Development of therapeutic chemokine-based formulations

开发基于趋化因子的治疗制剂

• 批准号: 8148261
• 负责人: Arya Biragyn
• 金额: $ 27.37万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8148261
• 关键词: CCR5 gene; CD8B1 gene; Cytosol; Development; Disease; Drug Formulations; Gene Expression; Genes; MicroRNAs; RNA Recognition Motif; Small Interfering RNA; base; chemokine; in vivo; therapeutic development

1) We have developed two novel technologies for depletion of unwanted subsets of cells at will. The strategy is based on the nature of chemokines to deliver antigens to the cytosol of cells expressing respective chemokine receptors. One technology designated chemotoxin, a chimeric chemokine fused with toxic moieties, can specifically kill cells expressing respective chemokine receptors. TARC-chemotoxin efficiently eradicates established leukemia (CEM cells) generating almost 100% tumor- free mice . We propose that the strategy may be applied for treatment of human T cell malignant diseases when patients immune system is severely immunocompromised. The results of this study have been recently published (Baatar et al, 2007a). We have also utilized this technology to specifically deplete Tregs in mice to augment immune responses to cancer vaccines. Part one of this ongoing project demonstrates that TARC-chemotoxin-mediated depletion of Tregs enhance CD8+ T cell responses to melanoma antigen gp100. This is an encouraging result, although its clinical significance remains to be evaluated. This technology is being successfully used to understand chemokine/chemokine receptor axis in regulation of metastasis (see AG000443-02). It was also used in our recent study on the mechanism of Treg-mediated suppression of resting T cells (Baatar et al.,2009). 2) Despite significant potency and attractiveness of siRNA and shRNA-mediated gene silencing methods, the technology can not be efficiently used in vivo. The major problem is that there is almost no practical way that delivers/targets oligonucleotides to cells of interest. The inability to deliver siRNA into primary cells precludes its utilization in vivo. To solve this, we have hypothesized that this can be achieved by generating modified chemokines that contain RNA binding domains (RBD). Although this technology designated chemoarp is still under development, we have successfully demonstrated proof of principle by delivering siRNA and knocking off expression of genes in various cells, including tumor cells. To date, we have generated three different chemoarps which deliver siRNA by targeting chemokine receptors, such as CCR2, CCR4 and CCR5. However and importantly, this technology enables delivery of siRNA into primary immune cells inactivating their genes of interest. We have also established a simplified but efficient yeast production system and created chemoarp purification methods. Overall, this truly novel project, which was recently applied for patenting from NIA, is not only progressing well, but also has attracted significant interest from commercial enterprises such as ECI, Inc., Japan.

1)我们已经开发了两种新的技术，可以随意去除不需要的细胞亚群。该策略基于趋化因子将抗原递送至表达相应趋化因子受体的细胞的胞质溶胶的性质。一种称为化学毒素的技术，一种与毒性部分融合的嵌合趋化因子，可以特异性地杀死表达相应趋化因子受体的细胞。TARC化学毒素有效根除已建立的白血病（CEM细胞），产生几乎100%无肿瘤的小鼠。我们建议，该策略可能适用于治疗人类T细胞恶性疾病时，患者的免疫系统是严重的免疫功能低下。这项研究的结果最近发表（Baatar等人，2007年a）。我们还利用这种技术专门消耗小鼠的TcR，以增强对癌症疫苗的免疫反应。这个正在进行的项目的第一部分证明了TARC-化学毒素介导的TcB耗竭增强了CD 8 + T细胞对黑色素瘤抗原gp 100的应答。这是一个令人鼓舞的结果，尽管其临床意义仍有待评估。该技术已成功用于了解转移调节中的趋化因子/趋化因子受体轴（参见AG 000443 -02）。它也用于我们最近关于Treg介导的静息T细胞抑制机制的研究（Baatar et al.，2009年）。 2)尽管siRNA和shRNA介导的基因沉默方法具有显著的效力和吸引力，但该技术不能有效地用于体内。主要问题是几乎没有将寡核苷酸递送/靶向至感兴趣的细胞的实用方法。不能将siRNA递送到原代细胞中排除了其在体内的利用。为了解决这个问题，我们假设这可以通过产生含有RNA结合结构域（RBD）的修饰趋化因子来实现。虽然这项名为chemoarp的技术仍在开发中，但我们已经成功地证明了通过递送siRNA并敲除包括肿瘤细胞在内的各种细胞中的基因表达的原理。到目前为止，我们已经产生了三种不同的chemoarp，它们通过靶向趋化因子受体（如CCR 2、CCR 4和CCR 5）来递送siRNA。然而，重要的是，该技术能够将siRNA递送到原代免疫细胞中，使其感兴趣的基因失活。我们还建立了一个简化但有效的酵母生产系统，并创建了chemoarp纯化方法。总的来说，这个最近向NIA申请专利的真正新颖的项目不仅进展顺利，而且还吸引了ECI，Inc.，日本