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Optimal transcorneal electrical stimulation parameters for preserving photoreceptors in a mouse model of retinitis pigmentosa

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Neural Regeneration Research 2024年第11期19卷 2543-2552页

作者： Sam Enayati Karen Chang Anton Lennikov Menglu Yang Cherin Lee Ajay Ashok Farris Elzaridi Christina Yen Kasim Gunes Jia Xie kin-sang cho Tor Paaske Utheim Dong Feng Chen Schepens Eye Research Institute of Massachusetts Eye and Ear Department of OphthalmologyHarvard Medical SchoolBostonMAUSA Institute of clinical medicine Faculty of MedicineUniversity of OsloOsloNorway Department of Medical Biochemistry Oslo University HospitalUniversity of OsloOsloNorway Department of Ophthalmology Drammen HospitalVestre Viken Hospital TrustDrammenNorway Department of Histology and Embryology School of MedicineMarmara UniversityIstanbulTurkiye

Retinitis pigmentosa is a hereditary retinal disease that affects rod and cone photoreceptors,leading to progressive photoreceptor *** research supports the beneficial effect of electrical stimulation on photoreceptor *** study aims to identify the most effective electrical stimulation parameters and functional advantages of transcorneal electrical stimulation(tcES)in mice affected by inherited retinal ***,the study seeked to analyze the electric field that reaches the retina in both eyes in mice and post-mortem *** this study,we recorded waveforms and voltages directed to the retina during transcorneal electrical stimulation in C57BL/6J mice using an intraocular needle probe with rectangular,sine,and ramp *** investigate the functional effects of electrical stimulation on photoreceptors,we used human retinal explant cultures and rhodopsin knockout(Rho^(-/-))mice,demonstrating progressive photoreceptor degeneration with *** retinal explants isolated from the donors’eyes were then subjected to electrical stimulation and cultured for 48 hours to simulate the neurodegenerative environment in *** density was evaluated by rhodopsin *** vivo Rho^(-/-)mice were subjected to two 5-day series of daily transcorneal electrical stimulation using rectangular and ramp *** function and visual perception of mice were evaluated by electroretinography and optomotor response(OMR),*** was used to assess the morphological and biochemical changes of the photoreceptor and bipolar cells in mouse *** recordings indicated effective delivery of rectangular,sine,and ramp waveforms to the retina by transcorneal electrical stimulation,of which the ramp waveform required the lowest *** of the total conductive resistance of the post-mortem human compared to the mouse eyes indicated higher cornea-to-retina resistance in human *** tem

关键词： bipolar cells electrical stimulation neuroprotection photoreceptor degeneration retina retinal explants retinitis pigmentosa transcorneal electrical stimulation waveform

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Absence of ephrin-A2/A3 increases retinal regenerative potential for Müller cells in Rhodopsin knockout mice

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Neural Regeneration Research 2021年第7期16卷 1317-1322页

作者： Rui-Lin Zhu Yuan Fang Hong-Hua Yu Dong FChen Liu Yang kin-sang cho Department of Ophthalmology Peking University First HospitalBeijingChina Schepens Eye Research Institute Massachusetts Eye and EarHarvard Medical SchoolBostonMAUSA Department of Ophthalmology and Visual Science Eye&ENT HospitalShanghai Medical CollegeFudan UniversityShanghaiChina Guangdong Eye Institute Department of OphthalmologyGuangdong Provincial People’s HospitalGuangdong Academy of Medical SciencesGuangzhouGuangdong ProvinceChina

