Targeting Tumor Initiating Cell in Undifferentiated Pleomorphic Sarcoma

靶向未分化多形性肉瘤中的肿瘤起始细胞

基本信息

批准号：
9120229
负责人：
Benjamin Aaron Alman
金额：
$ 43.56万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9120229
关键词：
Ablation Accounting Adenoviruses Aftercare Alleles Allografting Antibodies Automobile Driving Biological Assay CSPG4 gene Cell Fraction Cell surface Cells Characteristics Chondroitin Sulfate Proteoglycan Clear Cell Color Data Deletion Mutation Development Diphtheria Toxin Disease Erinaceidae Exhibits Experimental Models Gene Expression Genetic Recombination Growth Human Immune Immunodeficient Mouse Immunotherapy In Situ Individual Infection Intramuscular Injections Label Lead Malignant Fibrous Histiocytoma Mediating Molecular Profiling Mus Mutant Strains Mice Mutation Neoplasm Metastasis Oncogenes Oncogenic Outcome Patients Population Population Dynamics Proteins Radiosurgery Recurrence Resistance Role Side Signal Pathway Soft tissue sarcoma Stable Populations TP53 gene Tamoxifen Testing Time Transgenic Mice Transplantation Undifferentiated Work Xenograft procedure base chemotherapy conventional therapy daughter cell effective therapy innovation insight knock-down mouse Cre recombinase mouse model neoplastic cell notch protein novel novel strategies novel therapeutic intervention outcome forecast prevent public health relevance recombinase sarcoma self-renewal small hairpin RNA therapeutic target tumor tumor eradication tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): Undifferentiated pleomorphic sarcoma (UPS) is a soft tissue sarcoma, with one of the worst prognosis. We will build on our discovery that UPS contains a small subpopulation of tumor initiating cells (TICs) that are enhanced for the ability t form tumors when transplanted into immune-deficient mice. Our proof of principle data showed that therapeutically targeting signaling pathways activated in the TIC population inhibits growth in UPS tumors established as xenografts in mice. In our preliminary data, we identified the chondroitin sulfate proteoglycan, NG2, as expressed in the TIC subpopulation in human and mouse UPS tumors. Our hypothesis is that the TIC fraction in UPS is responsible for maintaining UPS self-renewal and tumor growth, and that therapeutically targeting the TIC fraction can be developed into an effective UPS treatment. We will test this hypothesis by answering the following questions: 1) Will the eradication of TICs cause the degeneration of UPS? We found that deletion of Ng2 expressing cells in mouse UPS tumors, by driving Diphtheria Toxin in tumors established as allografts, will cause degeneration of tumors. Using flippase to activate oncogenic alleles to drive UPS development in mice, and cre-recombinase to drive Diphtheria Toxin in Ng2 expressing, we will be able to delete the TIC population in tumors in-situ, and determine if this causes degeneration of UPS tumors, prevents recurrence, or inhibits metastatic capacity. 2) Are individual clones within the Ng2 expressing population stable over time and resistant to conventional therapies? Ng2 expressing cells could be composed of cells that survive long term in tumors, giving rise to daughter cells that maintain the tumor over time; or they may represent different cells, each having TIC characteristics at different points in time. To determine which is the case, we will use the Confetti allele to label Ng2 expressing cells in mouse UPS tumors with different colored fluorescent proteins labeling individual clones. How these cell populations behave as the tumors grow, with serial transplantation, and after treatment using conventional therapies will determine which one of these possibilities is the case. 3) Can targeting NG2 be used to treat UPS? To determine how targeting Ng2 expression regulates tumor growth and self-renewal, NG2 will be knocked-down with shRNA in human UPS tumors established as xenografts in immunodeficient mice and NG2 will be deleted in primary mouse UPS tumors initiated by Cre recombinase that drives expression of oncogenes and recombines conditional Ng2 null alleles. Immunotherapy to NG2 will be examined in a similar manner in UPS tumors established as xenografts in immunodeficient mice. In addition to proving important biologic insight into sarcomas, these data will determine if short or long term therapeutic targeting of the TIC population will be more effective in UPS treatment, and inform a novel approach to develop a therapy based on targeting the UPS TIC.

描述（由申请人提供）：未分化的多形性肉瘤（UPS）是一种软组织肉瘤，是最严重的预后之一。我们将基于发现，UPS包含肿瘤启动细胞（TIC）的少量亚群，这些细胞（TICS）在移植到免疫缺陷型小鼠中时会增强肿瘤的能力t形成肿瘤。我们的原则数据证明表明，在TIC种群中激活的治疗靶向信号通路会抑制小鼠异种移植物确定为异种移植物的UPS肿瘤的生长。在我们的初步数据中，我们确定了硫酸软骨蛋白蛋白聚糖NG2，如人和小鼠UPS肿瘤中的TIC亚群中所示。我们的假设是，UPS中的TIC分数负责维持UPS自我更新和肿瘤的生长，并且可以将靶向TIC分数的靶向靶向有效的UPS治疗。我们将通过回答以下问题来检验这一假设：1）消除抽动会导致UPS变性吗？我们发现，通过驱动作为同种异体移植物确定的肿瘤中的白喉毒素来删除NG2表达细胞，将导致肿瘤退化。使用Flippase激活致癌等位基因，以推动小鼠的UPS发育，并在NG2表达中驱动白喉毒素的CRE聚集酶，我们将能够在原位删除肿瘤中的TIC群体，并确定这会导致UPS肿瘤的肿瘤变性，可预防复发能力，或者会导致复发能力或抑制转移。 2）NG2中的单个克隆是否表达人口随着时间的流逝稳定并抵抗常规疗法？ NG2表达细胞可以由在肿瘤中长期存活的细胞组成，从而产生随着时间的推移维持肿瘤的子细胞。否则它们可能表示不同的单元，每个细胞在不同时间点具有TIC特征。为了确定哪种情况，我们将使用五彩纸屑等位基因标记NG2在小鼠UPS肿瘤中具有不同颜色的荧光蛋白标记单个克隆的细胞。这些细胞群体如何随着肿瘤的生长，序列移植以及使用常规疗法的治疗后如何表现，将确定这些可能性中的哪一种。 3）靶向NG2可以用来治疗UPS吗？ To determine how targeting Ng2 expression regulates tumor growth and self-renewal, NG2 will be knocked-down with shRNA in human UPS tumors established as xenografts in immunodeficient mice and NG2 will be deleted in primary mouse UPS tumors initiated by Cre recombinase that drives expression of oncogenes and recombines conditional Ng2 null alleles.对NG2的免疫疗法将在免疫缺陷小鼠的异种移植物中以类似的方式进行检查。除了证明对肉瘤的重要生物学见解外，这些数据还将确定短期或长期治疗靶向TIC种群是否在UPS治疗方面更有效，并为基于靶向UPS TIC的疗法提供新的方法。