Developing diagnostic and therapeutic stem cells for cancer therapy

开发用于癌症治疗的诊断和治疗干细胞

基本信息

批准号：
8048074
负责人：
Khalid A Shah
金额：
$ 35.44万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-03-18 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8048074
关键词：
ADP ribosylation Affect Angiogenesis Inhibitors Apoptosis Apoptotic Asses Attenuated Binding Sites Bioluminescence Boston Brain Brain Neoplasms Caspase Cell Death Cell Death Inhibition Cell Line Cell Proliferation Cell Survival Cell Therapy Cells Cessation of life Cetuximab Clinical Data Clinical Trials Collaborations Combined Modality Therapy Cytotoxic Chemotherapy Cytotoxic agent Death Domain Death Receptor 5 Deposition Diagnostic Diphtheria Toxin Down-Regulation EGF gene Effectiveness Embryo Engineering Epidermal Growth Factor Receptor Exotoxins G Cells Glioblastoma Glioma Half-Life Homing Hospitals Human Human Engineering Image Immunotoxins Implant In Vitro Interleukin-13 Laboratories Lentivirus Vector Ligands Malignant - descriptor Mammalian Cell Mediating Mesenchymal Stem Cells Modeling Molecular Weight Monoclonal Antibodies Mus Nature Netherlands Pathology Pathway interactions Peptide Elongation Factor 2 Pharmacodynamics Plasmids Primary Brain Neoplasms Proliferating Protein Biosynthesis Protein Isoforms Proto-Oncogene Proteins c-akt Pseudomonas Radiation therapy Receptor Signaling Research Resistance Signal Pathway Signal Transduction Site Stem cells TNFRSF10A gene TNFSF10 gene Testing Therapeutic Therapy Clinical Trials Time Toxin Transplantation Treatment Efficacy Tumor Necrosis Factor-alpha Universities Variant Work adult stem cell base brain tissue cancer therapy cell killing combinatorial cytokine cytotoxic death receptor-4 design human TNFRSF10A protein in vivo innovation interleukin-13 receptor intravital fluorescence microscopy intravital microscopy killings mouse model mutant neoplastic cell nerve stem cell neurosurgery novel optical imaging overexpression pre-clinical public health relevance receptor stem cell fate stem cell therapy targeted delivery therapeutic protein time use tomography tumor tumor growth

项目摘要

DESCRIPTION (provided by applicant): Recent evidence indicates that both embryonic and adult stem cells have enormous therapeutic potential for cell therapy. In prior research, we have established that: a) therapeutically engineered stem cells migrate extensively to tumors and to infiltrating deposits in the brain and have apoptotic and antiangiogenic effects when transplanted into mouse models of glioma; and b) the dynamics of receptor targeted anti-tumor therapies and fate of stem cells can be visualized in real time in vivo. In this proposal, we will create toxin resistant human mesenchymal stem cells (MSC) for on-site delivery of targeted nanobodies and cytotoxic agents to simultaneously block proliferation and induce killing of tumor cells without affecting the normal brain. Specifically, MSC will be engineered to express targeted therapeutic proteins directed against overexpressed EGFR and specifically expressed IL13Ra2 and death receptors (DR)4/5 in glioma cells. In close collaboration with Henegouwen lab (Utrecht University, The Netherlands) which is leading efforts in developing low molecular weight, highly soluble EGFR specific nanobodies (EGFR-NB), we have recently shown that mammalian cells can be employed to express secretable bivalent EGFR nanobodies. We will initially express EGFR-NB in MSC and study the effect of EGFR-NB on EGFR signaling and cell proliferation in a panel of primary glioma cells and CD133+ primary brain tumor cells in an ongoing collaboration with Settleman lab (MGH, Boston). A number of studies have shown synergistic anti-tumor effects when EGFR signaling antagonists are combined with cytokines. Two different cytotoxic therapies based on selectively targeting glioma cells will be tested. In the first approach we will create toxin resistant MSC in an ongoing collaboration with Rich lab (B&W Hospital, Boston) and engineer MSC expressing EGFR-NB and IL13R-targeted Diphtheria toxin (DT) which is known to induce cell death by inhibition of protein synthesis through ADP-ribosylation of elongation factor-2 (EF-2). In the second approach we will create MSC expressing EGFR-NB and S-TRAIL, which we have extensively characterized and shown to selectively induce apoptosis via up-regulated death receptors (DR)-4/5 in proliferating glioma cells. Both approaches will be tested for their efficacy in culture and in vivo in established glioma lines. Based on these findings, we will utilize the most efficient therapeutic MSC in a highly invasive primary mouse model of glioma. We hypothesize that on site delivery of therapeutic MSC will result in simultaneous down-regulation of cell survival pathways and activation of death pathways thus resulting in enhanced eradication of gliomas. The integration of genetically engineered fluorescent and bioluminescent imaging markers and in vivo imaging in close collaboration with Weissleder lab (MGH, Boston) will allow us to follow delivery and fate of MSC and to asses their therapeutic efficacy in vivo These studies are expected to have a major impact in developing novel stem cell therapies that will eventually be compatible with clinical trials. PUBLIC HEALTH RELEVANCE: In this proposal we will engineer human mesenchymal stem cells (MSC) to express targeted therapeutic proteins directed against overexpressed receptors in glioma cells. Two different combinatorial therapy approaches based on targeting cell proliferation and death pathways in glioma cells will be explored. The developed agents and strategies will be designed to be clinically translatable and should have a major impact in developing efficient therapies for brain tumors.