Müller cells(MC) are considered dormant retinal progenitor cells in *** studies demonstrated ephrin-As act as negative regulators of neural progenitor cells in the retina and *** remains unclear whether the lack of ephrin-A2/A3 is sufficient to promote the neurogenic potential of *** we investigated whether the MC is the primary retinal cell type expressing ephrin-A2/A3 and their role on the neurogenic potential of Müller *** this study, we showed that ephrin-A2/A3 and their receptor EphA4 were expressed in retina and especially enriched in *** level of ephrin As/EphA4 expression increased as the retina matured that is correlated with the reduced proliferative and progenitor cell potential of ***, we investigated the proliferation in primary MC cultures isolated from wild-type and A2~(–/–) A3~(–/–) mice by 5-ethynyl-2′-deoxyuridine(EdU) *** detected a significant increase of EdU~+ cells in MC derived from A2~(–/–) A3~(–/–) ***, we investigated the role of ephrin-A2/A3 in mice undergoing photoreceptor degeneration such as Rhodopsin knockout(Rho~(–/–)) *** further evaluate the role of ephrin-A2/A3 in MC proliferation in vivo, EdU was injected intraperitoneally to adult wild-type, A2~(–/–) A3~(–/–), Rho~(–/–) and Rho~(–/–) A2~(–/–) A3~(–/–) mice and the numbers of EdU~+ cells distributed among different layers of the *** As/EphA4 expression was upregulated in the retina of Rho~(–/–) mice compared to the wild-type *** addition, cultured MC derived from ephrin-A2~(–/–) A3~(–/–) mice also expressed higher levels of progenitor cell markers and exhibited higher proliferation potential than those from wild-type ***, we detected a significant increase of EdU~+ cells in the retinas of adult ephrin-A2~(–/–) A3~(–/–) mice mainly in the inner nuclear layer;and these EdU~+ cells were co-localized with MC marker, cellular retinaldehyde-binding protein, suggesting some proliferating cells are from

关键词： endogenous stem cell EphA4 ephrin-A2 ephrin-A3 ephrins Müller cell photoreceptor cell regeneration retinal degeneration retinal regeneration retinal stem cell

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Non-invasive electrical stimulation as a potential treatment for retinal degenerative diseases

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Neural Regeneration Research 2021年第8期16卷 1558-1559页

作者： Karen Chang Sam Enayati kin-sang cho Tor P.Utheim Dong Feng Chen Schepens Eye Research Institute of Massachusetts Eye and Ear Department of OphthalmologyHarvard Medical SchoolBostonMAUSA Department of Medical Biochemistry Oslo University HospitalOsloNorway Department of Ophthalmology Drammen HospitalNorway

More than 38.5 million people are estimated to be affected by blindness worldwide in 2020(Flaxman et al., 2017). Among the diseases that cause visual impairment, refractive errors are typically corrected with glasses and cataract with surgery. The leading causes of irreversible blindness nowadays are usually degenerative diseases related to the retina and its extending optic nerves.

关键词： glasses treatment typically

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Bcl-2转基因小鼠的视神经再生

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中华眼科杂志 2005年第9期41卷 832-836页

作者：杨柳 kin-sang cho 北京大学第一医院眼科 100034 美国哈佛医学院Schepens眼科研究所

目的探讨新生小鼠Bcl2高表达是否足以使视神经损伤后再生,并探讨再生的神经轴突能否到达脑内正常的靶组织。方法对出生后3d的C57BL/6J野生小鼠和Bcl2转基因小鼠进行视神经夹闭损伤,1～4d后处死小鼠,做视神经和脑组织切片,利用神经轴突... 详细信息

目的探讨新生小鼠Bcl2高表达是否足以使视神经损伤后再生,并探讨再生的神经轴突能否到达脑内正常的靶组织。方法对出生后3d的C57BL/6J野生小鼠和Bcl2转基因小鼠进行视神经夹闭损伤,1～4d后处死小鼠,做视神经和脑组织切片,利用神经轴突顺行性标记物—带有荧光的霍乱毒素B和GAP43免疫荧光染色检查并评估视神经的再生。结果C57BL/6J野生小鼠视神经损伤后不能再生,损伤的轴突在24h内退缩,而Bcl2转基因小鼠的视神经活跃地再生了相当长的距离,并于4d后到达脑内的靶组织,但大部分再生的神经轴突沿着对侧眼的视路到达损伤同侧的脑内的靶组织,只有很少的神经轴突到达对侧。结论在出生后3d,Bcl2高表达足以使小鼠视网膜神经节细胞损伤后再生,并且再生的神经轴突可以长距离生长至中脑的靶组织,但神经轴突沿着已存在的视路进入同侧的脑内。

