cell transplantation as a gateway to the spinal cord: clinical needs, biomaterials, and imaging technologies.

ABSTRACT

The prospects for cell replacement in spinal cord diseases are impeded by inefficient stem cell delivery. The deep location of the spinal cord and complex surgical access, as well as densely packed vital structures, question the feasibility of the widespread use of multiple spinal cord punctures to inject stem cells. Disorders characterized by disseminated pathology are particularly appealing for the distribution of cells globally throughout the spinal cord in a minimally invasive fashion. The intrathecal space, with access to a relatively large surface area along the spinal cord, is an attractive route for global stem cell delivery, and, indeed, is highly promising, but the success of this approach relies on the ability of cells (1) to survive in the cerebrospinal fluid (CSF), (2) to adhere to the spinal cord surface, and (3) to migrate, ultimately, into the parenchyma. Intrathecal infusion of cell suspension, however, has been insufficient and we postulate that embedding transplanted cells within hydrogel scaffolds will facilitate reaching these goals. In this review, we focus on practical considerations that render the intrathecal approach clinically viable, and then discuss the characteristics of various biomaterials that are suitable to serve as scaffolds. We also propose strategies to modulate the local microenvironment with nanoparticle carriers to improve the functionality of cellular grafts. Finally, we provide an overview of imaging modalities for in vivo monitoring and characterization of biomaterials and stem cells. This comprehensive review should serve as a guide for those planning preclinical and clinical studies on intrathecal stem cell transplantation.

The Differentiation of Rat Oligodendroglial Cells Is Highly Influenced by the Oxygen Tension: In Vitro Model Mimicking Physiologically Normoxic Conditions.

Int J Mol Sci. 2018 Jan 24;19(2). pii: E331. doi: 10.3390/ijms19020331.

Janowska J1, Ziemka-Nalecz M2, Sypecka J3.

1 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, 02-106 Warsaw, Poland. This email address is being protected from spambots. You need JavaScript enabled to view it..

2 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, 02-106 Warsaw, Poland. This email address is being protected from spambots. You need JavaScript enabled to view it..

3 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego Street, 02-106 Warsaw, Poland. This email address is being protected from spambots. You need JavaScript enabled to view it..

The Differentiation of Rat Oligodendroglial Cells Is Highly Influenced by the Oxygen Tension: In Vitro Model Mimicking Physiologically Normoxic Conditions.

ABSTRACT

Oligodendrocyte progenitor cells (OPCs) constitute one of the main populations of dividing cells in the central nervous system (CNS). Physiologically, OPCs give rise to mature, myelinating oligodendrocytes and confer trophic support to their neighboring cells within the nervous tissue. OPCs are known to be extremely sensitive to the influence of exogenous clues which might affect their crucial biological processes, like survival, proliferation, differentiation, and the ability to generate a myelin membrane. Alterations in their differentiation influencing their final potential for myelinogenesis are usually the leading cause of CNS dys- and demyelination, contributing to the development of leukodystrophic disorders. The evaluation of the mechanisms that cause oligodendrocytes to malfunction requires detailed studies based on designed in vitro models. Since OPCs readily respond to changes in local homeostasis, it is crucial to establish restricted culture conditions to eliminate the potential stimuli that might influence oligodendrocyte biology. Additionally, the in vitro settings should mimic the physiological conditions to enable the obtained results to be translated to future preclinical studies. Therefore, the aim of our study was to investigate OPC differentiation in physiological normoxia (5% O₂) and a restricted in vitro microenvironment. To evaluate the impact of the combined microenvironmental clues derived from other components of the nervous tissue, which are also influenced by the local oxygen concentration, the process of generating OPCs was additionally analyzed in organotypic hippocampal slices. The obtained results show that OPC differentiation, although significantly slowed down, proceeded correctly through its typical stages in the physiologically relevant conditions created in vitro. The established settings were also conducive to efficient cell proliferation, exerting also a neuroprotective effect by promoting the proliferation of neurons. In conclusion, the performed studies show how oxygen tension influences OPC proliferation, differentiation, and their ability to express myelin components, and should be taken into consideration while planning preclinical studies, e.g., to examine neurotoxic compounds or to test neuroprotective strategies.

