Three-dimensional Model of Human Ewing Sarcoma

人类尤文肉瘤三维模型

• 批准号: 9130807
• 负责人: Fred Kurtis Kasper
• 金额: $ 32.27万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-11 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9130807
• 关键词: Affect; Antineoplastic Agents; Architecture; Biomechanics; Biomimetics; Bioreactors; Blood Vessels; Bone neoplasms; Carcinoma; Cell Communication; Cell Differentiation process; Cell Survival; Cells; Characteristics; Chondrosarcoma; Clinical; Clinical Trials; Coculture Techniques; Colon Carcinoma; Complement; Complex; Cues; Deposition; Dimensions; Drug Targeting; Drug resistance; Endothelial Cells; Engineering; Ewings sarcoma; Exhibits; Extracellular Matrix; FRAP1 gene; Feedback; Goals; Health; Human; Hybrids; IGF1 gene; Institutional Review Boards; Insulin-Like Growth Factor I; Invaded; Laboratories; Left; Ligands; Malignant Neoplasms; Malignant neoplasm of lung; Mesenchymal Stem Cells; Microscopic; Modeling; Molecular Abnormality; Nature; Neoplasm Metastasis; Nutrient; Oxygen; Pathway interactions; Patients; Pattern; Perfusion; Polymers; Pre-Clinical Model; Proteomics; Publishing; Reporting; Research; Resistance; Role; SDZ RAD; Sampling; Signal Pathway; Signal Transduction; Specimen; Testing; Tissue Engineering; Tumor Cell Line; Tumor-Derived; Ursidae Family; Vascular Endothelial Growth Factors; bone; caprolactone; clinical investigation; clinically relevant; drug testing; in vivo; innovation; mTOR Inhibitor; malignant breast neoplasm; monolayer; neoplastic cell; novel; novel strategies; osteogenic; osteosarcoma; protein expression; resistance mechanism; scaffold; shear stress; stem; targeted treatment; therapeutic target; three dimensional cell culture; three-dimensional modeling; tumor; tumor growth; tumor microenvironment; tumorigenic; vasculogenesis

DESCRIPTION (provided by applicant): The overall goal of the proposed research is to apply static and flow perfusion bioreactor culture of bone sarcoma cells, grown upon a tissue-engineered polymer/extracellular matrix (ECM) hybrid model that reliably mimics key features of the bone tumor niche, to advance our understanding of the IGF-1R/mTOR cancer- related pathway and its clinically-relevant resistance mechanisms. Our laboratory has reported in vivo-like IGF- 1R/mTOR expression patterns, closely related to those observed in human Ewing sarcoma tumors (ES), when established ES cell lines are grown upon innovative biologically inert 3D electrospun poly(�-caprolactone) (PCL) microfiber scaffolds rather than upon traditional plastic monolayers. The present proposal seeks to elucidate the precise mechanisms by which cell-cell and cell-ECM interactions stimulate an activated IGF- 1R/mTOR signaling state within this engineered 3D bone tumor microenvironment, as those interactions are critically important in initiating and maintaining ES. In parallel with determining the influence of those parameters under static cell culture, 3D scaffolds and culture conditions will be adapted to better emulate the native bone microenvironment: (a) varied flow perfusion rates will be used to assess the effect of shear stress upon cell retention while facilitating uniform distribution of Ewing cells within the scaffold, (b) PCL scaffolds will incorporate IGF-1 to mimic the high concentration of IGF-1 naturally released as tumors invade surrounding bone, (c) the effect of an osteogenic ECM upon the IGF-1R/mTOR signaling cascade will be determined using decellularized scaffolds upon which heterotypic mesenchymal stem cells, differentiated toward an osteoblastic lineage, are first grown, and (d) ES cells will be co-cultured with endothelial cells (EC) to determine how heterotypic cells interact within a fabricated 3D bone tumor model to elicit viable ES tumors. Finally, to determine the mechanism(s) by which Ewing sarcoma evades sensitivity to combined mTOR/IGF-1R targeted therapy, freshly-derived tumor specimens (obtained from Ewing sarcoma patients treated with Medi- 573/everolimus in an IRB-approved clinical trial), will be grown in primary culture within 3D scaffolds and compared to 2D culture and patient-derived tumor explants (PDTX) by proteomic analysis of the IGF- 1R/mTOR pathway and putative resistance mechanisms. This novel approach of studying Ewing sarcoma within an ex vivo preclinical model of the bone microenvironment presents tremendous potential for understanding chemotherapeutic efficacy and for determining resistance mechanisms to biologically targeted therapies.

描述（由适用提供）：拟议的研究的总体目标是应用骨肉瘤细胞的静态和流动灌注生物反应器培养物，生长在组织工程化的聚合物/细胞外基质（ECM）混合模型上，以可靠地模拟我们对骨骼肿瘤的关键特征，以提高我们对IGF-1ER/MIGF CANCER CANCER CANCER-ig-1r/MTOR CANCER cANCER的关键特征机制。我们的实验室报告说，当建立的ES细胞系生长在创新的生物学上的3D静电纺丝poly（PCL）微生物纤维纤维丝球上的微型塑料塑料杂种时，与传统的塑料怪物相比，在人类ewing肉瘤肿瘤（ES）中观察到的类体类体内IGF-1R/MTOR表达模式与人类尤因肉瘤肿瘤（ES）密切相关。本提案旨在阐明细胞细胞和细胞ECM相互作用刺激该工程3D骨肿瘤微环境中激活的IGF-1R/mTOR信号传导状态的精确机制，因为这些相互作用在启动和维持ES中至关重要。 In parallel with determining the influence of those parameters under static cell culture, 3D scaffolds and culture conditions will be adapted to better emulate the native bone microenvironment: (a) varied flow perfusion rates will be used to assess the effect of shear stress upon cell retention while supporting uniform distribution of Ewing cells Within the scaffold, (b) PCL scaffolds will incorporate IGF-1 to mimic the high concentration of IGF-1自然释放为肿瘤侵入周围的骨骼，（c）成骨ECM对IGF-1R/MTOR信号传导级联的影响将使用脱细胞的脚手架确定，在该链球脚架上，在该型型中型间充质干细胞在该型中，朝着异质细胞的细胞与成骨细胞相关的细胞，并将其与Es eS eS-Debilect（d）确定（D）的细胞（d）。在制造的3D骨肿瘤模型中相互作用，以引起可行的ES肿瘤。最后，为了确定ewing肉瘤对MTOR/IGF-1R靶向治疗的敏感性的敏感性，新鲜衍生的肿瘤标本（从IRB批准的IRB临床试验中使用Medim-573/Everolimus治疗的Ewing Sarcom患者获得3D培养物和efulter cruitative to tod tags cruitative and frow 2d canters cruitatist和eufter-Effient to flust and frutive and frutive sarcome cruitative and Cruity-effert thauders cruitate-2D （PDTX）通过蛋白质组学分析IGF-1R/MTOR途径和推定的抗性机制。在骨微环境的离体临床前模型中研究EWING肉瘤的新方法为理解化学治疗效率以及确定对生物学靶向疗法的耐药机制提供了巨大的潜力。