Nano Fibers for ADME/Toxicology

描述（由申请人提供）： 在美国，用于药物发现和开发的年度研究和开发（R&D）支出约为600亿美元。使用细胞培养的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，在传统的2D培养中培养的细胞的行为与其在体内的3D对应物之间存在许多差异，导致可预测性低。 我们假设A）3D细胞支架构建体上级用于ADME/Tox研究的2D细胞培养物（H1），和B）模拟胶原蛋白的纳米纤维结构和表面化学的支架增强肝细胞粘附、存活/生长和分化功能（H2）。因此，我们建议开发一种三维纳米纤维支架，用于肝细胞粘附，维持分化，并可能增殖。这些细胞-支架构建体（或工程化组织）应保持3D细胞相互作用，应充当体内组织/器官的更接近的生理模拟物，并且将在药物研究和开发的ADME/Tox研究中提供更可预测的结果。 具体目标1。开发具有可操纵的大孔结构和孔表面形态的3D纳米纤维支架，用于多孔培养板中的细胞生长和组织形成。 具体目标2。使用体外肝细胞培养测定优化支架的大孔结构，并证明纳米纤维孔壁结构作为肝细胞形成组织模拟物的3D支架上级对照孔壁结构。 具体目标3。赋予仿生表面性能的内部孔隙的支架，以最佳的肝细胞粘附和功能。 具体目标4。展示含有表面改性3D的多孔培养板的优势 纳米纤维支架用于ADME/Tox研究的高通量效率、稳健性和有效性。 通过实现上述具体目标，我们将进一步了解支架设计以实现最佳肝细胞功能，并证明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.