Researcher:
Ünlü, Eda Kuşan

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Eda Kuşan

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Ünlü

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Ünlü, Eda Kuşan

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2020 - 20223

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    Investigation of mitophagy biomarkers in corneal epithelium of keratoconus patients
    (Taylor and Francis Inc, 2022) Gumus, Koray; Sarac, Ozge Ilhan; Cagil, Nurullah; N/A; N/A; N/A; N/A; N/A; N/A; Yıldız, Erdost; Aydemir, Dilara; Zibandeh, Noushin; Ünlü, Eda Kuşan; Karslıoğlu, Melisa Zişan; Şahin, Afsun; PhD Student; PhD Student; Researcher; Researcher; Doctor; Faculty Member; Graduate School of Health Sciences; Graduate School of Health Sciences; N/A; Graduate School of Health Sciences; N/A; School of Medicine; N/A; N/A; Koç University Research Center for Translational Medicine (KUTTAM) / Koç Üniversitesi Translasyonel Tıp Araştırma Merkezi (KUTTAM); N/A; Koç University Hospital; N/A; N/A; N/A; N/A; N/A; N/A; 171267
    Purpose: The pathological mechanisms of keratoconus (KC) have not been elucidated yet. Mitophagy is an important mechanism that eliminates damaged mitochondria under oxidative stress, and it could be one of the leading pathological causes of KC. This study aimed to find out the role of mitophagy in the keratoconic corneal epithelium. Methods: The corneal epithelia were collected from the 103 progressive KC patients and the 46 control subjects. The real-time quantitative PCR was performed for PTEN-putative kinase-1 (PINK1), PARKIN, p62, and BNIP3 gene expressions in 31 KC and 9 control subjects. Western blot analyses were performed to investigate the protein expressions of PINK1, PARKIN, LC3B, ATG5, and BECLIN in the remaining 109 corneal epithelium samples from 72 patients and 37 control subjects. Results: mRNA and protein expressions of PINK1 decreased significantly in the corneal epithelium of KC patients compared to the control subjects. No significant change was found in mRNA levels of PARKIN, p62, and BNIP3 in KC patients. The protein expression of PARKIN, LC3B, ATG5, and Beclin did not significantly differ between KC patients and control subjects. Gene expression levels of mitophagy biomarkers were not affected by the KC grade. Conclusions: PINK1/PARKIN-dependent mitophagy is affected in the keratoconic corneal epithelium. We found significant decreases in both mRNA and protein expressions of PINK1 in the keratoconic corneal epithelium. However, we did not observe any other significant change in mitophagy markers. Mitochondrial stress-related mitophagy pathways could be interrupted by the decreased levels of PINK1 in the keratoconic corneal epithelium, but solely PINK1 dysregulation is not likely to induce KC pathogenesis.
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    The regulatory effect of IFN-gamma or IL-17a primed limbal mesenchymal stem cells on lymphocytes immune response
    (association for Research in Vision and Ophthalmology (aRVO), 2021) N/A; N/A; N/A; Zibandeh, Noushin; Ünlü, Eda Kuşan; Şahin, Afsun; Resercher; PhD Student; Faculty Member; Koç University Research Center for Translational Medicine (KUTTAM) / Koç Üniversitesi Translasyonel Tıp Araştırma Merkezi (KUTTAM); Koç University Research Center for Translational Medicine (KUTTAM) / Koç Üniversitesi Translasyonel Tıp Araştırma Merkezi (KUTTAM); Koç University Research Center for Translational Medicine (KUTTAM) / Koç Üniversitesi Translasyonel Tıp Araştırma Merkezi (KUTTAM); N/A; Graduate School of Health Sciences; School of Medicine; N/A; N/A; 171267
    Purpose : The cornea is a transparent, avascular tissue that covers the front portion of the eye. In pathologic and highly inflammatory conditions, the entire ocular surface is at risk for persistent scarring and visual loss. Despite all the treatment strategies, no effective solution has been found for patients with severe corneal injuries. Mesenchymal stem cells (MSCs) are of particular interest for the treatment of immune-related diseases owing to their immunosuppressive properties. Priming with proinflammatory cytokines improve the immunosuppressive function of MSCs. The purpose of our study is to identify the immunosuppressive effect of interferon (IFN)-γ and Interleukin (IL)17A primed Limbal- MSCs (LMSCs) on lymphocyte response. Methods : LMSCs were isolated from cadaveric corneoscleral rims and cultured in DMEM with penicillin-streptomycin and fetal bovine serum. LMSCs were characterized and differentiated in passage 3. The lymphocytes were isolated from peripheral venous