Exposure to the complement C5b-9 complex sensitizes 661W photoreceptor cells to both apoptosis and necroptosis

Shi, Hui; Williams, Jennifer A. E.; Guo, Li; Stampoulis, Dimitrios; Cordeiro, M. Francesca; Moss, Stephen E.

doi:10.1007/s10495-015-1091-7

Cited by 15 publications

(11 citation statements)

References 35 publications

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“…It is known that depositions of MAC on the cell membrane induces apoptosis (Sato et al, 1999; Hughes et al, 2000). Shi et al reported that MAC promoted apoptosis in their used cone photoreceptor cell line (Shi et al, 2015). In an OHT glaucoma model, the depletion of the complement system reduced apoptotic RGCs (Jha et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

Intravitreal Therapy Against the Complement Factor C5 Prevents Retinal Degeneration in an Experimental Autoimmune Glaucoma Model

et al. 2019

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In glaucoma, studies revealed an involvement of the complement system. In an experimental autoimmune glaucoma model, immunization with an optic nerve homogenate antigen (ONA) led to retinal ganglion cell (RGC) loss, while intraocular pressure (IOP) remained unchanged. Here, we investigated the therapeutic effect of a complement system inhibition in this model. Hence, rats were immunized with ONA and compared to controls. In one eye of the ONA animals, an antibody against complement factor C5 was intravitreally injected (15 μmol: ONA+C5-I or 25 μmol: ONA+C5-II) before immunization and then every two weeks. IOP was measured weekly. After 6 weeks, spectral-domain optical coherence tomographies (SD-OCT), electroretinograms (ERG), immunohistochemistry, and quantitative real-time PCR analyses were performed. IOP and retinal thickness remained unchanged within all groups. The a-wave amplitudes were not altered in the ONA and ONA+C5-I groups, whereas a decrease was noted in ONA+C5-II animals (p < 0.05). ONA immunization provoked a significant decrease of the b-wave amplitude (p < 0.05), which could be preserved in ONA+C5-I, but not in ONA+C5-II animals. ONA animals showed a loss of RGCs (p = 0.001), while ONA+C5-I and ONA+C5-II retinae had similar cell counts as controls. A significant downregulation of apoptotic Bax/Bcl2 mRNA was noted in ONA+C5-I retinae (p = 0.02). Significantly more C3+ and MAC+ cells were observed in ONA animals (p < 0.001). The amount of C3+ cells in both treatment groups was significantly increased (p < 0.01), while the number of MAC+ cells in the treated retinas did not differ from controls. The number of activated microglia cells remained unchanged in ONA animals, but was increased in the treatment groups (p < 0.05). Recoverin+ cells were diminished in ONA animals (p = 0.049), but not in treated ones. Rho mRNA was downregulated in ONA and in ONA+C5-II retinas (both p = 0.014). Less opsin+ cones were observed in ONA animals (p = 0.009), but not in the treated groups. Our results indicate that the C5 antibody inhibits activation of the complement system, preventing the loss of retinal function as well as RGC, cone bipolar, and photoreceptor loss. Therefore, this approach might be a suitable new treatment for glaucoma patients, in which immune dysregulation plays an important factor for the development and progression of glaucoma.

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Section: Discussionmentioning

confidence: 99%

Intravitreal Therapy Against the Complement Factor C5 Prevents Retinal Degeneration in an Experimental Autoimmune Glaucoma Model

et al. 2019

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“…This is different from the present study, in which 661W and RGC-5 cells developed an average of 3.0 ± 0.1 and 3.1 ± 0.3 primary neurites per cell, respectively, when exposed to 316 nM staurosporine, the concentration which was optimal for differentiation. [ 17 ]. It should be also noted that that study and the present study only assessed morphological features of neuronal differentiation, and a more extensive study of cell-surface markers and electrophysiological activity would be needed to confirm a neuronal phenotype, as was done previously [ 1 ].…”

Section: Discussionmentioning

confidence: 99%

Induction of Neuronal Morphology in the 661W Cone Photoreceptor Cell Line with Staurosporine

