Institute of Fundamental Technological Research
Polish Academy of Sciences

Staff

Paulina Koza, PhD

Department of Biosystems and Soft Matter (ZBiMM)
Division of Modelling in Biology and Medicine (PMBM)
position: assistant professor
telephone: (+48) 22 826 12 81 ext.: 423
room: 328
e-mail:

Doctoral thesis
2016-06-24 The effects of TDP-43 depletion on neuronal plasticity in Syn-TDP-43WT transgenic rat model  (IBD PAN)
supervisor -- Leszek Kaczmarek, IBD PAN
 

Recent publications
1.  Zdioruk M., Want A., Mietelska-Porowska A., Laskowska-Kaszub K., Wojsiat J., Klejman A., Użarowska E., Koza P., Olejniczak S., Pikul S., Konopka W., Golab J., Wojda U., A new inhibitor of tubulin polymerization kills multiple cancer cell types and reveals p21-mediated mechanism determining cell death after mitotic catastrophe, Cancers, ISSN: 2072-6694, DOI: 10.3390/cancers12082161, Vol.12, No.8, pp.2161-1-21, 2020

Abstract:
Induction of mitotic catastrophe through the disruption of microtubules is an established target in cancer therapy. However, the molecular mechanisms determining the mitotic catastrophe and the following apoptotic or non-apoptotic cell death remain poorly understood. Moreover, many existing drugs targeting tubulin, such as vincristine, have reduced efficacy, resulting from poor solubility in physiological conditions. Here, we introduce a novel small molecule 2-aminoimidazoline derivative-OAT-449, a synthetic water-soluble tubulin inhibitor. OAT-449 in a concentration range from 6 to 30 nM causes cell death of eight different cancer cell lines in vitro, and significantly inhibits tumor development in such xenograft models as HT-29 (colorectal adenocarcinoma) and SK-N-MC (neuroepithelioma) in vivo. Mechanistic studies showed that OAT-449, like vincristine, inhibited tubulin polymerization and induced profound multi-nucleation and mitotic catastrophe in cancer cells. HeLa and HT-29 cells within 24 h of treatment arrested in G2/M cell cycle phase, presenting mitotic catastrophe features, and 24 h later died by non-apoptotic cell death. In HT-29 cells, both agents altered phosphorylation status of Cdk1 and of spindle assembly checkpoint proteins NuMa and Aurora B, while G2/M arrest and apoptosis blocking was consistent with p53-independent accumulation in the nucleus and largely in the cytoplasm of p21/waf1/cip1, a key determinant of cell fate programs. This is the first common mechanism for the two microtubule-dissociating agents, vincristine and OAT-449, determining the cell death pathway following mitotic catastrophe demonstrated in HT-29 cells.

Keywords:
cancer, chemotherapeutic, microtubule-poison, vincristine, mitotic catastrophe, non-apoptotic cell death, p21, p53

Affiliations:
Zdioruk M. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Want A. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Mietelska-Porowska A. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Laskowska-Kaszub K. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Wojsiat J. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Klejman A. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Użarowska E. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Koza P. - other affiliation
Olejniczak S. - OncoArendi Therapeutics (PL)
Pikul S. - OncoArendi Therapeutics (PL)
Konopka W. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Golab J. - Medical University of Warsaw (PL)
Wojda U. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
2.  Koza P., Przybyś J., Klejman A., Olech-Kochańczyk G., Konopka W., Generation of transgenic rats using a lentiviral vector approach, Journal of Visualized Experiments, ISSN: 1940-087X, DOI: 10.3791/60570, Vol.159, pp.e60570-1-8, 2020

Abstract:
Transgenic animal models are fundamentally important for modern biomedical research. The incorporation of foreign genes into early mouse or rat embryos is an invaluable tool for gene function analysis in living organisms. The standard transgenesis method is based on microinjecting foreign DNA fragments into a pronucleus of a fertilized oocyte. This technique is widely used in mice but remains relatively inefficient and technically demanding in other animal species. The transgene can also be introduced into one-cell-stage embryos via lentiviral infection, providing an effective alternative to standard pronuclear injections, especially in species or strains with a more challenging embryo structure. In this approach, a suspension that contains lentiviral vectors is injected into the perivitelline space of a fertilized rat embryo, which is technically less demanding and has a higher success rate. Lentiviral vectors were shown to efficiently incorporate the transgene into the genome to determine the generation of stable transgenic lines. Despite some limitations (e.g., Biosafety Level 2 requirements, DNA fragment size limits), lentiviral transgenesis is a rapid and efficient transgenesis method. Additionally, using female rats that are mated with a fertile male strain with a different dominant fur color is presented as an alternative to generate pseudopregnant foster mothers.

