Development of Ets2-Inhibitors

Ets2抑制剂的开发

• 批准号: 7847347
• 负责人: DEV PRIYA ARYA
• 金额: $ 17.52万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-03 至 2011-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7847347
• 关键词: Abbreviations; Affinity; Aminoglycoside Antibiotics; Aminoglycosides; Antineoplastic Agents; Antisense Oligonucleotides; Area; Binding; Biological Assay; Biology; Breast; Breast Cancer Cell; Calorimetry; Cancer Etiology; Cause of Death; Cell Proliferation; Cells; Charge; Chemicals; Chemistry; Chromatin; Chromatin Structure; Circular Dichroism Spectroscopy; Classification; Clinic; Coffee; Collaborations; Commit; Complement; DNA; DNA-Directed RNA Polymerase; Development; Doctor of Medicine; Dominant-Negative Mutation; Electrophoretic Mobility Shift Assay; Face; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Recombination; Genetic Transcription; Genomics; Goals; Growth; Health; Healthy People 2010; Human; Institutes; Interdisciplinary Study; Investigation; Joints; Kinetics; Knowledge; Laboratories; Life; Link; Luciferases; Malignant Neoplasms; Malignant neoplasm of prostate; Mammalian Cell; Measurement; Mediating; Methods; Molecular; Mutagenesis; Neomycin; Neoplastic Cell Transformation; Nuclear; Nucleic Acid Binding; Nucleic Acids; Oligonucleotides; Organic Synthesis; Outcome; Pathogenesis; Permeability; Pharmaceutical Preparations; Phase; Plasmids; Play; Preparation; Principal Investigator; Property; Prostate; Proteins; RNA; RNA analysis; Regulation; Reporter; Research; Role; Scanning; Single-Stranded DNA; Site; Small Interfering RNA; Solid; Solutions; Structure; Switzerland; Technology; Testing; Therapeutic; Thiourea; Titrations; Training; Transcription Initiation; Transfection; United States; United States Public Health Service; Work; anticancer activity; anticancer research; antigene; base; biophysical chemistry; c-myc Genes; cancer cell; cancer therapy; cost; design; disability; drug development; ds-DNA; gene function; gene repression; graduate student; health application; improved; inhibitor/antagonist; innovation; interest; methyl sulfate; microcalorimetry; multidisciplinary; novel; novel strategies; novel therapeutics; oncology; phosphodiester; phosphorothioate; programs; promoter; research study; success; transcription factor; transmission process; triple helix; triplex DNA; tumor progression; ultraviolet; uptake

