Research Initiation Award: Molecular Mechanisms for DCLK1 Tumorigenesis Revealed by Pathway Analysis using RNA Sequencing Data

研究启动奖：利用 RNA 测序数据进行通路分析揭示 DCLK1 肿瘤发生的分子机制

• 批准号: 2100805
• 负责人: Lianna Li
• 金额: $ 30万
• 依托单位: Tougaloo College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2100805&HistoricalAwards=false
• 关键词: Research; Initiation; Award; Molecular; Mechanisms

Research Initiation Awards provide support for junior and mid-career faculty at Historically Black Colleges and Universities who are building new research programs or redirecting and rebuilding existing research programs. It is expected that the award helps to further the faculty member's research capability and effectiveness, improve research and teaching at the home institution, and involves undergraduate students in research experiences. The award to Tougaloo College proposes a study of stem cell markers which play a role in the growth and chemical resistance of cancer cells. The proposed work will study the molecular mechanisms of different forms of these cells to better understand the origin of tumors. The research will provide a research opportunity for Tougaloo College undergraduates. Doublecortin-like kinase 1 (DCLK1) is a putative cancer stem cells marker with 5 isoforms. DCLK1 is up regulated and plays critical roles in the initiation and progression of multiple cancers. However, which isoform of DCLK1 is more closely correlated with tumorigenesis and chemoresistance, and the underlying molecular mechanisms are unclear. The long-term goal of the lab is to develop a comprehensive understanding of DCLK1. The overall objectives of this proposal are to: 1) reveal and compare association of different DCLK1 isoforms with stemness of cancer cells; and 2) discover molecular mechanisms of different DCLK1 isoforms in tumorigenesis and chemoresistance using RNA seq technology. The current hypothesis is that different DCLK1 isoforms may correlate differently with tumorigenesis and chemoresistance of cancers. The rational is that DCLK1 protein was up-regulated in clinical cancer samples, but the promoter region of DCLK1 isoform 1 (DCLK1-I1) was epigenetically imprinted. The discrepancy of protein and gene expression strongly suggests that specific DCLK1 isoform(s) are more closely associated with tumorigenesis and chemoresistance of cancers. To achieve our goals, 1): isogenic DCLK1 isoform (I1- I5) over-expressing cells will be established using colorectal cancer cell lines with different genetic background. Association of each isoform with stemness of cells will be determined; 2) modifications of transcriptome profiles in the isogenic DCLK1 isoform (I1- I5) over-expressing cells will be determined using RNA Seq technology. Differentially expressed genes will be identified and used for pathway analysis to establish the molecular networks of individual DCLK1 isoforms; and 3) cells will be treated with 5-Fu and transcriptome profiles will be determined using RNA seq technology. Differentially expressed genes will be identified and used for pathway analysis to reveal the molecular mechanism for chemoresistance. With successful completion of the proposal, it is expected that specific DCLK1 isoform(s) which contributes to the tumorigenesis and chemoresistance of cancer cells will be identified.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

研究启动奖为历史上黑人学院和大学的初级和中级职业教师提供支持，他们正在建立新的研究项目或重新定向和重建现有的研究项目。预计该奖项将有助于进一步提高教师的研究能力和效率，改善家庭机构的研究和教学，并使本科生参与研究经验。授予Tougaloo学院的奖项提出了一项干细胞标记物的研究，这些标记物在癌细胞的生长和化学抗性中发挥作用。拟议的工作将研究这些细胞不同形式的分子机制，以更好地了解肿瘤的起源。该研究将为Tougaloo学院的本科生提供一个研究机会。双皮质素样激酶1（DCLK 1）是一种公认的癌症干细胞标志物，具有5种亚型。DCLK 1被上调，在多种癌症的发生和发展中起着关键作用。然而，DCLK 1的哪种亚型与肿瘤的发生和化疗耐药关系更密切，其潜在的分子机制尚不清楚。该实验室的长期目标是全面了解DCLK 1。该提案的总体目标是：1）揭示和比较不同DCLK 1亚型与癌细胞干细胞性的关联;以及2）使用RNA seq技术发现不同DCLK 1亚型在肿瘤发生和化学抗性中的分子机制。目前的假设是，不同的DCLK 1亚型可能与肿瘤发生和癌症的化疗耐药性不同。其理论依据是DCLK 1蛋白在临床肿瘤中表达上调，但DCLK 1亚型1（DCLK 1-I1）的启动子区是表观遗传印记的。蛋白质和基因表达的差异强烈表明特定的DCLK 1亚型与癌症的肿瘤发生和化疗耐药性更密切相关。为了实现我们的目标，1）：使用具有不同遗传背景的结直肠癌细胞系建立过表达DCLK 1同种型（I1- I5）的同基因细胞。将确定每种同种型与细胞干性的关联; 2）将使用RNA Seq技术确定过表达等基因DCLK 1同种型（I1- I5）的细胞中转录组谱的修饰。差异表达的基因将被鉴定并用于途径分析，以建立单个DCLK 1亚型的分子网络;以及3）细胞将用5-Fu处理，并将使用RNA seq技术确定转录组谱。差异表达的基因将被鉴定并用于途径分析，以揭示化学抗性的分子机制。随着该项目的成功完成，预计将确定有助于肿瘤发生和癌细胞耐药性的特定DCLK 1亚型。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。