Nano Fibers for ADME/Toxicology

描述(由申请人提供)： 美国每年用于药物发现和开发的研发(R&D)支出约为600亿美元。利用细胞培养进行ADME/TOX(吸收、分布、代谢、排泄和毒理学)研究在促进药物发现和开发过程中发挥了重要作用。然而，传统2D培养中培养细胞的行为与体内3D培养细胞的行为存在许多差异，导致可预测性较低。我们假设A)3D细胞-支架结构在ADME/TOX研究中优于2D细胞培养(H1)，以及B)模拟胶原的纳米纤维结构和表面化学的支架可增强肝细胞的黏附、存活/生长和分化功能(H2)。因此，我们建议开发一种3D纳米纤维支架，用于肝细胞的黏附、维持分化和可能的增殖。这些细胞-支架结构(或工程组织)将保持3D细胞相互作用，应该更接近体内组织/器官的生理模拟，并将在ADME/Tox研究中为药物研究和开发提供更可预测的结果。 具体目标1.开发具有可操控的大孔结构和孔表面形态的三维纳米纤维支架，用于多孔培养板中细胞生长和组织形成。 具体目的2.通过体外肝细胞培养实验优化支架的大孔结构，证明纳米纤维孔壁结构优于可控孔壁结构作为肝细胞形成组织模拟物的三维支架。 具体目标3.赋予支架内部毛孔仿生表面特性，以实现最佳的肝细胞黏附和功能。 具体目标4.展示含有表面修饰3D的多孔培养板的优势 用于ADME/Tox研究的高通量效率、坚固性和有效性的纳米纤维支架。 通过实现上述具体目标，我们将加深对支架设计以优化肝细胞功能的理解，并展示3D纳米纤维支架作为更具预测性的临床前ADME/毒理学研究的新平台的优势。

DESCRIPTION (provided by applicant): The annual research and development (R&D) spending for drug discovery and development in the United States is approximately sixty billion dollars. ADME/Tox (absorption, distribution, metabolism, excretion and toxicology) studies using cell cultures have played important roles in facilitating the drug discovery and development process. However, there are many differences between the behavior of cultured cells in traditional 2D culture and their 3D counterparts in vivo, resulting in low predictability. We hypothesize that A) 3D cell-scaffold constructs are superior to 2D cell cultures for ADME/Tox studies (H1), and B) scaffolds that mimic the nano-fibrous architecture and surface chemistry of collagen enhance hepatocyte adhesion, survival/growth, and differentiated function (H2). We, therefore, propose to develop a 3D nano-fibrous scaffold for hepatocytes to adhere, maintain differentiation, and possibly proliferate. These cell-scaffold constructs (or engineered tissues), shall maintain 3D cellular interaction, should serve as closer physiological mimics of tissues/organs in vivo, and will afford more predictable outcomes in ADME/Tox studies for pharmaceutical research and development. Specific Aim 1. Develop 3D nano-fibrous scaffolds with manipulatable macro-pore architecture and pore surface morphology for cell growth and tissue formation in multi-well culture plates. Specific Aim 2. Optimize macro-pore structure of the scaffolds using in vitro hepatocyte culture assays, and demonstrate that nano-fibrous pore wall architecture is superior to control pore wall architecture as a 3D scaffold for hepatocytes to form tissue mimics. Specific Aim 3. Impart biomimetic surface properties to the internal pores of the scaffolds for optimal hepatocyte adhesion and function. Specific Aim 4. Demonstrate the advantages of multi-well culture plates containing surface-modified 3D nano-fibrous scaffolds for high-throughput efficiency, robustness and effectiveness for ADME/Tox studies. By accomplishing the above specific aims, we will advance our understanding of scaffold design for optimal hepatocyte function, and demonstrate the advantages of the 3D nano-fibrous scaffolds as a novel platform for more predictive preclinical ADME/Toxicology studies.