Engineering Brain Cancer in a Dish: Hydrogel-based 3D in vitro Models for Pediatric Brain Tumor

在培养皿中改造脑癌：基于水凝胶的小儿脑肿瘤 3D 体外模型

基本信息

批准号：
10284928
负责人：
Sauradeep Sinha
金额：
$ 3.98万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10284928
关键词：
3-Dimensional Address Adherent Culture Adhesions Adhesives Adult Affect Animal Model Automobile Driving Biochemical Biomedical Engineering Biomimetics Brain Brain Neoplasms Brain Stem Cancer Biology Cancer Model Cell Communication Cells Cessation of life Child Childhood Childhood Brain Neoplasm Clinical Coculture Techniques Collaborations Complex Cues Custom Development Diffuse intrinsic pontine glioma Discipline Disease Dose Drug resistance Engineering Environment Extracellular Matrix Faculty Future Glioblastoma Goals Histone Deacetylase Histones Hydrogels In Vitro Integrins Lead Ligands Malignant Childhood Neoplasm Malignant neoplasm of brain Mentorship Modeling Molecular Mus Neoplasms Oncogenic Outcome Pathway interactions Pharmaceutical Preparations Phenotype Physicians Play Pontine structure Positioning Attribute Proteins RNA Receptor Cell Reporting Resistance development Role Scientist Signal Transduction Site Solid Neoplasm Survival Rate Testing Therapeutic Time Training Treatment outcome Tumor Cell Invasion Work Yang anticancer research base cancer cell career development cell behavior cost disease phenotype effective therapy efficacy validation improved improved outcome in vitro Model in vivo inhibitor materials science mechanical signal migration mouse model neoplastic cell nerve stem cell neurosurgery novel novel therapeutic intervention novel therapeutics receptor response therapeutic candidate therapeutic target three-dimensional modeling tumor growth tumor microenvironment

项目摘要

Diffuse intrinsic pontine gliomas (DIPG) are a highly aggressive pediatric brain tumor of the ventral pons (brain stem), with a five-year survival rate of less than 1% and a median survival of only 9 months [1,2]. While significant improvement in survival has been achieved in treating other forms of pediatric cancer, survival rate for DIPG has not changed in over three decades [1]. While the brain tumor niche itself is a 3D, multi-factorial environment, previous attempts have relied on standard 2D monolayer culture or animal models to mimic the disease phenotype. However, increasing evidence has shown that cancer cell behavior in 2D differs substantially from the in vivo phenotype [3]; whereas animal models are costly, lengthy to produce, and often cumbersome for mechanistic studies. Furthermore, previous studies were done almost exclusively with adult brain tumor cells, whereas adult and pediatric brain tumors have been shown to demonstrate distinct phenotypes in their sites of origin, clinical presentations and molecular mechanisms [4]. Through working at the interface of bioengineering, materials science, cancer biology, neurosurgery, and animal models, the goals of this proposal are to develop hydrogels with optimized niche cues to support DIPG proliferation and invasion in 3D, and to harness such in vitro model for elucidating the role of integrin receptors and cell-cell interactions in driving DIPG progression. The efficacy of blocking specific integrin receptors for inhibiting DIPG progression will be further validated in vivo using our established mouse models. I hypothesize that blocking DIPG adhesion through specific integrin receptors would inhibit DIPG proliferation and invasion in 3D. Furthermore, there is a need to find and advance combinational therapeutic strategies since DIPG has been shown to ultimately develop resistance even to promising single targeting regimes like HDAC inhibition [7,8]. I hypothesize that blocking integrin receptor would synergize with HDAC inhibition to further improve treatment outcome of DIPG by disrupting two distinct oncogenic pathways. I further hypothesize that 3D co-culture of DIPG with neural progenitor cells (NPCs) in 3D would enhance DIPG invasion, a phenotype that mimics the in vivo response. To test these hypotheses, I propose to: (1) Develop 3D hydrogels with brain-mimicking stiffness and optimized adhesive ligands that support DIPG proliferation, invasion and drug responses in 3D; (2) Evaluate the effects of blocking specific integrin receptors required for DIPG adhesion in inhibiting DIPG invasion using our 3D hydrogels models, and validate the efficacy using a mouse DIPG model; and (3) Develop a 3D co-culture model to recapitulate NPC-induced DIPG invasion, and identify key signals impacted by NPC/DIPG interactions using RNA microarray. The outcomes of the proposed work would lead to the development of a bioengineered 3D in vitro model for DIPG with controlled cell-matrix and cell-cell interactions that mimics in vivo phenotype. Under the mentorship of a team of basic and physician scientists, with complimentary expertise, I will gain valuable interdisciplinary trainings and be uniquely positioned to carry out the proposed work.

弥漫性内在桥脑胶质瘤 (DIPG) 是一种高度侵袭性的脑桥腹侧儿童脑肿瘤（脑干），五年生存率低于1%，中位生存期仅为9个月[1,2]。尽管在治疗其他形式的儿科癌症时，生存率已取得显着改善，生存率 DIPG 三十多年来没有改变[1]。虽然脑肿瘤生态位本身是一个 3D 的、多因素的环境，之前的尝试依赖于标准的 2D 单层培养或动物模型来模仿疾病表型。然而，越来越多的证据表明，二维癌细胞的行为存在很大差异来自体内表型 [3]；而动物模型成本高昂，制作时间长，而且通常很麻烦用于机理研究。此外，之前的研究几乎都是针对成人脑肿瘤细胞进行的，而成人和儿童脑肿瘤已被证明在其部位表现出不同的表型起源、临床表现和分子机制[4]。通过在生物工程、材料科学、癌症生物学、神经外科和动物模型，该提案的目标是开发具有优化利基线索的水凝胶以支持 DIPG 3D 中的增殖和侵袭，并利用此类体外模型来阐明整合素受体的作用以及驱动 DIPG 进展的细胞间相互作用。阻断特定整合素受体的功效抑制 DIPG 进展将使用我们建立的小鼠模型在体内进一步验证。我假设通过特定整合素受体阻断 DIPG 粘附将抑制 DIPG 增殖和侵袭 3D。此外，由于 DIPG 已被广泛应用，因此需要寻找和推进联合治疗策略。事实证明，即使是对 HDAC 抑制等有希望的单一靶向方案，最终也会产生耐药性 [7,8]。我假设阻断整合素受体将与 HDAC 抑制协同作用以进一步改善治疗 DIPG 的结果是通过破坏两种不同的致癌途径。我进一步假设 3D 共培养 3D 状态下的 DIPG 与神经祖细胞 (NPC) 会增强 DIPG 侵袭，这是一种模仿体内神经祖细胞 (NPC) 的表型体内反应。为了检验这些假设，我建议：（1）开发具有模拟大脑硬度的 3D 水凝胶以及支持 DIPG 增殖、侵袭和 3D 药物反应的优化粘附配体； (2) 评估阻断 DIPG 粘附所需的特定整合素受体在抑制 DIPG 侵袭中的作用我们的 3D 水凝胶模型，并使用小鼠 DIPG 模型验证功效； (3) 开发 3D 共培养模型可重现 NPC 诱导的 DIPG 入侵，并识别受 NPC/DIPG 相互作用影响的关键信号使用RNA微阵列。拟议工作的成果将导致生物工程的发展 DIPG 3D 体外模型，具有模拟体内表型的受控细胞-基质和细胞-细胞相互作用。在基础科学家和医学科学家团队的指导下，在专业知识的补充下，我将获得有价值的跨学科培训，并具有独特的优势来开展拟议的工作。