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

• 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
• 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

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)是死亡率第二高的癌症，患者的存活率从90%下降到14% 癌症转移到远处的器官。在这里，我们建议研究一个长的非编码RNA的作用 (LncRNA)在失巢抵抗和结直肠癌转移中的作用。转移是一个多步骤的过程，也是关键之一 步骤是获得失巢抵抗，以求在脱离初级部位后存活。因此，理解 参与失巢凋亡过程和转移的分子参与者可能对提高肺癌的存活率至关重要。 结直肠癌患者。尿路上皮癌相关基因LncRNA的异常表达 (UCA1)在结直肠癌中已被发现，但其在转移过程中的作用尚未明确。我们的 锚定非依赖性生长(阿尼基模型)的初步结果显示，lncRNA UCA1增加 葡萄糖转运蛋白1(Glut1)蛋白表达。此外，lncRNA UCA1的过表达导致 导致结直肠癌细胞高Glut1表达和高葡萄糖摄取，这表明其潜在的机制作用 IncRNA UCA1在失巢抵抗中的作用。在本研究中，我们建议阐明UCA1及其相关基因S的作用 失巢抵抗过程中的信号通路。我们假设lncRNA UCA1的过度表达 通过失巢诱导耐药相关信号通路促进结直肠癌转移。我们将利用同源基因 大肠癌细胞系SW480(致癌)和SW620(转移)对失巢凋亡调控机制的研究 抵抗。目的1阐明lncRNAUCA1参与失巢失巢的相关分子机制。 耐药与结直肠癌转移。我们建议用以下方法研究lncRNA UCA1在失巢抵抗中的作用 慢病毒转导稳定过表达(SW480+UCA1//GFP)和敲除(SW620+CRISPRgUCA1) 细胞系利用细胞周期、促存活、抗凋亡、干性、葡萄糖代谢、激酶磷酸化 免疫沉淀法(IP)、共IP法和邻位连接分析法(PLA)。在目标2中，我们建议调查 UCA1连接蛋白、RNA和与失巢耐药相关的通路。我们将使用稳定的同源基因 LncRNA UCA1(OE和KD)细胞系模型及无偏鉴别新的UCA1相关/连锁方法 应用生物素-ReCLP(可逆交联沉淀法)的蛋白质和RNA，体内RNA反义 蛋白质组学(IRAP)和RNAseq分析，并通过RT-PCR、ddPCR、IP、co-IP和PLA研究验证它们。 虽然AIM 3被建议使用转移性小鼠来评估LncRNA UCA1表达的功能影响 模特。为了达到这个目的，将向SCID成年雄性/雌性小鼠注射有效稳定表达的荧光素酶 同基因人结直肠癌细胞系(UCA1、OE和KD，如AIM1)经门静脉途径及其转移 潜力将通过生物发光成像进行评估。这项研究将为结直肠癌的转移提供新的见解。 并将帮助开发创新的疗法，以提高患者的存活率，为未来的NIH产生数据 在德克萨斯大学格兰德河谷为服务不足的西班牙裔少数族裔学生提供建议和培训 (UTRGV)，这是南得克萨斯州里奥格兰德山谷地区一家著名的拉美裔服务机构。

项目成果

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.

A Case Report of a Patient on Therapeutic Warfarin Who Died of COVID-19 Infection with a Sudden Rise in D-Dimer.

• DOI: 10.3390/biomedicines9101382
• 发表时间: 2021-10-03
• 期刊: Biomedicines
• 影响因子: 4.7
• 作者: Agarwal RN;Aggarwal H;Verma A;Tripathi MK
• 通讯作者: Tripathi MK