DESCRIPTION (provided by applicant): Nucleic acids are central molecules in the transmission, expression and conservation of genetic information. Recognition of duplex DNA by oligonucleotides, forming DNA triple helices, provides a promising approach to a chemical solution for DNA recognition. These triplex forming oligonucleotides (TFOs) have aroused significant interest as potential inhibitors of the expression of particular genes. The ets2 gene encodes a transcription factor that plays a critical role in controlling cell proliferation and differentiation and also has a key role in the pathogenesis and progression of breast and prostate cancer. In the Catapano (co- PI) lab, triplex formation has been shown to sequence-specifically inhibit transcription at specific sites in the ets2 promoter sequences and also directly inhibit transcription by blocking RNA polymerase. Triplex technology however remains to be improved before health applications to these specific areas can be made. The need for an improved target affinity in the therapeutic regulation of specific gene expression in cancer cells remains, as does the delivery of these TFOs and all oligonucleotides in cells. Our preliminary results show the ability of neomycin to stabilize key triplex forming targets in Ets2 gene and aid in delivery of oligonucleotides. The objective of this project is to develop neomycin-TFOs that directly target the ets2 gene, found amplified in breast and prostate cancer. The proposed work will overcome important challenges such as TFO affinity to the duplex and delivery to the cells, that traditional TFOs face. The hypothesis :} Neomycin-mediated delivery of TFO-neomycin conjugates can be used to develop sequence-specific anticancer agents with improved delivery properties and enhanced duplex affinity.} We propose to develop neomycin conjugated TFOs that (1) enhance the binding affinity of the TFO to its target duplex, (2) improve the delivery and uptake of the TFO to the cancer cells. Using the recent findings from our labs, we will synthesize neomycin-TFO conjugates for triplex formation with sites of therapeutics interest in Ets-2 gene. We will then use neomycin- TFO conjugates to determine their efficacy in stabilizing the triplexes on Ets-2. Appropriate neomycin-TFO conjugates will be synthesized on a DNA synthesizer, and evaluated for anti- transcriptional and anticancer activity in binding to the Ets-2 targets of interest. The cellular and nuclear uptake of neomycin-conjugated TFOs is being done in collaboration with Carlo Catapano, M.D., Director, Laboratory of Experimental Oncology, Oncology Institute of Southern Switzerland (IOSI) Bellinzona, Switzerland. This testing will involve studies focused on the stability of the conjugate within cancer cells, examination of the sequence and target selectivity of the conjugate, systematic determination of the number of neomycin molecules per oligonucleotide needed for optimal delivery and anti-gene activity, the ability of the neomycin-TFO to discriminate for target sequences and test the effects of these high binding TFOs on endogeneous gene expression and growth of human breast cancer cells. The development of neomycin-TFO conjugates has the potential for bringing the much needed improvements in cancer therapy by vastly improving the permeability of TFOs and deregulating expression of ets2 gene in proliferation of breast and prostate cancer cells. The success of this approach will open up new avenues of TFO development and delivery not only as they relate to cancer, but in other oligonucleotide based therapies as well. Health Relevance Statement One of the challenges of research in oncology is to find ways to use the increasing knowledge of the mechanisms underlying neoplastic transformation and tumor progression to develop novel therapeutic strategies for cancer. Targeting specific genes, such as Ets2 or c-myc, which are involved in proliferation and survival of cancer cells is a promising approach. Our preliminary results show the ability of neomycin to stabilize key triplex forming targets in Ets2 gene and aid in delivery of oligonucleotides. This property of neomycin will now be extended to develop aminoglycoside-TFO conjugates to target Ets2 promoter sequences that bind with high affinity and can be delivered without external transfection agents. These conjugates can then be used to down regulate gene expression and inhibit proliferation of cancer cells. Proposed studies in our labs, using undergraduate and graduate students, and collaborative efforts with the Catapano lab will further help establish the efficacy of this approach. The work proposed here, a multidisciplinary effort encompassing organic synthesis, biophysical chemistry, and cancer research describes the development and neomycin mediated delivery/anticancer activity of novel positively charged TFOs (neomycin-TFOs). In particular, this work proposes to initiate a multidisciplinary team that leads to the creation and use of TFO conjugates that have novel properties and functions because of their conjugation with neomycin. The success of the proposed work would be a significant addition to currently available approaches in breast and prostate cancer drug development and TFO (oligonucleotide) delivery. We propose using 25 base Ets2 target sequences to design neomycin-TFO conjugates that can be employed to inhibit cancer growth, opening possibilities for the development of small sequence- designed anti-cancer agents. Using neomycin's ability to stabilize triplexes and deliver TFOs, neomycin-TFO (PS) conjugates will be developed as triplex-forming anticancer agents.

