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:

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