KEYWORDS:

cell proliferation; culture density; hippocampal organotypic slices; myelin protein amounts; myelinogenesis; oligodendrocyte maturation myelin; oligodendrocyte progenitor cells; physiological normoxia; serum-free culture

Impact of neonatal hypoxia-ischaemia on oligodendrocyte survival, maturation and myelinating potential.

J Cell Mol Med. 2018 Jan;22(1):207-222. doi: 10.1111/jcmm.13309. Epub 2017 Aug 7.

Ziemka-Nalecz M1, Janowska J1, Strojek L1, Jaworska J1, Zalewska T1, Frontczak-Baniewicz M2, Sypecka J1.

Author information

1 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

2 Electron Microscopy Platform, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

 

Impact of neonatal hypoxia-ischaemia on oligodendrocyte survival, maturation and myelinating potential.

ABSTRACT

Hypoxic-ischaemic episodes experienced at the perinatal period commonly lead to a development of neurological disabilities and cognitive impairments in neonates or later in childhood. Clinical symptoms often are associated with the observed alterations in white matter in the brains of diseased children, suggesting contribution of triggered oligodendrocyte/myelin pathology to the resulting disorders. To date, the processes initiated by perinatal asphyxia remain unclear, hampering the ability to develop preventions. To address the issue, the effects of temporal hypoxia-ischaemia on survival, proliferation and the myelinating potential of oligodendrocytes were evaluated ex vivo using cultures of hippocampal organotypic slices and in vivo in rat model of perinatal asphyxia. The potential engagement of gelatinases in oligodendrocyte maturation was assessed as well. The results pointed to a significant decrease in the number of oligodendrocyte progenitor cells (OPCs), which is compensated for to a certain extent by the increased rate of OPC proliferation. Oligodendrocyte maturation seemed however to be significantly altered. An ultrastructural examination of selected brain regions performed several weeks after the insult showed however that the process of developing central nervous system myelination proceeds efficiently resulting in enwrapping the majority of axons in compact myelin. The increased angiogenesis in response to neonatal hypoxic-ischaemic insult was also noticed. In conclusion, the study shows that hypoxic-ischaemic episodes experienced during the most active period of nervous system development might be efficiently compensated for by the oligodendroglial cell response triggered by the insult. The main obstacle seems to be the inflammatory process modulating the local microenvironment.

KEYWORDS:

electron microscopy; gelatinases; hippocampal organotypic slices; myelin structure; myelinogenesis; neonatal hypoxia-ischaemia; oligodendrocyte progenitor cells; oxygen and glycose deprivation; perinatal asphyxia

Imaging of extracellular vesicles derived from human bone marrow mesenchymal stem cells using fluorescent and magnetic labels

Int J Nanomedicine. 2018 Mar 19;13:1653-1664. doi: 10.2147/IJN.S159404. eCollection 2018.

Dabrowska S1, Del Fattore A2, Karnas E3,4, Frontczak-Baniewicz M5, Kozlowska H6, Muraca M7, Janowski M1,8, Lukomska B1.

1 NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

2 Multifactorial Disease and Complex Phenotype Research Area, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.

3 Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.

4 Malopolska Centre of Biotechnology, Krakow, Poland.

5 Electron Microscopy Platform, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

6 Laboratory of Advanced Microscopy Techniques, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.

7 Department of Women's and Children's Health, University of Padua, Padua, Italy.

8 Russel H. Morgan Department of Radiology and Radiological Science, Division of MR Research, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.

Imaging of extracellular vesicles derived from human bone marrow mesenchymal stem cells using fluorescent and magnetic labels.

 

ABSTRACT

Mesenchymal stem cells have been shown therapeutic in various neurological disorders. Recent studies support the notion that the predominant mechanism by which MSCs act is through the release of extracellular vesicles (EVs). EVs seem to have similar therapeutic activity as their cellular counterparts and may represent an interesting alternative standalone therapy for various diseases. The aim of the study was to optimize the method of EV imaging to better understand therapeutic effects mediated by EVs.

