Engineering 3D Osteosarcoma Models to Elucidate Biology and Inform Drug Discovery

工程 3D 骨肉瘤模型以阐明生物学并为药物发现提供信息

基本信息

批准号：
10564801
负责人：
Eric Alejandro Sweet-Cordero
金额：
$ 66.62万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10564801
关键词：
3-Dimensional ATAC-seq Acceleration Adherent Culture Adopted Adoption Affect Automobile Driving Bar Codes Biocompatible Materials Biology Bone Tissue Cancer Model Cell Line Cell Proliferation Cell model Cells Cellular Morphology Child Combination Drug Therapy Combined Modality Therapy Communities Cues DNA Data Development Disease Drug resistance Engineering Evaluation Exhibits Experimental Models Extracellular Matrix Gelatin Gene Amplification Genomics Goals Heterogeneity Hydroxyapatites Knowledge Lead Legal patent Malignant Bone Neoplasm Mediating Mind Minerals Modeling Mus Nanoporous Oncogenes Osteogenesis Outcome Pathology Patients Pharmaceutical Preparations Phenotype Physiological Positioning Attribute Receptor Protein-Tyrosine Kinases Regulatory Pathway Reporting Research Resistance Resources Role Scientist Signal Transduction Survival Rate Testing Therapeutic Tissue Engineering Treatment outcome Work Xenograft Model angiogenesis anticancer research bone cancer cell chemotherapy cost design disease heterogeneity drug candidate drug discovery drug response prediction effective therapy established cell line high dimensionality high throughput screening high-throughput drug screening improved in vitro Model in vivo invention mineralization mouse model new therapeutic target novel novel therapeutics osteosarcoma patient derived xenograft model porous hydrogel pre-clinical primary bone cancer resistance mechanism response scaffold screening single-cell RNA sequencing soft tissue stemness targeted treatment three dimensional cell culture three-dimensional modeling tool transcriptome sequencing tumor young adult

项目摘要

Osteosarcoma (OS) is an aggressive primary bone cancer that mainly affects children and young adults, and is characterized by high genomic complexity. Current treatment relies on chemotherapy, yet many patients exhibit resistance or develop metastatic disease. Current experimental models for OS research rely primarily on 2D monolayer culture or xenograft models. However, 2D cultures culture generally fail to retain tumor phenotypes and drug response in vivo, whereas mouse models are costly and impractical for high-throughput drug screening. Recently tissue engineered 3D cancer models have emerged as new cancer research tools, which better recapitulate in vivo tumor signaling and drug responses than 2D cultures. However, most tissue engineered cancer models to date are limited to soft tissues. Unlike soft tissues, bone is characterized by a highly- mineralized extracellular matrix (ECM) comprised of 70% minerals such as hydroxyapatite (HA) crystals. However, the role of bone mineral in driving OS progression and drug response remains largely unknown. Furthermore, previous OS studies rely on a narrow set of cell lines that have been in culture for decades, which may no longer reflect the biology and drug response in vivo The overall goal of this proposal is to integrate a scalable and physiologically relevant 3D OS model with high-dimensional sequencing tools to elucidate OS genomic heterogeneity and drug resistance, as well as screening novel combination therapies using multiple patient-derived OS cell lines. Our 3D models is specifically designed with high-throughput screening in mind, and leverages on a patented microribbon (µRB)-based scaffold invented by the Yang (PI) lab. This multi-PI application will bring together expertise in biomaterials design and 3D tumor models (Yang lab/Stanford) with expertise in patient-derived xenograft (PDX) cell lines, genomics and preclinical therapeutics of OS (Sweet- Cordero lab/UCSF). We hypothesize that OS signaling and drug responses in 3D culture can be modulated by tuning the type and size of mineral cues 3D gelatin µRB scaffolds to better mimic the in vivo phenotype, and combinational therapies that target identified signaling using 3D OS model will lead to better treatment outcomes for OS in vivo. To test these hypotheses, we will carry out the following aims. Aim 1: Develop 3D OS models with optimized niche cues for deep characterization of OS signaling and heterogeneity using multiple OS PDX cell lines and compare results to mouse orthotopic OS models. AIM 2: To harness 3D OS models to determine the regulatory pathways involved in mediating receptor tyrosine kinase expression in OS and identify lead drug candidates by screening a panel of targeted drug therapies. Aim 3: To identify novel combination therapies for PDX OS cell lines and elucidate potential drug resistance mechanisms using 3D OS models. This study will pioneer integrating 3D OS model with PDX cell lines and high-dimensional sequencing expertise. The outcomes would significantly advance the understanding of OS biology and heterogeneity, identifying drug resistance mechanisms, and accelerate discovery of combination therapies that cannot be achieved using existing tools.

骨肉瘤 (OS) 是一种侵袭性原发性骨癌，主要影响儿童和年轻人，其特点是基因组高度复杂，目前的治疗依赖于化疗，但许多患者表现出这种情况。当前 OS 研究的实验模型主要依赖于 2D。单层培养或异种移植模型然而，二维培养通常无法保留肿瘤表型。和体内药物反应，而小鼠模型对于高通量药物筛选来说成本高昂且不切实际。最近，组织工程 3D 癌症模型已成为新的癌症研究工具，它可以更好地然而，大多数组织工程技术比二维培养物更能概括体内肿瘤信号传导和药物反应。迄今为止的癌症模型仅限于软组织，与软组织不同，骨骼的特点是高度敏感。矿化细胞外基质 (ECM) 由 70% 的矿物质组成，例如羟基磷灰石 (HA) 晶体。然而，骨矿物质在驱动 OS 进展和药物反应中的作用仍然很大程度上未知。此外，之前的 OS 研究依赖于一组已培养了数十年的狭窄细胞系，这些细胞系可能不再反映体内的生物学和药物反应该提案的总体目标是整合可扩展且生理相关的 3D OS 模型，具有高维测序工具来阐明 OS 基因组异质性和耐药性，以及使用多种药物筛选新的联合疗法我们的 3D 模型专为高通量筛选而设计，并利用由 Yang (PI) 实验室发明的基于微带 (μRB) 的专利支架。该应用程序将汇集生物材料设计和 3D 肿瘤模型（Yang 实验室/斯坦福大学）方面的专业知识患者来源的异种移植（PDX）细胞系、基因组学和 OS 临床前治疗方面的专业知识（Sweet- Cordero 实验室/UCSF）我们勇敢地认为 3D 培养中的 OS 信号传导和药物反应可以通过以下方式进行调节：调整矿物质线索 3D 明胶 µRB 支架的类型和大小，以更好地模拟体内表型，以及使用 3D OS 模型针对已识别信号的联合疗法将带来更好的治疗结果为了测试这些假设，我们将实现以下目标：开发 3D OS 模型。使用多个 OS PDX 细胞优化利基线索，深入表征 OS 信号传导和异质性目标 2：利用 3D OS 模型来确定参与介导 OS 中受体酪氨酸激酶表达的调节途径并确定先导药物通过筛选一组靶向药物疗法来寻找候选者。目标 3：确定新的联合疗法。本研究将使用 3D OS 模型来研究 PDX OS 细胞系并阐明潜在的耐药机制。将 3D OS 模型与 PDX 细胞系和高维测序专业知识相结合的先驱。将显着促进对操作系统生物学和异质性的理解，识别耐药性机制，并加速发现使用现有工具无法实现的联合疗法。