Identification and therapeutic application of miRNA-drivers in lung cancer

肺癌中 miRNA 驱动因子的鉴定和治疗应用

• 批准号: 8566534
• 负责人: Andrea L Kasinski
• 金额: $ 9.96万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-13 至 2013-11-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8566534
• 关键词: 14-3-3 Family; Accounting; Address; Adenocarcinoma; Agar; Animal Model; Animals; Antineoplastic Agents; Area; Award; BCL2 gene; Binding; Biochemical; Biological Assay; Biology; Cancer Biology; Cancer Patient; Cancer cell line; Cancerous; Carcinoma in Situ; Cell Culture Techniques; Cell Line; Cell Survival; Cells; Cellular biology; Cessation of life; Chemicals; Clinical Trials; Code; Collaborations; Complement; Critiques; Curcumin; Data; Development Plans; Developmental Biology; Disease; Disease Progression; Epithelial Cells; Faculty; Gene Targeting; Genes; Genetic; Goals; Government; Growth; Health; Human; Hyperplasia; Individual; Instruction; Investigation; KRAS2 gene; Kinetics; Knowledge; Lead; Life Expectancy; Lobular Neoplasia; Lung; Lung Adenocarcinoma; Lung Neoplasms; Malignant Neoplasms; Malignant neoplasm of lung; Mentors; MicroRNAs; Molecular Biology; Molecular Conformation; Molecular Genetics; Mus; Mutation; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Oncogenic; Operative Surgical Procedures; Pathway interactions; Patients; Permeability; Pharmacology; Postdoctoral Fellow; Preclinical Drug Evaluation; Principal Investigator; Process; Proteins; Public Health; RNA-Binding Proteins; RNA-Protein Interaction; Radiation; Reporting; Repression; Research; Research Personnel; Resistance; Series; Signal Transduction; Survival Rate; TP53 gene; Techniques; Testing; Therapeutic; Toxic effect; Training; Transcriptional Regulation; Universities; Woman; Work; Writing; Xenograft procedure; analog; base; cancer initiation; cancer therapy; carcinogenesis; career development; chemotherapy; clinical application; combinatorial; design; high throughput screening; in vivo; in vivo Model; inhibitor/antagonist; innovation; lung tumorigenesis; meetings; member; men; metaplastic cell transformation; molecular marker; molecular pathology; mouse model; mutant; nanoparticle; neoplastic cell; novel; novel therapeutics; overexpression; prevent; public health relevance; research study; response; screening; small molecule; therapeutic development; therapeutic target; tumor; tumor progression; tumorigenic; unpublished works

DESCRIPTION (provided by applicant): The proposed work focuses on identifying novel miRNA drivers in cancer and utilizes this and other knowledge to develop miRNA-based therapeutics. This "Pathway to Independence" award application includes a mentored career development plan for the transition of the candidate, Dr. Andrea Kasinski, into an independent investigator, as well an accompanying research plan describing the proposed experiments on discovering miRNAs that can potentiate KRAS-driven lung adenocarcinoma and exploring novel therapeutic strategies for re-expressing tumor-suppressive miRNAs, which includes a high-throughput screen to identify a small molecule that can restore miRNA processing and combinatorial miRNA therapies for sensitizing tumors to conventional chemotherapies. The candidate, Dr. Kasinski, is a postdoctoral fellow at Yale, in the lab of Dr. Frank Slack in the Department of Molecular, Cellular, and Developmental Biology. The work leading to her graduate degree in Genetics and Molecular Biology at Emory University was conducted in the lab of Dr. Haian Fu in the Department of Pharmacology and focused on targeting cell survival signaling for therapeutic development. In the Fu lab, Dr. Kasinski performed two very distinct projects that were interrelated based on the ultimate goal of developing targeted therapeutics: in one project Dr. Kasinski utilized biochemical assays to identified IKK-¿ as a direct target of EF24, an analogue of curcumin that ultimately made its way into clinical trials. For the second project a series of genetic and molecular techniques were used to evaluate the transcriptional regulation of an oncogenic 14-3-3 family member, with the hope that this knowledge might spearhead subsequent studies to block 14-3-3 expression. The mentoring and career development plan will supplement her background, which is evenly split between genetic, molecular biology, whole animal studies and small molecule screening, with training and instruction in each, and in the particular areas that this project involves: murine biology, cancer biology, cell culture technique and high-throughput drug screening. Dr. Kasinski's goal is to become a faculty member in an interdisciplinary biosciences, cancer biology, or similar department at an academic, private, or government facility, in which she can research the biology of miRNAs in cancer and work to advance miRNA-based therapies. This research on miRNAs involvement in cancer requires that an innovative and selective screen be performed which Dr. Kasinski is actively pursuing first in cell culture and will advance into animal models. This assay in soft agar is selecting for miRNAs that can specifically cause normal human lung bronchial epithelial cells to become transformed. MiRNAs identified from this assay will be evaluated further in cell culture and ultimately in vivo. The proposed study involving miRNA therapeutics is two fold. The first takes into account the interaction of let-7 and LIN-28. Dr. Kasinski is in an active collaboration with Dr. Haian Fu, Co-Director of the Chemical Biology and Drug Screening Center at Emory University, to perform a high throughput screen (HTS) to identify inhibitors of this protein-RNA interaction. Hits from this HTS will be evaluated for cell permeability, kinetics and in vivo therapeutic potential, by several independent avenues of investigation. Finally building on Dr. Kasinski's current findings that miR-34 and let-7 represent valid therapeutic options for non-small cell lung cancer, she will evaluate these miRNA therapies in combination with currently used chemotherapies and targeted-therapeutics in cell culture, xenografts and in the Kras;p53 double mutant. This work is novel, timely and has clear and significant implications for human health and survival. The identification of miRNAs and targeted-therapies to perturb miRNA imbalance that occurs in cancer is invaluable to the fields of cancer and miRNA biology, and has direct clinical application.

