Regulation of breast cancer growth by activation peptide

激活肽调节乳腺癌生长

• 批准号: 7126474
• 负责人: VACLAV VETVICKA
• 金额: $ 20.46万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7126474
• 关键词: affinity chromatography; aspartic endopeptidases; athymic mouse; biological signal transduction; breast neoplasms; cathepsin D; cell growth regulation; complementary DNA; enzyme biosynthesis; enzyme mechanism; growth factor; growth factor receptors; metastasis; mitogens; neoplasm /cancer chemotherapy; neoplasm /cancer pharmacology; nucleotides; prognosis; ribozymes; synthetic peptide; tissue /cell culture; transfection; yeast two hybrid system; zymogens

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）：长期目标是开发一种基于阻断组织蛋白原D的自分泌生长因子活性的乳腺癌新疗法。乳腺癌细胞分泌组织蛋白酶原D，该酶原通过去除活化肽（APpCD）产生天冬氨酸蛋白酶组织蛋白酶D。组织蛋白酶D原已被确定为乳腺癌的独立预后因子。在初步实验中，发现前组织蛋白酶D作为乳腺癌衍生细胞的特异性自分泌生长因子，但不适用于任何其他测试的细胞类型。这些作用是通过乳腺癌细胞系上表达的一种新的、以前未知的特异性受体部分介导的。负责其促有丝分裂活性的组织蛋白原D的区域位于APCD序列的36-44位。成熟酶组织蛋白酶D没有显示出生长因子活性。所提出的具体目标是基于中心假设，即组织蛋白原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。为此，假设组织蛋白原D的过度产生导致乳腺肿瘤细胞转移潜能的增加。低转移性人乳腺癌细胞系将用人组织蛋白酶原D cDNA转染，使得细胞将分泌组成型不同量的组织蛋白酶原D。将在体外和体内评价每种转染细胞系的转移潜力与组织蛋白酶原D分泌量的关系。此外，使用特异性构建的核酶将抑制pCD的合成。将尝试确定确切的网站在前组织蛋白酶D负责乳腺癌细胞生长因子的活动。将制备代表APpCD片段的合成肽。最具活性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。对于目的2，将尝试鉴定组织蛋白酶原D的膜受体。代表前组织蛋白酶D的结合位点结构域的合成肽将用于分离候选受体分子。对于目的3，假设抑制APCD与其受体的相互作用将导致抑制癌细胞生长。肽类似物或互补肽将与D-氨基酸一起制备，以在体外和体内阻断癌细胞的生长和恶性。总体目标是基于阻断pCD的自分泌生长因子活性来产生用于乳腺癌的药理学药剂。

项目成果

Ribozyme-targeting procathepsin D and its effect on invasion and growth of breast cancer cells: an implication in breast cancer therapy.

靶向核酶的组织蛋白酶原 D 及其对乳腺癌细胞侵袭和生长的影响：对乳腺癌治疗的影响。

• 发表时间: 2007
• 期刊: International journal of oncology
• 影响因子: 5.2
• 作者: Vashishta,Aruna;Ohri,SujataSaraswat;Proctor,Mary;Fusek,Martin;Vetvicka,Vaclav
• 通讯作者: Vetvicka,Vaclav

Procathepsin D secreted by HaCaT keratinocyte cells - A novel regulator of keratinocyte growth.

HaCaT 角质形成细胞分泌的组织蛋白酶 D 原 - 角质形成细胞生长的新型调节剂。

• DOI: 10.1016/j.ejcb.2007.03.008
• 发表时间: 2007
• 期刊: European journal of cell biology
• 影响因子: 6.6
• 作者: Vashishta,Aruna;SaraswatOhri,Sujata;Vetvickova,Jana;Fusek,Martin;Ulrichova,Jitka;Vetvicka,Vaclav
• 通讯作者: Vetvicka,Vaclav

The propeptide of cathepsin D increases proliferation, invasion and metastasis of breast cancer cells.

• DOI: 10.3892/ijo.32.2.491
• 发表时间: 2008-02
• 期刊: International journal of oncology
• 影响因子: 5.2
• 作者: S. Ohri;A. Vashishta;M. Proctor;M. Fusek;V. Vetvicka