Therapeutic disruption of Connexin43-mediated microtubule regulation to target glioblastoma cancer stem cells

Connexin43 介导的微管调节对胶质母细胞瘤干细胞的治疗破坏

基本信息

批准号：
9346494
负责人：
Samy Lamouille
金额：
$ 22.5万
依托单位：
ACOMHAL RESEARCH, INC.
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2019-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9346494
关键词：
Adult Alkylating Agents Apoptosis Binding Biochemical Biochemistry Biodistribution Biological Assay Brain Neoplasms Cell Communication Cell Maintenance Cell Proliferation Cell Survival Cell physiology Cells Central Nervous System Diseases Characteristics Chemotherapy-Oncologic Procedure Clinic Clinical Trials Collaborations Communication Connexin 43 Connexins Cytotoxic Chemotherapy Data Dimensions Disease Drug Delivery Systems Encapsulated Excision FDA approved FOXP3 gene Failure Fostering Foundations Future Gap Junctions Glioblastoma Glioma Goals Heterogeneity Human Hypoxia In Vitro Ions Legal patent Letters Licensing Link Malignant Neoplasms Mediating Medical Microscopy Microtubules Mission Modeling Names Nature Newly Diagnosed Operative Surgical Procedures Optics Organoids Patient-Focused Outcomes Patients Peptides Pharmaceutical Preparations Physiological Polyanhydrides Population Propane Property Proteins Public Health Radiation Radiation therapy Recurrence Refractory Regulation Research Research Institute Research Personnel Resistance Resolution South Carolina Stem cells Techniques Testing Therapeutic Therapeutic Effect Therapeutic Intervention Treatment Protocols Tumor Initiators Tumorigenicity United States National Institutes of Health Universities Validation Virginia Work aggressive therapy base biomaterial compatibility cancer cell cancer stem cell cell behavior chemotherapy copolymer effective therapy experience human disease improved in vitro Assay in vivo in vivo Model innovation irradiation migration mouse model nanoparticle neoplastic cell neurosurgery new therapeutic target novel novel strategies novel therapeutic intervention novel therapeutics outcome forecast particle peptidomimetics pre-clinical prevent reconstruction sebacic acid self-renewal small molecule stem cell population temozolomide tool treatment strategy tumor tumor microenvironment tumor progression tumorigenic

项目摘要

Project Summary Glioblastoma (GBM) is one of the most lethal incurable human diseases. Even with aggressive therapies including surgical resection followed by radiotherapy and chemotherapy using temozolomide (TMZ), the median survival for GBM patients is only 14 months. In fact, GBM cancer cells are highly infiltrative and comprise a sub- population of glioma stem cells (GSCs) with tumorigenic properties regulated by the tumor microenvironment, and often resistant to chemotherapy and irradiation treatments. The remaining GSCs can then enter an active state of self-renewal, and asymmetric division that recapitulates the heterogeneous tumor. As a result, all treated GBM patients will experience tumor recurrence and subsequent surgeries and toxic radiotherapy and chemotherapy regimens are harmful for the patient and often remain insufficient. There is therefore an urgent need for new therapeutic drug to target GSCs and treat this devastating disease. Glioblastoma resistance to TMZ correlates with the expression of the gap junction protein Connexin43 (Cx43), a protein which enables communication between cells, and increased levels of Cx43 are observed in GSCs. The function of Cx43, is not limited to forming channels for the passage of ions and small molecules between cells, but also participates in cell proliferation, migration and apoptosis. Therefore, targeting Cx43 activity holds promise to treat GBM and prevent tumor recurrence. In this proposed research we use a novel Cx43 mimetic peptide named JM2 (juxtamembrane 2) that encompasses the microtubule binding sequence of Cx43. Our preliminary data show that JM2 alters Cx43 binding to microtubule, decreases the formation of Cx43 gap junctions, and inhibits cell- cell communication in GSCs. Most importantly, we reveal the therapeutic potential of JM2 in decreasing GSC survival in vitro and in vivo. With the goal of developing a new therapy based upon JM2 to target GSCs in GBM, our overall objective is to generate JM2-loaded polyanhydride biodegradable nanoparticles (JM2-NPs) for sustained delivery of JM2 to GSCs. We will use high-resolution microscopy techniques including stochastic optical reconstruction microscopy (STORM) and biochemistry assays to analyze the effect of JM2-NPs in GSCs derived from human primary GBM cells freshly isolated from dissected patient tumor. Finally, we will assess the therapeutic effect of JM2-NPs on GSCs ex vivo using a three dimensional patient GBM-derived organoid model, and in vivo using an orthotopic GBM mouse model. These results will validate the potent effect of JM2 peptide for treatment of high Cx43/chemoresistant GSCs and present a therapeutic opportunity to prevent GBM tumor recurrence. The proposed research is significant because this innovative approach will not only allow us to develop novel therapies for lethal GBM but also will lay foundation on potential clinical trials in newly diagnosed GBM patients in the near future. Finally, our new JM2-loaded nanoparticles may be scalable to other CNS diseases that could benefit from targeting Cx43-microtubule interaction.