关键词： Bcl-2转基因小鼠神经再生视神经视网膜神经节

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The potential of stem cell-based therapy for retinal repair

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Neural Regeneration Research 2014年第11期9卷 1100-1103页

作者： Honghua Yu Lin Cheng kin-sang cho Department of Ophthalmology General Hospital of Guangzhou Military Command of PLA Schepens Eye Research Institute Massachusetts Eye and Ear Infirmary Department of Ophthalmology Harvard Medical School20 Staniford St. Boston MA USA Department of Clinical Pharmacology Xiangya Hospital Central South University Institute of Clinical Pharmacology Central South University Hunan Key Laboratory of Pharmacogenetics

Like injured neurons in the brain or spinal cord, neurons in the retina are incapable to regenerate following injury and ultimately would lead to irreversible neuronal loss and vision impairment. Over decades, extensive effort has been made to develop strategies to protect retinal neurons from death; however, the outcome is limited （Pettmann and Henderson, 1998; Bahr, 2000; Lagali and Picketts, 2011）. Replacing the degenerated retinal neurons by newly generated and functional neurons would be an ideal scenario. The rapid development of stem cell biology has recently demonstrated that stem cells could be a potential source of cells for cell replace- ment therapy because these cells have the self-renewal capacity and could be differentiated into many cell types. This review will dis- cuss the therapeutic potential of stem cell-based therapy to retinal degenerative diseases.

关键词： stem cell The potential of stem cell-based therapy for retinal repair

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Optic neuropathy and increased retinal glial fibrillary acidic protein due to microbead-induced ocular hypertension in the rabbit

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International Journal of Ophthalmology(English edition) 2016年第12期9卷 1732-1739页

作者： Jun Zhao Tian-Hui Zhu Wen-Chie Chen Shi-Ming Peng Xiao Sheng Huang kin-sang cho Dong Feng Chen Guer-Sheung Liu School of Ophthalmology & Optometry Affiliated to Shenzhen University Shenzhen 518040 Guangdong Province China Shenzhen Eye Hospital Affiliated to Jinan University Shenzhen Key Laboratory of Ophthalmology Shenzhen 518040 Guangdong Province China Department of Ophthalmology Schepens Eye Research Institute Massachusetts Eye and Ear Harvard Medical School Boston 02114 USA Centre for Eye Research Australia Royal Victorian Eye and Ear Hospital East Melbourne 3002 Australia Ophthalmology Department of Surgery University of Melbourne East Melbourne 3002 Australia

AIM： To characterize whether a glaucoma model with chronic elevation of the intraocular pressure （IOP） was able to be induced by anterior chamber injection of microbeads in ***： In order to screen the optimal dose of microbead injection, IOP was measured every 3d for 4wk using handheld applanation tonometer after a single intracameral injection of 10 μL, 25 μL, 50 μL or 100 μL microbeads （5×10^6 beads/mL; n=6/group） in New Zealand White rabbits. To prolong IOP elevation, two intracameral injections of 50 μL microbeads or phosphate buffer saline （PBS） were made respectively at days 0 and 21 （n=24/group）. The fellow eye was not treated. At 5wk after the second injection of microbeads or PBS, bright-field microscopy and transmission electron microscopy （TEM） were used to assess the changes in the retina. The expression of glial fibrillary acidic protein （GFAP） in the retina was evaluated by immunofluorescence, quantitative real-time polymerase chain reaction and Western blot at 5wk after the second injection of ***： Following a single intracameral injection of 10 μL, 25 μL, 50 μL or 100 μL microbead, IOP levels showed a gradual increase and a later decrease over a 4wk period after a single injection of microbead into the anterior chamber of rabbits. A peak IOP was observed at day 15 after injection. No significant difference in peak value of IOP was found between 10 μL and 25 μL groups （17.13±1.25 mm Hg vs 17.63±0.74 mm Hg; P=0.346）. The peak value of IOP from 50 μL group （23.25±1.16 mm Hg） was significantly higher than 10 μL and 25 μL groups （all P〈0.05）. Administration of 100 μL microbead solution （23.00±0.93 mm Hg） did not lead to a significant increase in IOP compared to the 50 μL group （P=0.64）. A prolonged elevated IOP duration up to 8wk was achieved by administering two injections of 50 μL microbeads （20.48±1.21 mm Hg vs 13.60±0.90 mm Hg in PBS-injected group; P〈0.05）. The bright-field and TEM were used to assess the changes of retinal

