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，当患者死亡时，患者的存活率从90%下降到14%。 癌症转移到远处器官。在此，我们提出研究长非编码RNA的作用， （LncRNA）在失巢凋亡抵抗和CRC转移中的作用。转移是一个多步骤的过程， 步骤是获得抗失巢凋亡能力，以便在脱离原发部位后存活。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患者的生存率至关重要。 CRC患者。尿路上皮癌相关基因1的异常表达 UCA 1在CRC中已被鉴定;然而，其在转移过程中的作用尚未明确。我们 锚定非依赖性生长（失巢凋亡模型）的初步结果表明， 和葡萄糖转运蛋白1（GluT 1）蛋白表达。此外，lncRNA UCA 1的过表达导致了 CRC细胞中Glut 1的高表达和更高的葡萄糖摄取，这表明 lncRNA UCA 1在抗失巢凋亡中的作用在这项研究中，我们建议阐明UCA 1及其相关的作用。 失巢凋亡抵抗期间的信号通路。我们假设lncRNA UCA 1的过度表达 通过失巢凋亡抵抗相关信号通路增强CRC转移。我们将利用等基因 CRC细胞系SW 480（致癌）和SW 620（转移性），以了解失巢凋亡的机制调节 阻力AIM 1拟阐明lncRNA UCA 1相关的失巢凋亡分子机制 耐药和CRC转移。我们建议使用免疫组织化学方法研究lncRNA UCA 1在抗失巢凋亡中的作用。 慢病毒转导的稳定过表达（SW 480 + UCA 1//GFP）和敲减（SW 620 + CRISPRgUCA 1） 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 免疫沉淀（IP）、co-IP和邻位连接测定（PLA）测定。在AIM 2中，我们建议研究 UCA 1连接蛋白、RNA和与抗失巢凋亡相关的通路。我们将使用稳定的同基因 lncRNA UCA 1（OE和KD）细胞系模型和一种无偏倚的方法来鉴定新的UCA 1相关/连锁 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分析，并通过RT-PCR、ddPCR、IP、co-IP和PLA研究对其进行验证。 虽然AIM 3被提议使用转移性小鼠来评估lncRNA UCA 1表达的功能影响， 模型为此，SCID成年雄性/雌性小鼠将注射经验证的稳定表达荧光素酶的小鼠。 等基因人CRC细胞系（如aim 1中的UCA 1 OE和KD）通过门静脉， 将通过生物发光成像来评估潜力。这项研究将为CRC转移提供新的见解 并将有助于开发创新疗法，以提高患者的生存率，为未来的NIH提供数据， 提案，以及在德克萨斯大学格兰德河谷分校对西班牙裔少数民族学生的培训 （UTRGV），这是一个著名的西班牙裔服务机构在南德克萨斯州格兰德河谷地区。

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