Development of novel miRNA based novel therapeutics for metastatic colorectal cancer

开发基于 miRNA 的转移性结直肠癌新疗法

基本信息

批准号：
10512749
负责人：
JINGFANG JU
金额：
--
依托单位：
NORTHPORT VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10512749
关键词：
3-Dimensional Acute Apoptosis BCL2 gene BMI1 gene CD44 gene Cell Cycle Regulation Cell Line Cessation of life Chemoresistance Colon Colon Carcinoma Colorectal Cancer Complex Development Drug Kinetics Drug or chemical Tissue Distribution Early Diagnosis Enzymes Epigenetic Process Epithelial Cells Failure Female Fluorouracil Gene Expression Regulation Genes Goals Growth Hair Human Incidence Laboratories Life Location Magnetic nanoparticles MicroRNAs Modeling Modification Mus Organoids Pathway interactions Patients Play Population Proliferating Protein Biosynthesis Quality of life RNA Degradation Recurrent Malignant Neoplasm Resistance Role Specificity TYMS gene Therapeutic Thymidylate Synthase Toxic effect Treatment Efficacy Tumor Suppressor Proteins United States Untranslated RNA Uracil Veterans Weight Woman appetite loss base cancer cell cancer diagnosis cancer recurrence cancer stem cell chemotherapy colon cancer metastasis colorectal cancer metastasis delivery vehicle design fluoropyrimidine improved in vivo knock-down lipid nanoparticle loss of function mRNA Translation male men metastatic colorectal military veteran mouse model novel novel strategies novel therapeutics oligofectamine patient derived xenograft model posttranscriptional protein expression resistance mechanism stem cell growth subcutaneous synergism therapeutic development therapeutic miRNA therapeutic target treatment strategy tumor tumorigenesis

项目摘要

Project Summary/Abstract The goal of this application is to investigate the roles of miR-15a in colorectal cancer and to develop novel miR-15a mimic as potential therapeutics to treat advanced metastatic colorectal cancer in both male and female Veterans. Despite advancements in early detection and improved treatment strategies, there are still 50,260 deaths due to colorectal cancer in the United States, and its incidence is equally high in the Veteran population. Resistance to fluoropyrimidine-based chemotherapy is one of the major causes for the failure of treating advanced metastatic colorectal cancer. It has been recognized recently that epigenetic alterations play a key role in tumorigenesis and resistance to 5-fluorouracil (5-FU) based chemotherapy. Because colorectal cancer cells are highly heterogeneous, chemotherapy can be quite effective in eliminating most of the rapid proliferating cancer cells. However, a small population of slow proliferating cancer stem cells are highly resistant which leads to cancer recurrence. Although the mechanism of chemoresistance is complex and is often associated with elevated target enzyme thymidylate synthase (TS, TYMS), recent studies from our laboratory have shown that epigenetic alterations such as changes in expression of non-coding miRNAs are major contributors to such resistance mechanisms to 5-FU by providing acute changes in protein synthesis at the post-transcriptional and translational levels. miRNAs are a class of small non-coding RNAs with crucial regulatory functions. We have identified a number of miRNAs with tumor suppressive functions in colorectal cancer. In particular, we have demonstrated that miR-15a (hsa-miR-15a-5p) can overcome chemoresistance in colorectal cancer as a potent tumor suppressor by inhibiting the expression of several major therapeutic target genes (BMI1, BCL2, YAP1, DCLK1) and associated pathways. More importantly, we have recently developed a novel strategy to create modified miRNA mimics with enhanced stability and efficacy for eliminating 5-FU resistant colon cancer stem cells while retaining target specificity. miR-15a mimics were designed by modifying the target strand of miR-15a by replacing uracil (U) bases with 5- FU at various locations. The rationale behind this approach is that 5-FU modification of miR-15a will enhance stability, and also combining the power of 5-FU and multi-targeted miR-15a into one entity to create therapeutic synergy, as miR-15a will breakdown eventually to release 5-FU. A unique feature of the 5-FU modified miR- 15a is that it can be internalized by colon cancer cells without any delivery vehicle. This represents a major advancement and a paradigm shift in miRNA based therapeutic development. Our preliminary results show that such modification improves the potency and stability of the miR-15a mimic and enhances its ability to inhibit colon cancer metastasis in vivo without any observed toxicity. Specific Aim 1: We will define the direct targets and pathways of miR-15a and miR-15a mimic in colon cancer and characterize the effects of miR-15a mimic on apoptosis, cell cycle control, and chemoresistance in colon cancer cells and 3D organoids. Specific Aim 2: We will investigate the toxicity, pharmacokinetics and tissue distribution of miR-15a mimic in vivo colon cancer mouse models. Specific Aim 3: We will develop and characterize the therapeutic potential of miR-15a mimic in metastatic colorectal cancer using in vivo colon cancer mouse models.