Keywords:
retraction, issue 159, transgenic rat, lentiviral vectors, perivitelline space, foster mothers

Affiliations:
Koza P. - other affiliation
Przybyś J. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Klejman A. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Olech-Kochańczyk G. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Konopka W. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
3.  Koza P., Beroun A., Konopka A., Górkiewicz T., Bijoch Ł., Torres J.C., Bulska E., Knapska E., Kaczmarek L., Konopka W., Neuronal TDP-43 depletion affects activity-dependent plasticity, Neurobiology of Disease, ISSN: 0969-9961, DOI: 10.1016/j.nbd.2019.104499, Vol.130, pp.104499-1-12, 2019

Abstract:
TAR DNA-binding protein 43 (TDP-43) is a hallmark of some neurodegenerative disorders, such as frontotemporal lobar degeneration and amyotrophic lateral sclerosis. TDP-43-related pathology is characterized by its abnormally phosphorylated and ubiquitinated aggregates. It is involved in many aspects of RNA processing, including mRNA splicing, transport, and translation. However, its exact physiological function and role in mechanisms that lead to neuronal degeneration remain elusive. Transgenic rats that were characterized by TDP-43 depletion in neurons exhibited enhancement of the acquisition of fear memory. At the cellular level, TDP-43-depleted neurons exhibited a decrease in the short-term plasticity of intrinsic neuronal excitability. The induction of long-term potentiation in the CA3-CA1 areas of the hippocampus resulted in more stable synaptic enhancement. At the molecular level, the protein levels of an unedited (R) FLOP variant of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) GluR1 and GluR2/3 subunits decreased in the hippocampus. Alterations of FLOP/FLIP subunit composition affected AMPAR kinetics, reflected by cyclothiazide-dependent slowing of the decay time of AMPAR-mediated miniature excitatory postsynaptic currents. These findings suggest that TDP-43 may regulate activity-dependent neuronal plasticity, possibly by regulating the splicing of genes that are responsible for fast synaptic transmission and membrane potential.

Keywords:
TDP-43, AMPA receptors, FLOP/FLIP splice variants, PTZ model

Affiliations:
Koza P. - other affiliation
Beroun A. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Konopka A. - University of Warsaw (PL)
Górkiewicz T. - other affiliation
Bijoch Ł. - other affiliation
Torres J.C. - other affiliation
Bulska E. - other affiliation
Knapska E. - other affiliation
Kaczmarek L. - other affiliation
Konopka W. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
4.  Was H., Barszcz K., Czarnecka J., Kowalczyk A., Bernas T., Uzarowska E., Koza P., Klejman A., Piwocka K., Kaminska B., Sikora E., Bafilomycin A1 triggers proliferative potential of senescent cancer cells in vitro and in NOD/SCID mice, Oncotarget, ISSN: 1949-2553, DOI: 10.18632/oncotarget.14066, Vol.8, No.6, pp.9303-9322, 2017

Abstract:
Anticancer therapies that induce DNA damage tend to trigger senescence in cancer cells, a process known as therapy-induced senescence (TIS). Such cells may undergo atypical divisions, thus contributing to tumor re-growth. Accumulation of senescent cancer cells reduces survival of patients after chemotherapy. As senescence interplays with autophagy, a dynamic recycling process, we sought to study whether inhibition of autophagy interferes with divisions of TIS cells. We exposed human colon cancer HCT116 cells to repeated cycles of a chemotherapeutic agent - doxorubicin (doxo) and demonstrated induction of hallmarks of TIS (e.g. growth arrest, hypertrophy, poliploidization and secretory phenotype) and certain properties of cancer stem cells (increased NANOG expression, percentages of CD24+ cells and side population). Colonies of small and highly proliferative progeny appeared shortly after drug removal. Treatment with bafilomycin A1 (BAF A1), an autophagy inhibitor, postponed short term in vitro cell re-population. It was associated with reduction in the number of diploid and increase in the number of poliploid cells. In a long term, a pulse of BAF A1 resulted in reactivation of autophagy in a subpopulation of HCT116 cells and increased proliferation. Accordingly, the senescent HCT116 cells treated with BAF A1 when injected into NOD/SCID mice formed tumors, in contrast to the controls. Our results suggest that senescent cancer cells that appear during therapy, can be considered as dormant cells that contribute to cancer re-growth, when chemotherapeutic treatment is stopped. These data unveil new mechanisms of TIS-related cancer maintenance and re-population, triggered by a single pulse of BAF A1 treatment.

