ACTIVATION SPECIFIC C ERBB 2 DETECTION IN BREAST CANCER

大约50%的乳腺癌患者有淋巴结转移。 淋巴结阴性肿瘤 尽管此类患者的总体预后是 比那些有积极节点的人要好得多，30%的人仍然会 死于转移性疾病 淋巴结阳性疾病的经验 这表明，这个“假阴性”的患者队列可能会产生显着的 辅助全身治疗可提高生存率。 识别高- 因此，风险淋巴结阴性患者是一个密集的临床领域， 兴趣 c-erbB-2（neu，HER-2）受体的表达已被提出作为一种 预测人类乳腺癌自然史的标准，但 这种受体在早期疾病中预测性是一个问题， 很多辩论。 我和我的同事最近的观察可能有助于建立 c-erbB-2的真正发病和预后意义 表型。 我们已经证明c-erbB-2受体可能存在于两个 相互转化形式-活化的175kDa酪氨酸磷酸化形式 （p175）和无活性的185kDa酪氨酸去磷酸化形式（p185）。 的 这两种形式的存在（这是无法区分的常规 c-erbB-2检测技术）提供了一个可能的解释， 上述临床辩论。 我建议澄清临床 c-erbB-2在乳腺癌中表达意义 并使用一种新的实验方法在体内定量受体活化， approach. 该项目有三个目标： 第一个目的是制备抗p175和p185的多克隆抗血清 使用含有引发活化和灭活的c-erbB-2肽， 网站分别; 第二个目的是产生针对完整的 p175和p185分子通过表达突变全长人c- 杆状病毒erbB-2cDNA; 第三个目的是确定p175和p175蛋白的相对表达。 p185在正常、浸润前和浸润性乳腺组织中的表达， 确定这些c-erbB-2的生物学和预后意义 受体亚型在人乳腺癌中的作用。 鉴于我目前的临床表现，这些目标特别可行。 与研究所内的外科和病理学服务联络， 超灵敏同位素荧光成像仪实验室可用性 研究，无限供应的高亲和力磷酸酪氨酸抗体， 受体活性控制，我和我的同事的经验， 在生长因子/酪氨酸激酶领域。 实现这些 目标将为更雄心勃勃的赠款申请铺平道路， 我建议评估抗p175和/或抗p185的临床价值， 在接受辅助全身治疗的患者的前瞻性研究中， 淋巴结阴性的乳腺癌

Approximately 50% of patients presenting with breast cancer have lymph node-negative tumors. Although the overall prognosis for such patients is substantially better than for those with positive nodes, 30% will still succumb to metastatic disease. Experience with node-positive disease suggests that this 'false-negative' patient cohort may derive significant survival benefit from adjuvant systemic therapy. Identification of high- risk node-negative patients is therefore an area of intense clinical interest. Expression of the c-erbB-2 (neu, HER-2) receptor has been proposed as one criterion for predicting the natural history of human breast cancer, but the predictivity of this receptor in early-stage disease is an issue of much debate. Recent observations by my colleagues and I may help establish the true pathogenetic and prognostic significance of the c-erbB-2 phenotype. We have shown that c-erbB-2 receptors may exist in two interconvertible forms - an activated 175kDa tyrosine-phosphorylated form (p175) and an inactive 185kDa tyrosine-dephosphorylated form (p185). The existence of these two forms (which are indistinguishable by conventional c-erbB-2 detection techniques) provides a possible explanation for the clinical polemic mentioned above. I propose to clarify the clinical significance of c-erbB-2 expression in human breast cancer by identifying and quantifying receptor activation in vivo using a novel experimental approach. The project has three objectives: The first objective is to prepare polyclonal antisera against p175 and p185 using c-erbB-2 peptides containing the prime activation and inactivation sites respectively; The second objective is to generate monoclonal antibodies to the intact p175 and p185 molecules obtained by expressing mutant full-length human c- erbB-2 cDNAs in baculovirus; The third objective is to determine the relative expression of p175 and p185 in normal, preinvasive and invasive breast tissue, and thus to establish the biologic and prognostic significance of these c-erbB-2 receptor isoforms in human breast cancer. These objectives are particularly feasible given my current clinical liaison with surgical and pathology services within the Institute, the laboratory availability of an ultrasensitive PhosphorImager for isotope studies, an unlimited supply of high-affinity phosphotyrosine antibody for receptor activity controls, and the experience of myself and my colleagues in the growth factor/tyrosine kinase field. Achievement of these objectives will prepare the way for a more ambitious grant application in which I propose assessing the clinical value of anti-p175 and/or anti-p185 in a prospective study of patients undergoing adjuvant systemic treatment for node-negative breast cancer.