描述（由申请人提供）：最近的证据表明胚胎干细胞和成体干细胞都具有细胞治疗的巨大治疗潜力。在之前的研究中，我们已经确定：a) 治疗性工程干细胞广泛迁移至肿瘤并浸润大脑沉积物，当移植到神经胶质瘤小鼠模型中时，具有细胞凋亡和抗血管生成作用； b) 受体靶向抗肿瘤治疗的动态和干细胞的命运可以在体内实时可视化。在该提案中，我们将创建抗毒素人间充质干细胞（MSC），用于现场递送靶向纳米抗体和细胞毒性剂，以同时阻止肿瘤细胞增殖并诱导杀死肿瘤细胞，而不影响正常大脑。具体来说，MSC 将被设计为表达针对神经胶质瘤细胞中过度表达的 EGFR 和特异性表达的 IL13Ra2 和死亡受体 (DR)4/5 的靶向治疗蛋白。与 Henegouwen 实验室（荷兰乌得勒支大学）密切合作，该实验室在开发低分子量、高溶解性 EGFR 特异性纳米抗体 (EGFR-NB) 方面处于领先地位，最近我们发现哺乳动物细胞可用于表达可分泌的二价 EGFR 纳米抗体。我们将首先在 MSC 中表达 EGFR-NB，并与 Settleman 实验室（MGH，波士顿）持续合作，研究 EGFR-NB 对一组原代胶质瘤细胞和 CD133+ 原代脑肿瘤细胞中 EGFR 信号传导和细胞增殖的影响。多项研究表明，当 EGFR 信号拮抗剂与细胞因子联合使用时，可产生协同抗肿瘤作用。将测试两种基于选择性靶向神经胶质瘤细胞的不同细胞毒性疗法。在第一种方法中，我们将与 Rich 实验室（波士顿 B&W 医院）持续合作，创建抗毒素 MSC，并设计表达 EGFR-NB 和 IL13R 靶向白喉毒素 (DT) 的 MSC，已知白喉毒素可通过抑制蛋白质来诱导细胞死亡通过延伸因子 2 (EF-2) 的 ADP 核糖基化合成。在第二种方法中，我们将创建表达 EGFR-NB 和 S-TRAIL 的 MSC，我们对其进行了广泛的表征，并证明其通过上调的死亡受体 (DR)-4/5 在增殖的神经胶质瘤细胞中选择性诱导细胞凋亡。这两种方法都将在已建立的神经胶质瘤系中测试其在培养物和体内的功效。基于这些发现，我们将在高侵袭性原代小鼠神经胶质瘤模型中利用最有效的治疗性 MSC。我们假设，治疗性 MSC 的现场递送将导致细胞存活途径的同时下调和死亡途径的激活，从而增强神经胶质瘤的根除。与 Weissleder 实验室（MGH，波士顿）密切合作，将基因工程荧光和生物发光成像标记物与体内成像相结合，将使我们能够跟踪 MSC 的递送和命运，并评估其体内治疗效果。对开发最终与临床试验兼容的新型干细胞疗法产生重大影响。公共健康相关性：在这项提案中，我们将改造人类间充质干细胞（MSC）来表达针对神经胶质瘤细胞中过度表达的受体的靶向治疗蛋白。将探索基于靶向神经胶质瘤细胞的细胞增殖和死亡途径的两种不同的组合治疗方法。所开发的药物和策略将被设计为可临床转化，并且应该对开发脑肿瘤的有效疗法产生重大影响。