Augmented Expression of NOGO-A and Its Receptors in Human Retinal Pigment Epithelial Cells Following Treatment with Human Amniotic Fluid
Background: Nogo-A, a myelin-associated inhibitor for neurite outgrowth, has important role in visual system development. Trans-differentiation ability of human amniotic fluid (HAF) on human retinal pigment epithelial cells (hRPEs) towards neural progenitor cells has been observed in several studies. We aimed to investigate the expression of NOGO-A gene and its receptors as a marker of neural differentiation in HAF-treated hRPE cells.
Methods: hRPE cells were cultivated and immune characterized via RPE65 and cytokeratin 8/18 protein markers. Also, the cytotoxicity effect of 30% HAF on hRPE cells was evaluated using ELISA cell death assay. Finally, expression of NOGO-A and its receptors, RTN4R and LINGO1 was evaluated in the cells treated with HAF in comparison with FBS-treated cells using quantitative real-time PCR.
Results: Harvested cells showed immunoreactivity for cytokeratin 8/18 and RPE65, confirming the hRPE cell identity. Besides, HAF had no cytotoxic effect on hRPE cells compared with FBS-treated cells. Results showed that NOGO-A and its receptors were expressed in cultured hRPE cells. Besides, comparative gene expression analysis revealed significant increased expression of the investigated genes in HAF-treated hRPE cells compared to FBS-treated cells.
Conclusion: Augmented expression of NOGO-A and its receptors can support neural differentiation of hRPE when the cells are treated with HAF. Our outcomes provide more evidences on the trans-differentiation ability of HAF on hRPE cells into neural progenitors and retinal neural cells, but further studies are needed to elucidate the exact mechanism.
2. Sanie-Jahromi F, Ahmadieh H, Soheili Z-S, et al (2012). Enhanced generation of retinal progenitor cells from human retinal pigment epithelial cells induced by amniotic fluid. BMC Res Notes, 5:182.
3. Mishan MA, Kanavi MR, Shahpasand K, et al (2019). Pathogenic tau protein species: Promising therapeutic targets for ocular neurodegenerative diseases. J Ophthalmic Vis Res, 14(4):491-505.
4. Wu DM, Ji X, Ivanchenko MV, et al (2021). Nrf2 overexpression rescues the RPE in mouse models of retinitis pigmentosa. JCI insight, 6(2):e145029.
5. Ben M’Barek K, Monville C (2019). Cell therapy for retinal dystrophies: from cell suspension formulation to complex retinal tissue bioengineering. Stem Cells Intl, 2019:4568979.
6. Nair DSR, Seiler MJ, Patel KH, et al (2021). Tissue Engineering Strategies for Retina Regeneration. Appl Sci (Basel), 11(5):2154.
7. M’Barek KB, Habeler W, Monville C (2018). Stem cell-based RPE therapy for retinal diseases: engineering 3D tissues amenable for regenerative medicine. Adv Exp Med Biol, 1074:625-632.
8. Yan RT, Wang SZ (2000). Differential induction of gene expression by basic fibroblast growth factor and neuroD in cultured retinal pigment epithelial cells. Vis Neurosci, 17(2):157-64.
9. Liang L, Yan RT, Ma W, et al (2006). Exploring RPE as a source of photoreceptors: differentiation and integration of transdifferentiating cells grafted into embryonic chick eyes. Invest Ophthalmol Vis Sci, 47(11):5066-74.
10. Mao W, Yan RT, Wang SZ (2008). Reprogramming chick RPE progeny cells to differentiate towards retinal neurons by ash1. Mol Vis, 14:2309-20.
11. Wang S, Tukachinsky H, Romano FB, et al (2016). Cooperation of the ER-shaping proteins atlastin, lunapark, and reticulons to generate a tubular membrane network. Elife, 5:e18605.
12. Urade T, Yamamoto Y, Zhang X, et al (2014). Identification and characterization of TMEM33 as a reticulon-binding protein. Kobe J Med Sci, 60(3):E57-65.
13. Yamamoto Y, Yoshida A, Miyazaki N, et al (2014). Arl6IP1 has the ability to shape the mammalian ER membrane in a reticulon-like fashion. Biochem J, 458(1):69-79.
14. Schwab ME, Strittmatter SM (2014). Nogo limits neural plasticity and recovery from injury. Curr Opin Neurobiol, 27:53-60.
15. Sekine Y, Lindborg JA, Strittmatter SM (2020). A proteolytic C-terminal fragment of Nogo-A (reticulon-4A) is released in exosomes and potently inhibits axon regeneration. J Biol Chem, 295(8):2175-2183.
16. Petrinovic MM, Duncan CS, Bourikas D, et al (2010). Neuronal Nogo-A regulates neurite fasciculation, branching and extension in the developing nervous system. Development, 137(15):2539-50.
17. Schmandke A, Schmandke A, Schwab ME (2014). Nogo-A: multiple roles in CNS development, maintenance, and disease. Neuroscientist, 20(4):372-386.
18. Beliën AT, Paganetti PA, Schwab ME (1999). Membrane-type 1 matrix metalloprotease (MT1-MMP) enables invasive migration of glioma cells in central nervous system white matter. J Cell Biol, 144(2): 373–384.
19. Ineichen BV, Kapitza S, Bleul C, et al (2017). Nogo-A antibodies enhance axonal repair and remyelination in neuro-inflammatory and demyelinating pathology. Acta Neuropathol, 134(3):423-440.
20. Oertle T, Van Der Haar ME, Bandtlow CE, et al (2003). Nogo-A inhibits neurite outgrowth and cell spreading with three discrete regions. J Neurosci, 23(13):5393-406.
