Kinetic Biomarker for CLL Prognosis

描述(由申请人提供) 第一阶段癌症中需要可靠的生物标记物是显而易见的，包括预后、分层、治疗监测和药物发现和批准。癌症的根本障碍是细胞动力学改变(细胞增殖和/或死亡)。因此，癌症的理想生物标记物将直接测量这些细胞动力学事件。慢性淋巴细胞性白血病(CLL)是应用动态生物标记物的一种很好的临床癌症模型，因为它的可及性(作为液体肿瘤)和相对成熟的动力学基础(作为一种涉及抑制正常细胞凋亡过程的恶性范例)。然而，直到最近，还没有一种测量细胞动力学的方法可用，这种方法在技术上是可靠的，适用于人类。加州大学M.Hellerstein博士的实验室最近开发了一种细胞增殖和死亡的直接动力学生物标记物，并获得了专利，并在人类研究中广泛使用。这项技术的基础是稳定的(非放射性)同位素掺入细胞DNA(例如，重水，2/H20)，然后进行质谱分析，并独家授权给KineMed公司(KineMed，Inc.，一家为医学诊断和药物开发中的动力学测量商业化而成立的公司)。这一第一阶段应用的目标是在CLL中应用这种获得专利的动态生物标记物(KineMarker TM)取得技术进步。在B-CLL中产生了令人鼓舞的试点数据。在这里，B-CLL的纯度、组织采样的下限、重水剂量的时间和持续时间、潜在的超灵敏质谱仪的评估以及测试的其他技术方面将被确立。其具体目的是：(1)确认用简化方法从B-CLL患者的血液中分离的CD19+/CD5+细胞的纯度；(2)评估B-CLL患者10ml抽血的充分性；(3)比较B-CLL患者较短2H20标记方案和较长程序的结果；(4)评估用于测量B-CLL患者细胞动力学的潜在的高灵敏度的气相色谱/裂解同位素比率/质谱仪；以及(5)在该项目的第二阶段，建立KineMarker TM作为B-CLL的预测性测试的大规模临床研究的组织结构。确定了进入第二阶段的可衡量的里程碑。综上所述，该项目将试图为将稳定的同位素/质谱学技术应用于人类血液病CLL中细胞增殖的测量奠定基础，并随后用作CLL疾病进展的预测测试。

DESCRIPTION (provided by applicant) PHASE I The need for reliable biomarkers in cancer is clear, for areas including prognosis, stratification, therapeutic monitoring and drug discovery and approval. The fundamental disorder in cancer is altered cell kinetics (cell proliferation and/or death). The ideal biomarker for cancer would therefore measure these cellular kinetic events directly. Chronic lymphocytic leukemia (CLL) is an excellent clinical cancer model for applying a kinetic biomarker, by virtue of its accessibility (being a liquid tumor) and its relatively well-established kinetic basis (as a paradigm of malignancy involving inhibition of the normal apoptotic process). Until recently, no method for measuring cell kinetics had been available, however, that was technically reliable and applicable in humans. The laboratory of Dr M. Hellerstein at the University of California recently developed, patented, and has extensively used in human research a direct kinetic biomarker of cell proliferation and death. The technique is based on stable (non-radioactive) isotope incorporation (e.g. heavy water, 2/H20) into cellular DNA followed by mass spectrometric analysis and is exclusively licensed to KineMed, Inc. (a company formed to commercialize kinetic measurements in medical diagnostics and drug development). The objective of this Phase I applications to make technical advances toward the application of this patented kinetic biomarker (KineMarker TM) in CLL. Encouraging pilot data has been generated in B-CLL. Here, purity, lower limits on tissue sampling, timing and duration of heavy water dosing, evaluation of a potentially ultrasensitive mass spectrometric instrument, and other technical aspects of the test will be established for B-CLL. The specific aims are, (1) to confirm the purity of CD19+/CD5+ cells isolated by a simplified protocol from the blood of B-CLL patients; (2) to assess the adequacy of a 10 ml blood draw in B-CLL patients; (3) to compare results from short 2H20 labeling protocols to longer protocols in B- CLL patients; (4) to evaluate the potentially ultrasensitive gas chromatography/pyrrolysis-isotope ratio/mass spectrometric instrument for measuring cell kinetics in B-CLL patients; and (5) to establish the organizational structure for a large-scale clinical study of the KineMarker Tm as a predictive test in B-CLL, in phase II of this project. Measurable milestones for entry into Phase II are defined. In summary, this project will attempt to lay the foundation for applying stable isotope/mass spectrometric techniques to the measurement of cell proliferation in the hematologic cancer CLL in humans; for subsequent use as a predictive test of CLL disease progression.