Targeting ATAP-mediated Apoptosis in Treatment of Prostate Cancer

靶向 ATAP 介导的细胞凋亡治疗前列腺癌

• 批准号: 8523229
• 负责人: Zelin Sheng
• 金额: $ 15.61万
• 依托单位: TRIM-EDICINE, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-26 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8523229
• 关键词: Affect; Animal Model; Animals; Apoptosis; Apoptotic; BCL-2 Protein; BCL2 gene; BH3 peptide; Basic Science; Biotechnology; Bypass; Cell Death; Cells; Chemicals; Comparative Study; DU145; Data; Development; Dose; Drug Kinetics; Drug or chemical Tissue Distribution; Evaluation; Family member; Foundations; Future; Goals; Histology; Human; Induction of Apoptosis; Integrins; Intellectual Property; International; Investigational Drugs; Investigational New Drug Application; Laboratories; Lead; Legal patent; Malignant Neoplasms; Malignant neoplasm of prostate; Measures; Mediating; Membrane; Mitochondria; Mus; Names; Nude Mice; Outcome; PC3 cell line; Peptides; Permeability; Phase; Protein Family; Protein Inhibition; Proteins; RGD (sequence); Radiation therapy; Reagent; Regulation; Reporting; Research; Research Contracts; Resistance; Safety; Small Business Innovation Research Grant; Stimulus; Structure; TRIM Gene; Tail; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Toxic effect; Toxicology; Translating; Wood material; Xenograft procedure; anticancer research; bak protein; base; cancer cell; chemotherapy; cytochrome c; cytotoxic; design; drug development; efficacy testing; human disease; in vivo; intravenous injection; malignant breast neoplasm; medical schools; mimetics; mitochondrial membrane; mouse model; neoplastic cell; novel; novel therapeutics; overexpression; peptidomimetics; pro-apoptotic protein; protein expression; public health relevance; research and development; response; targeted delivery; tumor; tumor growth; tumor progression

DESCRIPTION (provided by applicant): Development of novel therapeutics that can induce apoptosis by directly targeting the Bcl-2 family proteins has been a major focus of cancer research. Several groups have developed BH3 peptides derived from pro- apoptotic Bcl-2 family proteins or produced chemical compounds that could act as BH3 peptidomimetics, which could induce apoptosis by releasing pro-apoptotic Bax and/or Bak protein from inhibition by the anti-apoptotic proteins. Applications of BH3 peptides or their derivatives produced some successful outcomes in treating cancers. However, this approach seems to have limitations when it is applied to cancers displaying overexpression of anti-apoptotic Bcl-2 proteins or reduced Bax and Bak levels, because aberrant expression of these proteins cause resistance to such BH3 approaches. Alternative strategies that bypass regulation by Bcl- 2 family proteins must be considered. Previously we reported that ATAP, a novel amphipathic tail-anchoring peptide of Bfl-1, triggered potent mitochondria-dependent cell death. ATAP can directly form cytochrome c- releasing pores in mitochondrial membranes, and the pro-apoptotic activity of ATAP does not require and is not influenced by either anti- or pro-apoptotic proteins. We also found that ATAP is more effective in inducing apoptosis than the various BH3-peptides in many cancer cells. We used the internalizing RGD peptide (iRGD) for selective tumor targeting and delivery of ATAP into cancer cells. This ATAP-iRGD fusion peptide can be effectively synthesized, and similar to the native ATAP, ATAP-iRGD can target mitochondria permeability for induction of apoptosis. In contrast to ATAP which causes cell death indiscriminately, the ATAP-iRGD peptide appears to selectively target tumor cells for apoptosis. Moreover, ATAP-iRGD was effective at inducing cell death in the DU145 human prostate cancer cell line that was resistant to the effects of BH3 peptide therapy. Preliminary studies with intravenous injection of ATAP-iRGD into xenograft nude mice with human cancer cells could reduce tumors size while not producing significant off-target toxicity. These findings lead us to believe that ATAP can be an effective therapeutic agent for treatment of cancer cells that are potentially resistant to the existing chemotherapeutic reagents. This SBIR-Phase I application will leverage our expertise in biologic drug development to pursue necessary studies for development of ATAP-iRGD as a treatment for prostate cancer in an effort to move towards the filing of an Investigational New Drug (IND) application with FDA. This effort will contain the following two specific aims. Aim 1, test the pharmacokinetic, tissue distribution and toxicological profile of ATAP- iRGD in a mouse model. While our preliminary results suggest limited toxicity associated with ATAP-iRGD, further toxicology studies to evaluate the safety profile of ATAP-iRGD are essential to enable IND filing. Completion of these evaluations will provide an essential guidance for our in vivo animal model studies testing the efficacy of ATAP-iRGD in treatment of prostate cancer. Aim 2, establish the efficacy of ATAP-iRGD in a xenograft mouse model of prostate cancer. Here we will test if various doses of ATAP-iRGD can affect tumor growth in xenograft mouse model with various human prostate cancer cell lines. These efforts will include comparative studies to the efficacy of BH3 peptides in reducing tumor growth. We will collect data on tumor size as well as use histology to grade the progression of the tumor following treatment with ATAP-iRGD or BH3 peptides. We will also conduct additional distribution studies using the xenograft mouse models. Fulfillment of the above studies will provide proof-of-principle data for our future extensive effot toward developing ATAP-iRGD as a novel therapeutic reagent for treatment of cancer cells that are resistant to the existing chemotherapeutic reagents due to the aberrant expression of Bcl-2 family proteins.

