In vivo imaging of encapsulated stem cells in mouse models of tumor resection

肿瘤切除小鼠模型中封装干细胞的体内成像

• 批准号: 8599446
• 负责人: Khalid A Shah
• 金额: $ 34.83万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8599446
• 关键词: Adjuvant; Adjuvant Chemotherapy; Adult; Aminolevulinic Acid; Apoptosis; Boston; Brain; Brain Neoplasms; Cancer Patient; Carmustine; Cell Survival; Cells; Clinical; Clinical Research; Collaborations; Deposition; Development; Disease; Drug Kinetics; Encapsulated; Engineering; Ensure; Epidermal Growth Factor Receptor; Excision; Extracellular Matrix; Fluorescence; Fluorescent Dyes; Future; Genetic Engineering; Glioblastoma; Glioma; Goals; Home environment; Homing; Human; Human Engineering; Image; In Vitro; Injection of therapeutic agent; Invaded; Laboratories; Lead; Ligands; Magnetic Resonance Imaging; Malignant - descriptor; Malignant neoplasm of brain; Mesenchymal Stem Cells; Microscopy; Modeling; Mus; Nature; Operative Surgical Procedures; Optics; Pathology; Patients; Phenotype; Play; Positron-Emission Tomography; Primary Brain Neoplasms; Primary Neoplasm; Progression-Free Survivals; Publishing; Radiation therapy; Recurrence; Research Personnel; Resected; Residual state; Resistance; Role; Safety; Simplexvirus; Simulate; Site; Solid; Stem cells; Surgically-Created Resection Cavity; TNFSF10 gene; Testing; Therapeutic; Thymidine Kinase; Time; Transplantation; Treatment Efficacy; Tumor Debulking; Tumor Necrosis Factor-alpha; Tumor Volume; Universities; Utah; Virus; antitumor agent; base; cell killing; cell motility; cell suicide; cellular engineering; chemotherapeutic agent; design; in vivo; in vivo imaging; intravital microscopy; killings; mortality; mouse model; nanobodies; neoplastic cell; novel; outcome forecast; preclinical study; prevent; public health relevance; standard care; suicide gene; therapeutic protein; tumor; tumor growth

DESCRIPTION (provided by applicant): The standard treatment for malignant glioblastoma multiforme (GBM) includes maximal surgical tumor resection followed by radiation therapy and adjuvant chemotherapy. However, recurrence rates of GBM and the associated patient mortality are nearly 100%. Despite the key role of tumor resection in clinical GBM therapy, most of the pre-clinical studies focus on treating solid intact intracranial GBM tumors without mimicking the clinical scenario of surgical resection. Therefore, implementation of tumor resection in mouse models that recapitulate the clinical disease features are critical in developing clinically translatable therapies for GBM. In the proposed studies, we will first create and characterize different GBM resection models using patient derived CD133+ GBMs based on 3 different phenotypes (invasive, semi-invasive and nodular). While resection of primary tumor has shown clinical benefit, systemically delivered chemotherapeutic agents or direct injection of viruses and placement of carmustine (BCNU) wafers in tumor resection cavities has provided very limited additional benefit. Based on our recent findings that encapsulation of mesenchymal stem cells (MSC) is necessary to prevent rapid "wash- out" of stem cells post-transplantation in the tumor resection cavity, we will encapsulate human MSC engineered to express in vitro and in vivo imaging markers into synthetic extracellular matrices (sECMs) and evaluate them for their retention, survival and tumor homing in mouse resection models of GBM with different phenotypes. Based on our preliminary studies which indicate that a novel anti-tumor agent consisting of a secretable version of epidermal growth factor receptor targeted nanobody fused to tumor necrosis factor apoptosis inducing ligand (Enb-TRAIL), induces GBM cell killing in both TRAIL resistant and sensitive GBMs, encapsulated MSC-Enb- TRAIL will be tested in different GBM models of resection. To ensure the safety of our approach, we will ultimately engineer human MSC-Enb-TRAIL to express HSV-thymidine kinase (TK), an activatable cellular suicide gene that will allow us to selectively eradicate MSC post-GBM treatment. The incorporation of genetically engineered fluorescent and bioluminescent imaging (BLI) markers into MSC and GBMs will allow us to follow GBM cell invasion, fate of MSC and pharmacokinetics of therapeutic proteins and their efficacy by in vivo BLI, intravital microscopy (IVM), magnetic resonance imaging (MRI) and positron emission tomography (PET) and thus to fine tune the proposed approaches. Once validated, we will initiate a clinical study in which at the time of brain tumor surgery, the main tumor mass will be removed and sECMs encapsulated MSC will be introduced to target a broad spectrum of remaining tumor cells and micro-invasive tumor deposits in the brain. This will have a major impact in saving the lives of many brain cancer patients.

描述（由申请人提供）：恶性多形性胶质母细胞瘤（GBM）的标准治疗包括最大限度的肿瘤切除术，然后进行放射治疗和辅助化疗。然而，GBM的复发率和相关的患者死亡率接近100%。尽管肿瘤切除术在临床GBM治疗中起着关键作用，但大多数临床前研究都集中在治疗实体完整的颅内GBM肿瘤，而不模拟手术切除的临床情况。因此，在重现临床疾病特征的小鼠模型中实施肿瘤切除对于开发GBM的临床可转化疗法至关重要。在拟定的研究中，我们将首先使用基于3种不同表型（侵袭性、半侵袭性和结节性）的患者来源的CD 133 + GBM创建和表征不同的GBM切除模型。虽然原发性肿瘤的切除已经显示出临床益处，但是全身递送的化疗剂或直接注射病毒和 在肿瘤切除腔中放置卡莫司汀（BCNU）薄片提供了非常有限的额外益处。基于我们最近的发现，即包封间充质干细胞（MSC）对于防止移植后干细胞在肿瘤切除腔中的快速“洗出”是必要的，我们将工程化以表达体外和体内成像标记物的人MSC包封到合成细胞外基质（sECM）中，并评估它们在具有不同表型的GBM的小鼠切除模型中的保留、存活和肿瘤归巢。基于我们的初步研究，其表明由与肿瘤坏死因子凋亡诱导配体（Enb-TRAIL）融合的表皮生长因子受体靶向纳米抗体的可分泌形式组成的新型抗肿瘤剂在TRAIL抗性和敏感GBM中诱导GBM细胞杀伤，将在不同的GBM切除模型中测试包封的MSC-Enb- TRAIL。为了确保我们方法的安全性，我们最终将改造人MSC-Enb-TRAIL以表达HSV-胸苷激酶（TK），这是一种可激活的细胞自杀基因，将使我们能够选择性地根除GBM治疗后的MSC。将基因工程荧光和生物发光成像（BLI）标记物掺入MSC和GBM将使我们能够通过体内BLI，活体显微镜（IVM），磁共振成像（MRI）和正电子发射断层扫描（PET）跟踪GBM细胞侵袭，MSC的命运和治疗性蛋白质的药代动力学及其功效，从而微调所提出的方法。一旦得到验证，我们将启动一项临床研究，在脑肿瘤手术时，将切除主要肿瘤块，并引入sECM包封的MSC，以靶向脑中广泛的剩余肿瘤细胞和微侵袭性肿瘤沉积物。这将对挽救许多脑癌患者的生命产生重大影响。