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型治疗剂的开发一直是癌症研究的重点。几个组已经开发了源自​​凋亡的Bcl-2家族蛋白或产生的化合物的BH3肽，这些化合物可以用作BH3肽症，这些化合物可以通过释放促凋亡的BAX和/或BAK蛋白来诱导凋亡，从而诱导凋亡。 BH3肽或其衍生物的应用在治疗癌症方面产生了一些成功的结果。但是，这种方法将其应用于表现出抗凋亡Bcl-2蛋白过度表达或降低的Bax和Bak水平的癌症时，似乎存在局限性，因为这些蛋白质的异常表达会导致对这种BH3方法的抗性。必须考虑绕开BCL-2家族蛋白的替代策略。以前，我们报道了ATAP是BFL-1的一种新型两亲性尾锚肽，它触发了有效的线粒体依赖性细胞死亡。 ATAP可以直接形成线粒体膜中的细胞色素c-释放毛孔，并且ATAP的促凋亡活性不需要，也不受抗凋亡蛋白或促凋亡蛋白的影响。我们还发现，与许多癌细胞中各种BH3肽相比，ATAP在诱导凋亡方面更有效。我们使用内部化RGD肽（IRGD）将ATAP靶向和递送到癌细胞中。该ATAP-IRGD融合肽可以有效合成，与天然ATAP相似，ATAP-IRGD可以靶向线粒体渗透性以诱导凋亡。与无与伦比的ATAP导致细胞死亡的ATAP相反，ATAP-IRGD肽似乎有选择地靶向肿瘤细胞进行凋亡。此外，ATAP-IRGD在DU145人类前列腺癌细胞系中有效诱导细胞死亡，该细胞对BH3肽治疗的作用有抵抗力。静脉注射ATAP-IRGD在具有人类癌细胞的异种移植小鼠中的初步研究可以降低肿瘤的大小，同时不产生明显的脱靶毒性。这些发现使我们相信ATAP可以成为治疗潜在抗性化学治疗试剂的癌细胞的有效治疗剂。 这种SBIR阶段的应用将利用我们在生物学药物开发方面的专业知识，以研究ATAP-IRGD的必要研究，以作为前列腺癌的治疗方法，以旨在向FDA提出研究新药（IND）应用。这项工作将包含以下两个具体目标。 AIM 1，测试小鼠模型中ATAP-IRGD的药代动力学，组织分布和毒理学特征。尽管我们的初步结果表明与ATAP-IRGD相关的毒性有限，但评估ATAP-IRGD安全性的进一步毒理学研究对于启用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基本的指导，以测试ATAP-IRGD治疗前列腺癌的功效。 AIM 2，建立ATAP-IRGD在前列腺癌的异种移植小鼠模型中的功效。在这里，我们将测试使用各种人类前列腺癌细胞系的异种移植小鼠模型中各种剂量的ATAP-IRGD会影响肿瘤的生长。这些努力将包括比较研究BH3肽在减少肿瘤生长中的功效。我们将收集有关肿瘤大小的数据，并使用组织学来对用ATAP-IRGD或BH3肽进行治疗后的肿瘤进展。我们还将使用异种移植小鼠模型进行其他分布研究。上述研究的履行将为我们的未来广泛的EFFOT提供原则数据，以开发ATAP-IRGD作为一种新型的治疗试剂，用于治疗由于Bcl-2家族蛋白异常表达而对现有的化学治疗试剂具有抗性的癌细胞。