et al. 2015

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PurposeRGC-5 cells undergo differentiation into a neuronal phenotype with low concentrations of staurosporine. Although the RGC-5 cell line was initially thought to be of retinal ganglion cell origin, recent evidence suggests that the RGC-5 line could have been the result of contamination with 661W mouse cone photoreceptor cells. This raised the possibility that a cone photoreceptor cell line could be multipotent and could be differentiated to a neuronal phenotype.Methods661W and RGC-5 cells, non-neuronal retinal astrocytes, retinal endothelial cells, retinal pericytes, M21 melanoma cells, K562 chronic myelogenous leukemia cells, and Daudi Burkitt lymphoma cells, were differentiated with staurosporine. The resulting morphology was quantitated using NeuronJ with respect to neurite counts and topology.ResultsTreatment with staurosporine induced similar-appearing morphological differentiation in both 661W and RGC-5 cells. The following measures were not significantly different between 661W and RGC-5 cells: number of neurites per cell, total neurite field length, number of neurite branch points, and cell viability. Neuronal-like differentiation was not observed in the other cell lines tested.Conclusions661W and RGC-5 cells have virtually identical and distinctive morphology when differentiated with low concentrations of staurosporine. This result demonstrates that a retinal neuronal precursor cell with cone photoreceptor lineage can be differentiated to express a neuronal morphology.

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“…Because of the difficulty of isolation and culture of primary photoreceptor cells, a photoreceptor cell line 661W has been widely used to study mechanisms of photoreceptor dysfunction and degeneration, and develop strategies to protect photoreceptors in a variety of conditions such as light damage, retinitis pigmentosa, diabetic retinopathy, retinal dystrophy, and retinal detachment (Aslanidis et al, 2015;Besirli et al, 2012;Byrne et al, 2016;Du et al, 2015;Izawa et al, 2016;Jackson et al, 2016;Krishnamoorthy et al, 1999;Lee et al, 2016;Rapp et al, 2014;Scholz et al, 2015;Shi et al, 2015;Tan et al, 2004;Zhang et al, 2014). To determine whether photoreceptor cells can produce chemokines CXCL10 and CCL2 under stress conditions, we treated 661W cells with thapsigargin (TG), which potently induces ER stress at 0.01-0.1 μM (Bartolome et al, 2012;Hamamura and Yokota, 2007) and is widely used to study ER stress-induced cellular responses (Bartolome et al, 2012;Hamamura and Yokota, 2007;Kim et al, 2008).…”

Section: Endoplasmic Reticulum (Er) Stress Induces Cxcl10 and Ccl2 Prmentioning

confidence: 99%

PERK and XBP1 differentially regulate CXCL10 and CCL2 production

Zhu

Liu

Sha

et al. 2017

Experimental Eye Research

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Inflammation plays a key role in the pathogenesis of many retinal degenerative diseases related with photoreceptor dysfunction/degeneration. However the involvement of photoreceptor cells in inflammatory reactions is largely unknown as they are not considered as inflammatory cells. In this study, we assessed whether photoreceptor cells can produce CCL2 and CXCL10, two important players in inflammation during endoplasmic reticulum (ER) stress. After photoreceptor 661W cells were treated with ER stress inducer thapsigargin (TG), induction of ER stress increased CXCL10 and CCL2 expression at both mRNA and protein levels, which was significantly blocked by an ER stress blocker 4-phenylbutyrate. ER stress contains three pathways: PERK, ATF6 and IRE1α. Knockdown of PERK attenuated TG-induced CXCL10 and CCL2 mRNA expression, associated with significant decreases in phosphorylation of NF-κB RelA and STAT3. In contrast to PERK, knockdown of XBP1, which is activated by IRE1α-mediated splicing, robustly enhanced TG-induced CXCL10 and CCL2 expression and phosphorylation of NF-κB RelA and STAT3. Blockade of NF-κB or STAT3 markedly diminished TG-induced CXCL10 and CCL2 expression. The specific roles of PERK and XBP1 in CXCL10 and CCL2 expression were further investigated by treating photoreceptor cells with advanced glycation end products (AGE) and high glucose (HG), two of the major contributors to diabetic complications. Similarly, AGE and HG induced CXCL10 and CCL2 expression in which PERK was a positive regulator while XBP1 was a negative regulator. These studies suggest that photoreceptors may be involved in retinal inflammation by expressing chemokines CXCL10 and CCL2. PERK and IRE1α/XBP1 in the unfolded protein response differentially regulate the expression of CXCL10 and CCL2 likely through modulation of ER stress-induced NF-κB RelA and STAT3 activation.

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Exposure to the complement C5b-9 complex sensitizes 661W photoreceptor cells to both apoptosis and necroptosis

Cited by 15 publications

References 35 publications

Intravitreal Therapy Against the Complement Factor C5 Prevents Retinal Degeneration in an Experimental Autoimmune Glaucoma Model

Intravitreal Therapy Against the Complement Factor C5 Prevents Retinal Degeneration in an Experimental Autoimmune Glaucoma Model

Induction of Neuronal Morphology in the 661W Cone Photoreceptor Cell Line with Staurosporine

PERK and XBP1 differentially regulate CXCL10 and CCL2 production

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