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Single-guide-RNA-directed Synergistic Activation Mediator, a novel strategy to sensitize Non-Hodgkin Lymphomas to therapeutic drugs.

单向导RNA引导的协同激活介质，一种使非霍奇金淋巴瘤对治疗药物敏感的新策略。

基本信息

批准号：
9899219
负责人：
Mustapha Kandouz
金额：
$ 13.4万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2022-03-31
项目状态：
已结题

项目摘要

PROJECT SUMMARY Objectives: Non-Hodgkin’s lymphomas (NHLs) are the 6th most common cancers in the US, of which 90% are B cells-type, and include 60% aggressive and 40% indolent. Although all types of NHL respond well to initial therapy, indolent NHLs are incurable: the disease relapses after the patient has achieved complete response (CR) to therapy. Also, despite efforts to develop effective therapies, many aggressive lymphomas recur and eventually 30% of these become incurable, highlighting the need to develop new therapies or to improve existing ones. In this proposal, we introduce a novel drug sensitization strategy for currently available anti-NHL drugs. The approach makes use of the sgRNA-SAM technology, a variant of the CRISPR tools, for effective, specific and sustained reactivation of the endogenous Ehd3 gene. The proposed approach is based on the fact that Ehd3 is preferentially down-regulated in a subpopulation of CD34+ cells isolated from NHL specimens and which show a reduced drug sensitivity. Therefore, our working hypothesis is that loss of Ehd3 expression in CD34+ cells is a driver event in the drug insensitivity of NHL cells. Consequently, restoring its expression will potentiate the cytotoxic effects of currently used drugs. Specific Aims: Aim1, will consist in a prospective study to assess Ehd3 expression in patient-derived NHL specimens. Here, we aim to identify specific subtypes of NHLs in which Ehd3 expression levels are predominantly decreased, and which could be more suitable for the proposed gene reactivation approach. In Aim2, we seek to perform in vitro development and test of the proposed sgRNA-SAM-directed Ehd3 transcriptional reactivation, in NHL established cell lines and patient-derived (PD) cells. We will also assess the power of the approach in potentiating the cells’ sensitivity to the cytotoxic effects of 2CdA-R and CHOP-R drugs in vitro. In Aim3, we will perform in vivo proof-of-principle of sgRNA-SAM approach as a drug sensitization strategy, under the CHOP-R and 2CdA-R regimens, using animal xenografts of NHL cells.

项目摘要目的：非霍奇金淋巴瘤（NHL）是美国第六大常见癌症，其中 90%为B细胞型，其中60%为侵袭型，40%为惰性型。虽然所有类型的NHL都对在初始治疗中，惰性NHL是不可治愈的：在患者已经达到完全缓解后，疾病复发。治疗反应（CR）。此外，尽管努力开发有效的治疗方法，许多侵袭性淋巴瘤复发，最终，其中30%成为不治之症，这突出了开发新疗法或改善现有的。在这个建议中，我们介绍了一种新的药物增敏策略，目前可用的抗NHL药物。该方法利用了sgRNA-SAM技术，这是CRISPR工具的一种变体，和内源性Ehd 3基因的持续再激活。所提议的方法基于以下事实： Ehd 3在从NHL标本分离的CD 34+细胞亚群中优先下调，其显示出降低的药物敏感性。因此，我们的工作假设是，CD 34+细胞中Ehd 3表达的缺失是CD 34+细胞中Ehd 3表达的驱动事件。 NHL细胞的药物不敏感性。因此，恢复其表达将增强其细胞毒性作用。目前使用的药物。具体目的：目的1，将包括一项前瞻性研究，以评估患者源性肿瘤中Ehd 3的表达。 NHL标本。在这里，我们的目的是确定特定亚型的NHL，其中Ehd 3的表达水平，主要减少，并且其可能更适合于所提出的基因再激活方法。在目的2，我们寻求进行体外开发和测试所提出的sgRNA-SAM指导的Ehd 3 在NHL建立的细胞系和患者来源的（PD）细胞中，转录再激活。我们亦会评估该方法在增强细胞对2CdA-R和CHOP-R的细胞毒性作用的敏感性方面的能力体外药物在Aim 3中，我们将进行sgRNA-SAM方法作为药物致敏的体内原理验证使用NHL细胞的动物异种移植物，在CHOP-R和2CdA-R方案下，使用免疫抑制策略。