Keywords:
colon cancer, chemotherapy, senescence, autophagy, angiogenesis

Affiliations:
Was H. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Barszcz K. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Czarnecka J. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Kowalczyk A. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Bernas T. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Uzarowska E. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Koza P. - other affiliation
Klejman A. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Piwocka K. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Kaminska B. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Sikora E. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
5.  Stefaniuk M., Gualda E.J., Pawlowska M., Legutko D., Matryba P., Koza P., Konopka W., Owczarek D., Wawrzyniak M., Loza-Alvarez P., Kaczmarek L., Light-sheet microscopy imaging of a whole cleared rat brain with Thy1-GFP transgene, Scientific Reports, ISSN: 2045-2322, DOI: 10.1038/srep28209, Vol.6, pp.28209-1-9, 2016

Abstract:
Whole-brain imaging with light-sheet fluorescence microscopy and optically cleared tissue is a new, rapidly developing research field. Whereas successful attempts to clear and image mouse brain have been reported, a similar result for rats has proven difficult to achieve. Herein, we report on creating novel transgenic rat harboring fluorescent reporter GFP under control of neuronal gene promoter. We then present data on clearing the rat brain, showing that FluoClearBABB was found superior over passive CLARITY and CUBIC methods. Finally, we demonstrate efficient imaging of the rat brain using light-sheet fluorescence microscopy.

Affiliations:
Stefaniuk M. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Gualda E.J. - Barcelona Institute of Science and Technology (ES)
Pawlowska M. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Legutko D. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Matryba P. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Koza P. - other affiliation
Konopka W. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Owczarek D. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Wawrzyniak M. - Nencki Institute of Experimental Biology, Polish Academy of Sciences (PL)
Loza-Alvarez P. - Barcelona Institute of Science and Technology (ES)
Kaczmarek L. - other affiliation
6.  Wiera G., Szczot M., Wojtowicz T., Lebida K., Koza P., Mozrzymas J.W., Impact of matrix metalloproteinase-9 overexpression on synaptic excitatory transmission and its plasticity in rat CA3-CA1 hippocampal pathway, JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY, ISSN: 0867-5910, Vol.66, No.2, pp.309-315, 2015

Abstract:
Metalloproteinases (MMPs) have been shown to play a crucial role in synaptic plasticity and cognitive processes. We recently reported that in the mossy fiber - CA3 hippocampal pathway, LTP maintenance required fine-tuned MMP-9 activity, as both MMP-9 excess and absence impaired LTP. Here we used acute brain slices from transgenic (TG) rats overexpressing MMP-9 to investigate the impact of excessive MMP-9 activity on the excitatory synaptic transmission in the CA3-CA1 projection. Using field potential recordings, we have demonstrated that MMP-9 overexpression increased the strength of basal synaptic transmission but had no effect on the short-term plasticity in comparison to the wild-type (WT) group. In attempt to shed light on mechanisms underlying this observation, miniature excitatory postsynaptic potentials (mEPSCs) were recorded from pyramidal CA1 neurons. We found that mEPSCs in the TG group had a significantly slower decaying phase than in WT but amplitudes and frequencies were similar. The lack of differences in mEPSC frequency and short-term plasticity between TG and WT groups suggests that MMP-9 overexpression effect on fEPSPs was mainly postsynaptic. Additionally, we have found that excess of MMP-9 in TG rats was associated with impaired late-phase of LTP in the considered pathway. It seems thus that augmented synaptic strength in TG rats occurred in expense of impaired long-term plasticity induced by tetanization. In conclusion, overexpression of MMP-9 leads to increase in the strength of basal excitatory synaptic transmission and impairs of LTP maintenance phase in the CA3-CA1 pathway in vitro.

Keywords:
hippocampus, metalloproteinase, high frequency stimulation, long-term potentiation, miniature excitatory postsynaptic potentials, field excitatory postsynaptic potentials, synapse

Affiliations:
Wiera G. - Wroclaw Medical University (PL)
Szczot M. - Wroclaw Medical University (PL)
Wojtowicz T. - Institute of Physics, Polish Academy of Sciences (PL)
Lebida K. - Wroclaw Medical University (PL)
Koza P. - other affiliation
Mozrzymas J.W. - Wroclaw Medical University (PL)

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