prikaz prve stranice dokumenta The cell-specific role of optineurin in TLR4- and ER stress-mediated activation of the NF-κB pathway
Embargo period 2030-08-26
master's thesis
The cell-specific role of optineurin in TLR4- and ER stress-mediated activation of the NF-κB pathway
2020. urn:nbn:hr:193:742363
Peradinović, Josip
University of Rijeka
Faculty of Biotechnology and Drug Development

Cite this document

Peradinović, J. (2020). The cell-specific role of optineurin in TLR4- and ER stress-mediated activation of the NF-κB pathway (Master's thesis). Rijeka: University of Rijeka. Retrieved from https://urn.nsk.hr/urn:nbn:hr:193:742363

Peradinović, Josip. "The cell-specific role of optineurin in TLR4- and ER stress-mediated activation of the NF-κB pathway." Master's thesis, University of Rijeka, 2020. https://urn.nsk.hr/urn:nbn:hr:193:742363

Peradinović, Josip. "The cell-specific role of optineurin in TLR4- and ER stress-mediated activation of the NF-κB pathway." Master's thesis, University of Rijeka, 2020. https://urn.nsk.hr/urn:nbn:hr:193:742363

Peradinović, J. (2020). 'The cell-specific role of optineurin in TLR4- and ER stress-mediated activation of the NF-κB pathway', Master's thesis, University of Rijeka, accessed 01 November 2024, https://urn.nsk.hr/urn:nbn:hr:193:742363

Peradinović J. The cell-specific role of optineurin in TLR4- and ER stress-mediated activation of the NF-κB pathway [Master's thesis]. Rijeka: University of Rijeka; 2020 [cited 2024 November 01] Available at: https://urn.nsk.hr/urn:nbn:hr:193:742363

J. Peradinović, "The cell-specific role of optineurin in TLR4- and ER stress-mediated activation of the NF-κB pathway", Master's thesis, University of Rijeka, Rijeka, 2020. Available at: https://urn.nsk.hr/urn:nbn:hr:193:742363

