The Primate Corticospinal Connectome and Transcriptome - Supplement

灵长类动物皮质脊髓连接组和转录组 - 补充

基本信息

批准号：
10876189
负责人：
MARK H. TUSZYNSKI
金额：
$ 14.9万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-15 至 2024-05-31
项目状态：
已结题

项目摘要

Abstract We recently published a report demonstrating that, remarkably, adult corticospinal neurons in the mouse revert to an embryonic transcriptional state after spinal cord injury (SCI) (Poplawski et al. Nature 2020). This state persists for two weeks, then reverts to an adult transcriptional state in which neurons are more refractory to regenerative therapies. This grant seeks to determine whether embryonic reversion of corticospinal neurons also occurs in primate corticospinal motor neurons after SCI, and if so, how long this state endures. This is critical information for planning human clinical trials because efforts to repair the injured spinal cord are most likely to be effective in the time period that corticospinal are in an embryonic state and most able to regenerate and respond to pro-growth therapies. Our results to date have shown that embryonic reversion of primate corticospinal neurons indeed appears to occur. We sampled time points of 3, 10 and 20 days after SCI. We find that embryonic reversion is present at Day 10 and is nearly gone by Day 20. We now seek to add an additional time point, 15 days, to this study. If embryonic reversion is still present 15 days after SCI, one could extend the duration of potential patient recruitment into clinical trials, representing an important improvement in increase the number of people eligible for pro-regenerative trials. This information is important not only for our stem cell program that we aim to clinically translate, but for any pro-regenerative treatment trial in humans. We will use techniques that have been successfully employed in this grant to date. We will perform RNAseq of corticospinal neurons in rhesus monkeys using retro-AAV vectors expressing HA- and FLAG- tagged polyribosomes. We then use translational Ribosomal Affinity Purification (TRAP) to obtain and sequence corticospinal-specific mRNA species. We will add to our current timeline a 15 days post injury group (n= 3) and one more monkey (n=1) to our 10 day post top strengthen statistical power and enable batch correction.

抽象的我们最近发表的一份报告表明，值得注意的是，小鼠中的成年皮质脊髓神经元恢复了脊髓损伤 (SCI) 后进入胚胎转录状态 (Poplawski et al. Nature 2020)。这种状态持续两周，然后恢复到成人转录状态，其中神经元更难以抵抗再生疗法。这笔赠款旨在确定皮质脊髓神经元的胚胎逆转是否也脊髓损伤后，灵长类皮质脊髓运动神经元会发生这种状态，如果是，这种状态会持续多久。这很关键规划人体临床试验的信息，因为修复受损脊髓的努力最有可能在皮质脊髓处于胚胎状态并且最有能力再生和再生的时期有效对促生长疗法有反应。我们迄今为止的结果表明，灵长类皮质脊髓神经元的胚胎逆转确实似乎发生。我们在 SCI 后 3、10 和 20 天的时间点进行采样。我们发现胚胎回复存在于第 10 天，到第 20 天就几乎消失了。我们现在寻求在这项研究中添加一个额外的时间点，即 15 天。如果 SCI 后 15 天胚胎回复仍然存在，可以延长潜在患者的持续时间临床试验招募，代表了增加合格人数方面的重要进步用于促再生试验。这些信息不仅对于我们的临床目标的干细胞计划很重要翻译，但适用于人类的任何促再生治疗试验。我们将使用迄今为止已在本次资助中成功采用的技术。我们将执行 RNAseq 使用表达 HA 和 FLAG 标记的逆转录 AAV 载体研究恒河猴的皮质脊髓神经元多聚核糖体。然后，我们使用翻译核糖体亲和纯化（TRAP）来获得并测序皮质脊髓特异性 mRNA 种类。我们将在当前的时间表中添加伤后 15 天组 (n= 3) 以及我们的 10 天帖子顶部又多了一只猴子 (n=1)，增强了统计能力并启用批量校正。