描述（由申请人提供）：拟议的工作重点是识别癌症中的新型miRNA驱动程序，并利用这一知识和其他知识开发基于miRNA的治疗方法。这个“独立之路”奖申请包括一个指导性的职业发展计划，用于候选人Andrea Kasinski博士转变为独立研究者，以及一个附带的研究计划，描述了发现可以增强KRAS驱动的肺腺癌的miRNAs的拟议实验，并探索重新表达肿瘤抑制miRNAs的新治疗策略，其包括高通量筛选以鉴定可以恢复miRNA加工的小分子和用于使肿瘤对常规化疗敏感的组合miRNA疗法。候选人Kasinski博士是耶鲁大学的博士后研究员，在分子，细胞和发育生物学系Frank Slack博士的实验室工作。导致她在埃默里大学遗传学和分子生物学研究生学位的工作是在药理学系Haian Fu博士的实验室进行的，重点是靶向细胞存活信号用于治疗开发。在Fu实验室，Kasinski博士进行了两个非常不同的项目，这些项目基于开发靶向治疗的最终目标而相互关联：在一个项目中，Kasinski博士利用生化分析将IKK-<$作为EF 24的直接靶点，EF 24是姜黄素的类似物，最终进入临床试验。对于第二个项目，一系列遗传和分子技术被用于评估致癌14-3-3家族成员的转录调控，希望这些知识可能会成为随后阻断14-3-3表达的研究的先锋。指导和职业发展计划将补充她的背景，这是平均分配之间的遗传，分子生物学，整个动物研究和小分子筛选，与培训和指导，在每一个，并在特定领域，该项目涉及：鼠生物学，癌症 生物学、细胞培养技术和高通量药物筛选。Kasinski博士的目标是成为跨学科生物科学，癌症生物学或学术，私人或政府机构的类似部门的教员，在那里她可以研究癌症中miRNA的生物学，并致力于推进基于miRNA的疗法。这项关于miRNAs参与癌症的研究需要进行创新和选择性的筛选，Kasinski博士首先在细胞培养中积极追求，并将进入动物模型。在软琼脂中的该测定选择可以特异性地引起正常人肺支气管上皮细胞转化的miRNA。将在细胞培养物中并最终在体内进一步评价从该测定中鉴定的miRNA。涉及miRNA治疗的拟议研究是双重的。第一个考虑let-7和LIN-28的相互作用。Kasinski博士与埃默里大学化学生物学和药物筛选中心的联合主任Haian Fu博士积极合作，进行高通量筛选（HTS）以鉴定这种蛋白质-RNA相互作用的抑制剂。将通过几种独立的研究途径，评价来自该HTS的命中的细胞渗透性、动力学和体内治疗潜力。最后，基于Kasinski博士目前的发现，即miR-34和let-7代表了非小细胞肺癌的有效治疗选择，她将评估这些miRNA疗法与目前使用的化疗和靶向疗法在细胞培养，异种移植和Kras中的组合;p53双突变体。这项工作是新颖的，及时的，对人类健康和生存有明确和重大的影响。鉴定miRNA和靶向治疗以扰乱癌症中发生的miRNA不平衡对于癌症和miRNA生物学领域是无价的，并且具有直接的临床应用。