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：
8211001
负责人：
Khalid A Shah
金额：
$ 35.43万
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/8211001
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 nanobodies 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，胚胎和成人干细胞都具有巨大的细胞治疗潜力。在先前的研究中，我们确定：a）治疗工程的干细胞广泛迁移到肿瘤并浸润大脑中的沉积物，并在移植到小鼠神经胶质瘤模型中时具有凋亡和抗血管生成作用； b）可以在体内实时可视化受体靶向抗肿瘤疗法和命运的动力学。在此提案中，我们将创建抗毒素的人间充质干细胞（MSC），以实现靶向纳米剂和细胞毒性剂的现场递送，以同时阻止增殖并诱导肿瘤细胞的杀死而不会影响正常大脑。具体而言，MSC将被设计为表达针对过表达EGFR的靶向治疗蛋白，并在神经胶质瘤细胞中特别表达IL13RA2和死亡受体（DR）4/5。在与Henegouwen Lab（荷兰Utrecht University）密切合作，该实验室正在领导开发低分子量，高度溶解的EGFR EGFR特异性纳米生物剂（EGFR-NB），我们最近表明哺乳动物可以用来表达出现的可分泌的Bectar-Biver bexent Begfr nananobodies。我们最初将在MSC中表达EGFR-NB，并研究EGFR-NB对原代胶质瘤细胞和CD133+原发性脑肿瘤细胞在与Settleman Lab（MGH，Boston）持续合作中的EGFR信号传导和细胞增殖的影响。许多研究表明，当EGFR信号拮抗剂与细胞因子结合使用时，协同的抗肿瘤作用。将测试基于选择性靶向神经胶质瘤细胞的两种不同的细胞毒性疗法。在第一种方法中，我们将在与Rich Lab（B＆W医院，波士顿）的持续合作中创建耐毒素的MSC，以及MSC表达EGFR-NB和IL13R靶向的白喉毒素（DT），该毒素（DT）已知通过抑制蛋白质通过ADP-ribibosylation抑制蛋白质诱导细胞死亡，通过ADP-ribibibosylation诱导细胞死亡。在第二种方法中，我们将创建表达EGFR-NB和S-TRAIL的MSC，我们在增殖的神经胶质瘤细胞中通过上调的死亡受体（DR）-4/5进行了广泛的表征和证明可以选择性地诱导凋亡。两种方法都将在既定的神经胶质瘤系中的培养和体内疗效进行测试。基于这些发现，我们将在高度侵入性的胶质瘤原发性小鼠模型中利用最有效的治疗MSC。我们假设在治疗MSC的现场传递时，将导致细胞存活途径的同时下调和死亡途径的激活，从而导致根除的根除。与Weissleder Lab（MGH，Boston）密切合作的基因工程荧光和生物发光成像标记和体内成像的整合将使我们能够遵循MSC的交付和命运，并评估其体内的治疗功效，这些研究预计这些研究最终会与临床相比，这将在开发新型的干细胞疗法中产生重大影响。公共卫生相关性：在此提案中，我们将设计人类间充质干细胞（MSC）表达针对胶质瘤细胞过表达受体的靶向治疗蛋白。将探索基于靶向细胞增殖和神经胶质瘤细胞死亡途径的两种不同的组合治疗方法。开发的药物和策略将被设计为可以在临床上翻译，并且在开发有效的脑肿瘤疗法方面应产生重大影响。