Cellular Transduction with Replication-Competent Retrovirus Vectors

使用具有复制能力的逆转录病毒载体进行细胞转导

基本信息

批准号：
7554139
负责人：
NORIYUKI KASAHARA
金额：
$ 48.19万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-03-12 至 2011-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7554139
关键词：
Adoptive Immunotherapy Adult Adverse effects Affect Animals Apoptosis Biodistribution Biological Assay Blood Circulation Bone Marrow Transplantation Brain Brain Neoplasms Cells Characteristics Clinical Clinical Trials Control Groups Cytolysis Cytosine deaminase Cytotoxic T-Lymphocytes DNA Sequence Rearrangement Development Diffusion Disease Emerging Technologies Engineering Environment Event Exhibits Extravasation Flucytosine Future Gene Transfer Genes Glioblastoma Glioma HLA Antigens Human Image Immune response Immune system Immunobiology Immunocompetent Immunodeficient Mouse Immunology Immunosuppressive Agents Immunotherapy Inbred F344 Rats Individual Injection of therapeutic agent Joints Kinetics Long Terminal Repeats Luciferases Malignant Glioma Malignant Neoplasms Mediating Methods Modeling Murine leukemia virus Mus Needles Neurosurgeon Normal Cell Normal tissue morphology Nude Mice Oncogenes Oncolytic Patients Penetration Phase Phase III Clinical Trials Preparation Primary Brain Neoplasms Principal Investigator Procedures Prodrugs Production Radiosurgery Rattus Recruitment Activity Research Personnel Retroviral Vector Retroviridae Risk Rodent Model Safety Serial Passage Site Solid Neoplasm Suicide Gene Therapy System Technology Testing Therapeutic Tissues Transgenes Translations Treatment Efficacy Tumor Suppression Viral Virus Xenograft Model Xenograft procedure Yeasts base brain tissue cancer cell cellular transduction chemotherapy clinical application cytokine design and construction dosage effective therapy follow-up gene therapy gene transfer vector genotoxicity immuno-gene therapy improved in vivo in vivo Model killings luminescence migration molecular imaging neoplastic cell novel outcome forecast programs quality assurance response subcutaneous suicide gene trafficking transduction efficiency tumor vector

项目摘要

Glioblastoma multiforme (GBM), the most common primary brain tumor in adults, is associated with a dismal prognosis of only 12-15 months despite aggressive surgery, radiation, and chemotherapy. The lack of effective treatment options has made this disease a target for new strategies such as gene therapy. However, the only major Phase III clinical trial of gene therapy, involving the use of conventional replication-defective retrovirus vectors in GBM patients, resulted in disappointingly low and therapeutically inadequate transduction levels on the order of only 0.02%. The inability of standard replication-defective retroviral vectors to achieve effective transduction of tumors in vivo is therefore a major obstacle to gene therapy for gliomas. The use of replication-competent vectors for gene transfer would be more efficient, as each tumor cell that is successfully transduced would itself become a virus-producing cell, sustaining further transduction events even after initial administration. We have previously demonstrated that direct intratumoral injection of murine leukemia virus (MLV)-based replication-competent retrovirus (RCR) vector preparations can achieve tremendously efficient suicide gene transfer in gliomas, with transduction stringently restricted to the actively dividing tumor cells without evidence of significant spread to extratumoral sites, and resulting in significantly prolonged survival upon prodrug administration, without detectable systemic side effects. Here we propose to further improve the efficiency of this approach by engineering alloreactive cytotoxic T lymphocytes (alloCTLs) to become RCR vector producer cells, which can then serve as motile cellular delivery platforms that can penetrate into the tumor mass and facilitate multifocal spread of the replicating vectors. Alloreactive CTL exhibit characteristics that provide unique advantages for this purpose: alloCTLs can move through tissue, can themselves kill tumor upon contact, and can produce cytokines that induce apoptosis or can initiate an endogenous immune response. Within CMS gliomas, an immunosuppressive environment within an immunologically privileged site, their own destruction by the immune system may be circumvented long enough for them to have a beneficial effect, but alloCTLs should be rapidly destroyed upon leakage into the general circulation. We will test their viability, biodistribution, safety, and utility as cellular delivery vehicles to enhance RCR vector spread and therapeutic efficacy, comparing immunodeficient and immunocompetent glioma models in vivo.

多形性胶质母细胞瘤 (GBM) 是成人中最常见的原发性脑肿瘤，与抑郁症相关尽管进行了积极的手术、放疗和化疗，但预后仅为 12-15 个月。缺乏有效的治疗方案使这种疾病成为基因治疗等新策略的目标。然而，基因治疗的唯一主要 III 期临床试验，涉及使用传统的复制缺陷型 GBM 患者中的逆转录病毒载体导致令人失望的低转导和治疗不足水平仅为 0.02%。标准复制缺陷型逆转录病毒载体无法实现因此，体内肿瘤的有效转导是神经胶质瘤基因治疗的主要障碍。使用用于基因转移的具有复制能力的载体将更加有效，因为每个成功转移的肿瘤细胞转导本身将成为病毒产生细胞，即使在初始转导之后也能维持进一步的转导事件行政。我们之前已经证明，直接瘤内注射鼠白血病病毒基于 MLV（MLV）的具有复制能力的逆转录病毒（RCR）载体制剂可以实现极高的效率神经胶质瘤中的自杀基因转移，转导严格限制于活跃分裂的肿瘤细胞没有证据表明显着扩散到肿瘤外部位，并导致生存期显着延长前药给药后，没有可检测到的全身副作用。在此我们建议进一步完善通过将同种异体反应性细胞毒性 T 淋巴细胞 (alloCTL) 改造为 RCR，提高了该方法的效率载体生产细胞，然后可以作为运动细胞递送平台，可以渗透到肿瘤块并促进复制载体的多灶性扩散。同种异体反应性CTL表现出的特点为此目的提供了独特的优势：alloCTL 可以穿过组织，本身可以杀死肿瘤接触后，可以产生诱导细胞凋亡或启动内源性免疫的细胞因子回复。在 CMS 神经胶质瘤中，免疫特权位点内的免疫抑制环境，免疫系统对自身的破坏可能会被规避足够长的时间，使他们能够获得有益的效应，但 alloCTL 在泄漏到全身循环后应迅速被破坏。我们将测试他们的活力、生物分布、安全性和作为细胞递送工具的实用性，以增强 RCR 载体的传播和治疗效果，体内比较免疫缺陷和免疫功能正常的神经胶质瘤模型。