Fluorescent Probes for Tracking Insulin Secretion in Therapeutic and Diagnostic Devices

 描述（由申请人提供）：意义：1型和2型糖尿病（T1 D和T2 D）正在成为社会经济资源的日益沉重的负担，全球患者数量激增。胰岛素依赖性治疗方案（如定期胰岛素注射）已得到充分确立，但对患者的生活质量产生负面影响，并伴有各种长期并发症。40多年来，寻找胰岛素非依赖性治疗方案一直是一个重要的，尽管难以捉摸的科学目标，特别是在胰岛移植领域取得了可喜的成果。在这里，移植整个尸体胰腺或由保护性胶囊中的胰岛素产生β细胞组成的包囊胰岛一直是研究的主要焦点。然而，可用的尸体胰腺组织很难获得，移植成功主要受到β细胞的细胞存活的限制。另一个需要解决的问题是如何根据胰岛素释放活性选择功能性胰岛，以实现更一致和更可预测的移植结果，并帮助患者在更长的时间内保持胰岛素独立性。迄今为止，还不存在可用于该监测过程以量化单个胰岛的胰岛素分泌的具有高功效的简单方法。此外，这种技术还可以适于获得诊断信息，以在确定糖尿病进展和病因时有效地监测胰岛素分泌。假设：我们假设使用我们的荧光成像剂，即ZIMIR，有能力解决这些治疗和诊断问题时，使用的技术，如荧光激活细胞分选（FACS）或ZIMIR功能化的光纤。初步数据：我们已经证明，ZIMIR代表了锌敏感荧光剂的先锋，具有标记β细胞和成像胰岛素分泌的能力，这两个基本特征用于筛选功能性胰岛、包封胰岛或β细胞。此外，我们表明，ZIMIR足够灵敏，可以定量Zn 2+浓度的增加，这可以通过门静脉处的荧光检测到，并与胰岛素分泌直接相关，这是设计可以定量体内胰岛素分泌的装置的关键先决条件。具体目标：第一阶段的主要目标是合成、纯化和表征大量的ZIMIR（目标1）和ZIMIR-马来酰亚胺（目标2）。后者将在第二阶段用于由二氧化硅组成的改性光学纤维的功能化。这些光纤将被用来构建光纤测光 仪器用于检测Zn 2+，在体外和体内验证。ZIMIR将用于I期临床前试验，以在随后的移植实验中使用FACS进行胰岛分选。总体影响：总之，VitalQuan开发的这些技术将证明ZIMIR在胰岛细胞分选应用中的实用性，以提高胰岛移植成功率，并将建立前所未有的体内胰岛素跟踪研究。这些应用将加速ZIMIR在糖尿病研究和治疗临床应用中的商业化。

 DESCRIPTION (provided by applicant): Significance: Type 1 and Type 2 Diabetes Mellitus (T1D and T2D) are becoming an increasing burden on socioeconomic resources with surging patient numbers all over the world. Insulin-dependent treatment regimen such as regular insulin injections are well established but have been associated with a negative impact on the patients' quality of life with a variety of long-term complications. The search for insulin-independent treatment options has been an important, albeit elusive, scientific goal for over 40 years with promising results particularly in the field of pancreatic islet transplantation. Here, grafting of either entire cadaveric pancreases or encapsulated islets comprised of insulin producing beta cells in a protective capsule has been the main focus of research. However, usable cadaveric pancreatic tissue is hard to acquire with graft success mostly limited by the cellular survival of the beta cells. Another issue that needs to be addressed is how to select functional islets based on their insulin release activity, in order to achieve a more consistent and more predictable transplant outcome, and to help patients maintain insulin independence over a longer period of time. No simple method with high efficacy exists to date that could be useful in this monitoring process to quantify insulin secretion of individual islets. Furthermore, such a technique can also be adapted to gain diagnostic information in an effort to monitor insulin secretion efficiently when determining diabetes progression and etiology. Hypothesis: We hypothesize that the use of our fluorescent imaging agent, namely ZIMIR, has the ability to solve these therapeutic and diagnostic problems when used in technologies such as fluorescence-activated cell sorting (FACS) or in ZIMIR-functionalized optical fibers. Preliminary Data: We have demonstrated that ZIMIR represents the vanguard of a zinc sensitive fluorescent agent with the ability to label beta cells and image insulin secretion, both essential features for the screening of functional islets, encapsulated islets, or beta cells. Furthermore, we showed that ZIMIR is sensitive enough to quantify a concentration increase of Zn2+ that can be detected by fluorescence at the portal vein and directly correlated to insulin secretion, a crucial prerequisite for the design of a devic that can quantify insulin secretion in vivo. Specific Aims: The key objectives in Phase I entail th synthesis, purification and characterization of a large amount of ZIMIR (Aim 1) and ZIMIR-maleimide (Aim 2). The latter will be used in Phase II for the functionalization of modified optica fibers comprised of silicon dioxide. These fibers will then be used to construct a fiber photometry instrument for the detection of Zn2+, validated in vitro and in vivo. ZIMIR will be used in Phase I for preclinical testing of the agent to find use in islet sorting using FACS with subsequent transplantation experiments. Overall Impact: Together, these technologies developed by VitalQuan will demonstrate the utility of ZIMIR in iselt cell sorting applications to increase isle graft success rates and will also establish unprecedented studies designed for in vivo insulin tracking. Taken in concert, these applications will accelerate the commercialization of ZIMIR for clinical use in diabetes research and treatment.