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

研究启动奖为历史悠久的黑人学院和大学的初级和职业生涯中期教师提供支持，这些教师正在建立新的研究计划或重新定向和重建现有的研究计划。预计该奖项有助于进一步提高教师的研究能力和有效性，改善所在机构的研究和教学，并让本科生参与研究经验。授予Tougaloo学院的奖项提议对干细胞标记物进行研究，干细胞标记物在癌细胞的生长和化学耐药性中发挥作用。这项拟议的工作将研究不同形式的这些细胞的分子机制，以更好地了解肿瘤的起源。这项研究将为Tougaloo学院的本科生提供一个研究机会。DCLK1(Doublecortin-like kinase1)是一种可能的肿瘤干细胞标志物，有5种亚型。DCLK1表达上调，在多种癌症的发生和发展中起关键作用。然而，DCLK1的哪种异构体与肿瘤的发生和化疗耐药更密切相关，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DCLK1。这项建议的总体目标是：1)揭示和比较不同DCLK1亚型与癌细胞干性的关系；2)利用RNA序列技术发现不同DCLK1亚型在肿瘤发生和化疗耐药中的分子机制。目前的假设是，不同的DCLK1亚型可能与肿瘤的发生和癌症的化疗耐药有不同的相关性。其原因是DCLK1蛋白在临床癌组织中表达上调，但DCLK1亚型1的启动子区域(DCLK1-I1)是表观遗传印记。蛋白质和基因表达的差异强烈表明，DCLK1的特异性亚型(S)与肿瘤的发生和化疗耐药有更密切的关系。为了实现我们的目标，1)：利用不同遗传背景的结直肠癌细胞系建立等基因DCLK1亚型(I1-I5)高表达细胞。将确定每种异构体与细胞茎的关联性；2)将使用RNA Seq技术确定同基因DCLK1异构体(I1-I5)高表达细胞中转录组图谱的修改。差异表达的基因将被识别并用于通路分析，以建立单个DCLK1亚型的分子网络；3)细胞将被5-FU处理，转录组图谱将使用RNA序列技术确定。差异表达的基因将被识别并用于通路分析，以揭示化疗耐药的分子机制。随着提案的成功完成，预计将识别出与癌细胞的肿瘤发生和化疗耐药有关的特定DCLK1亚型(S)。这一奖项反映了美国国家科学基金会的法定使命，并已通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.