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.

研究总结： 结直肠癌 (CRC) 是第二大致命癌症，患者生存率从 90% 下降至 14% 在此，我们建议研究长链非编码RNA的作用。 (LncRNA)在失巢抵抗和CRC转移中是一个多步骤的过程，也是关键之一。 步骤是获得失巢抵抗力，以便在脱离原发部位后生存。因此，理解。 参与失巢凋亡过程和转移的分子参与者对于提高肿瘤细胞的存活率至关重要 CRC 患者与尿路上皮癌相关的长非编码 RNA (lncRNA) 的异常表达 1 (UCA1) 已在结直肠癌中被发现；然而，其在转移过程中的作用尚未明确。 锚定独立生长（失巢模型）的初步结果表明 lncRNA UCA1 增加 此外，lncRNA UCA1 的过度表达导致了葡萄糖转运蛋白 1 (GluT1) 的表达。 CRC 细胞中高 Glut1 表达和更高的葡萄糖摄取，这表明 CRC 细胞中的潜在机制作用 lncRNA UCA1 在失巢凋亡抵抗中的作用 在这项研究中，我们建议阐明 UCA1 及其相关的作用。 我们勇敢地面对lncRNA UCA1的过度表达。 通过失巢凋亡抵抗相关信号通路增强结直肠癌转移。 CRC 细胞系 SW480（致癌）和 SW620（转移）了解失巢凋亡的机制调节 AIM 1 旨在阐明失巢凋亡中涉及的 lncRNA UCA1 相关分子机制。 我们建议利用lncRNA UCA1在失巢凋亡抵抗中的作用。 慢病毒转导稳定过表达 (SW480+UCA1//GFP) 和敲低 (SW620+CRISPRgUCA1) 使用细胞周期、促存活、抗凋亡、干性、葡萄糖代谢、激酶磷酸化的细胞系 在 AIM 2 中，我们建议研究免疫沉淀 (IP)、co-IP 和邻近连接分析 (PLA) 分析。 UCA1 连接的蛋白质、RNA 和与失巢凋亡抗性相关的途径我们将使用稳定的同基因。 lncRNA UCA1（OE 和 KD）细胞系模型以及识别新型 UCA1 相关/链接的无偏见方法 使用生物素-ReCLP（可逆交联沉淀）的蛋白质和 RNA，体内 RNA 反义 蛋白质组学 (iRAP) 和 RNAseq 通过 RT-PCR、ddPCR、IP、co-IP 和 PLA 研究进行分析和验证。 虽然 AIM 3 被提议使用转移性小鼠评估 lncRNA UCA1 表达的功能影响 在此模型中，SCID 成年雄性/雌性小鼠将被注射表达经过验证的稳定的荧光素酶。 通过门静脉的同基因人 CRC 细胞系（UCA1 OE 和 KD，如 Target1），及其转移性 这项研究将通过生物发光成像来评估其潜力，为结直肠癌转移提供新的见解。 并将帮助开发创新疗法以提高患者的生存率，为未来的 NIH 生成数据 德克萨斯大学里奥格兰德河谷分校西班牙裔服务不足的少数族裔学生的提案和培训 (UTRGV)，是德克萨斯州南部里奥格兰德河谷地区著名的西班牙裔服务机构。

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.

小批量初级保健诊所的 COVID-19 疫苗接种活动：挑战

• 发表时间: 2022-07-03
• 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

Long Non-Coding RNAs: New Insights in Neurodegenerative Diseases.

长非编码 RNA：神经退行性疾病的新见解。

• 发表时间: 2024-02-14
• 影响因子: 5.6
• 作者: Anilkumar, Adithya K;Vij, Puneet;Lopez, Samantha;Leslie, Sophia M;Doxtater, Kyle;Khan, Mohammad Moshahid;Yallapu, Murali M;Chauhan, Subhash C;Maestre, Gladys E;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.

一名接受华法林治疗的患者死于 COVID-19 感染且 D-二聚体突然升高的病例报告。

• 发表时间: 2021-10-03
• 影响因子: 4.7
• 作者: Agarwal, Reita N;Aggarwal, Hersheth;Verma, Ashmit;Tripathi, Manish K
• 通讯作者: Tripathi, Manish K