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（申请人提供）：核酸是遗传信息传递、表达和保存的中心分子。通过寡核苷酸识别双链DNA，形成DNA三螺旋，为DNA识别的化学解决方案提供了一种有前途的方法。这些三链体形成寡核苷酸（TFO）作为特定基因表达的潜在抑制剂引起了人们的极大兴趣。ets 2基因编码的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制和进展中也起关键作用。在Catapano（co-PI）实验室中，已显示三链体形成在ets 2启动子序列中的特定位点处序列特异性地抑制转录，并且还通过阻断RNA聚合酶直接抑制转录。然而，在将三重技术应用于这些特定领域之前，仍有待改进。在癌细胞中特异性基因表达的治疗性调节中仍然需要改进的靶亲和力，这些TFO和所有寡核苷酸在细胞中的递送也是如此。我们的初步结果显示了新霉素稳定Ets 2基因中的关键三链体形成靶标并有助于寡核苷酸递送的能力。该项目的目标是开发直接靶向ets 2基因的新霉素-TFO，该基因在乳腺癌和前列腺癌中被发现扩增。拟议的工作将克服传统TFO面临的重要挑战，如TFO对双链体的亲和力和向细胞的递送。假设：} TF 0-新霉素缀合物的新霉素介导的递送可用于开发具有改善的递送性质和增强的双链体亲和力的序列特异性抗癌剂。我们建议开发新霉素缀合的TFO，其（1）增强TFO与其靶双链体的结合亲和力，（2）改善TFO向癌细胞的递送和摄取。利用我们实验室的最新发现，我们将合成新霉素-TFO缀合物，用于形成具有Ets-2基因中的治疗学感兴趣位点的三链体。然后我们将使用新霉素- TFO缀合物来确定它们在稳定Ets-2上的三链体中的功效。将在DNA合成仪上合成适当的新霉素-TFO缀合物，并评价与感兴趣的Ets-2靶标结合的抗转录和抗癌活性。新霉素缀合的TFO的细胞和核摄取正在与Carlo Catapano，M.D.，瑞士贝林佐纳南部肿瘤研究所实验肿瘤学实验室主任。该测试将涉及集中于缀合物在癌细胞内的稳定性的研究、缀合物的序列和靶选择性的检查、最佳递送和抗基因活性所需的每个寡核苷酸的新霉素分子数目的系统测定，新霉素的能力TFO来区分靶序列并测试这些高结合TFO对人乳腺癌细胞的内源性基因表达和生长的影响。新霉素-TFO缀合物的开发具有通过极大地改善TFO的渗透性和解除乳腺癌和前列腺癌细胞增殖中ets 2基因表达的调节来带来癌症治疗中急需的改进的潜力。这种方法的成功将开辟TFO开发和递送的新途径，不仅因为它们与癌症有关，而且也与其他基于寡核苷酸的疗法有关。健康相关性声明肿瘤学研究的挑战之一是找到方法，利用对肿瘤转化和肿瘤进展机制的日益增长的知识，开发新的癌症治疗策略。靶向参与癌细胞增殖和存活的特定基因，如Ets 2或c-myc，是一种有前途的方法。我们的初步结果显示了新霉素稳定Ets 2基因中的关键三链体形成靶标并有助于寡核苷酸递送的能力。新霉素的这种性质现在将被扩展到开发氨基糖苷-TFO缀合物，以靶向Ets 2启动子序列，其以高亲和力结合，并且可以在没有外部转染剂的情况下递送。然后，这些缀合物可用于下调基因表达并抑制癌细胞的增殖。在我们的实验室中，使用本科生和研究生进行的拟议研究，以及与Catapano实验室的合作将进一步帮助建立这种方法的有效性。这里提出的工作，一个多学科的努力，包括有机合成，生物物理化学和癌症研究描述了新霉素介导的交付/抗癌活性的新型带正电荷的TFO（新霉素-TFO）的发展。特别是，这项工作建议发起一个多学科团队，导致TFO缀合物的创建和使用，由于其与新霉素的缀合而具有新的特性和功能。拟议工作的成功将是对乳腺癌和前列腺癌药物开发和TFO（寡核苷酸）递送目前可用方法的重要补充。我们提出使用25个碱基的Ets 2靶序列来设计可用于抑制癌症生长的新霉素-TFO缀合物，为开发小序列设计的抗癌剂打开了可能性。利用新霉素稳定三链体和递送TFO的能力，新霉素-TFO（PS）缀合物将被开发为形成三链体的抗癌剂。

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

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

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

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.