Academic interests
Academic background
- 2020 – Post-doctoral fellowship; Faculty of Medicine, University of Oslo, Norway.
- 2015 – 2019 PhD candidate in medicine and health sciences; Faculty of Medicine, University of Oslo, Norway. Disputation date: 18.02.2020
- 2013 – 2015 Master of Science in Biotechnology; Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Poland
- 2012 – 2013 Master of Science in Engineering, Environmental Engineering, specialization: Mineral Engineering; Faculty of Geology, Geophysics and Environmental Protection, AGH University of Science and Technology, Poland
- 2010 – 2013 Bachelor in Biotechnology; Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Poland
- 2008 – 2012 Bachelor (Engineer) in Geology and Mining; Faculty of Geology, Geophysics and Environmental Protection, AGH University of Science and Technology, Poland
Tags:
Immunology,
T cells,
CAR,
Receptor,
Cell signalling,
Molecular biology
Publications
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Borowicz, Pawel; Gjerstad, Vibeke Sundvold; Chan, Hanna; Abrahamsen, Greger; Kjelstrup, Hanna & Nyman, Tuula Anneli
[Show all 7 contributors for this article]
(2021).
Tyr192 Regulates Lymphocyte-Specific Tyrosine Kinase Activity in T Cells.
Journal of Immunology.
ISSN 0022-1767.
207(4),
p. 1128–1137.
doi:
10.4049/jimmunol.2001105.
Full text in Research Archive
Show summary
TCR signaling critically depends on the tyrosine kinase Lck (lymphocyte-specific protein tyrosine kinase). Two phosphotyrosines, the activating pTyr394 and the inhibitory pTyr505, control Lck activity. Recently, pTyr192 in the Lck SH2 domain emerged as a third regulator. How pTyr192 may affect Lck function remains unclear. In this study, we explored the role of Lck Tyr192 using CRISPR/Cas9-targeted knock-in mutations in the human Jurkat T cell line. Our data reveal that both Lck pTyr394 and pTyr505 are controlled by Lck Tyr192. Lck with a nonphosphorylated SH2 domain (Lck Phe192) displayed hyperactivity, possibly by promoting Lck Tyr394 transphosphorylation. Lck Glu192 mimicking stable Lck pTyr192 was inhibited by Tyr505 hyperphosphorylation. To overcome this effect, we further mutated Tyr505. The resulting Lck Glu192/Phe505 displayed strongly increased amounts of pTyr394 both in resting and activated T cells. Our results suggest that a fundamental role of Lck pTyr192 may be to protect Lck pTyr394 and/or pTyr505 to maintain a pool of already active Lck in resting T cells. This provides an additional mechanism for fine-tuning of Lck as well as T cell activity.
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Borowicz, Pawel; Chan, Hanna; Hauge, Anette & Spurkland, Anne
(2020).
Adaptor proteins: Flexible and dynamic modulators of immune cell signalling.
Scandinavian Journal of Immunology.
ISSN 0300-9475.
92(6).
doi:
10.1111/sji.12951.
Full text in Research Archive
Show summary
To maintain homeostasis, all cells respond to environmental cues via a multitude of surface receptors. In order to act appropriately in their environment, cells are dependent on the transduction of the incoming signal through tightly regulated and interconnected signalling pathways to the cell nucleus. In particular, cells implicated in the immune system greatly depend on such systems to respond in a flexible and dynamic manner to environmental challenges. One major group of intracellular proteins that are involved in these signalling pathways are adaptor proteins. Although adaptor proteins are essential for normal immune cell operation, the functional role of this group of signalling proteins remains to be fully appreciated. So far, research on adaptor proteins has revealed their unique potential in building transient complexes in a reversible, dynamic and inducible manner. In this review, we explore the roles of adaptor proteins – in space and time of intracellular signalling – and their associations with human disease. Examples of adaptor proteins expressed in hematopoietic cells highlight their crucial role in the immune system. Lastly, we present challenges faced in elucidating roles of adaptor proteins, as illustrated by the T cell–specific adaptor (TSAd) protein encoded by the SH2D2A gene.
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Borowicz, Pawel; Chan, Hanna; Medina, Daniel Talmo; Gumpelmair, Simon; Kjelstrup, Hanna & Spurkland, Anne
(2019).
A simple and efficient workflow for generation of knock-in mutations in Jurkat T cells using CRISPR/Cas9.
Scandinavian Journal of Immunology.
ISSN 0300-9475.
doi:
10.1111/sji.12862.
Full text in Research Archive
Show summary
CRISPR/Cas9 is a powerful gene‐editing tool allowing for specific gene manipulation at targeted sites in the genome. Here, we used CRISPR/Cas9‐mediated gene editing to introduce single amino acid mutations into proteins involved in T cell receptor signalling pathways. Knock‐in mutations were introduced in Jurkat T cells by homologous directed repair using single‐stranded oligodeoxynucleotides. Specifically, we aimed to create targeted mutations at two loci within LCK, a constitutively expressed gene, and at three loci within SH2D2A, whose expression is induced upon T cell activation. Here, we present a simple workflow that can be applied by any laboratory equipped for cell culture work, utilizing basic flow cytometry, Western blotting and PCR techniques. Our data reveal that gene editing may be locus‐dependent and can vary between target sites, also within a gene. In our two targeted genes, on average 2% of the clones harboured homozygous mutations as assessed by allele‐specific PCR and subsequent sequencing. We highlight the importance of decreasing the clonal heterogeneity and developing robust screening methods to accurately select for correct knock‐in mutations. Our workflow may be employed in other immune cell lines and acts as a useful approach for decoding functional mechanisms of proteins of interest.