关键词： microbead ocular hypertension optic neuropathy glial fibrillary acidic protein rabbit

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Primary retinal ganglion cells for neuron replacement therapy

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Eye Science 2016年第4期31卷 272-274页

作者： Karen Chang kin-sang cho Min-Huey Chen Dong Feng Chen Schepens Eye Research Institute Massachusetts Eye and Ear Department of Ophthalmology Harvard Medical School Boston MA 02114 USA Graduate Institute of Clinical Dentistry School of Dentistry National Taiwan University Taipei Taiwan

Optic nerve damage as a result of trauma, ischemia, glaucoma or other forms of optic neuropathy disease, leads to disconnection between the eye and brain and death of retinal ganglion cells(RGCs), causing permanent loss of vision. Therapeutic options for treating optic neuropathy are limited and represent a significant unmet medical need. Development of a regenerative strategy for replacement of lost RGCs lies at the core of the future cell-based therapy for these conditions. Successful long-term restoration of visual function depends on the type of cells for transplantation. Primary RGCs of neonatal mice are now reported to have the potential for serving such a purpose.

关键词：视网膜神经节细胞替代治疗神经元视神经损伤细胞治疗视觉功能青光眼永久性

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Ephrin-A2 and-A3 are negative regulators of the regenerative potential of Müller cells

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Chinese Medical Journal 2014年第19期127卷 3438-3442页

作者： Zhu Ruilin cho kin-sang Chen Dong Feng Yang Liu Department of Ophthalmology Peking University First Hospital KeyLaboratory of Vision Loss and Restoration Ministry of EducationBeijing China Schepens Eye Research Institute Massachusetts Eye and EarHarvard Medical School Boston MA USA

Background In a previous study, we demonstrated that ephrin-A2 and -A3 negatively regulate the growth of neural progenitor cells in the central nervous system. Adult mice deficient in ephrin-A2 and -A3 （A2＋A3＋） displayed active ongoing neurogenesis throughout the brain, and mice deficient in ephrin-A3 alone showed increased proliferation of ciliary epithelium derived retinal stem cells. This study aimed to detect that the increase in proliferation and neurogenic potential of MOiler cells is influenced by the absence of ephrin-A2 and -A3. Methods We assessed the retinal and MOiler cell expression of ephrin-As and their receptor and neural progenitor cell markers by immunostaining and real-time PCR. We cultured purified primary MOiler cells derived from wild-type and A2＋A3＋ mice in a defined culture medium that enables trans-differentiation of Mu11er cells into retinal neurons. To evaluate proliferating MOiler cells in vivo, we injected 5＇-ethylnyl-2-deoxiuridine （EdU） intraperitoneally to adult mice. Results Expression of ephrin-A2/A3 and their receptor EphA4 were detected in the retinas of adult mice, with EphA4 expression particularly enriched in MOiler cells. MOiler cells of A2＋A3＋ mice exhibited significantly elevated expression of retinal progenitor cell markers, Pax6 and Chx10, when compared with those from wild-type mice. Moreover, a higher percentage of Mu11er cells of A2＋A3＋ mice trans-differentiated and became recoverin＋ and β-Ⅲ-tublin＋ in the culture than those from wild type mice. Strikingly, an increased number of EdU＋ retinal cells was detected in the retinas of adult A2＋A3＋ mice as compared with wild-type mice. Conclusions Ephrin-A2 and -A3 are negative regulators of the proliferative and neurogenic potentials of Mu11er cells. Manipulating ephrin-A signaling may thus represent a novel strategy for stimulating neuroregeneration from endogenous progenitors to participate in retinal repair in case of disease or damage.

