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.

胶质母细胞瘤多形（GBM）是成年人中最常见的原发性脑肿瘤，与惨淡有关尽管进行了侵略性手术，放射和化学疗法，预后仅12-15个月。缺乏有效的治疗选择使该疾病成为基因治疗等新策略的靶标。然而，基因治疗的唯一主要III期临床试验，涉及使用常规复制缺陷 GBM患者的逆转录病毒载体导致令人失望的低和治疗性不足的转导不足仅0.02％的订单的水平。标准复制缺陷逆转录病毒载体无法实现因此，在体内有效转导肿瘤是神经胶质瘤基因治疗的主要障碍。使用基因转移的复制能力载体将更有效，因为每个肿瘤细胞成功转导本身将成为产生病毒的细胞，即使在初始之后也可以维持进一步的转导事件行政。我们以前已经证明了鼠白血病病毒的直接肿瘤内注射（MLV）基于复制功能的逆转录病毒（RCR）矢量制剂可以实现极高的效率神经胶质瘤中的自杀基因转移，并严格限制于主动分裂肿瘤细胞没有明显扩散到肿瘤部位的证据，并导致生存率显着延长给药后，没有可检测的全身副作用。在这里，我们建议进一步改善通过工程化同种反应性细胞毒性T淋巴细胞（同胞）来成为RCR，这种方法的效率矢量生产者细胞，然后可以用作运动蜂窝递送平台，可以渗透到肿瘤质量并促进复制向量的多灶性扩散。同种异体CTL表现出特征为此提供了独特的优势：同anctls可以通过组织移动，可以杀死肿瘤接触后，可以产生诱导细胞凋亡或可以启动内源性免疫的细胞因子回复。在CMS Gliomas中，在免疫特权场地内的免疫抑制环境，免疫系统的自己的毁灭可能会被绕过足够长的时间，以使他们拥有有益的效果，但是在泄漏到一般循环中时，应迅速破坏同层。我们将测试他们生存能力，生物分布，安全性和效用作为蜂窝递送车辆，以增强RCR矢量扩展和治疗功效，比较体内免疫缺陷和免疫能力的神经瘤模型。