Regulation of breast cancer growth by activation peptide

激活肽调节乳腺癌生长

基本信息

批准号：
6942739
负责人：
VACLAV VETVICKA
金额：
$ 20.59万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-09-01 至 2007-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The long-term objective is to develop a new treatment for breast cancer based on blockade of the autocrine growth factor activity of procathepsin D. Breast cancer cells secrete procathepsin D, the zymogen from which the aspartic proteinase cathepsin D is generated by removal of an activation peptide (APpCD). Procathepsin D has been identified as an independent prognostic factor in breast cancer. In preliminary experiments, procathepsin D was found to act as a specific autocrine growth factor for breast cancer-derived cells, but not for any other cell type tested. These effects were mediated through a new, previously unknown specific receptor moiety expressed on breast cancer cell lines. The region of procathepsin D responsible for its mitogenic activity was localized in position 36-44 of the APpCD sequence. No growth factor activity could be shown with the mature enzyme cathepsin D. The proposed specific aims are based on the central hypothesis that procathepsin D is involved in breast cancer via a specific receptor that mediates autocrine activation for increased metastatic growth. For Aim lit is hypothesized that the overproduction of procathepsin D results in an increase in the metastatic potential of breast tumor cells. A low metastatic human breast cancer cell line will be transfected with human procathepsin D cDNA such that the cells will secrete constitutively varying amounts of procathepsin D. The metastatic potential of each transfected cell line will be evaluated both in vitro and in vivo in relationship to the amount of procathepsin D secretion. In addition, the synthesis of pCD will be inhibited using specifically constructed ribozymes. Attempts will be made to determine the exact site in procathepsin D responsible for breast cancer cell growth factor activity. Synthetic peptides representing fragments of APpCD will be prepared. Amino acid substitutions in the most active peptide fragment will be used to map the essential amino acid contact sites for the receptor. For Aim 2 attempts will be made to identify the membrane receptor for procathepsin D. A synthetic peptide representing the binding site domain of procathepsin D will be used to isolate candidate receptor molecules. For Aim 3 it is hypothesized that inhibition of the APpCD interaction with its receptor will result in inhibition of cancer cell growth. Peptide analogs or complementary peptides will be prepared with D-amino acids to block the growth and malignancy of cancer cells both in vitro and in vivo. The overall goal is to generate a pharmacological agent for breast cancer based on blockage of the autocrine growth factor activity of pCD.

描述（由申请人提供）：长期目标是基于procathepsin的自分泌生长因子活性D.乳腺癌细胞分泌procathepsin d的乳腺癌的新治疗方法。 Procathepsin d已被确定为乳腺癌的独立预后因素。在初步的实验中，发现protathepsin d是乳腺癌衍生细胞的特定自分泌生长因子，但对于任何其他测试的细胞类型而言，没有。这些作用是通过在乳腺癌细胞系上表达的新的，未知的特异性受体部分介导的。负责其有丝分裂活性的Procathepsin d的区域位于APPCD序列的36-44位。成熟的酶组织蛋白酶D无法显示生长因子活性。提出的特定目的是基于核心假设，即prothepsin d通过介导自分泌激活以增加转移性生长的特定受体参与乳腺癌。对于AIM点亮，假设Procathepsin d的过量生产导致乳腺肿瘤细胞的转移潜力增加。低转移性人类乳腺癌细胞系将被人类的procathepsin d cDNA转染，以便细胞会分泌组成型变化的procathepsinD。每个转染的细胞系的转移潜力将在体外和体内评估与procathepsin d分泌的数量。另外，使用特定构建的核酶抑制PCD的合成。将尝试确定负责乳腺癌细胞生长因子活性的Procathepsin d中的确切位置。将制备代表APPCD片段的合成肽。最活跃的肽片段中的氨基酸取代将用于绘制受体的必需氨基酸接触位点。对于AIM 2，将尝试确定ProcathepsinD的膜受体D。代表Procathepsin d的结合位点结构域D的合成肽将用于隔离候选受体分子。对于目标3，假设抑制APPCD与其受体的相互作用将导致癌细胞生长的抑制。肽类似物或互补肽将用D-氨基酸制备，以阻止体外和体内癌细胞的生长和恶性肿瘤。总体目标是基于对PCD的自分泌生长因子活性的阻塞生成乳腺癌的药理学剂。