MARKERS OF BLADDER CANCER AND THEIR MODULATION BY DFMO

作为最近资助的合同（N01 CN-25434 - 01）的配套合同， 表浅性移行细胞癌（TCC）的化学预防 膀胱，我们建议调查TCC的中间分子标志物 以及它们通过二氟甲基鸟氨酸（DFMO）的调节， 多胺（PA）合成酶的抑制剂，鸟氨酸 脱羧酶（ODC）。 在N01 CN-25434中，40例TCC和20例非TCC患者 将确定2剂DFMO（或安慰剂）的生物学效应 在相关靶（恶性和正常尿路上皮细胞和组织， 和尿）以确定在治疗中使用的生物活性DFMO剂量。 安慰剂对照、III期、预防性试验，在240例 彻底切除浅表TCC。 目前的调查将 利用这些研究中的主题，重点关注相互作用的分子 已知在TCC和/或ODC阻断中发生改变的途径/PA 耗尽 这些包括：1）Pa代谢; 2） 尿表皮生长因子（EGF）及其受体（EGF-R） 已知其在TCC细胞中诱导ODC活性;和3）蛋白激酶 蛋白激酶C（PKC）是EGF信号传导和ODC的组成部分 activation. 尿路上皮组织和尿液中的乙酰化和未结合PA水平 （通过HPLC测定）将与尿路上皮ODC活性相关 （放射性生物测定）表达（免疫组织学） TCC和那些正在接受和没有接受DFMO的人。 尿[EGF]（RIA） 和尿路上皮EGF-R表达（免疫组织学/免疫细胞学）在这些 患者将与影响因素（例如尿 pH值），或由EGF/EGF-R（例如增殖[Ki67染色]）引起 相互作用，并与PA/ODC参数。 这些也将相互关联 主要PKC亚型的组织分布（免疫组织学）， 已知在正常细胞和恶性细胞中表达不同 在非尿路上皮组织中。 通过比较TCC患者和非TCC患者的这些参数， 特别是通过以纵向方式重复测量， 疾病的结果正在监测，疾病的影响及其 复发，以及ODC阻断将被更好地定义。 这些 研究应提供重要的临床信息， 恶性尿路上皮转化标志物、PA代谢和DFMO 生物学效应以及对TCC生物学的了解 发育、生长和复发。

As a companion to a recently funded contract (N01 CN-25434-01) for the chemoprevention of superficial transitional cell cancer (TCC) of the bladder, we propose to investigate intermediate molecular markers of TCC and their modulation by difluoro-methylornithine (DFMO), an irreversible inhibitor of the polyamine (PA) synthesizing enzyme, ornithine decarboxylase (ODC). In N01 CN-25434, 40 TCC and 20 non-TCC patients will have the biologic effects of 2 doses of DFMO (or placebo) determined in relevant targets (malignant and normal urothelial cells and tissues, and urine) to determine a biologically active DFMO dose to use in a placebo-controlled, Phase III, preventative trial in 240 patients with completely resected superficial TCC. The present investigation will utilize subjects in these studies to focus on interacting molecular pathways which are known to be altered in TCC and/or with ODC blockade/PA depletion. These include: 1) Pa metabolism; 2) the interaction of urinary epidermal growth factor (EGF) and its urothelial receptor (EGF-R) which is known to induce ODC activity in TCC cells; and 3) protein kinase C (PKC) which is an integral component of EGF signaling and ODC activation. Acetylated and unconjugated PA levels in urothelial tissues and urine (determined by HPLC) will be correlated with urothelial ODC activity (radiobioassay) expression (immunohistology) in patients with and without TCC and in those who are and are not receiving DFMO. Urinary [EGF] (RIA) and urothelial EGF-R expression (immunohistology/immunocytology) in these patients will be correlated with factors that influence (e.g. urinary pH), or result from (e.g. proliferation [Ki67 staining]) the EGF/EGF-R interaction, and with PA/ODC parameters. These will also be correlated with tissue profiles of predominant PKC isoforms (immunohistology) which are known to be differentially expressed in normal versus malignant cells in non-urothelial tissues. By comparing these parameters in patients with and without TCC, and particularly by repeating measurements in a longitudinal fashion in which disease outcome is being monitored, the effects of disease and its recurrence, as well as of ODC blockade will be better defined. These studies should provide important clinical information about molecular markers of malignant urothelial transformation, PA metabolism, and DFMO's biological effects as well as insights into the biology of TCC development, growth, and recurrence.