Breast Cancer Therapeutic Agents Based on Telomerase Misfunction

基于端粒酶功能障碍的乳腺癌治疗剂

• 批准号: 8182301
• 负责人: ELIZABETH H BLACKBURN
• 金额: $ 21.69万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8182301
• 关键词: Address; Amplifiers; Antibodies; Antineoplastic Agents; Apoptosis; Apoptotic; Archives; Biological; Biological Assay; Biological Markers; Biopsy Specimen; Breast; Breast Cancer Cell; Breast Cancer Model; Cancer cell line; Catalytic RNA; Cell Death; Cell Line; Cells; Characteristics; Chest Wall Disorder; Clinic; Clinical; Clinical Trials; Data; Development; Drug Formulations; ERBB2 gene; Freezing; Functional disorder; Funding; Genes; Genomics; Goals; Gray unit of radiation dose; Growth; Immunoliposome; In Vitro; Laboratories; Laboratory Research; Length; Libraries; Malignant Neoplasms; Mammary Neoplasms; Measures; Methods; Modeling; Molecular; Molecular Profiling; Monitor; Neoplasm Metastasis; Paraffin Embedding; Pathway interactions; Patients; Pattern; Performance; Principal Investigator; RNA; RNA Interference; Regulation; Reproduction spores; Rodent; Rodent Model; Sampling; Science; Series; Small Interfering RNA; Staging; Stem cells; System; TP53 gene; Telomerase; Telomerase RNA Component; Telomere Maintenance; Testing; Therapeutic; Therapeutic Agents; Tissue Sample; Toxic Actions; Toxicology; Translating; Translations; Validation; Work; Xenograft Model; Xenograft procedure; anticancer treatment; base; cancer cell; cancer therapy; clinical application; drug sensitivity; functional status; high throughput screening; in vitro testing; in vivo; killings; malignant breast neoplasm; manufacturing scale-up; mouse model; mutant; nanoparticle; neoplastic cell; programs; research clinical testing; response; seal; stem; telomere; therapeutic development; tumor; vector

The goal of UCSF Breast Cancer SPORE Project #4 is to develop and assess the translational potential of agents we have developed that force telomerase interference in breast cancer. This Project focuses on exploitation for clinical use of a new strategy: to turn the action of active telomerase against the breast cancer cells. In this current funding cycle, we have successfully demonstrated that a low threshold of expression of mutant-template telomerase RNA (MT-hTer) genes in human breast cancer cells is sufficient for a potent killing and growth inhibitory effect on these cells. The telomeres that result from MT-hTer action are "toxic" to cells, inducing a robust apoptotic response. Additionally, during the previous SPORE funding period, new science arising from the Blackburn laboratory's research on telomerase also led to two] unanticipated discoveries: first, that simply decreasing the endogenous telomerase level by ribozyme or RNA targeting methods rapidly decreased cancer potential. Specifically, we found that lowering overall telomerase diminishes the metastatic potential of cancer cells in vivo, and rapidly inhibited the growth of breast and other cancer cells in vitro. Second, cell death induced by MT-hTer expression is dominant and does not require the p53 or pRb checkpoint pathways. Based on these findings, we then showed that combining the expression of MT-hTer with small interfering RNA directed against the endogenous WT-hTER of cancer cells synergistically increases the potency of the MT-hTer effects in killing cancer cells. The following Specific Aims, which have the goal of bringing this work to the clinic, are to: #1 Further test and characterize the previously developed immunoliposome ("ILS") constituted with Her2-targeting antibody¿developed in SPORE Project 3¿containing the MT-hTer/anti-hTER siRNA construct ("MT-Rx" agent). In order to monitor MT-Rx efficacy we will use relevant biomarkers of response to the agent, suitable for early stage clinica trials. #2 Identify telomere/telomerase-based biomarker patterns predictive of apoptotic response t anticancer treatments and to specific MT-Rx therapy. We will identify the subset(s) of breast cancers that wil be most responsive to existing therapies and to "MT-Rx" using (i) a panel of 60 breast cancer cell line grouped by genomic and expression profiling, telomere maintenance status and other clinically relevan characteristics and (ii) patient-derived primary breast cancer cells, including stem/progenitor cell lines; tha targets the most sensitive patient subpopulation, as identified. #3. Validate and optimize the assays fo biomarkers of telomerase and telomere status on tumor and biopsy specimens, with the goal of validating] these assays per CLIA regulations in a CLIA certified laboratory such that the results can be used clinically.) Toward translation of MT-Rx, we will finalize the product configuration, perform initial manufacturing seal up, and evaluate initial toxicology targeted systemic delivery of MT-Rx agent in rodent models.

UCSF乳腺癌孢子项目#4的目标是开发和评估的翻译潜力， 我们已经开发出了一种药物，可以在乳腺癌中强制端粒酶干扰。本项目的重点是 探索一种新的临床应用策略：将活性端粒酶的作用转向乳腺癌 癌细胞在当前的融资周期中，我们成功地证明了低门槛 多模板端粒酶RNA（MT-hTer）基因在人乳腺癌细胞中的表达是足够的 用于对这些细胞的有效杀伤和生长抑制作用。由MT-hTer作用产生的端粒 对细胞是“有毒的”，诱导强烈的凋亡反应。此外，在此前的SPORE融资中， 期间，布莱克本实验室对端粒酶的研究产生的新科学也导致了两个] 意想不到的发现：首先，简单地通过核酶或RNA降低内源性端粒酶水平， 靶向方法迅速降低了癌症的可能性。具体来说，我们发现降低端粒酶的整体水平 降低了体内癌细胞的转移潜力，并迅速抑制乳腺癌和其他肿瘤的生长。 体外培养的癌细胞第二，由MT-hTer表达诱导的细胞死亡是显性的，并且不需要免疫抑制剂。 p53或pRb检查点通路。基于这些发现，我们随后表明， MT-hTer与针对癌细胞内源性WT-hTER的小干扰RNA的结合 协同增加MT-hTer杀伤癌细胞的效力。以下具体 目的是将这项工作带到临床，目的是：#1进一步测试和表征 先前开发的免疫脂质体（“ILS”）由Her 2靶向抗体构成， 含有MT-hTer/抗-hTER siRNA构建体（“MT-Rx”试剂）的孢子项目3。为了监测 MT-Rx疗效我们将使用对药物反应的相关生物标志物，适用于早期临床 审判#2识别基于端粒/端粒酶的生物标志物模式预测凋亡反应 抗癌治疗和特异性MT-Rx治疗。我们将确定乳腺癌的亚群， 使用（i）一组60个乳腺癌细胞系， 根据基因组和表达谱、端粒维持状态和其他临床相关因素分组， 特征和（ii）患者来源的原发性乳腺癌细胞，包括干/祖细胞系; 针对所确定的最敏感的患者亚群。#3.优化和优化了检测方法 肿瘤和活检标本上端粒酶和端粒状态的生物标志物，目的是验证] 在CLIA认证的实验室中按照CLIA法规进行这些测定，以便结果可用于临床。） 关于MT-Rx的翻译，我们将最终确定产品配置，执行初始制造密封 向上，并评估MT-Rx药剂在啮齿动物模型中的初始毒理学靶向全身给药。