INSPIRE: Neutrophil Delivery of Apoptosis-Inducing Anticancer Drugs

曼哈顿堪萨斯州立大学的INSPIRE奖部分由数学和物理科学理事会材料研究部的生物材料项目资助。另外两个跨学科项目是化学、生物工程、环境和运输系统部门的生物光子项目，以及土木、机械和制造创新部门的表面工程材料项目，这两个项目都来自工程理事会。有了这个奖项，靶向蛋白酶激活聚合物笼脂质体（PPCLs）将被设计和合成，用于从外周血中被中性粒细胞快速摄取。所制备的脂质体将装载金属结合抗癌药物(4,4-二甲基-1-（二（吡啶-2-基）亚甲基）（Dp44mT），该药物可使脂质体不稳定。这种金属螯合剂将与线粒体失活肽D[KLAKLAK]2偶联，然后将偶联物装入脂质体中。这种装载药物的新型脂质体有望在通过血液运输到脑肿瘤细胞（多形性胶质母细胞瘤）时在中性粒细胞中保持完整。一旦中性粒细胞到达目标细胞并发生凋亡，PPCLs将被半胱天冬酶激活并释放其有效载荷。预计两种药物在脂质体中的协同作用将在原发性肿瘤和转移性肿瘤中启动凋亡级联反应。值得注意的是，中性粒细胞能够穿过血脑屏障，并且能够杀死癌细胞，即使这些脑癌细胞很小，处于发展的早期阶段。所提出的基于细胞的癌症治疗策略可能优于使用纳米颗粒和其他药物载体的治疗策略，因为所提出的基于中性粒细胞的治疗方法可以治疗早期肿瘤，而且它们具有选择性、有效性和快速性。此外，自体细胞的使用有可能显著降低基于细胞的人类癌症治疗中常见的调节障碍。此外，该策略可以扩展到其他应用，使用不同的寡肽作为靶向序列靶向多种细胞类型。单一细胞类型的靶向原则将有可能演变成针对病毒、细菌和原生动物感染的靶向治疗，以及用于不同应用的生物光子方法。此外，INSPIRE项目的成功成果可能会在材料科学、表面加工、合成生物学、分子工程、纳米技术和生物光子学的界面上引领一种新的跨学科研究方法。作为教学、培训和推广活动的一部分，PI计划在堪萨斯州立大学开发和教授一门名为“治疗学”的课程。此外，PI和Co-PI将轮流主持一个关于提高写作技巧的研讨会，该研讨会将提供给所有本科生，研究生和博士后科学家。目前，项目负责人和联合负责人有5名女研究生、4名发展学者项目学生、1名本科生暑期研究机会项目学生和2名本科生在实验室进行研究体验。这笔奖金将扩大和加强这些招聘、指导和培训活动。这项研究旨在开发治疗脑肿瘤（胶质瘤或多形性胶质母细胞瘤）的新材料。将开发一种新的治疗概念，即利用患者自身的白细胞作为抗癌药物的“运输船”。一旦白细胞到达肿瘤，这些药物就会被释放。利用癌症患者的全血，该项目将针对白细胞装载抗癌药物，然后重新注入他们的血液。然后，白细胞会移动到肿瘤的部位。传统的脂质体不会被血液中各种类型的细胞选择性地吸收。它们要么在摄取后破裂，要么与细胞融合。无论哪种情况，本应留在脂质体内的药物都会过早释放，从而杀死运输细胞而不是肿瘤。然而，提出的新型蛋白酶激活聚合物笼脂质体（PPCLs）在白细胞内运输过程中预计是稳定的，并且一旦这些白细胞整合到肿瘤细胞中就会被激活。这些药物会导致程序性细胞死亡（细胞凋亡）。它们将能够杀死快速生长的肿瘤细胞和缓慢生长的癌症干细胞。后者是肿瘤复发和转移形成的原因。使用患者来源的细胞将为真正的个性化医疗提供一条途径，无需担心干细胞可能导致癌症，也无需担心伦理问题。在此项目期间，将开展三项教育重点：(i) PI将开发和教授一门名为“治疗学”的课程。本课程面向兽医学、化学、生物化学、生物学、物理学等专业的研究生和本科生开放；（ii）首席研究员和副首席研究员轮流主持撰写研究论文的工作坊，并向首席研究员研究团队的所有本科生、研究生和博士后科学家开放；（三）该研究还将为即将到来的本科生研究经验、暑期本科生研究机会计划和发展学者创造机会。堪萨斯州立大学的项目学生。PI和Co-PI将继续与来自代表性不足群体的研究生和本科生以及退伍军人合作。此外，这项研究将有助于扩大堪萨斯州立大学在材料化学、癌症治疗和诊断方面正在进行的合作努力。

This INSPIRE award to Kansas State University at Manhattan is partially funded by the Biomaterial program in the Division of Materials Research in the Directorate for Mathematical and Physical Sciences. The other two interdisciplinary programs that are partially funding this award are the Biophotonic program in the Division of Chemical, Bioengineering, Environmental, & Transport Systems, and the Materials for Surface Engineering program in the Division of Civil, Mechanical and Manufacturing Innovation, both from the Directorate for Engineering. With this award, targeted protease-activatable polymer-caged liposomes (PPCLs) will be designed and synthesized for rapid uptake by neutrophils from peripheral blood. The liposomes prepared will be loaded with a metal binding anticancer drug (4,4-dimethyl-1-(di(pyridine-2-yl)methylene) (Dp44mT), which destabilizes liposomes. This metal chelating agent will be conjugated with a mitochondria inactivating peptide D[KLAKLAK]2 before the conjugate is loaded in the liposomes. This new class of liposomes with loaded drugs are expected to remain intact in the neutrophil while being transported by blood to the brain cancer cells (glioblastoma multiforme). PPCLs will be activated by caspases and discharge their payload once the neutrophils have reached their target and undergo apoptosis. Synergy between the two drugs loaded in the liposomes is anticipated and that will start apoptotic cascades in primary tumors and metastases. It is important to note that neutrophils are able to cross the blood brain barrier, and will be able to kill cancer cells even if these brain cancer cells are small and in the early stages of development. The proposed cell-based cancer treatment strategies are potentially superior to the ones that use nanoparticles and other drug carriers, because the proposed neutrophil-based therapies can treat early tumors, and they are selective, efficient, and fast. Additionally, the use of autologous cells has the potential of significantly lowering the regulatory barriers generally seen with cell-based human cancer therapies. In addition, this strategy could be