Controlling Protein Release via Intermolecular Hybridization

蛋白类药物在治疗各种人类疾病方面有着巨大的前景。然而，蛋白质药物的安全高效递送一直是药物递送领域的难题。许多蛋白质递送系统仍然存在一些问题，包括蛋白质药物的快速释放，控制多种蛋白质释放的效率低下，以及在蛋白质递送系统制备过程中涉及有毒分子和/或恶劣条件。这个项目就是为了解决这些问题而提出的。智力优势：提出的研究目标是彻底了解互补寡核苷酸介导的蛋白质-适体解离的机制，并在此基础上，开发一种利用核酸适体、互补寡核苷酸和海藻酸盐水凝胶的新型蛋白质递送方法。我们的新假设是：1)适体可以有效地捕获海藻酸盐基质中的一个或多个蛋白质，因为它们具有高的结合亲和力和特异性；2)互补寡核苷酸可以作为分子触发器，通过与适体杂交来调节蛋白质释放；3)由于水凝胶制备的温和程序和适体的保护，蛋白质可以保持高水平的生物活性。为了验证这些假设，已经进行了初步研究，表明互补寡核苷酸能够加速蛋白质适体解离。在令人信服的初步结果的鼓舞下，我们将进行三项涉及实验和数学建模的任务。研究任务是：1)研究互补寡核苷酸介导的蛋白质适体解离；2)研究存在或不存在互补寡核苷酸时水凝胶中蛋白质的释放；3)建立数学模型来模拟蛋白质释放过程。我们预计这项研究的结果将是对分子识别的更深入理解和蛋白质传递的变革方法。更广泛的影响：拟议研究的成功将产生若干广泛的科学和经济影响。首先，它将为药物输送系统的发展开辟一条新的途径。二是丰富分子识别知识，为核酸研究提供有价值的信息。第三，蛋白质输送的成功将极大地改善各种人类疾病的治疗，并节省数十亿美元的医疗费用。这个项目的广泛影响也将体现在我们对教育和人力资源开发的坚定承诺上，这将对研究生、本科生和K-12学生产生直接影响。首先，这个项目将为学生提供一个独特的智力环境来学习药物传递、水凝胶合成、生物分子工程、动力学分析、细胞表征和数学建模。所有参与的学生都将有模块化的科学问题来学习，并将在定期小组会议上讨论他们的研究成果。学生不仅可以获得动手研究技能，还可以学习分析、沟通、协作和创新技能。此外，PI将把从拟议的研究中获得的结果纳入每年春季学期向研究生和大四本科生开设的药物输送课程。第二，我们将启动一项新的外展计划，与当地高中合作，在高级生物学课程中教授生物医学工程的概念。参与学生获得的结果将在访问期间呈现给高中生。此外，我们将继续参与康涅狄格大学已建立的外展项目。这些拓展工作将提高高中生对科学和工程的兴趣，并促进发展一支可行的、可持续的科学和工程劳动力队伍。第三，学生的研究成果将通过在同行评审的期刊上发表文章和在国内/国际会议上发表演讲的方式广泛传播。

0967512WangProtein drugs hold great promise for the treatment of various human diseases. However, efficient and safe delivery of protein drugs is a long-standing challenge in the field of drug delivery. Many protein delivery systems still suffer from problems including the rapid release of protein drugs, the inefficiency of controlling the release of multiple proteins, and the involvement of toxic molecules and/or harsh conditions during the preparation of protein delivery systems. This project is proposed to address these issues.Intellectual merits:The objectives of the proposed research are to thoroughly understand the mechanisms of complementary oligonucleotide-mediated protein-aptamer dissociation, and based on this understanding, to develop a novel protein delivery method using nucleic acid aptamers, complementary oligonucleotides, and alginate hydrogels. Our novel hypotheses are: 1) aptamers can efficiently entrap one or multiple proteins in the alginate matrix because of their high binding affinity and specificity; 2) complementary oligonucleotides can be used as a molecular trigger to modulate protein release via hybridization with the aptamers; and 3) proteins can maintain a high level of bioactivity due to the mild procedure for hydrogel preparation and the protection by the aptamers. To test these hypotheses, preliminary studies have been carried out, showing that complementary oligonucleotides are capable of accelerating protein-aptamer dissociation. Encouraged by the compelling preliminary results, we will perform three tasks involving both experiments and mathematical modeling. The tasks are: 1) to investigate complementary oligonucleotide-mediated protein-aptamer dissociation, 2) to investigate the release of proteins from hydrogels in the presence or absence of complementary oligonucleotides, and 3) to develop a mathematical model to simulate the protein release process. We anticipate the outcomes of this study will be a deeper understanding of molecular recognition and a transformative method for protein delivery.Broader impacts:The success of the proposed research will make several broad scientific and economical impacts. First, it will open a new avenue for the development of drug delivery systems. Second, it will enrich the knowledge of molecular recognition and provide valuable information for nucleic acid research. Third, the success of protein delivery will tremendously improve the treatments of various human diseases and save billions of dollars in healthcare costs. The broader impacts of this program will also be evident in our strong commitment to education and human resource development, which will have direct impacts on graduate, undergraduate, and K-12 students. First, this project will provide students with a unique intellectual environment to learn drug delivery, hydrogel synthesis, biomolecular engineering, kinetic analysis, cell characterization, and mathematical modeling. All participating students willhave modularized scientific questions to study and will discuss their research findings in regular group meetings. The students will be able to not only acquire hands-on research skills, but also learn analytical, communication, collaboration, and innovation skills. In addition, the PI will incorporate the results acquired from the proposed research into the Drug Delivery course thatis offered to both graduate and senior undergraduate students every spring semester. Second, we will initiate a new outreach program by collaborating with local high schools to teach the concepts of biomedical engineering in the Advanced Biology course. The results acquired by the participating students will be presented to high school students during the visits. In addition,we will continuously participate in the established outreach programs at UConn. These outreach efforts will raise high school students' interests in science and engineering and facilitate the development of a viable, sustainable science and engineering workforce. Third, the students' research findings will be widely disseminated through publications in peer-refereed journals and presentations at national/international conferences.