Novel DNAzyme sensors for lithium and sodium to understand cellular and molecular mechanisms of lithium treatment of bipolar disorder

新型锂和钠 DNAzyme 传感器可了解锂治疗双相情感障碍的细胞和分子机制

• 批准号: 9306205
• 负责人: Yi Lu
• 金额: $ 18.31万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9306205
• 关键词: Adverse effects; Affect; Area; Binding; Biochemical; Biology; Biosensor; Bipolar Disorder; Catalytic DNA; Catalytic RNA; Cell model; Cells; Cellular biology; Cleaved cell; Clinical; DNA Library; Depressive disorder; Detection; Disease; Event; Fibroblasts; Fluorescence; Fluorescence Resonance Energy Transfer; Goals; Image; In Vitro; Ions; Legal patent; Link; Lithium; Measures; Mental Health; Mental disorders; Metals; Methodology; Methods; Molecular; Monitor; Pathologic; Pathway interactions; Patient-Focused Outcomes; Patients; Physiological; Physiology; Play; Process; Reaction; Reporting; Ribonucleotides; Role; Signal Transduction; Sodium; Specificity; System; Therapeutic; Therapeutic Effect; Transfection; Vertebral column; base; biomaterial compatibility; biophysical techniques; clinical diagnostics; combinatorial; design; divalent metal; dosage; effective therapy; fluorophore; improved; innovation; insight; lymphoblast; metalloenzyme; novel; phosphodiester; ratiometric; response; scaffold; sensor; success; uptake; water solubility

Project Summary/Abstract Bipolar disorder (BD) and other depressive disorders are some of the most common mental illnesses affecting millions of people. The primary treatment, lithium (Li+), has remained unchanged for > 50 years. However, the therapeutic window for Li+ is very narrow (0.5-1.2 mM) and the side effects can be severe. Also, Na+ dysregulation is implicated in BD and its pathways are also involved in Li+ uptake, but very little is known about the basis of BD or the mechanism of action of Li+. A critical barrier to maximizing the therapeutic effect and minimizing the side effect is missing information of the concentrations and distributions of Li+ and Na+ in BD cells, because of lack of selective sensors for Li+ and Na+ in living cells. This R21 proposal seeks to explore and demonstrate proof-of-concept of a novel class of DNAzyme sensors with high specificity for either Li+ or Na+ for simultaneous detection of Li+ and Na+ in BD cells, in order to provide new insights into the cellular and molecular mechanisms of Li+ treatment of BD. Specifically, we propose to use an in vitro selection method to obtain Li+- or Na+-specific DNAzymes with high catalytic activity towards ribonucleotide cleavage. These sequences will be characterized and optimized for sensor applications, whereupon we will transform them into fluorescent sensors using the catalytic beacon method for simultaneous imaging of Li+ and Na+ in lymphoblast cell models of BD, including cells obtained from BD- and non-BD patients. These fluorescent sensors will further be improved upon by incorporating both a photolabile-caging group to protect against ribonucleotide cleavage during the cell transfection, and a FRET pair of fluorophores to enable ratiometric sensing to allow for better quantitation of ion concentrations. While most cellular sensors have been designed for detection of divalent metal ions (e.g. Ca2+), few effective cellular sensors have been developed for monovalent ions. By developing a novel class of DNAzyme sensors to provide a direct measure of Li+ and Na+ simultaneously in BD cells, this proposal provides a critical missing piece of information in providing insights into the cellular and molecular mechanisms for Li+ treatment of BD. In the process, we will demonstrate general methodologies for DNAzyme-based sensors applicable for any metal ion, including the use of in vitro selection incorporating negative selection to improve selectivity and use of the catalytic beacon system to transform metal binding into different fluorescence readouts. The methods demonstrated can be applied for detection and imaging of Li+ and Na+ for other BD cells and can be generalized to develop similar sensors to image many other metal ions, which will advance the fields of mental health, cell biology, and clinical diagnostics.

项目总结/文摘

项目成果

Evidence of a General Acid-Base Catalysis Mechanism in the 8-17 DNAzyme.

• DOI: 10.1021/acs.biochem.7b01096
• 发表时间: 2018-03-06
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Cepeda-Plaza M;McGhee CE;Lu Y
• 通讯作者: Lu Y

Discovery of and Insights into DNA "Codes" for Tunable Morphologies of Metal Nanoparticles.

• DOI: 10.1002/smll.201900975
• 发表时间: 2019-05
• 期刊: Small
• 影响因子: 13.3
• 作者: Nitya Sai Reddy Satyavolu;K. Y. Loh;L. H. Tan;Yi Lu

DNAzyme sensors for detection of metal ions in the environment and imaging them in living cells.

• DOI: 10.1016/j.copbio.2017.03.002
• 发表时间: 2017-06
• 期刊: Current opinion in biotechnology
• 影响因子: 7.7
• 作者: McGhee CE;Loh KY;Lu Y
• 通讯作者: Lu Y

Advancing Point-of-Care Diagnostics of Metabolites Through Engineering Semisynthetic Proteins.

通过工程半合成蛋白质推进代谢物的即时诊断。

• DOI: 10.1373/clinchem.2018.298836
• 发表时间: 2019
• 期刊: Clinical chemistry
• 影响因子: 9.3
• 作者: Zhang,JingJing;Lu,Yi
• 通讯作者: Lu,Yi

Optical Control of Metal Ion Probes in Cells and Zebrafish Using Highly Selective DNAzymes Conjugated to Upconversion Nanoparticles.

• DOI: 10.1021/jacs.8b09867
• 发表时间: 2018-11
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Zhenglin Yang;K. Y. Loh;Yu Chu;Ruopei Feng;Nitya Sai Reddy Satyavolu;Mengyi Xiong;Stephanie M Nakamata Huynh-Stephan