Targeting Tumor-Derived exRNA-Containing Microvesicles by High Throughput Screeni

通过高通量筛选靶向肿瘤衍生的含有 exRNA 的微泡

• 批准号: 8581989
• 负责人: Asim B Abdel-Mageed
• 金额: $ 49.99万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8581989
• 关键词: Adipose tissue; Animal Model; Animals; Binding; Biogenesis; Biological; Biological Assay; CD81 gene; Cancer Patient; Cell Culture Techniques; Cell Line; Cells; Chimeric Proteins; Chromosome abnormality; Clinical; Clinical Management; Collection; Conditioned Culture Media; Coupled; Data; Detection; Development; Disease; Dose; Drug Kinetics; Drug Targeting; Epithelial Cells; Figs - dietary; Flow Cytometry; Fluorescence Microscopy; Frequencies; Genetic; Goals; HRAS gene; Human; Immunoblotting; Immunocompromised Host; Implant; In Vitro; Individual; Intervention; Investigation; Label; Lead; Lesion; Libraries; Measures; Mediating; Messenger RNA; MicroRNAs; Modeling; Molecular; Monitor; Mus; Neoplasm Metastasis; Neoplastic Cell Transformation; Obesity; Oncogenic; Outcome; Patients; Pharmaceutical Preparations; Pharmacodynamics; Pharmacologic Substance; Phase; Preventive; Process; Property; Prostate; Prostatic Neoplasms; RNA; Recruitment Activity; Research; Reverse Transcriptase Polymerase Chain Reaction; Robotics; Role; Route; Safety; Scientific Advances and Accomplishments; Serum; Stem cells; Surface; TP53 gene; Technology; Testing; Therapeutic; Therapeutic Intervention; Tracer; Translational Research; Tumor Burden; Tumor Suppressor Proteins; Tumor-Derived; United States National Institutes of Health; Work; base; castration resistant prostate cancer; cell growth; clinical application; cost; density; drug discovery; epithelial to mesenchymal transition; extracellular; gene repression; high throughput screening; improved; in vivo; innovation; migration; mimicry; miniaturize; neoplastic; neoplastic cell; neovasculature; novel; preclinical study; public health relevance; research study; screening; small molecule libraries; tumor; tumor growth; tumor progression; tumor xenograft; tumorigenic; uptake; validation studies

DESCRIPTION (provided by applicant): Unraveling the underlying mechanisms of tumor growth and metastasis is critical to developing curative strategies against castration-resistant prostate cancer (CRPC). Recent findings demonstrate that tumor-recruited adipose stem cells (ASCs) can promote tumor growth, especially in obese patients. Unlike normal counterparts, our preliminary data showed that factors secreted by PC cells induce phenotypic genotypic changes in patient-derived ASCs (pASCs) and subvert them to undergo neoplastic transformation. Microvesicles (MVs) (50-200 nm) from PC cells (C4-2B and PC-3), but not normal prostate epithelial cells (RWPE1), primed tumor-tropic pASCs to form neoplastic lesions with cytogenetic aberrations reminiscent of the molecular features of prostate tumors and mesenchymal-to-epithelial transition (MET). The oncogenic 'reprogramming' of pASCs was associated with the transfer of a number of oncogenic miRNAs (miR-125b, miR-130b, miR-155) and oncogenic mRNAs (H-Ras and K-Ras), which in turn down-regulated several key tumor suppressors (TP53, PDCD4 and Lats2) in pASCs. Our data deciphered previously uncharacterized roles for tumor-derived extracellular RNAs (exRNAs) in promoting tumor growth via the release and uptake of MVs by the recipient pASCs. Cancer patient's sera contain high levels of circulating MVs in comparison to normal subjects; thus it is possible to speculate the transfer of oncogenic exRNA cargo by the tumor-derived MVs enable neoplastic transformation of pASCs in cancer patients. Accordingly, targeting MVs biogenesis and release by tumor cells and/or uptake by pASCs, rather than individual exRNAs, would be more efficacious in abrogating the transfer and tumor development by multiple oncogenic exRNAs in pASCs. We therefore hypothesize that a high throughput screening (HTS) of clinically approved compounds will enable us to select lead agents that suppress the biogenesis and release of MVs from tumor cells and/or the uptake of MVs by pASCs. We will corroborate our hypothesis by utilizing milestone-driven experiments in cell culture models and by proof-of-concept pre-clinical studies in animal models of PC. The UH2 (phase-I) will identify compounds that potently abrogate the biogenesis, release and/or uptake of MVs in vitro and in vivo. The UH3 (phase-II) will further validate the in vivo anti-tumor efficacy of these pharmacological leads and will support more rigorous milestone-driven pre-clinical studies in animal models. The following specific aims, to be executes through two phases, corroborate our hypothesis: Phase-I (UH2) (1) Demonstrate if circulating MVs procured from CRPC patients harbor oncogenic exRNAs and induce neoplastic transformation of pASCs. (2) Optimize HTS assays to identify lead compounds which inhibit the release of MVs by tumor cells, or their uptake by recipient pASCs, or reduce their oncogenic miRNA/RNA load. (3) Establish if lead compounds inhibit tumor cell release and/or uptake of MVs by pASCs in vivo. Phase II (UH3) (1) Examine the efficacy of lead compounds in inhibiting ASC-derived tumor development in vivo. (2) Clinical applicability of the lead compounds from the NCATS library. Although MVs have been implicated in cancer progression (neovacularization), their role in neoplastic transformation of stem cells in cancer patients has not been investigated. Our demonstration of tumor cell derived MVs in transfer of extracellular onco-RNAs and transformation of patient procured ASCs is novel and lend credence to their potential role in outgrowth and/or progression of metastatic disease in cancer patients. Accordingly, the proposed work is innovative, because it capitalizes not only in underpinning discovery of new functional roles for MV-mediated onco-miRs and onco-mRNAs, but also in identifying new lead therapeutic compounds to circumvent PC progression. By establishing preventive and/or therapeutic intervention strategies, the outcome of the proposed studies is expected to exert a positive impact on the clinical management of advanced PC.

