干神经变性的细胞疗法中共济失调眼球运动不能类型 2 和小脑性共济失调

 

干细胞疗法,神经退行性疾病 ,共济失调,眼球运动不能, 类型 2

共济失调动眼神经失用型 2 (AOA2) 是一种罕见的常染色体隐性小脑性共济失调. 最近的证据表明,在此综合征的蛋白缺陷, senataxin (SETX), 在RNA处理功能,以保护基因组的完整性.

至今, 只有患者来源的类淋巴母细胞, 成纤维细胞和SETX敲除细胞可用来调查AOA2. 在SETX基因的破坏近期并没有导致神经行为缺陷或神经退行性疾病, 因此很难研究AOA2的病因.

 

用干细胞治疗共济失调

干细胞thery相结合,与专业 治疗共济失调, 这不仅专注于帮助病人,以应付他们的症状, 但也把症状的直接原因, 促进创伤性脑损伤的愈合. We believe that comprehensive approach to ataxia stem cell therapy gives patients a high chance of improvement, 使他们能够改善他们的生活质量.

干细胞的治疗方案可以被应用于各种类型的共济失调的, 包括SCA1, SCA2, SCA3, SCA6, AOA1,AOA2, 共济失调弗雷德里克, 这是由颅脑损伤所致, 以及更多.

What are the potential improvements in the stem cell treatment?

治疗的目标是治愈疾病造成的脑损伤, 恢复神经功能.

有多种类型的改善我们的治疗后,和我们过去的患者有以下 *:

改善平衡和协调
减少疲劳
改进的语音
减少震颤
改进电机装置的功能
改善吞咽
减少神经性疼痛
提高警觉性

与MRI脑萎缩激动.

最近几年, 干细胞疗法已成为这样的接入点的疾病的治疗. 通过体外扩增容易地获得间充质干细胞可以有很多在体外长期来看. 培养过程中保持其在特定条件下的多能性可以能够在神经细胞分化和可分泌多种神经营养因子, 促进细胞的神经修复. 在干细胞治疗, 克服神经干细胞从危险和限制成人脑组织中的收购, 也 避免在伦理的存在胎脑移植, 免疫排斥反应, 问题的来源有限, 没有急性或慢性毒性和致瘤性在体内可以向受损神经的修复.

随着干细胞技术的帮助下,很少在临床医院使用.

早期的自体骨髓间充质干细胞中的血液或脂肪组织治疗脊髓损伤等神经损伤,并取得了积极成效. Now investigators use an alternative cord umbilical cord, 不仅获得足够数量的骨髓间充质干细胞也被冻结, 预应用的恢复, 很短的时间来组装足够数量在临床治疗中使用的干细胞的, 即使病人已经消除大脑疼痛的提取, 同时也缩短了细胞培养的等待时间.

而在临床实践中证明了它的可靠性和安全性, 而不是免疫反应. Clinical application can be obtained from autologous mesenchymal stem cells to treat the same effect.

案件 小脑性共济失调 患者症状脐蛛网膜下腔注射疗法中的间充质干细胞的神经变性疾病患者的治疗运动之后, 平衡, 语言, 写如具有不同的恢复水平的能力, 提高生活质量, 大规模后显著降低国际共济失调和ADL治疗分数 (P <0.05), 且无不良反应.

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细节 –

干乌克兰细胞疗法

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更多关于共济失调和科研的干细胞治疗

共济失调动眼神经失用型 2 (AOA2) 最初被描述 15 几年前,随后定位于染色体 9.

为了开发更相关的神经元模型来研究神经退行性疾病中AOA2, 研究者衍生的神经祖细胞从与AOA2患者和诱导多能干细胞的控制 (IPSC) 细胞重新编程. AOA2的iPSC和神经祖细胞显示出增加的氧化损伤的水平, DNA双链断裂, 增加的DNA损伤诱导的细胞死亡和R-环积累. 全基因组表达和加权基因共表达网络分析中鉴定的这些神经祖细胞两者先前报道的和新颖的受影响的基因和细胞途径与senataxin功能障碍和AOA2的病理生理学相关联, 提供进一步的深入了解senataxin的作用在上一个全基因组范围的基因表达调控. 这些数据表明,iPS细胞可以从患者中产生与常染色体隐性遗传性共济失调, AOA2, 分化成神经元, 而这两种细胞类型概括了AOA2细胞表型. 这代表了一种新的和适当的模型系统在此综合征的研究神经退行性疾病.

