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：
10539308
负责人：
Sauradeep Sinha
金额：
$ 4.07万
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/10539308
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 Development Diffuse intrinsic pontine glioma Discipline Disease Dose Engineering Environment Extracellular Matrix Faculty Future Glioblastoma Goals Histone Deacetylase Histone Deacetylation Hydrogels In Vitro Integrins Invaded 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 Proliferating Proteins RNA Receptor Inhibition Reporting Resistance development Role Scientist Signal Transduction Site Solid Neoplasm Survival Rate Testing Therapeutic Time Training Treatment outcome Tumor Cell Invasion Work anticancer research cancer cell career development cell behavior cost disease phenotype drug resistance development 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 synergism 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.

弥漫性内在的蓬托神经胶质瘤（DIPG）是高度攻击性的腹盆子的小儿脑肿瘤（脑干），五年生存率小于1％，中位存活率仅为9个月[1,2]。尽管在治疗其他形式的小儿癌，生存率方面，已经实现了生存率的显着提高因为DIPG在三十年中没有改变[1]。而脑肿瘤生态位本身是3D，多因素环境，以前的尝试依赖于标准的2D单层文化或动物模型来模仿疾病表型。但是，越来越多的证据表明，2D中的癌细胞行为有很大不同来自体内表型[3]；而动物模型的成本很高，生产冗长，而且通常很麻烦用于机械研究。此外，先前的研究几乎完全使用成人脑肿瘤细胞进行，而成人和小儿脑肿瘤已显示出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，并利用这种体外模型来阐明整合素受体的作用在驱动DIPG进展中的细胞 - 细胞相互作用。阻断特定整联蛋白受体的功效抑制DIPG进展将在体内进一步验证我们已建立的小鼠模型。我假设通过特定整联蛋白受体阻断DIPG粘附会抑制DIPG的增殖和侵袭 3D。此外，由于DIPG一直是证明甚至可以最终发展出对HDAC抑制等有希望的单个靶向方案的抵抗力[7,8]。我假设阻断整联蛋白受体将与HDAC抑制协同以进一步改善治疗 DIPG的结果通过破坏两种不同的致癌途径。我进一步假设3D共同文化在3D中与神经祖细胞（NPC）的DIPG会增强DIPG侵袭，这种表型模仿IN 体内响应。为了检验这些假设，我建议：（1）与脑模拟刚度开发3D水凝胶并优化了支持3D中DIPG增殖，侵袭和药物反应的粘合剂配体；（2）评估 DIPG粘附所需的特定特定整合素受体在抑制使用DIPG侵袭中的影响我们的3D水凝胶模型，并使用小鼠DIPG模型验证功效；（3）开发3D共同文化概括NPC诱导的DIPG入侵的模型，并确定受NPC/DIPG相互作用影响的关键信号使用RNA微阵列。拟议工作的结果将导致生物工程的发展 3D体外模型用于DIPG，具有模仿体内表型中的细胞 - 基质和细胞 - 细胞相互作用。在基本和医师科学家团队的指导下，拥有免费的专业知识，我将获得有价值的跨学科培训，并具有独特的位置，可以进行拟议的工作。