Role of lncRNA UCA1 in anoikis resistantce and colorectal cancer metastasis

lncRNA UCA1在失巢凋亡抵抗和结直肠癌转移中的作用

基本信息

批准号：
10689777
负责人：
Manish K Tripathi
金额：
$ 18.5万
依托单位：
UNIVERSITY OF TEXAS RIO GRANDE VALLEY
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：
2022-09-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10689777
关键词：
Adult Anchorage-Independent Growth Anoikis Antibodies Antisense RNA Apoptotic Biological Assay Biotin Cardiovascular system Cell Cycle Cell Line Cells Clinical Code Colon Colorectal Cancer Consumption Data Development Disease Distant Drops Female Freezing Future Glucose Transporter Goals Hispanic Hispanic-serving Institution Human Immunoprecipitation In Vitro Investigation Ligation Link Liver Luciferases Lymphatic System Malignant Neoplasms Minority Modeling Molecular Monitor Mus Neoplasm Metastasis Oncogenic Organ Paraffin Embedding Pathway interactions Patients Phosphorylation Phosphotransferases Portal vein structure Precipitation Process Proteins Proteome Proteomics RNA Regulation Research Resistance Reverse Transcriptase Polymerase Chain Reaction Role SW480 SW620 Signal Pathway Site Slide South Texas Students Survival Rate Testing Texas Therapeutic Time Tissue Embedding Tissues Training Transitional Cell Carcinoma Travel United States National Institutes of Health Universities Untranslated RNA Up-Regulation Western Blotting anticancer research bioluminescence imaging cancer cell colon cancer cell line colon cancer metastasis colon cancer patients colorectal cancer metastasis crosslink digital fighting glucose metabolism glucose uptake improved in vivo innovation insight knock-down lentivirally transduced link protein male metastatic colorectal minority student mouse model novel novel therapeutic intervention overexpression programs protein expression stemness sugar transcriptome sequencing trizol underserved minority

项目摘要

Research Summary: Colorectal cancer (CRC) is the second deadliest cancer, and patients’ survival rate drops from 90% to 14% when cancer metastasizes to distant organs. Herein, we proposed investigating the role of a long noncoding RNA (LncRNA) in anoikis resistance and CRC metastasis. Metastasis is a multistep process, and one of the key steps is to acquire anoikis resistance to survive after detachment from the primary sites. Thus, understanding the molecular players involved in the anoikis process and metastasis could be vital for improving the survival of CRC patients. The aberrant expression of a long noncoding RNA (lncRNA) urothelial carcinoma-associated 1 (UCA1) has been identified in CRC; however, its roles in metastasis processes are not yet well defined. Our preliminary results in the anchorage-independent growth (anoikis model) demonstrate increased lncRNA UCA1 and Glucose Transporter1 (GluT1) protein expression. Moreover, the overexpression of lncRNA UCA1 led to high Glut1 expression and higher glucose uptake in CRC cells, which indicates a potential mechanistic role of lncRNA UCA1 in anoikis resistance. In this study, we propose to elucidate the role(s) of UCA1 and its associated signaling pathways during anoikis resistance. We hypothesize that the overexpression of lncRNA UCA1 enhances CRC metastasis through anoikis resistance-associated signaling pathways. We will utilize Isogenic CRC cell lines SW480 (oncogenic) and SW620 (metastatic) to understand the mechanistic regulation of anoikis resistance. AIM 1 is proposed to elucidate lncRNA UCA1 associated molecular mechanisms involved in anoikis resistance and CRC metastasis. We propose to study the role of lncRNA UCA1 in anoikis resistance using Lentiviral transduced stable overexpression (SW480+UCA1//GFP) and knockdown (SW620+CRISPRgUCA1) cell lines using cell cycle, pro-survival, anti-apoptotic, stemness, glucose metabolism, kinase phosphorylation immunoprecipitation (IP), co-IP and Proximity Ligation Assay (PLA) assays. In AIM 2, we proposed investigating UCA1 linked proteins, RNAs, and pathways associated with anoikis resistance. We will use stable isogenic lncRNA UCA1 (OE and KD) cell line models and an unbiased approach to identify novel UCA1 associated/linked proteins and RNAs using Biotin-ReCLP (Reversible Cross-Linked Precipitation), in vivo RNA Antisense Proteomics (iRAP), and RNAseq analyses and validate them by RT-PCR, ddPCR, IP, co-IP and PLA studies. While AIM 3 is proposed to evaluate the functional impact of lncRNA UCA1 expression using a metastatic mouse model. In this aim, SCID adult male/female mice will be injected with Luciferase expressing validated stable isogenic human CRC cell lines (UCA1 OE and KD as in aim1) through the portal vein, and their metastatic potential will be evaluated by bioluminescence imaging. This study will provide new insights into CRC metastasis and will help in developing innovative therapeutics to improve patients’ survival, generate data for future NIH proposals, and training of Hispanic underserved minority students at the University of Texas Rio Grande Valley (UTRGV), which is a prominent Hispanic serving institution in South Texas Rio Grande Valley region.