blood of healthy controls (n=10). LMSCs were stimulated with IFN-γ or IL-17A for 48 hours. At the end of this period, peripheral blood mononuclear cells of healthy individuals were isolated and cultured with or without stimulated LMSCs for 72 hours. The cultures were stimulated with anti-CD3 and anti-CD28 antibodies. After 72 hours, lymphocytes were collected and analyzed for proliferation assay, cell viability assay with Annexinv/PI, Fas, FasL and CD4+CD25+FoxP3+T regulatory cell ratio via flow cytometry. Results : Our results demonstrated that IFN-γ or IL-17A stimulated LMSCs suppressed the proliferation of T lymphocytes compared to unstimulated LMSCs cultures and it was statistically significant (P<0.05, P<0.05, respectively). IFN-γ stimulated LMSCs suppressed the Annexin V and Fas expression compared to unstimulated LMSCs cultures (P<0.05) but IL-17A stimulated LMSCs did not reduce the AnnexinV and Fas expression significantly compared to unstimulated LMSCs cultures (P>0.05). IFN- γ stimulated LMSCs increased T regulatory cell frequency compared to unstimulated LMSCs cultures (P<0.05). Conclusions : Our data showed that stimulated LMSCs especially IFN-γ decreased lymphocyte proliferation and apoptosis while increased the T regulatory cell ratio. We believe that LMScs could potentially be a source for future treatment strategies of inflammatory conditions particularly corneal disease.
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    Does brimonidine tartarate have any effect on viability and/or proliferation of limbal mesenchymal limbal stem cells?
    (Assoc Research Vision Ophthalmology Inc, 2020) N/A; N/A; N/A; N/A; N/A; N/A; Karslıoğlu, Melisa Zişan; Zibandeh, Noushin; Aydemir, Dilara; Ünlü, Eda Kuşan; Kesim, Cem; Taş, Ayşe Yıldız; Şahin, Afsun; Doctor; Researcher; PhD Student; Researcher; Researcher; Doctor; Faculty Member; Faculty Member; Koç University Research Center for Translational Medicine (KUTTAM) / Koç Üniversitesi Translasyonel Tıp Araştırma Merkezi (KUTTAM); Koç University Hospital; N/A; Graduate School of Health Sciences; Graduate School of Health Sciences; School of Medicine; School of Medicine; School of Medicine; N/A; N/A; N/A; N/A; 387367; 200905; 171267
    Purpose : Mesenchymal stem cell transplantation is still one of the hot topics within translational medicine. The limbus, rim of the corneoscleral junction of human eye, is highly rich in terms of stem cells. Limbal mesenchymal stem cells (LMSCs) have been used in many ocular diseases. Brimonidine tartrate (BT) is selective alpha-2 adrenergic agonist that its neuroprotective effect had been proven. BT drops are preferred in glaucoma patients especially for neuroprotection. Our purpose is to evaluate the effect of BT on viability and/or proliferation of LMSCs in vitro. Methods : LMSCs were isolated from corneoscleral rim. The cells in third passage were characterized and differentiated. For MTT assay, cells were seeded in the density of 3x103 in 96 well plate and pretreatment with BT in 0.1, 1, 5,10 and 200 μM concentrations. After 48 hours, MTT solution was added to each wells and plate was read in 570nm absorbance. For scratch assay, cells were seeded in the density of 6x104 in 12 well plate and pretreatment with BT in pervious mentioned concentrations. For Annexin V/PI cell viability test, cells were seeded in the density of 2x105 in 6 well plate and pretreatment with BT in pervious mentioned concentrations. After 48 hours, cells were washed and stained with Annexin V The cells were analysed by flowcytometry. Results : Our experiments showed that pre-treated LMSCs with 10 μM BT increased cell viability significantly compared to untreated cells in MTT assay (p<0.05). In scratch assay pre-treated LMSCs with 5 μM and 10 μM BT displayed statistically significant rapid migration at 20th hours regarding to other untreated cells (p<0.05). According to cell viability results, pre-treated LMSCs with 10 μM BT decreased Annexin V expression compared to untreated group. And it is statistically significant (p<0.05). Conclusions : Since limbal mesenchymal stem cells and optic nerve share the same embryological origin (ectodermal), it is not surp rising that in our study BT enhanced the viability of limbal mesenchymal stem cells and improved proliferation. From this point of view, we hypothesized that intravitreally injection of brimonidine tartrate loaded limbal mesenchymal stem cells may not only protect axon loss, but also induce axon regeneration in traumatic optic nerve injuries. This preliminary study supported our hypothesis and we are in the process of planning future studies about this topic.