Novel bioengineered microRNA therapeutics for lung cancer

新型生物工程 microRNA 疗法治疗肺癌

基本信息

批准号：
10530617
负责人：
Aiming Yu
金额：
$ 33.91万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-12-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10530617
关键词：
Accounting Adverse effects Biologic Development Biological Biomedical Engineering Blood Blood Chemical Analysis Cancer Etiology Cancer cell line Cancerous Cell Proliferation Cell physiology Cells Cessation of life Chemical Engineering Development Disease Drug Kinetics Effectiveness Encapsulated Endotoxins Engineering Escherichia coli Excretory function Exhibits Foundations Gene Expression Genetic Transcription Genomics Goals Human Hybrids Immune response Immunocompetent Invaded Kidney Liposomes Malignant Neoplasms Malignant neoplasm of lung Metabolic Methods MicroRNAs Modeling Modification Molecular Mus Non-Small-Cell Lung Carcinoma Oligoribonucleotides Oncogenic Peripheral Blood Mononuclear Cell Pharmacologic Actions Phase I Clinical Trials Polynucleotides Prodrugs Production Proliferating Property Proteins Proteome RNA Regulation Replacement Therapy Research Ribonucleases STAT3 gene Safety Serious Adverse Event Serum Solid Structure of parenchyma of lung Techniques Technology Testing Therapeutic Transfer RNA United States Untranslated RNA Woman Xenograft procedure antagonist body system chemical synthesis combat cost effective cytokine cytokine release syndrome high risk immunogenicity improved in vivo innovation insight interest large scale production lung cancer cell men milligram mouse model nanocomplexes nanotherapeutic new technology novel novel strategies novel therapeutic intervention novel therapeutics overexpression patient derived xenograft model polypeptide posttranscriptional research and development restoration stemness success therapeutic RNA therapeutic miRNA therapeutically effective therapy outcome transcriptome translational potential tumor tumor growth tumor progression tumor xenograft

项目摘要

Lung cancer remains a leading cause of cancer death in both women and men in the United States. There is a clear need for developing new and more effective therapeutics for the treatment of lung cancer, especially the most common subtype non-small cell lung cancer (NSCLC). As master regulators of transcriptome and proteome dynamics, microRNAs (miRNAs or miRs) govern many critical cancer cellular processes including proliferation, invasion and stemness. Therefore, restoration of tumor suppressive miRNAs (e.g., miR-34a and miR-124) lost in NSCLC cells represents a new therapeutic strategy. However, current miRNA mimics for research and development are made by chemical synthesis and decorated with various and extensive artificial modifications. This is in sharp contrast to miRNA molecules produced in living cells that do not carry any modifications or just a few necessary posttranscriptional modifications. Indeed it has been well documented that chemically-engineered/synthesized oligoribonucleotides (e.g., miRNA mimics) are readily recognized as foreign RNA molecules and thus cause immunogenicity. To break this barrier, we have made large efforts to develop novel approach for large-scale production of biologic miRNA agents (BERAs) in living cells. Our identification of hybrid tRNA/pre-miRNA molecules stably expressed in E. coli opens up a new avenue for RNA bioengineering. Furthermore, we have demonstrated that target miRNAs (e.g., miR-34a) are selectively released from BERA “prodrugs” in human cells, and consequently regulate target gene expression, inhibit NSCLC cell proliferation, and suppress xenograft tumor growth while they do not induce severe immune responses. In addition, our studies have found that liposome-polyethylenimine (LPP) nanocomplex increases BERA stability in serum and improves delivery efficiency to lung tissues. Given these exciting preliminary findings, we hypothesize that BERAs can be engineered at higher levels and on large scale; and fully-humanized BERA/miR-34a and BERA/miR-124 may be delivered by LPP as novel therapeutics for the treatment of NSCLC. To test the hypothesis, we proposed to establish more stable ncRNA carriers and produce a set of full-humanized ready-to-use BERAs (Aim 1), delineate the molecular pharmacological actions of BERAs in the control of human NSCLC cellular processes critical for therapeutic outcomes (Aim 2), and define the effectiveness and safety profiles of LPP-loaded BERA/miR-34a and miR-124 in metastatic NSCLC xenograft and patient-derived xenograft (PDX) mouse models (Aim 3). The proposed research will establish a novel technology for the production of fully-humanized biologic miRNA agents and open up new directions for the development of biologic RNA therapeutics.

肺癌仍然是美国男性和男性癌症死亡的主要原因。有明确需要开发新的，更有效的治疗肺癌的治疗方法，尤其是最常见的亚型非小细胞肺癌（NSCLC）。作为转录组和蛋白质组动力学，microRNA（miRNA或miRS）控制着许多关键的癌细胞过程增殖，入侵和茎。因此，恢复肿瘤抑制miRNA（例如miR-34a NSCLC细胞中丢失的miR-124）代表了一种新的治疗策略。但是，当前的miRNA模拟用于研究和开发是由化学合成制造的，并用各种广泛的人工修饰。这与在活细胞中产生的miRNA分子形成鲜明对比的是进行任何修改或仅进行一些必要的转录后修改。确实很好记录到化学设计/合成的寡核苷酸（例如miRNA模拟物）很容易被公认为外源RNA分子，因此引起免疫原性。为了打破这个障碍，我们有为大规模生产生物miRNA剂而做出了巨大的努力（Beras）在活细胞中。我们在大肠杆菌中稳定表达的杂化tRNA/pre-miRNA分子的鉴定为RNA生物工程开辟了新的途径。此外，我们已经证明了目标miRNA （例如，miR-34a）从人类细胞中的贝拉“前药”中有选择地释放，因此受调节靶基因表达，抑制NSCLC细胞增殖并抑制Xenographic肿瘤的生长不影响严重的免疫反应。此外，我们的研究发现脂质体聚集甲基亚胺（LPP）纳米复合物可提高血清中的BERA稳定性，并提高对肺组织的递送效率。鉴于这些令人兴奋的初步发现，我们假设Beras可以在更高级别上设计大规模； LPP可以将完全人性化的BERA/MIR-34A和BERA/MIR-124作为 NSCLC治疗的新理论。为了检验假设，我们提议建立更稳定的 NCRNA载体并产生一组全人道的现成Beras（AIM 1），描绘了分子 BERA在控制人NSCLC细胞过程中的药理作用对治疗至关重要结果（AIM 2），并定义了负载LPP的BERA/miR-34a的有效性和安全性。转移性NSCLC特征和患者衍生的Xenographotic（PDX）小鼠模型中的miR-124（AIM 3）。这拟议的研究将建立一种新技术，用于生产完全人性化的生物学miRNA 代理并为开发生物RNA疗法开发新方向。