Rapid Label-Free Detection of Acute Promyelocytic Leukemia

急性早幼粒细胞白血病的快速无标记检测

• 批准号: 8032345
• 负责人: Lydia L Sohn
• 金额: $ 16.21万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-18 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8032345
• 关键词: Accident and Emergency department; Acute Myelocytic Leukemia; Acute Promyelocytic Leukemia; Acute leukemia; Address; Antibodies; Antigens; Arsenic Trioxide; Back; Blast Cell; Blood; California; Cell Line; Cell Size; Cell surface; Cells; Chicago; Clinical; Coagulation Process; Complete Blood Count; Computer software; Consultations; Detection; Detection of Minimal Residual Disease; Developing Countries; Device or Instrument Development; Devices; Diagnosis; Disease; Disease remission; Disseminated Intravascular Coagulation; Doctor of Medicine; Doctor of Philosophy; Emergency Situation; Equipment; Experimental Designs; Goals; Hand; Health Personnel; Home environment; Hospitals; Hour; Human; Human Cell Line; Human Resources; Immunophenotyping; Incidence; Individual; Institution; International; Label; Latin America; Lead; Leukemic Cell; Leukocytes; Life; Malignant - descriptor; Measures; Mechanics; Medical; Methods; Microfluidic Microchips; Microfluidics; Monitor; Newly Diagnosed; Organ; Outcome; Patients; Pharmaceutical Preparations; Physicians; Physicians' Offices; Preparation; Progranulocytes; Resistance; Sampling; Screening procedure; Shapes; Sorting - Cell Movement; Surface; System; Testing; Time; Training; Tretinoin; United States; United States National Institutes of Health; Universities; Whole Blood; base; clinical Diagnosis; clinically relevant; data acquisition; design; falls; innovation; leukemia; mortality; point of care; pre-clinical; technology development

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. PUBLIC HEALTH RELEVANCE: We propose to develop a rapid label-free method of screening cells to detect leukemic cells based on their cell-surface markers from patients with acute promyelocytic leukemia (APL), a disease that provides a pressing need for urgent diagnosis. Such a method is needed clinically, because about 7% of newly- diagnosed APL patients in the United States (13% in Latin America) die within the first 48 hours, often of disseminated intravascular coagulation (DIC). If APL were diagnosed quickly, then all-trans retinoic acid (ATRA), a drug which causes differentiation of APL cells and induces hematologic remissions initially on its own at a rate of >90%, could be administered. Our method would enable quick and accurate diagnosis of APL and could ultimately be could be applied to virtually any leukemia and easily adaptable for pre-clinical diagnosis or the detection of minimal residual disease.

描述(由申请人提供)：我们建议开发一种微流控设备，能够根据急性早幼粒细胞白血病(APL)患者的细胞表面标记检测白血病细胞。在美国，大约7%的新诊断的APL患者在最初的48小时内死亡，通常死于弥漫性血管内凝血(DIC)。在拉丁美洲，APL是急性髓系白血病(AML)最常见的亚型，死亡率要高得多：前五天内死亡率为13%，总存活率约为50%[1]。这一结果是不幸的，因为它是可以预防的：如果在患者第一次向医生提出诊断时立即做出APL诊断，那么可以立即使用全反式维甲酸(ATRA)。如果对APL患者迅速给予ATRA是拯救生命的，因为DIC的发生率在治疗后几个小时内下降。全反式维甲酸诱导APL细胞分化，最初以90%的比率单独诱导血液学缓解。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表型分析。当APL患者在深夜或周末到当地医生或急诊室(ER)就诊时，他们通常不能立即得到诊断，因为那时通常无法获得血液学咨询和临床免疫表型鉴定。许多医院甚至没有能力做出急性白血病的诊断，因为血液病理学专业知识不足，或者缺乏必要的设备或训练有素的技术人员。因此，许多患者并没有立即接受挽救生命的全反式维甲酸治疗。我们建议用一种能够诊断APL的护理点式设备来满足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人造毛孔为中心，使用特定的抗体进行功能化，以筛选大小、形状和特定的细胞表面标记的无标记细胞。这种简单的设备允许真正的护理，因为它只需要很少的样本；几乎不需要所有的样本准备；具有低功耗要求；并在&lt；5分钟内提供直接、准确的结果。结果可以很容易地适应手持格式，可以在屏幕上显示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)。我们的设备将进行完整的血细胞计数，并检测对原代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对里程碑的仔细安排，拟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，以进行快速临床监测。皮莉迪亚·L·孙(Pi Lydia L.Sohn，Assoc.机械工程系教授。加州大学伯克利分校将领导这一NIH R21项目，并制造/测试该设备。共同派露西·A·戈德利，医学博士，博士，协助。芝加哥大学的教授将在样本选择、实验设计和临床相关性之间提供接口。加州大学伯克利分校的高级人员乔治·安瓦尔博士将领导该设备的软件和硬件设计以及包装。 公共卫生相关性：我们建议开发一种快速无标记的细胞筛选方法，根据急性早幼粒细胞白血病(APL)患者的细胞表面标记检测白血病细胞，APL是一种迫切需要紧急诊断的疾病。这种方法在临床上是必要的，因为在美国，大约7%的新诊断的APL患者(拉丁美洲的13%)在最初的48小时内死亡，通常死于弥漫性血管内凝血(DIC)。如果APL被迅速诊断出来，那么可以使用全反式维甲酸(ATRA)，这是一种导致APL细胞分化并最初以90%的比率单独诱导血液系统缓解的药物。我们的方法将能够快速准确地诊断APL，最终可以应用于几乎任何白血病，并很容易适用于临床前诊断或微小残留病的检测。