项目摘要胶质母细胞瘤（GBM）是最致命的人类疾病之一。即使有积极的疗法包括手术切除，然后进行放疗和使用替莫唑胺（TMZ）的化学疗法，中位数 GBM患者的生存率仅为14个月。实际上，GBM癌细胞具有很高的渗透性，包括胶质瘤干细胞（GSC）的种群具有由肿瘤微环境调节的致瘤特性，并且通常抗化疗和辐照治疗。其余的GSC可以进入活动自我更新状态和不对称分裂，概括了异质肿瘤。结果，所有人都接受了 GBM患者将经历肿瘤复发，随后的手术以及毒性放疗以及化学疗法方案对患者有害，并且通常不足。因此有紧急的需要新的治疗药物来靶向GSC并治疗这种毁灭性疾病。胶质母细胞瘤的抗性 TMZ与间隙连接蛋白连接的表达相关（CX43），该蛋白可以启用在GSC中观察到细胞之间的通信和CX43水平的增加。 CX43的功能不是仅限于形成细胞之间离子和小分子通过的通道，但也参与细胞增殖，迁移和凋亡。因此，针对CX43活动有望治疗GBM和预防肿瘤复发。在这项拟议的研究中，我们使用了一种新型的CX43模拟肽JM2 （Juxtammbrane 2）包含CX43的微管结合序列。我们的初步数据显示 JM2改变了CX43与微管结合，减少CX43间隙连接的形成，并抑制细胞 - GSC中的细胞通信。最重要的是，我们揭示了JM2降低GSC的治疗潜力体外和体内生存。为了开发基于JM2的新疗法以靶向GBM中的GSC，我们的总体目的是生成JM2负载的多丙二醇可生物降解纳米颗粒（JM2-NP）持续将JM2传递到GSC。我们将使用包括随机的高分辨率显微镜技术光学重建显微镜（Storm）和生物化学测定法分析JM2-NP在GSC中的影响源自从解剖的患者肿瘤新鲜分离的人类原代GBM细胞。最后，我们将评估使用三维患者GBM衍生的类器官模型，JM2-NP对GSC的治疗效应，并使用原位GBM鼠标模型进行体内。这些结果将验证JM2肽的有效作用用于治疗高CX43/化学耐药性GSC，并提供预防GBM肿瘤的治疗机会复发。拟议的研究很重要，因为这种创新的方法不仅可以使我们能够开发致命GBM的新型疗法，但也将基础在新诊断的潜在临床试验上 GBM患者在不久的将来。最后，我们加载的新的JM2纳米颗粒可能可扩展到其他CNS 靶向CX43微管相互作用可能受益的疾病。