BRIGE: Engineering a BioMatrix Library Derived from Induced Pluripotent Stem Cells

技术描述：人类诱导多能干细胞（hiPSC）是成体细胞（例如，来自活检的皮肤细胞），通过被迫表达对维持胚胎干细胞的定义特性重要的基因和因子，已被遗传重编程为胚胎干细胞样状态。 它们在药物筛选、疾病建模和最终人类细胞治疗方面具有巨大的潜力。诱导多能干细胞（iPSC）在理解疾病和开发治疗方法方面具有巨大潜力。 目前用于iPSC自我更新和分化的基质未能概括谱系特化期间的复杂信号传导网络。 为了解决这个问题，一个新的生物基质库将在拟议的研究中产生，以研究干细胞发育过程中的复杂信号传导。 所提出的Biomatrix文库的独特之处在于在组织发育和形态发生期间维持广泛的信号传导能力和信号传导网络。 该研究的具体任务包括：1）具有各种生化和生物学特性的Biomatrix文库的生成和高含量表征; 2）用于iPSCs自我更新和神经分化的Biomatrix文库的高通量筛选和验证。 将基于Biomatrix的功能性神经分化转化为生物反应器，以确认筛选的预测效果。 本研究的长期目标是获得对细胞-生物基质相互作用在调控iPSC的谱系特异性分化中的全面理解。更广泛的意义和重要性：人工来源于成体细胞的诱导多能干细胞（例如，皮肤细胞）为研究提供了胚胎干细胞的替代方案。 这一2012年诺贝尔奖获得者的发现代表了科学研究的一个重要进展，因为它们使研究人员能够获得多能干细胞，这些干细胞具有成为体内任何细胞类型的能力，而无需使用有争议的胚胎。 使用完善的iPSC细胞系，拟议的研究将确定细胞，基质和生长因子之间的关键相互作用，这些相互作用可以调节细胞命运决定。 研究成果将指导仿生材料设计和大规模产生功能性组织细胞，用于药物筛选，疾病建模和最终的细胞治疗。 该提案详细介绍了一项变革性的研究计划，以实现iPSCs的高效分化，由工程生物基质调节，将通过创新的高含量表征，高通量筛选和功能分析进行鉴定。 获得的知识可以直接转化为生物制药和生物技术行业的技术商业化。 扩大代表性不足的群体在工程中的参与扩大参与活动的目标人群在拟议的努力包括非洲裔美国人，西班牙裔，和妇女的学生在佛罗里达A M大学，一个历史上的黑人大学，佛罗里达州立大学，一个主要的研究型大学与大量的女性入学，和塔拉哈西社区。 在全球范围内，PI将利用全球科学、工程和技术教育外联门户网站为土耳其女学生提供辅导和领导力培训。 PI将在课程中纳入行业所需的技能，如当前的良好生产规范和设计质量。 这些活动将弥合教育与应用之间的差距，并吸引来自代表性不足群体的学生留在科学和工程领域。 拟议的研究和教育工作的结果将分发给莱昂县学区和挑战者学习中心的K-12学生，生物医学工程学会的本科生和全国黑人工程师学会的学生。 特别是，辅导活动将扩大妇女在当地和全球，特别是在塔拉哈西和土耳其参与科学和工程，这项研究是通过工程学扩大参与研究启动赠款征集资助的，这是工程教育和中心司工程学扩大参与方案的一部分。

Technical Description:Human Induced Pluripotent Stem Cells (hiPSC) are adult cells (for example, skin cells derived from a biopsy) that have been genetically reprogrammed to an embryonic stem cell-like state by being forced to express genes and factors important for maintaining the defining properties of embryonic stem cells. They have great potential for drug screening, disease modeling, and eventually cell therapy for humans. Induced pluripotent stem cells (iPSCs) have great potential to understand diseases and develop therapeutics. Current matrices that are used for iPSC self-renewal and differentiation fail to recapitulate the complex signaling network during lineage specification. To address this issue, a novel Biomatrix Library will be generated in the proposed research to study the complex signaling during stem cell development. The unique feature of the proposed Biomatrix Library is the maintenance of the broad signaling capacity and preservation of signaling network during tissue development and morphogenesis. The specific tasks in the proposed study include: 1) generation and high-content characterization of a Biomatrix Library with various biochemical and biological properties; 2) high-throughput screening and validation of the Biomatrix Library for self-renewal and neural differentiation of iPSCs. The Biomatrix-based functional neural differentiation will be translated into a bioreactor to confirm the predicative effect from the screening. The long-term objective of this research is to obtain comprehensive understanding of cell-biomatrix interactions in regulating lineage-specific differentiation of iPSCs.Broader Significance and Importance:Induced pluripotent stem cells that are artificially derived from adult cells (e.g., skin cells) provide an alternative to embryonic stem cells for research. This 2012 Nobel Prize winning discovery represents an important advance in scientific research as they allow researchers to obtain pluripotent stem cells, which have the ability to become any cell type in the body, without the controversial use of embryos. Using well-established iPSC cell lines, the proposed study will identify the key interactions of cells, matrices and growth factors that could regulate cell fate decision. The research outcome will guide the biomimetic material design and the large scale generation of functional tissue cells for drug screening, disease modeling, and eventually cell therapy. This proposal details a transformative research plan to achieve highly efficient differentiation of iPSCs, regulated by the engineered Biomatrix that will be identified through innovative high content characterization, high-throughput screening, and functional analysis. The gained knowledge can be directly transformed to technology commercialization in the biopharmaceutical and biotechnology industries. Broadening Participation of Underrepresented Groups in EngineeringThe target populations of broadening participation activities in the proposed efforts include the African-American, Hispanic, and women students in Florida A&M University, a historically black college, Florida State University, a major research university with a large female enrollment, and Tallahassee community. Globally, the PI will provide mentoring and leadership training for Turkish women students using Global Educational Outreach in Science, Engineering and Technology portal. The PI will incorporate the skills required in industry such as current Good Manufacturing Practices and Quality by Design in the curriculum. These activities will bridge the gap between education and application, and also attract the students from under-represented groups to stay in science and engineering. The results from the proposed research and education efforts will be disseminated to K-12 students in Leon County School District and in Challenger Learning Center, to undergraduates in Biomedical Engineering Society, and to students in National Society of Black Engineers. In particular, the mentoring activities will broaden the participation of women in science and engineering locally and globally, especially in Tallahassee and Turkey.This research has been funded through the Broadening Participation Research Initiation Grants in Engineering solicitation, which is part of the Broadening Participation in Engineering Program of the Engineering Education and Centers Division.