Mechanisms of Bladder Cancer Progression

描述（由申请人提供）：鉴定调节膀胱癌（BCa）进展的“分子决定因素”可以改善BCa患者的治疗和复发监测。 透明质酸（HA）是一种促进肿瘤转移的糖胺聚糖。 透明质酸酶（HAase）是将HA降解成血管生成片段的酶。 HYAL 1是在膀胱肿瘤（BT）细胞中表达的主要HAase。 它在体外和BT异种移植物中调节BCa生长和侵袭。 虽然HYAL 1野生型（wt）具有酶活性，仅在高级别BCa中表达，但5种HYAL 1变体无酶活性，在正常和低级别BCa组织中表达。 在BT组织中，肿瘤细胞和间质都产生HA，然而，HAS 1型HA合酶仅在BT细胞中表达。 尿HA和HAase水平的测定（HA-HAase试验）在检测BCa方面具有较高的准确性。 该提案旨在研究HYAL 1、HYAL 1剪接变体和HAS 1在BCa进展中的治疗和预后潜力。 此外，在一项多中心试验中，将评估HA-HAase检测（单独或与其他尿液检测联合）是否可用于监测BCa复发。 为了确定HYAL 1在BCa生长和进展中的功能，将在使用病毒系统递送HYAL 1反义cDNA或通过用HAase抑制剂治疗后，在BT异种移植物中测试抗HAase治疗的功效。 HYAL 1的作用机制将通过分析细胞周期调节因子、基质降解酶和血管生成因子的改变来研究（Aim 1）。 通过用HYAL 1剪接变体cDNA转染表达HYAL 1 wt的BT细胞来检查HYAL 1变体对BCa生长和侵袭的可能的中和作用。 BT组织中HYAL 1变体的差异表达将与BT预后相关（目的2）。 将通过使用HAS 1正义和HAS 1反义构建体转染表达HYAL 1或在HYAL 1产生中被阻断的BCa细胞来评价HAS 1在BT生长和进展中的功能。 在BT组织中HAS 1及其变体（HAS 1v）的表达将与BCa预后相关（Aim 3）。 在一项多中心试验中，将检查HA-HAase检测、尿细胞学检查、BTA-Stat、NMP 22检测（单独和联合）在100至150例BCa患者中监测肿瘤复发的精确度。 将结果与临床结果进行比较（目标4）。 这项研究将揭示HA、HYAL 1和相关分子在BT进展中的功能、治疗和/或预后潜力。 此外，它将确定HA-HAase测试或其与其他测试的组合是否可以精确地监测BCa复发。

DESCRIPTION (provided by applicant): Identification of "molecular determinants" that regulate bladder cancer (BCa) progression could improve treatment and recurrence monitoring for BCa patients. Hyaluronic acid (HA) is a glycosaminoglycan that promotes tumor metastasis. Hyaluronidase (HAase) is an enzyme that degrades HA into angiogenic fragments. HYAL1 is the major HAase expressed in bladder tumor (BT) cells. It regulates BCa growth and invasion both in vitro and in BT xenografts. While HYAL1 wild type (wt) is enzymatically active and is exclusively expressed in high-grade BCa, 5 HYAL1 variants are enzymatically inactive and expressed in normal and low-grade BCa tissues. In BT tissues, both tumor cells and the stroma produce HA, however, HAS1 type HA-synthase is exclusively expressed in BT cells. The measurement of urinary HA and HAase levels (HA-HAase test) has high accuracy in detecting BCa. This proposal is designed to investigate the therapeutic and prognostic potentials of HYAL1, HYAL1 splice variants, and HAS1 in BCa progression. Furthermore, in a multi-center trial whether the HA-HAase test, either alone or in combination with other urine tests, can be used for monitoring BCa recurrence will be evaluated. To define HYAL1 function(s) in BCa growth and progression, the efficacy of anti-HAase therapy will be tested in BT xenografts, following delivery of HYAL1-antisense cDNA using a viral system or by treatment with a HAase inhibitor. The mechanism of HYAL1 action will be examined by analyzing alterations in cell cycle regulators, matrix degrading enzymes, and angiogenic factors (Aim 1). A possible neutralizing effect of HYAL1 variants on BCa growth and invasion will be examined by cDNA transfection of HYAL1wt expressing BT cells with HYAL1 splice variant cDNAs. Differential expression of HYAL1 variants in BT tissues will be correlated with BT prognosis (Aim 2). HAS1 function in BT growth and progression will be evaluated by transfecting BCa cells which either express or are blocked in HYAL1 production, using HAS1-sense and HAS1-antisense constructs. Expression of HAS1 and its variant (HAS1v) in BT tissues will be correlated with BCa prognosis (Aim 3). In a multi-center trial, the utility of the HA-HAase test, urine cytology, BTA-Stat, NMP22 tests, individually and in combination, will be examined for precision to monitor tumor recurrence in 100 to 150 BCa patients. The results will be compared to clinical findings (Aim 4). The proposed study will reveal the function, therapeutic and/or prognostic potentials of the HA, HYAL1 and related molecules in BT progression. Furthermore, it will establish whether the HA-HAase test or its combination with other tests can precisely monitor BCa recurrence.