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Metadata
TitleThe cell-specific role of optineurin in TLR4- and ER stress-mediated activation of the NF-κB pathway
Title (croatian)Stanično-specifična uloga optineurina u TLR4 ili ER stresom posredovanoj aktivaciji NF-κB puta
AuthorJosip Peradinović
MentorIvana Munitić (mentor)
MentorJelena Ban (komentor)
Committee memberAntonija Jurak Begonja (predsjednik povjerenstva)
Committee memberRozi Andretić Waldowski (član povjerenstva)
Committee memberIvana Munitić (član povjerenstva)
Committee memberJelena Ban (član povjerenstva)
GranterUniversity of Rijeka
(Faculty of Biotechnology and Drug Development)
Rijeka
Defense date and country2020-08-26, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease, which leads to motor neuron death in the brain and the spinal cord. In 2010 the first optineurin mutations were found in the ALS patients, and the initial biochemical analyses in cell lines suggested that optineurin truncation or mutation in the ubiquitin binding region result in exaggerated nuclear factor κB (NF-κB) activation. Such excessive NF-κB activation and subsequent inflammation were reported to result in neuronal cell death. However, several follow-up studies in primary innate immune cells from optineurin deficiency or insufficiency models could not reproduce these findings. To further test this issue, we decided to induce NF-κB activation with paclitaxel [Toll-like receptor 4 (TLR4) agonist] and tunicamycin [endoplasmic reticulum (ER) stress inducer] to observe if optineurin will exert its role as a negative regulator. These stimuli could mimic some of the stresses that central nervous system (CNS) cells are exposed to during the neurodegenerative process. Microglial (BV2) and neuronal (N2A) optineurin wild-type (WT) and knock-out (KO) cells were used in this research. Optineurin KO BV2 cells exhibited a diminished p65 phosphorylation and TNF-α secretion upon paclitaxel treatment but stronger inhibitor of κB-α (IκB-α) degradation, while N2A KO cells exhibited slightly higher NF-κB activation as detected by p65 phosphorylation. Tunicamycin treated optineurin KO BV2 cells also revealed slightly higher IκB-α degradation and diminished p65 phosphorylation, but TNF-α secretion was not different. Notably, optineurin KO N2A cells also exhibited a slight tendency of stronger NF-κB activation as evidenced by p65 phosphorylation. Results obtained from N2A cells are in correlation with previous studies done on neuronal cell lines, while the ones obtained in BV2 cells differ from results done on primary microglia. Taken together, we propose that optineurin has cell-specific effects, acting as a negative regulator of NF-κB activation in BV2 and N2A cells in response to paclitaxel. It must be noted that these results are obtained in cell lines so they should be used as a preliminary information for subsequent studies on primary cells or in vivo models.
Abstract (croatian)
Amiotrofična lateralna skleroza (ALS) je fatalna neurodegenerativna bolest koja dovodi do smrti motoričkih neurona u mozgu i leđnoj moždini. Prve mutacije optineurina pronađene su u pacijenata oboljelih od ALS-a 2010. godine. Prvotne biokemijske analize u staničnim linijama pokazale su da trunkacija ili mutacija ubikvitin vežuće regije optineurina dovodi do pojačane aktivacije puta nuklearnog čimbenika κB (NF-κB). Pretjerana NF-κB aktivacija te posljedična upala mogu dovesti do neuralne stanične smrti. Međutim, nekoliko studija napravljenih na primarnim stanicama urođene imunosti izoliranih iz modela deficijencije ili insuficijencije optineurina nisu mogle reproducirati rezultate dobivene na staničnim linijama. U svrhu ispitivanja tih kontradikcija, odlučili smo inducirati NF-κB aktivaciju s paklitakselom [agoinst receptora sličnom Toll-u (TLR4)] i tunikamicinom [izaziva stres endoplazmatskog retikuluma (ER)] kako bismo ispitali hoće li optineurin djelovati kao negativan regulator. Navedeni stimulusi mogu imitirati stresore kojem su stanice središnjeg živčanog sustava izložene prilikom procesa neurodegeneracije. U ovom istraživanju koristili smo migroglijalne (BV2) i neuronalne (N2A) stanične linije divljeg tipa (eng. wild-type, WT) i optineurin deficijentne (eng. knock-out, KO). BV2 KO stanice su pokazale smanjenu fosforilaciju p65 aktivaciju i sekreciju TNF-α, no jaču razgradnju inhibitora κB-α (IκB-α) nakon tretmana s paklitakselom. U N2A KO stanicama tretman paklitakselom rezultirao je blago pojačanom NF-κB aktivacijom što je zabilježeno fosforilacijom p65. BV2 KO stanice tretirane s tunikamicinom su pokazale jaču razgradnju IκB-α i smanjenu fosforilaciju p65, no razlika u izlučivanju TNF-α nije zabilježena. N2A KO stanice pokazale su tendenciju blago pojačane NF-κB aktivacije nakon tretmana tunikamicinom što je praćeno fosforilacijom p65. Rezultati dobiveni na N2A stanicama se slažu s prethodnim studijama provedenim na neuronalnim staničnim linijama. Međutim, rezultati dobiveni na BV2 stanicama se ne slažu s onim dobivenim na primarnoj mikrogliji. Uzimajući sve rezultate u obzir, predlažemo da optineurin ima stanično-specifični, te da ima potencijalnu ulogu kao negativan regulator NF-κB aktivacije u BV2 i N2A stanicama. Moralo bi se uzeti u obzir kako su ovi rezultati dobiveni iz staničnih linija te bi se pritom trebali koristiti kao preliminarna informacija za buduće studije koje bi se provele na primarnim stanicama ili u in vivo modelima.
Keywords
Keywords (croatian)
Languageenglish
URN:NBNurn:nbn:hr:193:742363
Study programmeTitle: Biotechnology in medicine
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra biotehnologije u medicini (magistar/magistra biotehnologije u medicini)
Type of resourceText
File originBorn digital
Access conditionsEmbargoed access
Embargo expiration date: 2030-08-26
Terms of useLicence In copyright icon
Created on2020-08-27 08:56:32