SYSTEMIC ANTIANGIOGENIC GENE THERAPY FOR BREAST CANCER

乳腺癌的全身抗血管生成基因治疗

基本信息

批准号：
6514117
负责人：
ROBERT J DEBS
金额：
$ 31.24万
依托单位：
CALIFORNIA PACIFIC MED CTR RES INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-07-23 至 2004-10-31
项目状态：
已结题

项目摘要

Our goal is to develop effective cationic liposome:DNA complex (CLDC)-based systemic gene therapy for metastatic human breast cancer. Tumor metastasis is the major cause of cancer death. Thus, meaningful reductions in cancer mortality require effective, systemic therapy. Our primary hypothesis is that new, more efficient cationic liposomes and expression vectors have rendered CLDC-based intravenous (iv) gene delivery both an effective anti-metastatic therapy, and a powerful tool for elucidating the regulation of tumor angiogenesis. We have already shown that CLDC-based iv gene delivery can i) significantly reduce both metastatic spread and tumor angiogenesis in tumor-bearing mice, ii) transfect very large numbers of metastatic tumor cells, endothelial cells and macrophages, iii) maintain therapeutic levels of gene expression for greater than or equal to 2 months, and iv) then efficiently re-express the gene following repeated re-injection in immunocompetent mice. We will test this iv CLDC gene delivery approach in two metastatic tumor models: C3H/HeN mice bearing the syngeneic mouse mammary carcinoma line, C3L5 and in SCID/beige mice bearing the human breast cancer xenograft line, MDA-MB-435. Our five specific aims are: Aim 1: Determine the anti-metastatic, anti-angiogenic and pro-apoptotic effects produced by a variety of anti-tumor genes, following their CLDC-based iv gene delivery into tumor-bearing mice. Aim 2: Increase anti-tumor activity by identifying CLDC co-delivered genes that produce synergistic anti-cancer activity in mice. Aim 3: Use our novel, durably- expressing EBV-based plasmid vector to attempt to: 3A: Produce sustained eradication of metastatic breast cancer, and 3B: Provide long-term anti-metastatic protection in mice bearing localized breast tumors. AIM 4: Increase therapeutic activity and reduce host toxicity by using two cell type-specific promoters to target the expression of CLDC-delivered genes: 4A: A 900 bp sequence of the Von Willibrand Factor (VWF)-3 promoter element incorporated into our expression plasmid to specifically target expression of CLDC delivered anti-angiogenic genes to vascular endothelial cells. 4B: A 694 bp sequence of the human mucin-like glycoprotein promoter DF3 incorporated into our expression plasmid to target expression of iv, CLDC-delivered genes to breast cancer cells that overexpress DF3. Aim 5: Use both combination gene delivery and promoter-based targeting studies to better characterize the genetic and cellular factors that regulate the angiogenic phenotype in tumor-bearing mice. These studies should both improve system anti-cancer gene therapy and identify specific genes and cell types that can control tumor angiogenesis in tumor-bearing animals.

我们的目标是开发有效的阳离子脂质体：基于DNA复合物（CLDC）的全身基因治疗，用于转移性人类乳腺癌。肿瘤转移是癌症死亡的主要原因。因此，癌症死亡率有意义的降低需要有效的全身治疗。我们的主要假设是，新的，更有效的阳离子脂质体和表达载体已经使基于CLDC的静脉内（IV）基因递送既是有效的抗转移疗法，又是阐明肿瘤血管生成调节的有效工具。我们已经表明，基于CLDC的IV基因递送可以显着降低肿瘤性小鼠中的转移性扩散和肿瘤血管生成，ii）转染非常大量的转移性肿瘤细胞，内皮细胞和巨噬细胞，iii）维持治疗水平的基因表达水平更大或等于2个月，并在2个月内进行了反复反应，并及时地及时地进行了反复的反复反应。老鼠。我们将在两个转移性肿瘤模型中测试这种IV CLDC基因递送方法：携带合成小鼠乳腺癌系的C3H/HEN小鼠，C3L5，以及带有人类乳腺癌的SCID/米色小鼠MDA-MB-435。我们的五个具体目的是：目标1：确定各种抗肿瘤基因产生的抗中转移，抗血管生成和促凋亡作用，其基于CLDC的IV基因递送到肿瘤小鼠中。 AIM 2：通过鉴定CLDC共同传递的基因来增加抗肿瘤活性，从而在小鼠中产生协同抗癌活性。 AIM 3：使用我们的新型，持久表达基于EBV的质粒载体的尝试：3A：产生转移性乳腺癌的持续根除，3B：在带有局部乳腺肿瘤的小鼠中提供长期的抗转移性保护。目标4：通过使用两个细胞类型特异性启动子来靶向CLDC传递基因的表达：4A：von Willibrand因子（VWF）-3启动子元件的900 bp序列，将900 bp的序列纳入我们的表达中的900 bp序列，以靶向CLDC抗体性基因的表达中，von Willibrand因子（VWF）-3启动子元素均拟合到我们的表达中的900 bp序列，从而提高治疗活性并降低宿主毒性。 4B：人粘蛋白样糖蛋白启动子DF3的694 bp序列掺入我们的表达质粒中，以靶向IV的表达，CLDC传递基因对过表达DF3的乳腺癌细胞。目标5：使用组合基因递送和基于启动子的靶向研究来更好地表征调节肿瘤小鼠血管生成表型的遗传和细胞因子。这些研究应改善系统抗癌基因治疗，并确定可以控制肿瘤动物中肿瘤血管生成的特定基因和细胞类型。