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Borowicz, Pawel; King, Carolyn G; Dustin, Michael L; Wherry, E John; Koretzky, Gary A & Spurkland, Anne
(2024).
The future of immunology: a Lofoten perspective.
Immunology and Cell Biology.
ISSN 0818-9641.
102(9),
p. 760–765.
doi:
10.1111/imcb.12805.
Full text in Research Archive
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Visser, Johan Georg; Borowicz, Pawel; Chan, Hanna; Hauge, Anette; Kjelstrup, Hanna & Spurkland, Anne
(2023).
Lck-SH2 domain superbinder for characterization of phosphotyrosine signaling in T-cell receptor pathway.
Full text in Research Archive
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Gilmour, Brian Christopher; Chan, Hanna; Borowicz, Pawel; Phuyal, Santosh; Lossius, Andreas & Spurkland, Anne
(2023).
β2, or not β2? Connecting the Lck-adaptor TSAd to integrin β2 via a combined wet lab & bioinformatic approach.
Full text in Research Archive
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Gilmour, Brian Christopher; Chan, Hanna; Borowicz, Pawel; Georg, Visser Johan; Lossius, Andreas & Spurkland, Anne
(2022).
Applying bioinformatics to pick-apart elusive protein functions: Focusing on TSAd/SH2D2A, an immune kinase adaptor.
Full text in Research Archive
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Gilmour, Brian Christopher; Chan, Hanna; Borowicz, Pawel; Kjelstrup, Hanna & Spurkland, Anne
(2021).
Application of bioinformatics to outstanding questions in immunology: TSAd (T cell adapter protein).
Full text in Research Archive
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Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger; Kjelstrup, Hanna; Nyman, Tuula Anneli & Spurkland, Anne
(2018).
Lck’s interactome is influenced by phosphorylation of Tyr192 in the Lck SH2 domain.
Full text in Research Archive
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Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger; Kjelstrup, Hanna; Nyman, Tuula Anneli & Spurkland, Anne
(2018).
Lck’s interactome is influenced by phosphorylation of Tyr192 in the Lck SH2 domain.
Full text in Research Archive
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Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger; Kjelstrup, Hanna; Nyman, Tuula Anneli & Spurkland, Anne
(2018).
Lck’s interactome is influenced by phosphorylation of Tyr192 in the Lck SH2 domain.
Full text in Research Archive
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Borowicz, Pawel; Gopalakrishnan, Ramakrishna Prabhu; Souza, Gustavo Antonio De; Abrahamsen, Greger; Sundvold-Gjerstad, Vibeke & Spurkland, Anne
(2017).
The interactome of TSAd phosphopeptides in T cells.
Full text in Research Archive
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Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger; Kjelstrup, Hanna & Spurkland, Anne
(2017).
The regulatory role of Lck Tyr192 in T cell receptor signalling.
Full text in Research Archive
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Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger & Spurkland, Anne
(2016).
Generation of CRISPR/Cas9 mutants in
order to study TCR signalling pathway.
Full text in Research Archive
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Borowicz, Pawel & Spurkland, Anne
(2020).
Regulation of T cell activation by two conserved phosphotyrosines in Lck and its adapter protein TSAd.
Universitetet i Oslo.
Full text in Research Archive
Show summary
T cells are major players in the immune system. These cells can recognize dangerous antigens through their specific receptors. Harnessing the power of T cells would be beneficial for not only fighting numerous infectious diseases, but also for treating cancer or preventing autoimmune diseases. However, the intracellular signalling involved in the T-cell receptor (TCR) pathway is remarkably complex. It is necessary to understand it more fully in order to control the T cells behaviour. We focused on two proteins involved in TCR signalling. First, lymphocyte-specific protein tyrosine kinase (Lck) starts a phosphorylation cascade downstream of TCR stimulation. Lck itself is tightly controlled by the phosphorylation of two tyrosines: Tyr394 and Tyr505. Recently a third tyrosine, Tyr192, emerged as an important regulatory site. Second, T cell specific adaptor protein (TSAd) is best known for its interaction with Lck. Its main function is to facilitate binding between other proteins. TSAd itself contains a number of phosphotyrosines. One of the most interesting is Tyr290, as it is highly conserved, but it is not predicted to be a prototypic binding site. The aim of this thesis was to understand the function of two phosphotyrosines, Tyr192 in Lck and Tyr290 in TSAd, in order to better comprehend the intracellular signalling of the TCR. The study was based on the knock-in cell lines generated with CRISPR/Cas9 genome editing. The mutated cell lines were studied using a wide spectrum of molecular biology techniques, including mass spectrometry, western blotting, immunoprecipitation and flow cytometry. Our findings revealed that Lck Tyr192 could control and stabilize Lck activity, both positively and negatively. TSAd Tyr290 could also control the protein’s activity. Our results suggest that this tyrosine most likely facilitates the proteins conformation. We have shown that both phosphotyrosines could influence the whole TCR signalling pathway.
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Published
May 25, 2017 12:28 AM
- Last modified
Oct. 28, 2021 5:08 PM
