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Chiro-optical modulation for NURR1 production from stem cells Madhumita Patel, Hyo Jung Moon, Ja Hye Hong, and Byeongmoon Jeong ACS Chem. Neurosci., Just Accepted Manuscript • Publication Date (Web): 28 Apr 2017 Downloaded from http://pubs.acs.org on May 1, 2017
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ACS Chemical Neuroscience
Chiro-optical Modulation for NURR1 Production from Stem Cells
Madhumita Patel, Hyo Jung Moon, Ja Hye Hong, and Byeongmoon Jeong*
Department of Chemistry and Nanoscience, Ewha Womans University, 52 Ewhayeodae-gil, Seodaemun-gu, Seoul, 03760 Korea
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ABSTRACT: Nuclear receptor related 1 (NURR1) is an essential protein for maintenance of dopaminergic neurons in adult midbrain of which deficiency leads to Parkinson’s disease. To enhance the NURR1 production of neural cells, various approaches are under investigation. Here we report that the NURR1 is highly expressed in stem cells by exposure to an Lpolarized blue light emitting diode (LED). Compared to stem cells cultured in the absence of a LED, under polarized green and red LEDs, the stem cells exposed to a polarized blue LED significantly enhanced neuronal biomarkers such as neurofilament M (NFM) and neuron specific enolase (NSE) at both mRNA and protein levels. In particular, NURR1 was selectively enhanced by the stem cells exposed to the L-polarized blue LED. Stem cells exposed to the L-polarized blue LED increased mitochondrial ATP and intracellular calcium ions, which support neuronal differentiation of the stem cells. This study suggests that chirooptical treatments by using polarized light with a specific wavelength can be used for engineering of stem cells with enhanced specific biochemicals, which may open a new method for a specific disease. Keywords: NURR1 • polarized light • light emitting diode • neuronal differentiation
Neurodegenerative diseases including Alzheimer disease and Parkinson’s disease become more and more important as the old population increases.1,2 Selective production of pharmacologically important molecules by using cells, cell engineering, can be a promising tool to cope with the diseases.3 NURR1 is an essential protein for maintenance of dopaminergic neurons in adult midbrain of which deficiency leads to Parkinson’s disease.4-6 To enhance the NURR1 production in brain cells, various approaches including agonistic drug treatments and gene delivery are under investigation.7,8 Here, we report that production
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of NURR1 is preferentially enhanced in stem cells by simple exposure of the stem cells to a L-polarized blue light. A light emitting diode (LED) is a safe and cheap source of light, and its biological applications are based on the principle that certain molecules in living systems are able to absorb photons and trigger signaling pathways in response to the light.9,10 In comparison to pharmacobiological stimulation through chemicals and growth factors, optical stimulation of cells has its advantages. Note the simplicity and selectivity of focusing light on an individual cell without disturbing the other cells. However, there are limited studies on bio modulations of stem cells using LED lights. A red LED light affects osteogenic differentiation,11,12 whereas a blue LED light, widely used as an antibacterial or germicidal tool,13 induces neuronal differentiation from embryonic stem cells.14-16 Polarized light is an electromagnetic wave in which vibration occurs in a single plane. Various applications of the polarized light for chemical reactions such as enantioselective polymerization of diacetylene,17 control of supramolecular chirality of poly(diacetylene) derivatives,18 chiral induction of an azobenzene methacrylate polymer,19 and switching of helicity of a polymer20 have been reported. On the other hand, previous studies for cells or stem cells employed unpolarized light as a single source of stimulation,11-16 and the effects of a polarized light on stem cell stimulation have not been reported yet. In this study, we investigated the effects of L-polarized (PL) and R-polarized (PR) LED lights of blue (460 nm), green (540 nm) and red (630 nm) on tonsil-derived mesenchymal stem cells (TMSCs) by exposing the TMSCs to 0.14 mW/mm2 polarized LED lights for 24 hours during the neuronal differentiation of the stem cells.
■ RESULTS AND DISCUSSION
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TMSCs were isolated from the tonsil tissue of an 11 years old female donor after her tonsillectomy,21 and kindly donated from the Ewha Womans University, Mokdong Hospital. TMSCs were also cultured under the same conditions without LED exposure (-L) as a case of the traditional culture system. All the cells were viable after the exposure to the polarized light as exhibited by the green color in the live (green) and dead (red) images, and the cells developed similar fibrous morphologies, irrespective of the presence of L- and R-polarized LED lights (Supporting Information, Figure S1). This fact suggests that 24 hour exposure of the stem cells to an L- or R-polarized LED light used in this study did not invoke any detrimental effects on the stem cells. Expressions of neuronal biomarkers including NURR1, neuron specific enolase (NSE), and neurofilament M (NFM) were compared at the mRNA level in the –L, PL, and PR systems (Figure 1). NURR1 mRNA expression exhibited 14 and 4 times enhancements for TMSCs exposed to L- and R-polarized blue LED lights, respectively, compared to the unexposed cells to an LED light. In particular, the expression of NURR1 mRNA in the PL system is significantly greater (about 3.5 times) than the PR system, suggesting that TMSCs differentiate the L- polarized blue LED light from the R-polarized blue LED light. About 2 times of the enhancements of NURR1 mRNA expression were observed in the cells exposed to L-and R-polarized green LED lights than unexposed cells. Compared to the –L system, the expression of NSE mRNA enhanced 20–25 times in both PL and PR systems. There was no significant difference between PL and PR systems for the expression of NSE mRNA. The enhancement of NSE mRNA expression was significant only for the polarized blue LED lights, but not for the polarized green and red LED lights. Expression of NFM mRNA enhanced 2–3 times for the blue PL and PR systems, compared to the –L system. Again, the enhancement in NFM mRNA expression was significant only for the polarized blue LED lights, but not for the polarized green and red LED lights. Protein level was assayed for the
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Figure 1. mRNA expressions of NURR1 (a) NSE (b), and NFM (c) assayed by RT-PCR. The systems in absence of light (-L), cells exposed to L-polarized light (PL) and R-polarized light (PR) are compared. * and ** indicate p