干细胞与间充质起源的细胞最初出现在宫内生活的早期阶段为细胞,其分化成所有其他细胞. 他们还认为,在早期阶段负责血细胞生成. 在胎儿阶段所形成的不同的功能系统的细胞是干细胞的变换前. 细胞就像成软骨细胞, 神经细胞, 和成骨细胞浮现在脑海中.

 

此病症的特征在于进行性小脑萎缩, 周围神经病变, 眼球运动不能在患者和升高的α胎蛋白的水平〜50%与发病之间的年龄 10 和 20 年份 .

在AOA2有缺陷的基因被确定为SETX编码senataxin, 一个 2667 包含该超家族的高度保守的C-末端七基序结构域的氨基酸的蛋白 1 的DNA / RNA解旋酶和一个N-末端结构域对蛋白质 - 蛋白质相互作用的重要 .

Using lymphoblastoid cells and fibroblasts from AOA2 patients and SETX-RNAi-depleted cells as model systems, roles for senataxin have been uncovered that encompass (一世) protection against DNA damage , (二) transcription regulation including transcription termination, splicing efficiency of specific mRNAs and alternate splice site selection and R-loop resolution and (iii) localization at the interface of transcription and replication .

R-loops are RNA/DNA hybrids that form over transcription pause sites by interaction with an ssDNA template behind an elongating RNA Pol II complex, which are potentially harmful and can cause genomic instability if left unresolved. A role for senataxin in transcription elongation and termination is further supported by a report, showing that cells with senataxin knockdown display an increase in RNA readthrough and RNA Pol II density downstream of the Poly(一个) site and also exhibit increased levels of R-loop formation . Senataxin was found to localize to distinct nuclear foci in S/G2 phase cells, and the number of these foci increased in response to impaired replication, suggesting that senataxin localizes to collision sites between the transcription apparatus and components of the replisome .

此外, a SUMO-dependent interaction between senataxin and Rrp45, a core component of the exosome, was also shown to co-localize in nuclear foci corresponding to sites of R-loops, suggesting that senataxin connects transcription, DNA damage and RNA surveillance .

Generation and characterization of AOA2 iPSC stem cells
In order to optimize conditions and to reduce the risk of chromosomal instability, used early passage (P < 5) fibroblasts for reprogramming. Following transfection with pEP4EO2SCK2MEN2L and pEP4EO2SET2K episomal plasmids, stepwise adapted the cells to knockout serum replacement (KOSR) medium over the first 4–5 days of iPSC generation, as direct replacement with the KOSR medium was found to lead to extensive death of the AOA2 fibroblasts.

后 2 周, transduced AOA2 patient and matched control fibroblasts gave rise to colonies of small round cells with a high nucleus-to-cytoplasm ratio typical of pluripotent human stem cells

Although data show that it was possible to reprogram AOA2 fibroblasts, the efficiency was somewhat reduced compared with that of controls. Thirteen colonies from the AOA2 homozygote patient who expressed the TRA-1-60 stem cell surface marker

were expanded for further analysis. Two of these AOA2 clones, AOA2(C7) and AOA2(C8), were selected for further analysis as these clones displayed robust expression of the pluripotency markers TRA-1-60, TRA-1-81, Nanog and Oct4

Control clones were screened similarly as described earlier and conformed to the same criteria. The presence of the c.6109 A>G homozygous missense mutation in AOA2 iPSC was confirmed at the mRNA levels by sequencing , and a normal karyotype was observed for AOA2(C7) and AOA2(C8) iPS clones

OCT4, NANOG and SOX2 expression in the AOA2 iPSC was not derived from integrated or persistent reprogramming plasmids as (一世) polymerase chain reaction (PCR) analysis of genomic DNA revealed no amplicons following 36 rounds of PCR using IRES-anchored primers designed to amplify the reprogramming genes OCT4, SOX2, LIN28, KLF4 and c-MYC (plasmids used as positive controls) 和 (二) reverse transcriptase (RT)–PCR analysis of RNA isolated from the AOA2 and control iPSC showed an absence of transgene expression (human fibroblasts transiently transfected with the reprogramming plasmids used as a positive control)

 

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细节 –

干乌克兰细胞疗法

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干细胞治疗