21. Fournier AE, GrandPre T, Strittmatter SM (2001). Identification of a receptor mediating Nogo-66 inhibition of axonal regeneration. Nature, 409(6818):341-6.
22. Mi S, Lee X, Shao Z, et al (2004). LINGO-1 is a component of the Nogo-66 receptor/p75 signaling complex. Nat Neurosci, 7(3):221-8.
23. Shao Z, Browning JL, Lee X, et al (2005). TAJ/TROY, an orphan TNF receptor family member, binds Nogo-66 receptor 1 and regulates axonal regeneration. Neuron, 45(3):353-9.
24. Wang KC, Kim JA, Sivasankaran R, et al (2002). P75 interacts with the Nogo receptor as a co-receptor for Nogo, MAG and OMgp. Nature, 420(6911):74-8.
25. Yamashita T, Fujitani M, Yamagishi S, et al (2005). Multiple signals regulate axon regeneration through the Nogo receptor complex. Mol Neurobiol, 32(2):105-11.
26. Pernet V (2017). Nogo-A in the visual system development and in ocular diseases. Biochim Biophys Acta Mol Basis Dis, 1863(6):1300-1311.
27. Mdzomba JB, Joly S, Rodriguez L, et al (2020). Nogo-A-targeting antibody promotes visual recovery and inhibits neuroinflammation after retinal injury. Cell Death & Disease, 11:1-16.
28. Feizi S, Soheili ZS, Bagheri A, et al (2014). Effect of amniotic fluid on the in vitro culture of human corneal endothelial cells. Exp Eye Res, 122:132-40.
29. Pierce J, Jacobson P, Benedetti E, et al (2016). Collection and characterization of amniotic fluid from scheduled C-section deliveries. Cell Tissue Bank, 17(3):413-25.
30. Davari M, Soheili ZS, Ahmadieh H, et al (2013). Amniotic fluid promotes the appearance of neural retinal progenitors and neurons in human RPE cell cultures. Mol Vis, 19: 2330–2342.
31. Ghaderi S, Soheili ZS, Ahmadieh H, et al (2011). Human amniotic fluid promotes retinal pigmented epithelial cells' trans-differentiation into rod photoreceptors and retinal ganglion cells. Stem Cells Dev, 20(9):1615-25.
32. Heidari R, Soheili ZS, Samiei S, et al (2015). Alginate as a cell culture substrate for growth and differentiation of human retinal pigment epithelial cells. Appl Biochem Biotechnol, 175(5):2399-412.
33. Liu Y, Ma C, Li H, et al (2019). Nogo-A/Pir-B/TrkB signaling pathway activation inhibits neuronal survival and axonal regeneration after experimental intracerebral hemorrhage in rats. J Mol Neurosci, 69(3):360-370.
34. Wang L, Wang J, Ma D, et al (2016). Isoform‐specific localization of Nogo protein in the optic pathway of mouse embryos. J Comp Neurol, 524(11):2322-34.
35. Wälchli T, Pernet V, Weinmann O, et al (2013). Nogo-A is a negative regulator of CNS angiogenesis. Proc Natl Acad Sci U S A, 110(21):E1943-52.
36. Pernet V, Joly S, Christ F, et al (2008). Nogo-A and myelin-associated glycoprotein differently regulate oligodendrocyte maturation and myelin formation. J Neurosci, 28(29):7435-44.
37. Ahmed Z, Suggate EL, Brown ER, et al (2006). Schwann cell-derived factor-induced modulation of the NgR/p75NTR/EGFR axis disinhibits axon growth through CNS myelin in vivo and in vitro. Brain, 129(Pt 6):1517-33.
38. Huo Y, Yin XL, Ji SX, et al (2013). Inhibition of retinal ganglion cell axonal outgrowth through the Amino-Nogo-A signaling pathway. Neurochem Res, 38(7):1365-74.
39. Pernet V, Joly S, Dalkara D, et al (2012). Neuronal Nogo-A upregulation does not contribute to ER stress-associated apoptosis but participates in the regenerative response in the axotomized adult retina. Cell Death Differ, 19(7):1096-108.
40. Iketani M, Yokoyama T, Kurihara Y, et al (2016). Axonal branching in lateral olfactory tract is promoted by Nogo signaling. Sci Rep, 6:39586.
41. Hu F, Strittmatter SM (2008). The N-terminal domain of Nogo-A inhibits cell adhesion and axonal outgrowth by an integrin-specific mechanism. J Neurosci, 28(5):1262-1269.
42. Koprivica V, Cho KS, Park JB, et al (2005). EGFR activation mediates inhibition of axon regeneration by myelin and chondroitin sulfate proteoglycans. Science, 310(5745):106-10.
43. Shao Z, Lee X, Huang G, et al (2017). LINGO-1 regulates oligodendrocyte differentiation through the cytoplasmic gelsolin signaling pathway. J Neurosci, 37(12):3127-3137.
44. Lv J, Xu Rx, Jiang XD, et al (2010). Passive immunization with LINGO-1 polyclonal antiserum afforded neuroprotection and promoted functional recovery in a rat model of spinal cord injury. Neuroimmunomodulation, 17(4):270-8.
45. Mathis C, Schröter A, Thallmair M, et al (2010). Nogo-a regulates neural precursor migration in the embryonic mouse cortex. Cereb Cortex, 20(10):2380-90.
|Issue||Vol 51 No 7 (2022)|
|Retinal pigment epithelial cells Human amniotic fluid Ophthalmology|
|Rights and permissions|
|This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.|