描述(由申请人提供)：通过直接靶向Bcl-2家族蛋白来诱导细胞凋亡的新疗法的发展一直是癌症研究的主要焦点。有几个小组已经开发出来源于促凋亡蛋白的BH3多肽或产生的化合物可以作为BH3多肽模拟物，通过释放促凋亡的Bax和/或Bak蛋白来诱导细胞凋亡。BH3多肽或其衍生物的应用在治疗癌症方面取得了一些成功的结果。然而，这种方法似乎有局限性，当它应用于肿瘤表现出过表达的抗凋亡的Bcl-2蛋白或降低Bax和Bak水平，因为这些蛋白的异常表达导致对这种BH3方法的抵抗。必须考虑绕过Bcl2家族蛋白调控的替代策略。先前我们报道了BFL-1的一种新的两亲性尾部锚定多肽ATAP可以引发强大的线粒体依赖性细胞死亡。ATAP可以直接在线粒体膜上形成细胞色素C释放孔，并且ATAP的促凋亡活性不需要也不受抗或促凋亡蛋白的影响。我们还发现，在许多癌细胞中，ATAP在诱导细胞凋亡方面比各种BH3-肽更有效。我们使用内化RGD多肽(IRGD)来选择性靶向肿瘤并将ATAP输送到癌细胞中。这种ATAP-IRGD融合肽可以被有效地合成，并且与天然ATAP类似，ATAP-IRGD可以靶向线粒体通透性诱导细胞凋亡。与不分青红皂白地导致细胞死亡的ATAP不同，ATAP-IRGD多肽似乎选择性地针对肿瘤细胞进行凋亡。此外，ATAP-IRGD能有效地诱导抵抗BH3多肽治疗的DU145人前列腺癌DU145细胞死亡。初步研究表明，静脉注射ATAP-IRGD给裸鼠移植人癌细胞可以缩小肿瘤体积，同时不会产生明显的脱靶毒性。这些发现使我们相信，ATAP可以成为一种有效的治疗药物，用于治疗对现有化疗药物具有潜在耐药性的癌细胞。这一SBIR-第一阶段申请将利用我们在生物药物开发方面的专业知识，为开发ATAP-IRGD作为前列腺癌治疗药物进行必要的研究，努力向FDA提交研究新药(IND)申请。这一努力将包含以下两个具体目标。目的1、检测ATAP-IRGD在小鼠体内的药代动力学、组织分布和毒理学特征。虽然我们的初步结果表明与ATAP-IRGD相关的毒性有限，但进一步的毒理学研究以评估ATAP-IRGD的安全性对于实现IND申请至关重要。这些评估的完成将为我们的体内动物模型研究提供必要的指导，以测试ATAP-IRGD在前列腺癌治疗中的有效性。目的：建立ATAP-IRGD对小鼠前列腺癌移植瘤模型的疗效。在这里，我们将测试不同剂量的ATAP-IRGD是否可以影响不同人前列腺癌细胞系异种移植模型小鼠的肿瘤生长。这些努力将包括对BH3多肽在减少肿瘤生长方面的有效性进行比较研究。我们将收集有关肿瘤大小的数据，并使用组织学方法对ATAP-IRGD或BH3多肽治疗后的肿瘤进展进行分级。我们还将使用异种移植小鼠模型进行额外的分布研究。上述研究的完成将为我们未来开发ATAP-IRGD作为一种新的治疗试剂的广泛效果提供原理数据，该试剂用于治疗由于Bcl-2家族蛋白的异常表达而对现有化疗药物产生耐药性的癌细胞。