描述（由申请人提供）：核酸是遗传信息传播，表达和保存中的中央分子。通过寡核苷酸对双链DNA的识别，形成DNA三螺旋，为DNA识别的化学溶液提供了有希望的方法。这些形成寡核苷酸（TFO）的三环形成，作为特定基因表达的潜在抑制剂引起了重大兴趣。 ETS2基因编码转录因子在控制细胞增殖和分化中起关键作用，并且在乳腺癌和前列腺癌的发病机理和进展中也具有关键作用。在Catapano（Co-PI）实验室中，已证明三环形成可以特异性抑制ETS2启动子序列中特定位点的转录，并通过阻断RNA聚合酶直接抑制转录。然而，在对这些特定领域的健康应用之前，三元技术仍有待改进。在癌细胞中特异性基因表达的治疗调节中，需要改善靶标亲和力，这些TFO和所有寡核苷酸在细胞中的递送也是如此。我们的初步结果表明，新霉素能够稳定ETS2基因中的关键三心形成靶标并有助于寡核苷酸的递送。该项目的目的是开发直接靶向ETS2基因的新霉素-TFO，发现在乳腺癌和前列腺癌中被放大。拟议的工作将克服传统TFO面临的TFO亲和力，例如对双链体的TFO亲和力，并将其传递给细胞。假设：}新霉素介导的TFO-神经结合物可用于开发具有提高递送特性并增强双工亲和力的序列特异性抗癌剂的序列特异性的抗癌药。}我们建议开发新霉素共轭TFOS，以增强（1）增强其目标癌症的结合性，并增强其目标的结合，并提高其靶化典型（2）（2）（2）（2）（2）（2）（2）2）（2）（2）（2）（2）（2）细胞。使用实验室的最新发现，我们将合成新霉素-TFO结合物，以与ETS-2基因的疗法兴趣相结合。然后，我们将使用新霉素偶联物来确定它们在稳定ETS-2上的三重速度方面的功效。适当的新霉素-TFO偶联物将在DNA合成器上合成，并在与ETTS-2靶标结合时评估抗转录和抗癌活性。新霉素偶联的TFO的细胞和核吸收与Carlo Catapano，M.D。合作，瑞士南部瑞士肿瘤学研究所（IOSI）Bellinzona实验性肿瘤学实验室主任。该测试将涉及研究集中在癌细胞内结合物的稳定性的研究，检查结合物的序列和靶标选择性，对每个寡核苷酸所需的新霉素分子的数量进行系统的确定，以最佳递送和抗基因活性，抗抗蛋白-TFO的能力，以鉴于这些较高的序列的TF效​​应的效果，以抗性的能力和抗基因的能力效应，以抗性的能力。乳腺癌细胞。新霉素-TFO偶联物的发展具有通过极大地提高TFO的渗透性以及Eets2基因在乳腺癌和前列腺癌细胞增殖中的过度调节表达来实现巨大必要改善的癌症治疗。这种方法的成功将为TFO开发和分娩的新途径不仅与癌症有关，而且在其他基于寡核苷酸的疗法方面也为它们提供了。健康相关性陈述肿瘤学领域的挑战之一是找到方法来利用对肿瘤转化和肿瘤进展的机制越来越多的知识，以开发新的癌症治疗策略。靶向涉及癌细胞增殖和存活的特定基因，例如ETS2或C-MYC是一种有前途的方法。我们的初步结果表明，新霉素能够稳定ETS2基因中的关键三心形成靶标并有助于寡核苷酸的递送。现在将扩展新霉素的这种特性，以开发氨基糖苷-TFO偶联物，以与高亲和力结合的靶向ETS2启动子序列，并且可以在没有外部转染药物的情况下进行交付。然后，这些结合物可用于降低基因表达并抑制癌细胞的增殖。在我们的实验室，使用本科生和研究生以及与Catapano Lab的协作工作的拟议研究将进一步帮助确定这种方法的功效。这里提出的工作是一项涵盖有机合成，生物物理化学和癌症研究的多学科工作，描述了新型带正电TFO（Neomycin-TFOS）的发育和新霉素介导的递送/抗癌活性。特别是，这项工作建议启动一个多学科团队，该团队导致创建和使用TFO共轭物，这些结合物具有新颖的特性和功能，因为它们与Neomycin结合了。拟议工作的成功将是当前在乳腺癌和前列腺癌药物开发和TFO（寡核苷酸）递送方面的可用方法中的重要补充。我们建议使用25个基本ETS2目标序列设计新霉素-TFO结合物，这些结合物可用于抑制癌症的生长，为开发小序列设计的抗癌剂的开发可能性。使用新霉素稳定三倍体并传递TFO的能力，Neomycin-TFO（PS）缀合物将作为形成三型抗癌剂的剂发展。

项目成果

Neomycin-neomycin dimer: an all-carbohydrate scaffold with high affinity for AT-rich DNA duplexes.

• DOI: 10.1021/ja108118v
• 发表时间: 2011-05-18
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Kumar S;Xue L;Arya DP
• 通讯作者: Arya DP

Multivalency in the recognition and antagonism of a HIV TAR RNA-TAT assembly using an aminoglycoside benzimidazole scaffold.

• DOI: 10.1039/c5ob02016f
• 发表时间: 2016-02-14
• 期刊: Organic & biomolecular chemistry
• 影响因子: 3.2
• 作者: Kumar S;Ranjan N;Kellish P;Gong C;Watkins D;Arya DP
• 通讯作者: Arya DP

Selective Inhibition of Bacterial Topoisomerase I by alkynyl-bisbenzimidazoles.

• DOI: 10.1039/c4md00140k
• 发表时间: 2014-06-01
• 期刊: MedChemComm
• 作者: Ranjan N;Fulcrand G;King A;Brown J;Jiang X;Leng F;Arya DP
• 通讯作者: Arya DP

Shape readout of AT-rich DNA by carbohydrates.

• DOI: 10.1002/bip.22448
• 发表时间: 2014-07
• 期刊: BIOPOLYMERS
• 影响因子: 2.9
• 作者: Kumar, Sunil;Spano, Meredith Newby;Arya, Dev P.
• 通讯作者: Arya, Dev P.

Recognition of HIV-TAR RNA using neomycin-benzimidazole conjugates.

• DOI: 10.1016/j.bmcl.2013.08.014
• 发表时间: 2013-10-15
• 期刊: BIOORGANIC & MEDICINAL CHEMISTRY LETTERS
• 影响因子: 2.7
• 作者: Ranjan, Nihar;Kumar, Sunil;Watkins, Derrick;Wang, Deyun;Appella, Daniel H.;Arya, Dev P.
• 通讯作者: Arya, Dev P.