 

KEYWORDS:

MRI; cell tracking; extracellular vesicles; fluorescent dye; iron oxide; mesenchymal stem cells

Glycogen metabolism in brain and neurons - astrocytes metabolic cooperation can be altered by pre- and neonatal lead (Pb) exposure.

Toxicology. 2017 Sep 12;390:146-158. doi: 10.1016/j.tox.2017.09.007. 

Baranowska-Bosiacka I1, Falkowska A2, Gutowska I3, Gąssowska M4, Kolasa-Wołosiuk A5, Tarnowski M6, Chibowska K2, Goschorska M2, Lubkowska A7, Chlubek D2.
Author information
Glycogen metabolism in brain and neurons - astrocytes metabolic cooperation can be altered by pre- and neonatal lead (Pb) exposure.
1 Department of Biochemistry and Medical Chemistry, Powstańców Wlkp. 72 St., 70-111 Szczecin, Pomeranian Medical University in Szczecin, Poland. Electronic address: This email address is being protected from spambots. You need JavaScript enabled to view it..
2 Department of Biochemistry and Medical Chemistry, Powstańców Wlkp. 72 St., 70-111 Szczecin, Pomeranian Medical University in Szczecin, Poland.
3 Department of Biochemistry and Human Nutrition, Broniewskiego 24 St., 71-460 Szczecin, Pomeranian Medical University in Szczecin, Poland.
4 Department of Cellular Signalling, Mossakowski Medical Research Centre, Polish Academy of Sciences, Pawińskiego 5, 02-106 Warsaw, Poland.
5 Department of Histology and Embriology, Powstańców Wlkp. 72 St., 70-111 Szczecin, Pomeranian Medical University in Szczecin, Poland.
6 Department of Physiology, Powstańców Wlkp. 72 St., 70-111 Szczecin, Pomeranian Medical University in Szczecin, Poland.
7 Department of Functional Diagnostics and Physical Medicine, Pomeranian Medical University in Szczecin, Żołnierska 48 Str, 71-210 Szczecin, Poland.

ABSTRACT

Lead (Pb) is an environmental neurotoxin which particularly affects the developing brain but the molecular mechanism of its neurotoxicity still needs clarification. The aim of this paper was to examine whether pre- and neonatal exposure to Pb (concentration of Pb in rat offspring blood below the "threshold level") may affect the brain's energy metabolism in neurons and astrocytes via the amount of available glycogen. We investigated the glycogen concentration in the brain, as well as the expression of the key enzymes involved in glycogen metabolism in brain: glycogen synthase 1 (Gys1), glycogen phosphorylase (PYGM, an isoform active in astrocytes; and PYGB, an isoform active in neurons) and phosphorylase kinase β (PHKB). Moreover, the expression of connexin 43 (Cx43) was evaluated to analyze whether Pb poisoning during the early phase of life may affect the neuron-astrocytes' metabolic cooperation. This work shows for the first time that exposure to Pb in early life can impair brain energy metabolism by reducing the amount of glycogen and decreasing the rate of its metabolism. This reduction in brain glycogen level was accompanied by a decrease in Gys1 expression. We noted a reduction in the immunoreactivity and the gene expression of both PYGB and PYGM isoform, as well as an increase in the expression of PHKB in Pb-treated rats. Moreover, exposure to Pb induced decrease in connexin 43 immunoexpression in all the brain structures analyzed, both in astrocytes as well as in neurons. Our data suggests that exposure to Pb in the pre- and neonatal periods results in a decrease in the level of brain glycogen and a reduction in the rate of its metabolism, thereby reducing glucose availability, which as a further consequence may lead to the impairment of brain energy metabolism and the metabolic cooperation between neurons and astrocytes.

KEYWORDS:

Brain glycogen metabolism; Glycogen phosphorylase brain isoform (PYGB); Glycogen phosphorylase kinase (PHKB); Glycogen phosphorylase muscle isoform (PYGM); Glycogen synthase (Gys1); Lead (Pb) neurotoxicity