Rapid Label-Free Detection of Acute Promyelocytic Leukemia

描述（由申请人提供）：我们建议开发一种微流体装置，能够根据急性早幼粒细胞白血病（APL）患者的细胞表面标记物检测白血病细胞。在美国，大约7%的新诊断APL患者在最初48小时内死亡，通常死于弥散性血管内凝血（DIC）。在拉丁美洲，APL是急性髓系白血病（AML）最常见的亚型，其死亡率明显更高：前5天死亡率为13%，总生存率约为50%。这种结果是不幸的，因为它是可以预防的：如果在患者第一次就诊时立即做出APL诊断，那么可以立即给予全反式维甲酸（ATRA）治疗。如果迅速给予APL患者ATRA可以挽救生命，因为DIC的发病率在治疗后数小时内就会下降。ATRA可引起APL细胞分化，并在初期自行诱导血液学缓解，缓解率约为90%。不幸的是，APL的诊断需要专家的血液病理学检查和仔细的免疫分型。当APL患者在深夜或周末到当地医生或急诊室就诊时，他们通常不能立即得到诊断，因为那时通常无法进行血液学咨询和临床免疫表型分型。由于血液病专业知识不足或缺乏所需设备或训练有素的技术人员，许多医院甚至没有能力做出急性白血病的诊断。因此，许多患者没有立即接受挽救生命的ATRA治疗。我们建议通过一种能够诊断APL的即时护理设备来解决这一未满足的患者需求，从而允许快速启动适当的治疗。我们的设备以芯片上的人工孔为中心，这些孔具有特异性抗体的功能，可以筛选细胞的大小，形状和特定的细胞表面标记。简单的设备允许真正的点护理，因为它需要很少的样品；几乎消除了所有的样品制备；具有低功耗要求；并在<5分钟内提供直接、准确的结果。很容易适应手持格式，结果可以显示在屏幕上或无线传输到医生办公室（如果在患者家中进行）或医院（如果在医生办公室或当地急诊室进行）。我们的设备将进行全血细胞计数并检测对原发性APL细胞免疫表型重要的单个抗原。考虑到我们目前的技术发展和里程碑的仔细放置，提议的两年设备开发时间表是可行的。最终，我们的设备可以适应许多其他临床情况，用于快速临床监测。PI Lydia L. Sohn助理机械工程教授加州大学伯克利分校的教授将领导这个NIH R21项目，并制造/测试该设备。Lucy A. Godley，医学博士和助理。芝加哥大学教授将提供样本选择、实验设计和临床相关性之间的接口。高级人员，加州大学伯克利分校的乔治·安瓦尔博士将领导该设备的软硬件设计和包装。

DESCRIPTION (provided by applicant): We propose to develop a microfluidics device capable of detecting leukemic cells based on their cell- surface markers from patients with acute promyelocytic leukemia (APL). Approximately 7% of newly- diagnosed APL patients in the United States die within the first 48 hours, often of disseminated intravascular coagulation (DIC). In Latin America, where APL is the most frequent subtype of acute myeloid leukemia (AML), the mortality rate is significantly higher: 13% mortality within the first five days and an overall survival of about 50% [1]. This outcome is unfortunate, because it is preventable: if an APL diagnosis were made immediately when a patient first presents to a physician, then all-trans retinoic acid (ATRA) could be administered immediately. ATRA is life saving if given quickly to APL patients, because the incidence of DIC falls within hours of therapy. ATRA causes differentiation of APL cells and induces hematologic remissions initially on its own at a rate of >90%. Unfortunately, APL diagnosis requires expert hematopathologic review and careful immunophenotyping. When APL patients present late at night or on the weekends to their local doctor or emergency room (ER), they are often not diagnosed immediately, since hematologic consultation and clinical immunophenotyping are often unavailable then. Many hospitals do not even have the capability to make an acute leukemia diagnosis, due to insufficient hematopathologic expertise or lack of required equipment or trained technologists. Thus, many patients do not receive life-saving ATRA therapy right away. We propose to address this unfilled patient need with a point-of-care device capable of diagnosing APL, thereby allowing rapid initiation of appropriate therapy. Our device centers on-chip artificial pores that are functionalized with specific antibodies to screen, label- free, cells for size, shape, and specific cell-surface markers. The simple device allows true point-of-care, as it requires very little sample; virtually eliminates all sample preparation; has low-power requirements; and provides direct, accurate result in <5 minutes. Easily adaptable to a hand-held format, the results can display on a screen or be transmitted wirelessly to a physician's office (if performed at the patient's home) or hospital (if performed in a physician's office or local ER). Our device will perform complete blood counts and detect individual antigens important to immunophenotyping primary APL cells. The proposed two-year time line for device development is feasible given our current technology development and careful placement of milestones. Ultimately, our device could be adapted to many other clinical situations for rapid clinical monitoring. PI Lydia L. Sohn, Assoc. Prof. of Mechanical Eng. at the University of California, Berkeley will lead this NIH R21 project and fabricate/test the device. Co-PI Lucy A. Godley, M.D., Ph.D., and Assist. Prof. at The University of Chicago will provide interfacing among sample choice, experimental design, and clinical relevance. Senior Personnel, George Anwar, Ph.D. at UC Berkeley will lead in the software and hardware design and packaging of the device for point-of-care.