Investigating the Biogenesis and Function of piRNAs in Germline Development

研究 piRNA 在种系发育中的生物发生和功能

基本信息

批准号：
8202553
负责人：
Heng-Chi Lee
金额：
$ 5.13万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8202553
关键词：
Address Animal Model Animal Testing Apoptosis Binding Biochemical Biogenesis Biological Markers Caenorhabditis elegans Cell Count Cell Maintenance Cells Cleaved cell Complex Development Disease Family Fertility Functional RNA Gene Expression Gene Expression Regulation Gene Silencing Genes Genetic Screening Genets Germ Cell Cancers Germ Cells Infertility Lead Link Malignant Neoplasms Mediating Messenger RNA Molecular Molecular Genetics Mutation Names Nematoda Outcome Pathway interactions Play Protein Family Proteins Proteomics RNA RNA Interference RNA Sequences Reporter Role Small Interfering RNA Small RNA Staging Stem cells Sterility T-cell acute lymphocytic leukemia 1 protein Tissues Transcript Transgenic Organisms cancer cell chromatin immunoprecipitation genome-wide malignant breast neoplasm member mutant novel novel therapeutic intervention novel therapeutics outcome forecast overexpression research study treatment strategy tumor

项目摘要

DESCRIPTION (provided by applicant): Small RNAs play remarkable roles in regulating gene expression by binding to Argonaute family proteins and guiding them to recognize their targets. A specific subfamily of Argonaute, named PIWI, binds PIWI-interacting RNA (piRNA) and promotes fertility in all animals tested to date. piRNAs and PIWI proteins are highly enriched in germ cells, and PIWI mutations lead to sterility and dramatically reduced germ-cell numbers. In addition, PIWI-like proteins are overexpressed in several cancer tissues and this overexpression has been linked to poor prognosis. However, the biogenesis of piRNAs, as well as the mechanism by which piRNAs regulate germ cell and cancer cell, remain largely unknown. In this proposal I will use specific experiments to address three fundamental questions about piRNA pathways using nematode C. elegans as model organism: What is the molecular mechanism of piRNA-induced gene silencing? What are the genes regulated by piRNAs? Finally, what are the components involved in piRNA biogenesis and downstream silencing? The understanding of this evolutionally conserved pathway has potential to provide novel therapeutic approaches for treatment of infertility and certain cancers. PUBLIC HEALTH RELEVANCE: Small RNAs play remarkable roles in regulating gene expression by binding to Argonaute family of proteins and guiding them to recognize their targets1. A specific subfamily of Argonaute, named PIWI, binds small RNA (piRNA) and has been linked to functions in stem cell maintenance and tumorgenesis of various cancers2-4. Here we propose to apply molecular, genetic and biochemical approaches to understand the biogenesis and mechanism of PIWI / piRNAs-mediated gene regulation, which have the potential to provide novel therapeutic strategies for treatment of diseases such as infertility and certain cancers. Reference 1. Ghildiyal, M., and Zamore, P. D. (2009). Small silencing RNAs: an expanding universe. Nat. Rev. Genet 10, 94-108. 2. Thomson, T., and Lin, H. (2009). The biogenesis and function of PIWI proteins and piRNAs: progress and prospect. Annu. Rev. Cell Dev. Biol 25, 355-76. 3. Lee, J. H., Sch|tte, D., Wulf, G., F|zesi, L., Radzun, H. J., Schweyer, S., Engel, W., and Nayernia, K. (2006). Stem-cell protein Piwil2 is widely expressed in tumors and inhibits apoptosis through activation of Stat3/Bcl-XL pathway. Hum. Mol. Genet 15, 201-11. 4. Liu, J. J., Shen, R., Chen, L., Ye, Y., He, G., Hua, K., Jarjoura, D., Nakano, T., Ramesh, G. K., Shapiro, C. L., et al. (2010). Piwil2 is expressed in various stages of breast cancers and has the potential to be used as a novel biomarker. Int J Clin Exp Pathol 3, 328-37.

描述（由申请人提供）：小型RNA通过与Argonaute家族蛋白结合并指导它们识别其靶标，在调节基因表达中起着显着的作用。 Argonaute的特定亚科，名为Piwi，结合了PIWI相互作用的RNA（PIRNA），并促进迄今为止所有测试的动物的生育能力。 PIRNA和PIWI蛋白在生殖细胞中高度富集，PIWI突变导致不育和大幅度降低的种系数量。此外，在几种癌症组织中，PIWI样蛋白过表达，这种过表达与预后不良有关。然而，PIRNA的生物发生以及PIRNA调节生殖细胞和癌细胞的机制，在很大程度上尚不清楚。在此提案中，我将使用特定的实验来解决有关PIRNA途径的三个基本问题，该问题使用线虫C. elegans作为模型生物：PIRNA诱导的基因沉默的分子机制是什么？ PIRNA调节的基因是什么？最后，PiRNA生物发生和下游沉默涉及哪些组成部分？对这一进化保守的途径的理解有可能提供新颖的治疗方法来治疗不育和某些癌症。公共卫生相关性：小型RNA在调节基因表达方面发挥着出色的作用，通过与Argonaute蛋白质家族结合并指导它们认识到其靶标1。 Argonaute的特定亚科，称为PIWI，结合了小RNA（PIRNA），并与各种Cancers2-4的干细胞维持和肿瘤发生的功能有关。在这里，我们建议采用分子，遗传和生化方法来了解PIWI / PIRNAS介导的基因调控的生物发生和机制，这些基因调节有可能提供新颖的治疗策略来治疗不育症和某些癌症等疾病。参考文献1。Ghildiyal，M。和Zamore，P。D.（2009）。小型沉默RNA：一个不断扩大的宇宙。纳特。基因10，94-108。 2。Thomson，T。和Lin，H。（2009）。 PIWI蛋白质和PIRNA的生物发生和功能：进步和前景。安努。 Rev. Cell Dev。生物25，355-76。 3。Lee，J。H.，Sch | Tte，D.，Wulf，G.，F | Zesi，L.，Radzun，H。J.，Schweyer，S.，Engel，W。和Nayernia，K。（2006）。干细胞蛋白piwil2在肿瘤中广泛表达，并通过激活STAT3/BCL-XL途径抑制凋亡。哼。摩尔。基因15、201-11。 4。Liu，J.J.，Shen，R.，Chen，L.，Ye，Y.，He，G.，Hua，K.，Jarjoura，D.，Nakano，T.，Ramesh，G.K.，Shapiro，C。L.等。（2010）。 Piwil2以乳腺癌的各个阶段表达，并且有可能用作新型生物标志物。 Int J Clin Exp Pathol 3，328-37。