关键词： Muller cell stem cell retinal regeneration

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Intrinsic determinants of optic nerve regeneration

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Chinese Medical Journal 2013年第13期126卷 2543-2547页

作者： ZHU Rui-lin cho kin-sang GU0 Chen-ying CHEW Justin CHEN Dong-feng YANG Liu Department of Ophthalmology Peking University First Hospital Key Laboratory of Vision Loss and Restoration Ministry of Education Beijing 100034 China Schepens Eye Research Institute Massachusetts Eye and Ear Department of Ophthalmology Harvard Medical School Boston MA 02114 United States

Objective To review the functions of these intracellular signals in their regulation of retinal ganglion cell （RGC） axon regeneration. Data sources Relevant articles published in English or Chinese from 1970 to present were selected from PubMed. Searches were made using the terms ＂intrinsic determinants, axon regeneration, RGC, optic nerve regeneration, and central nervous system axon regeneration.＂ Study selection Articles studying the mechanisms controlling RGC and central nervous system （CNS） axon regeneration were reviewed. Articles focusing on the intrinsic determinants of axon regeneration were selected. Results Like other CNS neurons of mammals, RGCs undergo a developmental loss in their ability to grow axons as they mature, which is a critical contributing factor to the failure of nerve regeneration and repair after injury. This growth failure can be attributed, at least in part, by the induction of molecular programs preventing cellular overgrowth and termination of axonal growth upon maturation. Key intracellular signals and transcription factors, including B cell lymphoma/leukemia 2, cyclic adenine monophosphate, mammalian target of rapamycin, and Kr^Jppel-like transcription factors, have been identified to play central roles in this process. Conclusions Intense effort and substantial progress have been made to identify the various intrinsic growth pathways that regulate RGC axon regeneration. More work is needed to elucidate the mechanisms of and the interrelationship between the actions of these factors and to successfully achieve regeneration and repair of the severed RGC axons.

关键词： nerve regeneration retinal ganglion cells, intrinsic determinants cAMP roTOR PTEN, SOCS3

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Clinical factors affecting rejection rates in liver transplantation

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Hepatobiliary & Pancreatic Diseases International 2015年第4期14卷 367-373页

作者： kin Pan Au See-Ching Chan Kenneth Siu-Ho chok William Wei Sharr Wing-Chiu Dai Sui-Ling Sin Tiffany cho-Lam Wong Chung-Mau Lo Department of Surgery the University of Hong Kong Medical Center Queen Mary Hospital

With improvements in survival, liver trans- plant recipients now suffer more morbidity from long-term immunosuppression. Considerations were given to develop individualized immunosuppression based on their risk of re- jection. METHOD： We retrospectively analyzed the data of 788 liver transplants performed during the period from October 1991 to December 2011 to study the relationship between acute cel- lular rejection （ACR） and various clinical factors. RESULTS： Multivariate analysis showed that older age （P=0.04, OR=0.982）, chronic hepatitis B virus infection （P=0.005, OR= 0.574）, living donor liver transplantation （P=0.02, OR=0.648） and use of interleukin-2 receptor antagonist on induction （P〈0.001, OR=0.401） were associated with fewer ACRs. Patients with fulminant liver failure （P=.004, OR=4.05） were more likely to develop moderate to severe grade ACR. CONCLUSIONS： Liver transplant recipients with older age, chronic hepatitis B virus infection, living donor liver trans- plantation and use of interleukin-2 receptor antagonist on in- duction have fewer ACR. Patients transplanted for fulminant liver failure are at higher risk of moderate to severe grade ACR. These results provide theoretical framework for developing individualized immunosuppression.

关键词： liver transplantation acute rejection immunosuppression

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