expanded to other applications for targeting numerous cell types using different oligopeptides as targeting sequences. The targeting principles of a single cell-type would have the potential to evolve into targeted therapies for viral, bacterial and protozoal infections, and biophotonic approaches for different applications. In addition, a successful outcome from this INSPIRE project could lead to a new interdisciplinary research approach at the interface of materials science, surface processing, synthetic biology, molecular engineering, nanotechnology and biophotonics. As part of teaching, training and outreach activities, the PI plans to develop and teach a course entitled 'Theranostics' at Kansas State University. In addition, the PI and Co-PI will alternately lead a workshop on developing skills in writing manuscripts, and this workshop will be offered to all undergraduates, graduate students and postdoctoral scientists in their groups. Currently, the PI and Co-PI have five female graduate students, four Developing Scholars' Program students, one Summer Undergraduate Research Opportunity Program student and two Research Experience of Undergraduates students in their laboratories. These recruiting, mentoring and training activities will be enlarged and enhanced with this award.This research is directed at developing new materials for the treatment for brain tumors (glioma or glioblastoma multiforme). A new treatment concept will be developed, which utilizes the patients' own white blood cells as "transport ships" for anticancer drugs. These drugs will be released once the white blood cells have reached the tumor. Using whole blood from cancer patients, this project will target the white blood cells to load them with anticancer drugs, and then re-inject their blood. The white blood cells will then travel to the sites of tumors. Classic liposomes are not taken up selectively by the various cell types in blood. They either burst after uptake or the fuse with the cells. In either case, the drugs that are supposed to stay within the liposomes are released prematurely, thus killing the transport cells and not the tumors. However, the proposed novel protease-activatable polymer-caged liposomes (PPCLs) are expected to be stable during transport within white blood cells, and become activated once these white cells become integrated into the tumor cells. The drugs will then cause programmed cell death (apoptosis). They will be able to kill the fast growing tumor cells and the slower growing cancer stem cells. The latter are responsible for the reappearance of tumors and the formation of metastases. The use of patient derived cells will provide a pathway to truly personalized medicine, free of concerns about the potential of stem cells causing cancer, and also free of ethical concerns. Three education thrusts will be carried out during the period of this project: (i) the PI will develop and teach a block course entitled 'Theranostics'. This course will be open to all graduate and undergraduate students in veterinary medicine, chemistry, biochemistry, biology and physics; (ii) the PI and Co-PI will alternately lead a workshop on writing research papers, and will be offered to all undergraduates, graduate students and postdoctoral scientists in the PIs' research team; and (iii) the research will also create opportunities for incoming Research Experience of Undergraduates, Summer Undergraduate Research Opportunity Program and Developing Scholars? Program students at Kansas State University. Both PI and Co-PI will continue to work with graduate and undergraduate students from underrepresented groups, as well as veterans. In addition, this research will help extend the efforts to ongoing collaborative efforts in materials chemistry, cancer therapy and diagnostics at Kansas State University.