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)是成人细胞(例如，来自活组织检查的皮肤细胞)，通过被迫表达对维持胚胎干细胞的决定性特性至关重要的基因和因子，已在基因上重新编程为类似胚胎干细胞的状态。它们在药物筛选、疾病建模以及最终用于人类的细胞治疗方面具有巨大的潜力。诱导多能干细胞(IPSCs)在认识疾病和开发治疗方面具有巨大的潜力。目前用于IPSC自我更新和分化的矩阵在谱系指定过程中不能概括复杂的信号网络。为了解决这个问题，在拟议的研究中将建立一个新的Biomatrix文库来研究干细胞发育过程中的复杂信号转导。建议的Biomatrix文库的独特之处在于在组织发育和形态发生过程中保持了广泛的信号能力和信号网络的保存。本研究的具体任务包括：1)构建具有多种生化和生物学特性的Biomatrix文库并进行高含量鉴定；2)对Biomatrix文库进行高通量筛选和验证，用于IPSCs的自我更新和神经分化。基于Biomatrix的功能神经分化将被转化到生物反应器中，以确认筛选的预测效果。这项研究的长期目标是全面了解细胞-生物基质相互作用在调节IPSC谱系特异性分化中的作用。广泛的意义和重要性：从成体细胞(例如皮肤细胞)人工来源的诱导多能干细胞为研究提供了一种替代胚胎干细胞的方法。2012年诺贝尔奖获得者的这一发现代表着科学研究的一项重要进步，因为它们使研究人员能够获得多能干细胞，这种干细胞具有成为体内任何细胞类型的能力，而不需要使用有争议的胚胎。利用成熟的IPSC细胞系，这项拟议的研究将确定可能调节细胞命运决定的细胞、基质和生长因子之间的关键相互作用。研究结果将指导仿生材料的设计和功能组织细胞的大规模生成，用于药物筛选、疾病建模，并最终用于细胞治疗。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，该计划由工程Biomatrix调控，将通过创新的高含量表征、高通量筛选和功能分析来确定。所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。扩大未得到充分代表的群体在工程方面的参与扩大参与活动的目标人群包括佛罗里达农工大学、历史上的黑人学院、佛罗里达州立大学和塔拉哈西社区的非裔美国人、西班牙裔和女性学生。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指导和领导力培训。PI将在课程中融入行业所需的技能，如当前良好的制造实践和设计质量。这些活动将弥合教育和应用之间的差距，并吸引来自代表性不足群体的学生留在科学和工程专业。拟议的研究和教育工作的结果将传播给里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协会的学生。特别是，辅导活动将扩大妇女在当地和全球科学和工程领域的参与，特别是在塔拉哈西和土耳其。这项研究的资金来源是工程教育和中心司扩大工程参与方案的一部分--工程征集中的扩大参与研究启动补助金。

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.