研究摘要：结直肠癌（CRC）是第二个致命的癌症，患者的生存率从90％下降到14％癌症转移到遥远的器官。在此，我们提出了研究长未编码RNA的作用（lncrna）在抗阳极抗性和CRC转移中。转移是一个多步过程，也是关键之一步骤是获得与主要部位脱离后生存的抗阳极抗性。那，理解参与Anoikis过程和转移的分子参与者对于改善生存至关重要 CRC患者。长的非编码RNA（LNCRNA）尿路癌相关的异常表达1 （UCA1）已在CRC中鉴定出来；但是，其在转移过程中的作用尚未得到很好的定义。我们的初步的锚定生长（Anoikis模型）的初步结果表明LNCRNA UCA1增加和葡萄糖转运蛋白1（GLUT1）蛋白表达。此外，lncRNA UCA1 LED的过表达在CRC细胞中，高GLUT1表达和较高的葡萄糖摄取，这表明 lncRNA UCA1在抗阳极抗性中。在这项研究中，我们建议阐明UCA1及其相关的作用在抗阳极抗性期间的信号通路。我们假设lncRNA UCA1的过表达通过与抗抗性相关的信号通路增强CRC转移。我们将利用等源性 CRC细胞系SW480（致癌）和SW620（转移性），以了解Anoikis的机械调节反抗。提出了AIM 1来阐明与Anoikis相关的LNCRNA UCA1相关的分子机制抗性和CRC转移。我们建议研究使用LncRNA UCA1在使用Anoikis抗性中的作用慢病毒翻译稳定的过表达（SW480+UCA1 // GFP）和敲低（SW620+CRISPRGUCA1）使用细胞系的细胞系使用细胞周期，促生存，抗凋亡，干性，葡萄糖代谢，激酶磷酸化免疫沉淀（IP），CO-IP和接近连接测定法（PLA）测定法。在AIM 2中，我们提出了调查 UCA1连接了与抗Anoikis抗性相关的蛋白质，RNA和途径。我们将使用稳定的同源性 lncRNA UCA1（OE和KD）细胞系模型和一种无偏的方法来识别新型UCA1相关/链接蛋白质和RNA使用Biotin-reclp（可逆的交联沉淀），体内RNA反义蛋白质组学（IRAP）和RNASEQ通过RT-PCR，DDPCR，IP，CO-IP和PLA研究对其进行验证。虽然提出AIM 3使用转移小鼠评估LNCRNA UCA1表达的功能影响模型。在此目标中，SCID成年雄性/雌性小鼠将注入表达经过验证稳定的荧光素酶通过门静脉和它们的转移性电势将通过生物发光成像评估。这项研究将为CRC转移提供新的见解并将有助于开发创新的治疗剂以改善患者的生存，生成未来NIH的数据德克萨斯大学里奥格兰德分校的西班牙裔少数族裔学生的建议和培训（UTRGV），这是南德克萨斯州里奥格兰德谷地区的著名西班牙裔服务机构。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

COVID-19 Vaccination Drive in a Low-Volume Primary Care Clinic: Challenges & Lessons Learned in Using Homegrown Self-Scheduling Web-Based Mobile Platforms.

DOI：
10.3390/vaccines10071072
发表时间：
2022-07-03
期刊：
VACCINES
影响因子：
7.8
作者：
Agarwal, Reita N.;Aggarwal, Rajesh;Nandarapu, Pridhviraj;Aggarwal, Hersheth;Verma, Ashmit;Haque, Absarul;Tripathi, Manish K.
通讯作者：
Tripathi, Manish K.

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