Kinetic Biomarker for CLL Prognosis

描述（由申请人提供） 第一阶段癌症中对可靠生物标志物的需求是明确的，包括预后，分层，治疗监测和药物发现和批准。癌症中的基本病症是改变的细胞动力学（细胞增殖和/或死亡）。因此，癌症的理想生物标志物将直接测量这些细胞动力学事件。慢性淋巴细胞白血病（CLL）是用于应用动力学生物标志物的极好的临床癌症模型，这是由于其可及性（作为液体肿瘤）和其相对良好建立的动力学基础（作为涉及抑制正常凋亡过程的恶性肿瘤的范例）。然而，直到最近，还没有技术上可靠且适用于人类的测量细胞动力学的方法。M博士的实验室。加州大学的赫勒斯坦最近开发了一种细胞增殖和死亡的直接动力学生物标志物，并申请了专利，并广泛用于人类研究。该技术基于稳定（非放射性）同位素掺入（例如重水，2/H2O）到细胞DNA中，然后进行质谱分析，并独家授权给KineMed，Inc.。（一家旨在将医学诊断和药物开发中的动力学测量商业化的公司）。第一阶段申请的目的是使这项专利动力学生物标志物（KineMarker TM）在CLL中的应用取得技术进步。在B-CLL中已经产生了令人鼓舞的试验数据。在这里，纯度，组织取样的下限，重水给药的时间和持续时间，潜在的超灵敏质谱仪器的评估，以及测试的其他技术方面将为B-CLL建立。具体的目的是，（1）确认通过简化方案从B-CLL患者的血液中分离的CD 19 +/CD 5+细胞的纯度;（2）评估在B-CLL患者中抽取10 ml血液的适当性;（3）比较在B- CLL患者中短2 H20标记方案与长方案的结果;（4）评价用于测定B-CLL患者细胞动力学的潜在超灵敏气相色谱/吡咯裂解-同位素比率/质谱仪器;以及（5）在该项目的第二阶段中，建立KineMarker Tm作为B-CLL预测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。总之，该项目将尝试为应用稳定同位素/质谱技术测量人类血液癌症CLL中的细胞增殖奠定基础;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.