CTCF peptides - a new therapeutic alternative to target HIV in the brain.

CTCF 肽 - 一种针对大脑中 HIV 的新治疗替代方案。

基本信息

批准号：
8320910
负责人：
Malgorzata Simm
金额：
$ 39.84万
依托单位：
ST. LUKE'S-ROOSEVELT INST FOR HLTH SCIS
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-30 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8320910
关键词：
AIDS Dementia Complex Acquired Immunodeficiency Syndrome Affinity Amino Acid Sequence Antiviral Agents Antiviral Response Antiviral resistance Applications Grants Astrocytes Binding Binding Sites Biological Biological Assay Biological Response Modifier Therapy Brain CCCTC-binding factor CD4 Positive T Lymphocytes Cell Culture Techniques Cell Line Cell Surface Receptors Cells Complex DNA DNA Binding DNA Probes Data Dose Evaluation Extracellular Space Foundations Future Genetic Transcription Genome Goals HIV HIV-1 Highly Active Antiretroviral Therapy Human In Vitro Incidence Infection Interleukin-1 Kinetics Length Ligation Maintenance Mediating Messenger RNA Methods Mus N-terminal Names Neurotoxins Nitric Oxide Nuclear Nuclear Extract Nucleotides Oligonucleotides Organ Pathway interactions Patients Penetration Peptides Pharmaceutical Preparations Phase Phenotype Process Proteins Protocols documentation Reaction Recombinant Proteins Recombinants Refractory Reporter Genes Resistance Small Interfering RNA TNF gene TNFRSF5 gene Testing Therapeutic Agents Time Toxic effect Transmembrane Transport Virus Virus Replication base brain tissue cytokine deletion library dimer extracellular fetal in vitro activity in vivo macrophage mutant novel novel therapeutics prevent promoter protein complex research study resistance factors response therapeutic target tool uptake

项目摘要

Virus-infected macrophages are the major therapeutic target in the brain tissue, even though the productive virus replication in these cells is necessary but not sufficient to cause HIV Associated Dementia (HAD). HAART is still generally recommended for patients with AIDS and in general this treatment has been effective in decreasing the incidence of HAD; however a significant group of patients remains refractory to HAART even when its CNS penetration is enhanced by supplementing drugs. For this reason alone, the activation of native cellular mechanisms inducing anti-viral resistance in cells migrating to various organs including CNS should be considered a viable alternative to add to the existing antiviral approaches for HAD. We have induced such responses mediated through soluble molecule [previously named HRF (HIV-1 Resistance Factor)] able to prevent virus replication in CD4+ T cells. Recently we identified the HRF as a truncated form of a potent transcription suppressor - CTCF (tCTCF). The biological activity of tCTCF was confirmed by several experimental methods: (1) the siRNA silencing of CTCF mRNA in HRF(+) cells reversed the HIV-1 resistance phenotype and reduced the biological activity of their cell culture supernatants; (2) depletion of tCTCF from HRF(+) cell culture supernatant abolished its antiviral activity; while (3) the affinity purified tCTCF induced antiviral activity in human macrophages; and (4) the affinity purified tCTCF inhibited the transcription of HIV-1 LTR promoted expression of reporter gene. Our preliminary data showed that upon HIV-1 exposure, CTCF from nuclear extracts of HRF(+) but not HRF(-) control cells bound to DNA probe comprising NF-¿B, Sp1 and YB-1 binding sequences, possibly through its interaction with YB-1 protein. We hypothesize that since NF-¿B and YB-1 binding sites are separated by only 6 nucleotides the CTCF/YB1 complex bound to the virus promoter might obstruct the formation of NF- ¿B/DNA binding, thus halting transcription of virus genome. Taken together we discovered a novel form of CTCF protein that mediates innate responses to HIV-1 in human macrophages. The main goal of this application is to prepare foundation for future delivery of tCTCF as a biotherapeutic for treatment of HIV-1 in CNS. In this grant application we propose: (1) To study the mechanism of tCTCF mediated cellular responses to HIV-1; (2) To determine the minimum tCTCF amino-acid sequence required for the induction of antiviral responses and efficacy assessment of tCTCF treatment in primary human macrophages; and (3) To determine the minimal CTCF toxicity and brain entry in vivo.

病毒感染的巨噬细胞是脑组织中的主要治疗靶标尽管这些细胞中的产物病毒复制是必要的，但不足导致艾滋病毒相关的痴呆症（患有）。 Haart通常仍然建议使用艾滋病患者，总的来说，这种治疗方法已有效减少 HAD的发生率；但是，一组大量的患者对Haart仍然难治性即使通过补充药物来增强其中枢神经系统的渗透。为此原因单独，天然细胞机制的激活引起细胞中的抗病毒抗性迁移到包括CN在内的各种器官应被视为可行的替代品加上现有的抗病毒方法。我们引起了这样的回应通过固体分子介导[先前命名为HRF（HIV-1抗性因子）] 可以预防CD4+ T细胞中的病毒复制。最近，我们确定了人力资源管理潜在转录抑制器-CTCF（TCTCF）的截断形式。生物学通过几种实验方法证实了TCTCF的活性：（1）siRNA HRF（+）细胞中CTCF mRNA的沉默逆转了HIV-1抗性表型和降低了其细胞培养上清液的生物学活性；（2）TCTCF的耗竭来自HRF（+）细胞培养上清液废除了其抗病毒活性。而（3）亲和力纯化的TCTCF诱导人类巨噬细胞的抗病毒活性；（4）亲和力纯化的TCTCF抑制了HIV-1 LTR的转录促进了报告基因的表达基因。我们的初步数据表明，HIV-1暴露后，核CTCF HRF（+）的提取物，但不提供HRF（ - ）控制细胞，该探针包含NF-®B， SP1和YB-1结合序列，通过与YB-1蛋白的相互作用可能。我们假设，既然NF- - B和YB-1结合位点仅被6个核苷酸分开与病毒启动子结合的CTCF/YB1复合物可能会阻碍NF-的形成 B/DNA结合，因此停止了病毒基因组的转录。总的来说我们发现了一种新型的CTCF蛋白质形式，它介导了对HIV-1的先天反应人类巨噬细胞。该应用程序的主要目标是为将TCTCF作为CNS中HIV-1治疗的生物疗法的未来递送。在这个我们提出的授予应用程序：（1）研究TCTCF介导的机制细胞对HIV-1的反应；（2）确定最小TCTCF氨基酸诱导抗病毒反应所需的顺序和有效评估原代人巨噬细胞中的TCTCF治疗；（3）确定 CTCF最小的毒性和体内大脑进入。