描述（由申请人提供）：揭示肿瘤生长和转移的潜在机制对于制定针对去势抵抗性前列腺癌（CRPC）的治疗策略至关重要。最近的研究结果表明，肿瘤招募的脂肪干细胞（ASC）可以促进肿瘤生长，特别是在肥胖患者中。与正常细胞不同，我们的初步数据表明，PC 细胞分泌的因子会诱导患者来源的 ASC (pASC) 发生表型基因型变化，并颠覆它们进行肿瘤转化。来自 PC 细胞（C4-2B 和 PC-3）而非正常前列腺上皮细胞 (RWPE1) 的微泡 (MV) (50-200 nm) 引发亲肿瘤性 pASC 形成具有细胞遗传学畸变的肿瘤病变，让人想起前列腺肿瘤和间充质到上皮转化 (MET) 的分子特征。 pASC 的致癌“重编程”与许多致癌 miRNA（miR-125b、miR-130b、miR-155）和致癌 mRNA（H-Ras 和 K-Ras）的转移有关，这反过来又下调 pASC 中的几种关键肿瘤抑制因子（TP53、PDCD4 和 Lats2）。我们的数据破译了肿瘤来源的细胞外 RNA (exRNA) 在通过受体 pASC 释放和摄取 MV 来促进肿瘤生长方面的先前未表征的作用。与正常人相比，癌症患者的血清中含有高水平的循环MV；因此，可以推测肿瘤来源的 MV 转移致癌 exRNA 货物能够使癌症患者中的 pASC 发生肿瘤转化。因此，靶向MV的生物发生和肿瘤细胞的释放和/或pASC的摄取，而不是单个exRNA，将更有效地消除pASC中多种致癌exRNA的转移和肿瘤发展。因此，我们假设对临床批准的化合物进行高通量筛选 (HTS) 将使我们能够选择抑制肿瘤细胞中 MV 的生物发生和释放和/或 pASC 摄取 MV 的先导药物。我们将通过在细胞培养模型中利用里程碑驱动的实验以及在 PC 动物模型中进行概念验证临床前研究来证实我们的假设。 UH2（I 期）将鉴定能够在体外和体内有效消除 MV 的生物发生、释放和/或摄取的化合物。 UH3（II期）将进一步验证其体内抗肿瘤作用 这些药理学先导化合物的功效，并将支持在动物模型中进行更严格的里程碑驱动的临床前研究。以下具体目标将通过两个阶段执行，证实我们的假设： 第一阶段 (UH2) (1) 证明从 CRPC 患者获得的循环 MV 是否含有致癌 exRNA 并诱导 pASC 的肿瘤转化。 (2) 优化 HTS 测定，以确定抑制肿瘤细胞释放 MV 或受体 pASC 摄取 MV 或减少其致癌 miRNA/RNA 负载的先导化合物。 (3) 确定先导化合物是否在体内抑制肿瘤细胞释放和/或 pASC 对 MV 的摄取。 II 期 (UH3) (1) 检查先导化合物在体内抑制 ASC 衍生肿瘤发展的功效。 (2) NCATS 库中先导化合物的临床适用性。尽管 MV 与癌症进展（新生卵泡化）有关，但其在癌症患者干细胞肿瘤转化中的作用尚未得到研究。我们对肿瘤细胞衍生的 MV 在细胞外癌 RNA 转移和患者获得的 ASC 转化中的演示是新颖的，并证实了它们在癌症患者转移性疾病的生长和/或进展中的潜在作用。因此，所提出的工作具有创新性，因为它不仅有助于发现MV介导的onco-miR和onco-mRNA的新功能作用，而且还有助于识别新的主要治疗化合物来规避PC进展。通过制定预防和/或治疗干预策略，拟议研究的结果预计将对晚期 PC 的临床管理产生积极影响。