项目摘要/摘要本申请的目标是研究 miR-15a 在结直肠癌中的作用癌症并开发新型 miR-15a 模拟物作为治疗晚期转移性结直肠癌的潜在疗法男性和女性退伍军人都患有癌症。尽管在早期检测和改善治疗方面取得了进步策略，美国仍有 50,260 人死于结直肠癌，其发病率在退伍军人人口中同样高。对以氟嘧啶为基础的化疗的耐药性是主要的原因之一晚期转移性结直肠癌治疗失败的原因。最近人们认识到表观遗传改变在肿瘤发生和对 5-氟尿嘧啶 (5-FU) 的耐药性中发挥关键作用化疗。由于结直肠癌细胞具有高度异质性，化疗非常有效消除大部分快速增殖的癌细胞。但数量少、增殖缓慢癌症干细胞具有高度抵抗力，导致癌症复发。虽然其机制化疗耐药性很复杂，通常与靶酶胸苷酸合成酶（TS、 TYMS），我们实验室最近的研究表明，表观遗传改变，例如非编码 miRNA 的表达是 5-FU 耐药机制的主要贡献者，通过提供转录后和翻译水平上蛋白质合成的急剧变化。 miRNA 是一类具有重要调节功能的小非编码RNA。我们已经鉴定出许多与肿瘤相关的 miRNA 结直肠癌的抑制功能。特别是，我们已经证明 miR-15a (hsa-miR-15a-5p) 可以通过抑制表达作为有效的肿瘤抑制剂来克服结直肠癌的化疗耐药性几个主要治疗靶基因（BMI1、BCL2、YAP1、DCLK1）和相关途径的研究。更多的重要的是，我们最近开发了一种新策略来创建具有增强功能的修饰 miRNA 模拟物消除 5-FU 耐药性结肠癌干细胞同时保留靶标特异性的稳定性和功效。 miR-15a 模拟物是通过用 5- 取代尿嘧啶 (U) 碱基来修饰 miR-15a 的靶链而设计的。 FU 在不同地点。这种方法背后的基本原理是 miR-15a 的 5-FU 修饰将增强稳定性，并将 5-FU 和多靶点 miR-15a 的力量结合到一个实体中以创造治疗效果协同作用，因为 miR-15a 最终会分解释放 5-FU。 5-FU 修饰的 miR-的独特功能 15a的特点是它可以被结肠癌细胞内化，无需任何递送载体。这代表着一个重大的基于 miRNA 的治疗开发的进步和范式转变。我们的初步结果显示这种修饰提高了 miR-15a 模拟物的效力和稳定性，并增强了其在体内抑制结肠癌转移，且没有观察到任何毒性。具体目标 1：我们将定义直接 miR-15a 和 miR-15a 模拟物在结肠癌中的靶点和通路，并表征 miR-15a 的作用模拟结肠癌细胞和 3D 类器官的细胞凋亡、细胞周期控制和化疗耐药性。具体的目标2：我们将研究miR-15a模拟物在体内结肠的毒性、药代动力学和组织分布癌症小鼠模型。具体目标 3：我们将开发并表征 miR-15a 的治疗潜力使用体内结肠癌小鼠模型模拟转移性结直肠癌。